Complete Abstract Book - HIV Research For Prevention

Transcription

HIV Research for
Prevention 2014
AIDS Vaccine, Microbicide and
ARV-based Prevention Science
Cape Town, South Africa
28–31 October 2014
Cape Town, South Africa 28–31 October 2014
www.hivr4p.org
ARV Exposure and Efficacy in the Genital Tract (OA13)
Level 01, Auditorium 02
Host Factors: Injury, Acquisition and Infection (OA14)
Level 02, Meeting Room 2.40
PrEP and Microbicide Adherence in Women (OA15)
Level 02, Roof Terrace Room
Animal Model Studies of Microbicides and Injectables (OA3)
Innate Immunity (OA4)
Vaccine, Viral Latency and Cure (OA5)
Mucosal Target and Effector Cells (OA17)
Evaluation of Novel Biomedical Interventions (OA18)
Good Participatory Practices in HIV Prevention (OA19)
Reproductive Hormones and HIV Risk (OA20)
Risk and Prevention for Men who Have Sex with Men (OA7)
Correlates of Protection and Exposure (OA8)
Engaging, Recruiting and Retaining Trial Participants (OA9)
Bacterial Vaginosis and HSV-2: Impact on Genital Immunity (OA10)
Thursday, 30 October
Mucosal Barriers to Infection (SY8)
Sustaining Durability of Responses (SY9)
Novel Formulations and Sustained Delivery of Antiretrovirals (SY7)
Safer Sex in 2014: Has the Paradigm Shifted? (RT1)
Please visit
hivr4p.org/program/posters
to view poster presentations
by theme.
28–31 October 2014
Welcome Reception 17:00 – 19:00 Level 01, Ballroom E+W
Building Combination Prevention Trials (SY4)
Poster Session 01 and Reception 17:00 –18:30 Level 0, Hall 2
RT Roundtable
Policy, Advocacy and Modeling (PD6) Room C
Preclinical and Clinical Vaccine Trials (PD3) Room C
PD Poster Discussion
SY Symposium
Glycans and Antibody Effector Functions (PD5) Room B
OA Oral Abstract
Session
Behavioral and Social Sciences (PD4) Room A
Oral Poster Discussions, 17:15 – 18:05
PL Plenary Session
Key
For Satellite Sessions on
Monday, 27 October
and Friday 31 October, visit
http://hivr4p.org/program/
satellite-sessions to view
the program.
Afternoon Satellites
13:30 – 17:30
Mind the Gap: Bridging from
Trial Success to Access (PL04)
Closing Plenary Session 04
10:30 – 12:30
Refreshment Break 10:00 – 10:30,
Auditorium 01 Foyer
Correlates of Protection in Highly Exposed Seronegative People (PD2)
Room B
Community Engagement and Advocacy (PD1) Room A
Oral Poster Discussions, 17:15 – 18:05
Poster Session 02 and Reception 17:00 –18:30
Level 0, Hall 2
Research Where and With Whom it Matters (RT4)
Harnessing Antibody Effector Functions (SY6)
Treatment as Prevention: The Promise and the Perils (SY3)
Prevention of Mother-to-Child Transmission Revisited (RT2)
Diffusion of Innovation: Accelerating Along the Research to
Rollout Continuum (RT3)
The Role of Structure Based Design in Vaccine Development and
Immunoprophylaxis (SY5)
Non-Abstract Driven Sessions 15:30 – 17:00
Refreshment Break 15:00 – 15:30, Level 0, Hall 2
Antibody Functions and Protection (OA30)
T Cell Immunity (OA29)
Treating and Preventing: the Role of ARVs (OA28)
PrEP: Self-testing, Safety and Modeling (OA27)
Microbicides and Multipurpose Prevention Technologies (OA26)
Oral Abstract Sessions 13:30 – 15:00
Delegate and Networking Lunch 12:30 – 13:30, Level 0, Hall 2
Adjuvants and Immunogens (OA25)
Overcoming Barriers to Broadly Neutralizing Antibody Induction (SY1)
Advances in Animal Models (SY2)
Non-Abstract Driven
Sessions 08:30 – 10:00
Friday, 31 October
PROGRAM-AT-A-GLANCE
Multipurpose Technologies
(RT5)
Poster Session 02 I Refreshment Break 10:00 – 11:00, Level 0, Hall 2 Level 01, Meeting Room 1.60
Challenges of Biomedical
HIV Prevention Trials (SY10)
Viral Transmission Studies (OA21)
Emerging Areas in Immunity
Cell and Tissue Models of ARVs for Prevention (OA22)
(SY11)
Pregnancy Intentions, Safe Conception and PMTCT (OA23)
Differential Use of Antibodies
in Prevention (SY12)
Mucosal Responses (OA24)
Importance of Mucosa in Prevention Research (PL03)
Plenary Session 03 08:30 – 10:00
Refreshment Break 15:00 – 15:30, Level 0, Hall 2
Novel Vaccine Concepts (OA16)
B Cell Repertoires for Protection (OA6)
Refreshment Break 15:00 – 15:30, Level 01, Ballroom Foyer
Delegate and Networking Lunch 12:30 – 13:30, Level 0, Hall 2
Towards Broadly Neutralizing Antibody Induction (OA12)
Microbicides: Male Partner Engagement and Sexual Behaviors (OA2)
Delegate and Networking Lunch 12:30 – 13:30, Level 01, Ballroom E+W
Vaccine Development: Emerging Insights (OA11)
B Cell Immunogen Design (OA1)
Poster Session 01 I Refreshment Break 10:00 – 11:00, Level 0, Hall 2
Refreshment Break 10:30 – 11:00, Level 01, Ballroom Foyer
Targeting Biomedical Preventions to Different At-risk Populations (PL02)
State of the Art: Biomedical Prevention in 2014 (PL01)
Wednesday, 29 October
Plenary Session 02 08:30 – 10:00
Opening Plenary Session 01 08:15 – 10:30
Tuesday, 28 October
HIV Research for Prevention 2014: AIDS Vaccine, Microbicide and ARV-based Prevention Science
ABSTRACT BOOK
28 – 31 October 2014
Cape Town, South Africa 28–31 October 2014
Abstracts will be published as an online supplement at www.liebertpub.com/aid at the conclusion of the conference on Friday, 31 October.
HIV Research for Prevention 2014 | HIV R4P
Contents
Program at a Glance . . . . . . . . . . . . . . . . . . . . Inside Front Cover
Conference Committees . . . . . . . . . . . . . . . . . . . . . . . . . . . 6–8
Award and Scholarship Recipients . . . . . . . . . . . . . . . . . . 9–14
Presentations
Plenary Sessions . . . . . . . . . . . . . . . . . . . . . . . . . .
Non-Abstract Driven Sessions . . . . . . . . . . . . . . . .
Oral Abstract Sessions: Tuesday, 28 October . . . .
Oral Abstract Sessions: Wednesday, 29 October .
Oral Abstract Sessions: Thursday, 30 October . . .
Poster Discussions: Wednesday, 29 October . . . .
Poster Discussions: Thursday, 30 October . . . . . .
Overview of Poster Sessions . . . . . . . . . . . . . . . . .
Posters: Wednesday, 29 October . . . . . . . . . . . . .
Posters: Thursday, 30 October . . . . . . . . . . . . . . .
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. . 15–18
. . 19–36
. . 37–66
. . 67–96
. 97–127
128–136
137–145
146–147
148–283
. 284–417
Author Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 419–435
Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 436–438
Certificate of Attendance . . . . . . . . . . . . . . . . . . . . . . . . . . . 439
Partner Recognition . . . . . . . . . . . . . . . . . . . . Inside Back Cover
Abbreviations
Late Breaker Abstract . . . . .
Level . . . . . . . . . . . . . . . . . .
New Investigator Awardee .
Oral Abstract. . . . . . . . . . . .
Plenary Session. . . . . . . . . .
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OA
PL
Poster. . . . . . . . . . . . .
Poster Discussion. . . .
Roundtable . . . . . . . .
Symposium Session. .
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CHICAGO
Satellites begin on Monday, 17 October
Tuesday, 18 October–Friday, 21 October
www.hivr4p.org
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Conference Committees
CONFERENCE CHAIRS
CONFERENCE COMMITTEES
Sharon Hillier
University of Pittsburgh, United States
Eric Hunter
Emory University, Atlanta, United States
Anatoli Kamali
Medical Research Council/UVRI Uganda Research Unit on AIDS, Entebbe, Uganda
Helen Rees
Wits Reproductive Health and HIV Institute (Wits RHI), Johannesburg, South Africa
Robin Shattock
Imperial College, London, United Kingdom
PROGRAM ORGANIZING COMMITTEE MEMBERS
Alash’le Abimiku
Institute of Human Virology, University of Maryland School of
Medicine, United States
Susan Barnett
Novartis, United States
Dan Barouch
Beth Israel Deaconess Medical Center, Ragon Institute of MGH
MIT and Harvard, United States
Stephen Becker
The Bill & Melinda Gates Foundation, United States
Gina Brown
Office of AIDS Research, National Institutes of Health,
United States
Susan Buchbinder
San Francisco Department of Public Health, United States
Elizabeth Bukusi
Kenya Medical Research Institute, Kenya
Dennis Burton
The Scripps Research Institute, United States
Nomita Chandhiok
Indian Council of Medical Research, India
Z. Mike Chirenje
University of Zimbabwe - University of California Collaborative
Research Programme, Zimbabwe
Myron Cohen
University of North Carolina, United States
David Cooper
Kirby Institute, University of New South Wales, Australia
Carl Dieffenbach
National Institute of Allergy and Infectious Diseases, National
Institutes of Health, United States
Patricia Fast
International AIDS Vaccine Initiative, United States
Glenda Gray
Perinatal HIV Research Unit and South African Medical Research
Council, South Africa
Scott Hammer
Columbia University, United States
Catherine Hankins
Amsterdam Institute for Global Health and Development,
Netherlands
Angela Kashuba
University of North Carolina, United States
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Milly Katana
John Snow, Inc., Uganda
Roger Le Grand
Commissariat à l’Énergie Atomique, France
Bonnie Mathieson
Office of AIDS Research, National Institutes of Health,
United States
Andrew McMichael
Oxford University, United Kingdom
Lynn Morris
National Institute for Communicable Diseases, South Africa
Thumbi Ndung’u
University of KwaZulu-Natal, South Africa
Julie Overbaugh
Fred Hutchinson Cancer Research Center, United States
Jim Pickett
International Rectal Microbicide Advocates, United States
Punnee Pitisuttithum
Vaccine Trial Centre, Mahidol University, Thailand
Gita Ramjee
HIV Prevention Research Unit, South African Medical Research
Council, South Africa
Merlin Robb
US Military HIV Research Program, United States
Nina Russell
Barbara Shacklett
University of California at Davis, United States
Yiming Shao
National Center for AIDS/STD Control and Prevention (NCAIDS),
Department of the Research on Virology and Immunology, and
Chinese Center fo Disease Control and Prevention, China
Bill Snow
Global HIV Vaccine Enterprise, United States
Morenike Ukpong
Obafemi Awolowo University; HIV Vaccine and Microbicide
Advocacy Society, Nigeria
Mitchell Warren
AVAC: Global Advocacy for HIV Prevention, United States
Carolyn Williamson
University of Cape Town, South Africa
Ntando Yola
Desmond Tutu HIV Foundation, South Africa
In addition to the Program Organizing Committee, HIV R4P would like to thank the following individuals for reviewing
abstracts.
Todd Allen
Wayne Koff
Galit Alter
Charles Lacey
Marcus Altfeld
Yves Levy
Rama Amara
George Lewis
Omu Anzala
Amapola Manrique
Jared Baeten
Jeanne Marrazzo
Prince Bahati
John Mascola
Ragon Institute of MGH, MIT and Harvard, United States
Heinrich Pette Institute, Germany
The Yerkes National Primate Research Center, Emory University,
United States
KAVI Institute of Clinical Research, University of Nairobi, Kenya
University of Washington, United States
International AIDS Vaccine Initiative, Kenya
University of York, United Kingdom
Vaccine Research Institute, France
Medicine, United States
Christian Brander
Robert Chen
CHU Sainte Justine and the Montreal Children’s Hospital, Canada
Josephine Cox
Fenway Health, United States
Cynthia Derdeyn
Population Council, United States
Charlene Dezzutti
Gustavo Doncel
Daniel Douek
International Partnership for Microbicides, United States
IrsiCaixa Institute for AIDS Research, HIVACAT, Spain
Centers for Disease Control and Prevention, United States
Emory University, United States
CONRAD/Eastern Virginia Medical School, United States
Benoit Masse
Kenneth Mayer
Barbara Mensch
Nelson Michael
Penny Moore
Jeremy Nuttall
Jean Patterson
Keith Fowke
Dorothy Patton
Nicole Frahm
Damian Purcell
David Friend
Harriet Robinson
Henry Gabelnick
Lisa Rohan
University of Manitoba, Canada
CONRAD, United States
Independent Consultant, United States
Office of AIDS Research, National Institutes of Health, United States
University of Melbourne, Australia
Geovax Inc., United States
J. Victor Garcia
Department of Pharmaceutical Sciences School of Pharmacy,
University of Pittsburgh, Magee-Women’s Research Institute,
United States
Paul Goepfert
Morgane Rolland
University of North Carolina at Chapel Hill, United States
University of Alabama at Birmingham, United States
Barney Graham
Vaccine Research Center, National Insittute of Allergy and
Infectious Dieases, National Institutes of Health, United States
US Military HIV Research Program, Henry M. Jackson Foundation,
United States
Joseph Romano
NWJ Group LLC, United States
Tomas Hanke
Jeffrey Safrit
Barton Haynes
Eric Sandström
Craig Hendrix
William Schief
University of Oxford, United Kingdom
Duke Human Vaccine Institute, Duke University, United States
Johns Hopkins University, United States
Walid Heneine
Centers for Disease Control and Prevention, United States
Betsy Herold
Albert Einstein College of Medicine, United States
Saidi Kapiga
London School of Hygiene and Tropical Medicine, Mwanza
Intervention Trials Unit, United Kingdom
Elizabeth Glaser Pediatric AIDS Foundation, United States
Karolinska Institutet, Sweden
The Scripps Research Institute, International AIDS Vaccine
Initiative, Ragon Institute of MGH, MIT and Harvard, United States
Alexandra Schuetz
Armed Forces Research Institute for Medical Science, United States
Olivier Schwartz
Institut Pasteur, France
Guido Silvestri
Stephen Kent
Jonathan Stadler
Jerome Kim
Alan Stone
University of Melbourne, Australia
Wits Reproductive Health and HIV Institute, South Africa
MEDSA Ltd., United Kingdom
www.hivr4p.org
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ABSTRACT REVIEW COMMITTEE
ABSTRACT REVIEW COMMITTEE (continued)
Jim Tartaglia
Lut Van Damme
Caroline Tiemessen
Ronald Veazey
Georgia Tomaras
Steven Wakefield
Alexandra Trkola
Anna-Lise Williamson
Jim Turpin
Allen Zhiwei Wu
Sanofi Pasteur, United States
Duke Human Vaccine Institute, Duke University, United States
University of Zurich, Germany
Tulane National Primate Research Center, United States
HIV Vaccine Trials Network, United States
Nanjing University, China
SECRETARIAT
Mark Aurigemma
Kristin Morrell
Jennifer Brunet
Kate Porter
Karlyanna Kopra
Nicole Santamaria
Communications Consultant, United States
Conference Solutions, United States
Amapola Manrique
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Scholars
NEW INVESTIGATOR AWARDEES
HIV R4P 2014 is proud to introduce this year’s New Investigator Awardees.
Siriwat Akapirat
Minlu Hu
Jinal Bhiman
Luca Schifanella
Armed Forces Research Institute of Medical Sciences, Department
of Retrovirology, Thailand
National Institute for Communicable Diseases and the University
of the Witwatersrand, South Africa
Magee-Womens Research Institute / University of Pittsburgh,
United States
National Cancer Institute, National Institutes of Health, United States
Elizabeth Byrne
Harvard University, United States
CONFERENCE SCHOLARS
Scholarships were made possible by generous donations from our partners and from registration funds.
Medical Research Council / Uganda Virus Research Institute,
Uganda
Nathlee Abbai
South African Medical Research Council, HIV Prevention Research
Unit, South Africa
Olatunji Adetokunboh
Stellenbosch University, South Africa
Wbeimar Aguilar
Grupo Inmunovirología, Facultad de Medicina, Universidad de
Antioquia, Colombia
Nurelign Ahmed
Projet San Francisco, Rwanda-Zambia HIV Research Group,
Rwanda
Seema Ajbani
National Institute for Research in Reproductive Health, India
Carolyne Akello
Makerere University - Johns Hopkins University Research
Collaboration, Uganda
Aderemi Oyekola Alagbe
Namibia Ministry of Health and Social Services, Namibia
Muriel Aldunate
Burnet Institute for Medical Research / Monash University,
Australia
Mallika Alexander
National AIDS Research Institute, India
Kabamba Alexandre
Council for Scientific and Industrial Research, South Africa
Mambo Amisi Modeste
Humanitarian Action for Health and Community Development,
Democratic Republic of the Congo
Obiajulu Amuamuziam
New HIV Vaccine and Microbicide Advocacy Society, Nigeria
Winnie Apidi
Eva Archer
Vaccine Research Center, National Institute of Allergy and
Infectious Diseases, National Institutes of Health, United States
Gershim Asiki
Uganda
Abdul Azeem
National AIDS Control Program, Pakistan
Veenu Bala
Central Drug Research Institute, India
Alejandro Balazs
Cameron Ball
Shaun Barnabas
Deborah Baron
Tahir Bashir Dar
Cheryl Baxter
AWARD / SCHOLARS
Andrew Abaasa
Centre for AIDS Programme of Research in South Africa,
South Africa
Mara Biasin
University of Milan, Italy
Patrick Bitangumutwenzi
Young Women’s Knowledge and Leadership Institute, Burundi
Saikat Boliar
Translational Health Science and Technology Institute, India
Jacqui Brener
Peter Medewar Building for Pathogen Research, University of
Oxford, United Kingdom
Charles Brown
Infectious Diseases Institute, AVAC, Uganda
William Brown, III
HIV Center for Clinical and Behavioral Studies, Columbia
University, United States
Omkar Chaudhary
All India Institute of Medical Sciences, India
Hannah Cheeseman
Imperial College London, United Kingdom
Yan Li Chen
National Center for AIDS/STD Control and Prevention, Chinese
Center for Disease Control and Prevention, China
Christopher Chianese
Fenway Health, United States
Clever Chilende
Treatment Advocacy and Literacy Campaign, Zambia
Jonah Chinga
Gay and Lesbian Coalition of Kenya, Kenya
Bhavna Chohan
Amy Chung
Daniel Claiborne
Núria Climent
AIDS Research Group, CELLEX-IDIBAPS-HIVACAT, Hospital Clinic i
Provincial de Barcelona, Spain
Sarah Cohen
Smritee Dabee
Institute of Infectious Disease and Molecular Medicine, University
of Cape Town, South Africa
Christine Dahlke
Universitätsklinikum Hamburg-Eppendorf, Germany
www.hivr4p.org
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Scholars
CONFERENCE SCHOLARS (continued)
José das Neves
Chiedu Ifekandu
Martin Deymier
Rosine Ingabire
Instituto de Engenharia Biomédica, Porto, Portugal, Portugal
Kathleen Doherty
Population Council, Nigeria
Rwanda
Vanderbilt University, Ragon Institute of MGH, MIT and Harvard,
United States
Desmond Iriaye
Clare Dott
Lakshmi Jagesur
Victor “Yimin” Du
Mariel Jais
South African Medical Research Council, South Africa
Zoe Duby
Milken Institute School of Public Health, The George Washington
University, United States
Juliane Etima
University of North Carolina and Center for AIDS Research,
Chapel Hill, United States
University of Cape Town, Desmond Tutu HIV Foundation,
South Africa
Kara Jensen
Sanne Skov Jensen
Abbey Evans
Vineet Joag
Yu Feng
Tsungai Ivai Jongwe
Susan Fetherston
Kadryn Kadasia
Jacqueline Flynn
Betty Kamira
SCHOLARS
University of Ulster, United Kingdom
Burnet Institute, Australia
Statens Serum Institut, Denmark
University of Toronto, Canada
Boston University, United States
Pascaline Fonteh
Alemju Fontu
Desmond Tutu HIV Foundation, South Africa
Anna Forbes
Rwanda Zambia HIV Research Group, Zambia
Joseph Francica
Rwanda Zambia HIV Research Group, United States
Ereshia Gabier
Innsbruck Medical University, Austria
University of Pretoria, South Africa
Association Camerouniase Pour le Marketing Social, Cameroon
Independent Consultant, United States
National Institutes of Health, United States
Brian Kanyemba
William Kilembe
Linda Kimaru
Janine Kimpel
South African National Bioinformatics Institute / University of the
Western Cape, South Africa
Deborah King
Tanuja Gengiah
Rose Kitawi
South Africa
Yanina Ghiglione
Instituto de Investigaciones Biomédicas en Retrovirus y Sida,
Argentina
Anna Gibbs
Karolinska Institutet, Sweden
Wesley Grimm
Northwestern University, United States
Anneke Grobler
South Africa
Tiffany Grooms-Williams
University of Louisville, United States
Ntombesizwe Nombasa Gxuluwe
World AIDS Campaign, South Africa
Antje Heit
Centre for Research in Therapeutic Sciences, Kenya
Nichole Klatt
Henrik Kløverpris
KwaZulu-Natal Research Institute for Tuberculosis and HIV,
South Africa
Rewa Kohli
National AIDS Research Institute, India
Catherine Kegakilwe Koofhethile
HIV Pathogenesis Programme, South Africa
Jennifer Kotlewski
Rwanda Zambia HIV Research Group, Zambia
Jean-Mari Kriek
Institute of Infectious Disease and Molecular Medicine, University
of Cape Town, South Africa
Sylvia Kusemererwa
Uganda
Tiffany Hensley-McBain
Zachary Kwena
Carolina Herrera
Rachel Kyeyune
Heather Hong
Jordan Kyongo
Centre for HIV and STIs, National Institute for Communicable
Diseases of the National Health Laboratory Service, and the
Faculty of Health Sciences, University of the Witwatersrand,
South Africa
Infectious Diseases Institute, Uganda
Institute of Tropical Medicine - Antwerp, Kenya
Faatima Laher
Elise Landais
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Scholars
KAVI-Institute of Clinical Research,University of Nairobi, Kenya
Melanie Merbah
Ria Lassauniere
U.S. Military HIV Research Program, Henry M. Jackson
Foundation, United States
Marc-André LeBlanc
South African National Bioinformatics Institute, South Africa
Boon Kiat Lee
University of Zimbabwe - University of California Collaborative
Research Programme, Zimbabwe
Duduzile Lembethe
Maureen Milanga
Maria Lemos
Lori Miller
Mildie Leuvennink
Brenda Gati Mirembe
Self-employed, Canada
AIDS Institute, The University of Hong Kong, Hong Kong
MatCH Research University of Witwatersrand, South Africa
International Partnership for Microbicides, South Africa
Clement Levin
INSERM U1135 CIMI, Paris, France
Haiying Li
Beth Israel Deaconess Medical Center, Harvard Medical School,
United States
Lenine Liebenberg
South Africa
Clint Mercuur
Nyaradzo Mgodi
AIDS Law Project, Kenya
London School of Hygiene & Tropical Medicine, United Kingdom
Nonhlanhla Mkhize
Kathryn Mngadi
South Africa
Daniela Monaco
Namal P.M. Liyanage
Jayajothi Moodley
Murray Logan
Jeeva Moodley
National Cancer Institute, National Institutes of Health,
United States
Evelyn Lumngwena
Alison Mahan
Mookho Malahleha
Setshaba Research Centre, South Africa
Thandi Maluka
SHIPP, South Africa
Jai Marathe
Boston University, United States
Elena Martinelli
Population Council, United States
Enrique Martin-Gayo
Yasuhiro Maruta
Matsushita Project Laboratory, Center for AIDS Research,
Kumamoto University, Kumamoto, Japan
Rose Masilo
Madibeng Centre for Research, South Africa
Flavia Matovu Kiweewa
Elizabeth Mbabazi Atuhurra
Medical Research Council, Uganda
Andrew McGuire
Seattle Biomed, United States
Eric Mcheka
National Association for People Living with HIV/AIDS in Malawi,
Malawi
Lyle McKinnon
South Africa
Meron Mengistu
Institute of Human Virology, United States
Sergey Menis
Unit, South Africa
Unit, South Africa
Nishila Moodley
Perinatal HIV Research Unit, South Africa
Neetha Morar
Unit, South Africa
Sara Morón-López
IrsiCaixa AIDS Research Institute - HIVACAT, Spain
Beatriz Mothe
Thandeka Moyo
Juliet Mpendo
Uganda Virus Research Institute, International AIDS Vaccine
Institute, Uganda
Sandra Mudhune
Peter Mugo
Kenya Medical Research Institute, Wellcome Trust, Kenya
Kenneth Mugwanya
Andrew Mujugira
University of Washington, Uganda
Wendy Murillo
Universidad Nacional Autonoma de Honduras, Honduras
Eva Muro
Kilimanjaro Clinical Research Institute, United Republic of
Tanzania
Petina Musara
University of Zimbabwe - University of California San Francisco
Collaborative Research Programme, Zimbabwe
Leonard Mutisya
The East African Sexual Health and Right Initiative, Kenya
Shem Mutuiri
Institute of Primate Research, Kenya
Rosemary Muwawu
www.hivr4p.org
11
SCHOLARS
Robert Langat
Scholars
Simon Mwangi
Alex Olvera
Lawrence Mwihaki
Shatha Omar
Bar Hostess Empowerment and Support Program, Kenya
Partners in Prevention Thika - University of Washington Site,
Kenya
Philip Mwimanzi
Simon Fraser University, Canada
Pumeza Mzizi
Perinatal HIV Research Unit, Wits Health Consortium,
South Africa
Sarita Naidoo
Unit, South Africa
Lillian Naigaga Mutengu
Teopista Nakyanzi
Sophie Clare Nanziri
Makerere University - John Hopkins University Research
SCHOLARS
Tashini Nayager
South African Medical Research Council, HIV Prevention Unit,
South Africa
Pepukai Ndadziyira
University of Zimbabwe - University of California San Francisco
Collaborative Research Programme, Zimbabwe
Zaza Ndhlovu
University of KwaZulu Natal, Nelson Mandela School of
Medicine, South Africa
Bongiwe Ndlovu
Evanson Ndung’u
Duduzile Ndwandwe
Unit, South Africa
Everlyne Ombati
Gloria Omosa-Manyonyi
University of Nairobi, Kenya
Simon Ondiek
HIV/AIDS Research & Advocacy Program, Kenya
Sylvia Onyango
Uganda
Chiara Orlandi
Institute of Human Virology - University of Maryland,
United States
Sophia Osawe
Institute of Human Virology, Nigeria
George Victor Owino
Patrick Owiti
Kenya Medical Research Institute / Family AIDS Care and
Education Services project, Kenya
Mickey Patel
Geisel School of Medicine at Dartmouth, United States
Arendevi Pather
Unit, South Africa
Shilpa Patil
Translational Health Science and Technology Institute, India
Sinazo Pato
Laura Pattacini
San Patten
San Patten and Associates, Inc., Canada
Robert Newells
Binghao Peng
Kenneth Ngure
Catia Perciani
AVAC PxROAR, United States
Jomo Kenyatta University of Agriculture and Technology College of Health Sciences, Kenya
Nadesh Ngechae Nji
Chantal Biya International Reference Center for Research on the
Prevention and Management of HIV/AIDS, Cameroon
University of Toronto, Canada
Capucine Phelip
Institut de Biologie et Chimie des Protéines, LBTI, UMR 5305 CNRS/Université de Lyon, France
Jessica Phillip
Stella Njuguna
HIV Prevention Research Unit, Medical Research Council,
South Africa
Chanda Nsofwa
Anabela Picton
Family Health Trust, Zambia
Fanelesibonge Ntombela
South Africa
Julien Nyombayire
Rwanda
Ayodeji Oginni
Sandra Okala
Dismas Oketch
Moi University School of Medicine, Kenya
Kennedy Olango
Men Against AIDS Youth Group, Kenya
Catherine Oldenburg
Harvard School of Public Health, United States
South Africa
Anna Piddubna
Sumy State University, Ukraine
Diantha Pillay
University of KwaZulu-Natal, Centre for AIDS Programme of
Research in South Africa, South Africa
Anastasia Pokrovskaya
Federal Budget Institution of Science Central Scientific Research
Institute of Epidemiology of Rospotrebnadzor, Russian Federal
AIDS Centre, Russian Federation
Gloria Preza
University of Southern California, United States
Jessica Prince
Zakiya Qualls
Rutgers Biomedical and Health Sciences, United States
12
Scholars
Chitra Singh
Srinika Ranasinghe
South Africa
Mary-Jane Ratlhagana
Indian Institute of Technology - Madras, India
Krishnaveni Reddy
Sam Higginbottom Institute of Agriculture, Technology &
Sciences, India
International Training and Education for Health, South Africa
Wits Reproductive Health and HIV Institute (Wits RHI),
South Africa
Satya Prakash Singh
Udaya Pratap Singh
Magdalena Sips
KVR Reddy
Simone Richardson
AURUM Institute, South Africa
Tsakani Sithole
South Africa
Aida Sivro
Laura Richert Spuhler
Kieron Smith
Meika Richmond
Marta Rodriguez Garcia
Geisel School of Medicine at Dartmouth, United States
Maria Julia Ruiz
Universidad de Buenos Aires/CONICET, Argentina
Wiriya Rutvisuttinunt
AFRIMS, Thailand
Richard Rwanyonga
Uganda
Darpun Sachdev
Bridge HIV, San Francisco Department of Public Health and
Center for AIDS Prevention Studies, University of California San
Francisco, United States
Anwesha Sanyal
University of Pittsburgh Graduate School of Public Health,
United States
Kennedy Smart
Institute of Human Virology, Nigeria
University of Aberdeen, United Kingdom
Melissa Smith
Stacey Smith
Emory University, Yerkes National Primate Research Center,
United States
Olivia Snyder
University of North Carolina, Chapel Hill, United States
Devin Sok
Livingstone Ssali
The AIDS Support Organisation, TASO, Uganda
Derek Stein
Renee Street
Unit, South Africa
Bin Su
INSERM U1109, FMTS, Université de Strasbourg, France
Irma Saulle
Ibrahim Suleiman
Cathrine Scheepers
Zehua Sun
University of Milan, Italy
Center for HIV and STIs, National Institute for Communicable
Diseases and the University of the Witwatersrand, South Africa
AIDS Institute, The University of Hong Kong, Hong Kong
Nsubuga Supercharger Moses
Torben Schiffner
Joint Clinical Research Centre, Uganda
Veronika Schmid
Thai NGO Coalition on AIDS, Thailand
University of Oxford, United Kingdom
Niwat Suwanphatthana
Institute of Medical Microbiology and Hygiene, University of
Regensburg, Germany
Jessica Terlikowski
Jeffrey Schneider
Sydney Tetteh Hushie
Yanille Scott
Aime Marcel Tongo Passo
Adekemi Sekoni
Hung Trinh
Kwame Shanaube
Damien Tully
Remmy Shawa
Morenike Ukpong
Northwestern University, United States
University of Pittsburgh Graduate School of Public Health,
United States
University of Lagos, Nigeria
ZAMBART, Zambia
Sonke Gender Justice, South Africa
Mahesh Sherkar
AIDS Foundation of Chicago, United States
Global Youth Coalition on HIV and AIDS, Ghana
International Centre for Genetic Engineering and Biotechnology,
U.S. Military HIV Research Program, United States
Institute of Public Health, Obafemi Awolowo University; New HIV
Vaccine and Microbicide Advocacy Society, Nigeria
Shri Vivekanand Nursing Home Trust’s College of Pharmacy,
Rahuri, Ahmednagar, Maharashtra, India, India
DaShawn Usher
Stuart Sievers
Neliëtte Van Niekerk
Paola Silveira
Thomas Vazquez
California Institute of Technology, United States
Federal University of Rio de Janeiro, Brazil
SCHOLARS
Pranitha Ramchuran
Project ACHIEVE, United States
International Partnership for Microbicides, South Africa
I3 Laboratory - INSERM U959 - GHPS Paris, France
www.hivr4p.org
13
Scholars
Bellington Vwalika
Wendy Winnall
Dana Watnick
Xilin Wu
Huamian Wei
Wildeman Zapata
Zambia Emory HIV Research Project, Zambia
Albert Einstein College of Medicine, United States
State Key Laboratory for Infectious Disease Prevention and
Control, National Center for AIDS/STD Control and Prevention,
Chinese Center for Disease Control and Prevention, China
Dawn Renee Weinman
Tendayi Westerhof
Women’s Health HIV and AIDS Southern Africa, Zimbabwe
Lee Adam Wheeler
Harvard Medical School, United States
Constantinos Kurt Wibmer
South Africa
SCHOLARS
14
The University of Melbourne, Australia
The University of Hong Kong, Hong Kong
Universidad de Antioquia, Colombia
Susanne Ziegler
Heinrich Pette Institute, Leibniz Institute for Experimental
Virology, Germany
Michael Zulu
Doris Duke Medical Research Institute, HIV Pathogenesis
Programme, University of KwaZulu-Natal, South Africa
Tuesday, 28 October
Opening Plenary: State of the Art Biomedical Prevention in 2014
PL01.01
PL01.02
Prospects for an Antibody-based HIV Vaccine
Advances in Antiretroviral-Based Prevention
Research
National Institute for Communicable Diseases of the NHLS, Centre for
HIV & STI’s: HIV Virology, Johannesburg, South Africa, 2University of
the Witwatersrand, Johannesburg, South Africa
1
Intensive efforts to develop a preventive HIV vaccine have been
significantly bolstered by recent discoveries of broad and potent
neutralizing antibodies in some HIV infected humans. Many of these
antibodies have unusual genetic features that present a significant
challenge for conventional vaccine approaches. Nevertheless,
longitudinal studies are revealing how such lineages evolve and
enabling the identification of germline B cells that an HIV vaccine would
need to stimulate. Parallel viral genetic analysis further demonstrates
how viral evolution shapes these responses. Collectively such studies
in HIV- infected humans who develop broadly cross-neutralizing
antibodies are providing important clues for HIV vaccine design.
Jared Baeten1
University of Washington, Seattle, WA, United States
1
Antiretroviral-based HIV-1 prevention strategies — including
antiretroviral treatment (ART) to reduce the infectiousness of HIV-1
infected persons and oral, topical, and injectable antiretroviral preexposure prophylaxis (PrEP) for uninfected persons to prevent HIV1 acquisition — are groundbreaking new approaches for decreasing
HIV-1 spread. The past three years have seen substantial advances in
knowledge regarding ART and PrEP for HIV-1 prevention, including
definitive demonstration in randomized trials that both ART and
PrEP reduce HIV-1 risk and the development of normative guidance
for prescribing these HIV-1 prevention strategies. Ongoing research
into longer-acting PrEP agents could add new prevention options.
There are numerous parallels in the opportunities ahead for ART and
PrEP, including evaluating successful approaches to deliver these
interventions to realize maximum population benefits, developing
messaging that speaks to potential ART and PrEP users, understanding
how sufficiently high adherence can be sustained to achieve high
effectiveness, and integrating these strategies into health systems.
Achieving success in antiretroviral-based prevention will demand the
full force and breadth of prevention research and advocacy.
PL01.03
Comprehensive HIV Prevention: Synergy
Between Vaccine and Non-Vaccine Modalities
Anthony S. Fauci1
National Institute of Allergy and Infectious Diseases, National Institutes
of Health, Bethesda, MD, United States
1
www.hivr4p.org
15
PLENARY SESSIONS
Lynn Morris1,2
Plenary 02: Targeting Biomedical Preventions to Different At-Risk Populations
PL02.01
PL02.03
Tailoring Biomedical Preventive Interventions
for Key Populations: Towards Safety, Efficacy,
Effectiveness
Tailoring Interventions to Different Populations
Chris Beyrer
1
Pontiano Kaleebu1
Basic Sciences, MRC/UVRI Uganda Research Unit on AIDS, Entebbe,
Uganda
1
Johns Hopkins Bloomberg School of Public Health, Epidemiology,
Baltimore, MD, United States
1
PL02.02
Working with Special Populations within HIV
Prevention Intervention Programs and Trials
Bridget G. Haire1,2
University of New South Wales, School of Public Health and
Community Medicine, Sydney, Australia, 2University of Sydney, Centre
for Values, Ethics and the Law in Medicine, Sydney, Australia
1
PLENARY SESSIONS
It is well accepted that effective HIV prevention must target the high
risk populations in particular local epidemics, and that working
collaboratively with these affected populations to design and deliver
prevention interventions is an ethical requirement that can maximise
uptake and acceptability.
In 2014, we have a greater than ever range of interventions to prevent
HIV, but access, sustainability and adherence remain major issues, and
there is no established optimal formula for implementing combination
HIV prevention approaches in the specific population groups who need
them most. In addition, some of the social and political barriers to
effective implementation within key populations have become more
entrenched. This poses particular problems both for ongoing research
into new interventions and for programmatic implementation of proven
interventions.
This plenary will look at the evidence about the use of HIV prevention
in particular populations, and consider how HIV prevention research and
implementation can work within a broader human rights framework that
takes into account social and political barriers that affect vulnerability,
sustainability and adherence to prevention interventions.
16
UNAIDS estimates there were 6,300 new infections a day in 2012,
with 95% of these in low- and middle-income countries. Advances in
biomedical HIV prevention are giving us hope that the HIV epidemic can
be brought under control.
Since Biomedical, Behavioural and Structural interventions intersect, it is
important to understand and engage the different priority populations
and communities in order to tailor interventions for maximum benefit.
Social behavioural, health systems, community participation, epidemic
stage, and policy are some factors that will determine the benefits for
priority populations of the advances made in biomedical HIV prevention.
For example, in most generalized epidemics effective prevention
approaches are required for women who are at a disproportionate risk
of HIV acquisition for biological as well as social reasons. Mobilising and
meaningfully engaging key populations, such as people who inject drugs
(PWID), men who have sex with men (MSM), sex workers (SW), and the
fisherfolk (FF) in some countries will take different forms in local contexts
for these often hard to reach populations. For example, in our studies in
Uganda among SW, interventions are provided through special clinics,
staffed by skilled health workers with knowledge and understanding of
the sex work milieu, offer friendly services and involvement of peers in
mobilisation and follow up. For both SW and FF, localising clinics and
services nearer to their places of work is essential to their involvement.
For FF this includes providing services that are open hours when they
are not working. Examples of other tailored programmes engaging
populations such as MSM and adolescents will be discussed.
Investing resources and focusing national HIV prevention campaigns
on well-defined strategies for and with populations that are key to
the epidemic and key to the response will increase the potential for
biomedical interventions to slow the HIV epidemic, reducing new
infections.
Plenary 03: Importance of Mucosa in Prevention Research
PL03.01
PL03.02
Utilizing NHP Models to Understand Mucosal
HIV Transmission and Dissemination
Genital Inflammation and HIV Risk in
Prevention Research
Jake D. Estes1
Jo-Ann Passmore1,2,3
Leidos Biomedical Research, Inc., Frederick National Laboratory for
Cancer Research, AIDS & Cancer Virus Program, Frederick, MD, United
States
1
Over 80% of sexual HIV-1 transmissions originate from a single viral
variant, but the underlying basis for this transmission bottleneck
remains to be elucidated. Nonhuman primate models of mucosal virus
transmission allow opportunities to gain insight into the basis of this
mucosal bottleneck. This talk will focus on studies that utilize NHP
models to understand i) the host innate antiviral responses during the
earliest time points after mucosal SIV challenge and ii) the lymphatic
drainage pathways of viral dissemination in order to better elucidate the
earliest events of virus mucosal transmission and potential intervention
strategies.
University of Cape Town, Institute of Infectious Disease and Molecular
Medicine, Cape Town, South Africa, 2National Health Laboratory
Service, Cape Town, South Africa, 3CAPRISA, Durban, South Africa
PL03.03
Mucosal Immune Assays in HIV Vaccine Clinical
Trials
Omu A. Anzala1
KAVI - Institute of Clinical Research (KAVI-ICR), University of Nairobi,
College of Health Sciences, Nairobi, Kenya
1
Background: Mucosal immune responses and mucosal sampling from
genitourinary (GU) and gastrointestinal (GI) tracts are at an increased
focus for HIV vaccine research and development as well as other
HIV prevention and treatment strategies that are targeted at mucosal
surfaces. This is in realization that mucosal surface forms the major route
of HIV acquisition and transmission across the world.
Collection of mucosal samples during the conduct of clinical trials is
associated with significant operational challenges, expenses, as well as
some risk and discomfort to study participants. It is therefore critical
that appropriate measures are taken into account including, (clinical,
behavioral, and demographic characteristics) from study participants
so that factors that may influence mucosal immunology and thus the
interpretation of assay data are efficiently captured in parallel with
mucosal specimens during the conduct of clinical trials. KAVI-Institute
of Clinical Research at the University of Nairobi, and others have studied
the acceptability and tolerability of repeated mucosal sampling in clinical
trial participants as well as other participants at low risk of HIV infection.
To this end the samples obtained were also used to establish and
standardize the immune assays for adaptation in evaluation of mucosal
immune responses in clinical trials.
Discussion: Repeated mucosal sampling is achievable both in HIVinfected and in healthy adult HIV uninfected clinical trials participants.
The sampling methods that have been studied include saliva, oral fluids,
semen, cervical, vaginal, rectal, and gut. Participants consented to most
specimen collection methods with the exception of rectal sampling.
Samples obtained are of good quality for process, analysis and can be
standardized for both T/B cell as well antibody assays for adaptation
for use in evaluating mucosal immune responses in HIV vaccine clinical
trials in addition to peripheral samples.
www.hivr4p.org
17
PLENARY SESSIONS
1
Friday, 31 October
Closing Plenary : Mind the Gap: Bridging from Trial Success to Access
PL04.01
PL04.02
Translating “Controlled” Trial Results to the
Cities & Villages of Africa: Past, Present, & Future
Scaling-up HIV Prevention Science from the
Laboratory to the Village
Douglas N. Shaffer1, Robert W. Eisinger1, Deborah L. Birx1
Alex G. Coutinho1
Office of the U.S. Global AIDS Coordinator, U.S. Department of State,
Washington, DC, United States
1
1
Infectious Diseases Institute, Director, Kampala, Uganda
HIV prevention science requires a critical mass scale-up to have a
population impact. The talk will share experiences of scale-up with
medical male circumcision and PMTCT and seek to learn lessons that
can be utilized for emerging HIV prevention science.
PL04.03
Antiretrovirals for Prevention
Glenda Elisabeth Gray1
Department of Paediatrics, at the Perinatal HIV Research Unit,
University of the Witwatersrand, Johannesburg, South Africa
1
PLENARY SESSIONS
18
Tuesday, 28 October
Symposium 01: Overcoming Barriers to Broadly Neutralizing Antibody Induction
SY01.01
SY01.03
Prevention Strategies Based on HIV-1
Neutralizing Antibodies
HIV Envelope Interactions with the Progenitor
BCRs of Narrow and Broadly Neutralizing
Antibodies
National Institutes of Health, Vaccine Research Center, NIAID,
Bethesda, MD, United States
1
Effective vaccines often generate protective antibody responses that
are similar to those produced during natural infection. During HIV-1
infection, most individuals generate neutralizing antibodies that arise
too late to be protective. However, during the course of infection,
some individuals develop extraordinarily potent and broadly reactive
neutralizing monoclonal antibodies (bNAbs) that have been recently
isolated and studied in detail. Such antibodies, when isolated and
used for immunoprophylaxis in animal models, can completely
block HIV infection. Thus, bNAbs provided a template for the kind of
immune response we aim to elicit via immunization. The analysis of
their structural mode of recognition and how these antibodies develop
to attain effective functional neutralization; i.e. genetic pathways of
affinity maturation, can impact HIV vaccine design and prevention
efforts in several ways including 1) structure-based vaccine design, 2)
understanding antibody evolution to optimize immunization strategies
and 3) direct immunoprophylaxis by passive antibody or gene-based
vectors. Highlights from each of these areas will be discussed.
SY01.02
Broadly Neutralizing Antibody Responses in
HIV-1 Infection: Insights from a Large Cohort
and from Individuals
Pascal Poignard1
IAVI Neutralizing Antibody Center at The Scripps Research Institute, La
Jolla, CA, United States
1
Leonidas Stamatatos1
Seattle Biomedical Research Institute, Seattle, WA, United States
1
SY01.04
HIV-1 Mimicry of Host Antigens: Hiding in Plain
Sight to Evade Immunity
Garnett Kelsoe1
Duke University, Immunology and Human Vaccine Center, Durham,
NC, United States
1
Although controversial, it has been proposed that antibodies (Ab) that
neutralize multiple HIV-1 clades (bNAbs) are characteristically poly- or
reactive to host antigens (autoreactive). We have identified a number of
conserved mammalian autoantigens that are avidly bound by different
bnAbs, and have shown in mice that relaxation of immune tolerance
significantly elevates Ab responses to HIV-1 neutralizing epitopes. The
generation of knockin mice that express human bnAb variable regions
commonly has shown that the cross-recognition of host antigens is
sufficient to block B-cell development or maturation by known tolerance
mechanisms. Knockin strains that express non-neutralizing Ab variable
regions support normal B-cell development. There is, however, no broad
agreement as to whether HIV-1 mimicry of host determinants represents
a crucial limitation of protective immunity to HIV-1 or is an ancillary
property of Abs generated over the course of a chronic infection. I shall
demonstrate that poly- and autoreactivity are characteristic of HIV1 bnAbs but not high affinity, strain-specific or non-neutralizing HIV1 Abs generated in chronically infected individuals. As a class, bnAbs
are significantly more polyreactive and autoreactive than other HIV-1
Abs even though they are generated under similar conditions of chronic
infection and inflammation. Interestingly, four bnAbs specific for the
HIV-1 CD4 binding site (CD4bs) (VRC01, VRC02, CH106, and CH103)
bind a common human autoantigen, ubiquitin ligase E3A (UBE3A). The
UBE3A protein competitively inhibits gp120 binding to VRC01 and,
remarkably, avidity for UBE3A among these four bnAbs was correlated
with neutralization breadth. The absence of poly- or autoreactivity
suggests that non-neutralizing HIV-1 Abs may dominate responses
because they not be subject to control by immunological tolerance. The
rarity and delayed development of CD4bs bNAb in infected individuals
may be the consequence of viral mimicry of human UBE3A.
www.hivr4p.org
19
NON-ABSTRACT DRIVEN
John Mascola1
Tuesday, 28 October
Symposium 02: Advances in Animal Models
SY02.01
SY02.03
Non-Human Primate Models for New
Prevention Strategies of HIV Transmission
Prevention Studies in Nonhuman Primate
Models
Roger Le Grand1
Jeffrey Lifson1
CEA, IDMIT, Fontenay-aux-Roses, France
1
Nonhuman primate (NHP) studies significantly contribute to the
understanding of HIV sexual transmission mechanisms and to the
development of approaches aimed at preventing infection. In this
presentation we will review studies in NHP for assessing microbicides
and vaccines efficacy. Future directions for preventive strategies will
be discussed.
SY02.02
How Mice to Monkeys Inform Human B Cell
Repertoire Responses
National Institutes of Health, National Cancer Institute, Bethesda, MD,
United States
1
SY02.04
The Role of Humanized Mice in Advancing HIV
Vaccine and Eradication Studies
Todd M. Allen1
Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United
States
1
Gunilla B. Karlsson Hedestam1
Karolinska Institutet, Department of Microbiology, Tumor and Cell
Biology, Stockholm, Sweden
1
NON-ABSTRACT DRIVEN
Continued efforts to evaluate HIV-1 envelope glycoprotein (Env)-based
vaccines in vivo are needed to identify immunogens and immunization
regimens that elicit responses that protect against HIV-1 acquisition. HIV1 is a highly neutralization-resistant virus due to conformational and
glycan shielding of conserved Ab determinants making the development
of an effective vaccine extremely challenging. Understanding current
obstacles to elicit broadly neutralizing Abs that target poorly accessible
epitope on the native Env spike requires the use of both small animal
models and non-human primates (NHPs). Among small animals, mice
are most useful for basic mechanistic studies due to the ease by
which they can be manipulated, the availability of numerous knockout strains, shorter experimental cycles and lower costs. On the other
hand, NHPs are more similar to humans, both in terms of the genetics of
antibody gene segments and the characteristics of immune cell subsets.
NHPs are also more amenable to detailed studies of vaccine-induced
responses due to the larger sample volumes that can be obtained and
the possibility to perform SHIV challenge studies, but they are costly.
Importantly, the choice of animal model should be determined based
on the question to be addressed to ensure maximal information output
while yet adhering to the 3R principle. Over the past several years, we
have established unique methodology to characterize B cell responses
in both mice and non-human primates including the isolation of
monoclonal Abs from single-cell sorted memory B cells. These studies
confirm that an in-depth understanding of the principles that govern
B cell activation, differentiation and affinity maturation is needed to
accelerate the development of a vaccine capable of stimulating broadly
neutralizing antibody responses against HIV-1. Such studies will require
several types of animal models including both mice and monkeys.
20
Efforts to determine how human immune responses to HIV can be
induced or augmented would be greatly facilitated by the availability
of a suitable small animal model. There have been a number of recent
advances in the development of small animal models for HIV, including
the recent generation of humanized BLT (bone marrow, liver, thymus)
mice where implanted human fetal CD34+ hematopoietic stem cells
(HSCs) become educated within transplanted autologous human thymic
tissues. This model has dramatically improved the ability of humanized
mice to support productive HIV replication and CD4+ T cell depletion,
as well as recapitulate human cellular and humoral responses to HIV.
Here we discuss recent data illustrating the ability of this model to
support HIV-specific and vaccine-induced immunity to HIV. We will also
discuss applications of this model for elucidating critical aspects of HIV
transmission, dissemination, and HIV-specific immunity as well as the
opportunities and limitations of this model to support HIV vaccine and
eradication efforts.
Tuesday, 28 October
Symposium 03: Treatment as Prevention: The Promise and the Perils
SY03.01
SY03.03
No Time to Lose - The Importance of Early HIV
Infection
Treatment as Prevention - How Can We Predict
Success?
Kimberly Powers1
Deenan Pillay1,2, Till Barnighausen1, Tulio De Oliveira1, Kobus
Herbst1, Frank Tanser1
Detection of early HIV infection is important for individual and public
health. This talk will review opportunities and challenges related to
the detection of early infection, as well as the implications of ongoing
transmission during this critical window.
SY03.02
A Tale of Four Treatment-as-Prevention Trials in
Africa
Frank Tanser1,2, Collins Iwuji1,3
University of KwaZulu-Natal, Africa Centre for Health and Population
Studies, Mtubatuba, South Africa, 2University of KwaZulu-Natal, School
of Nursing and Public Health, Durban, South Africa, 3University College
London, London, United Kingdom
1
Africa Centre for Health and Population Studies, University of
KwaZulu-Natal, Mtubatuba, South Africa, 2University College London,
Division of Infection and Immunity, London, United Kingdom
1
With major trials now ongoing to explore the impact of a treatment as
prevention approach to HIV, it is critical that we consider the factors
which will predict and determine success, as defined by a reduction in
HIV incidence. We will consider three specific areas. Firstly, a prerequisite
for TasP impact is the linkage to and retention in care for those diagnosed
with HIV. The ability to deliver such linkage is critical to the success
of TasP. Secondly, we should expect an increase in absolute numbers
of individuals harbouring drug resistant viruses, commensurate with
an increasing coverage of antiretroviral treatment. On the one hand,
this may become a major impediment to maintaining a reduction in
incidence. By contrast, this could merely be a surrogate of treatment
coverage, with the benefits of therapy far outweighing the negative
impact of resistance mutants. Thirdly, within the now established era of
pathogen genomics, we have the capability to monitor the dynamics of
spread of HIV through a sampling frame for virus gene sequences. We
speculate that such approaches can predict a reduction of incidence in
the context of TasP, although this remains to be formally demonstrated.
SY03.04
TasP Promises… But What Is The Reality?
Valerie C Delpech1, Alison E Brown1
Public Health England, HIV & STI, London, United Kingdom
1
The clinical evidence of TasP is compeling and modelling studies are
inspirational but what is the reality? Can TasP reduce population level
incidence?
In this presentation we assess the impact of antiretroviral treatment on
HIV incidence, prevalence and undiagnosed infections using examples
from the UK, South Africa and other countries.
The importance of good HIV surveillance data including testing data,
HIV diagnoses by key groups, link to and retention in care, ART uptake
and viral load data are critical in monitoring our successes.
NON-ABSTRACT DRIVEN
The University of North Carolina at Chapel Hill, Department of
Epidemiology, Chapel Hill, NC, United States
1
www.hivr4p.org
21
Tuesday, 28 October
Symposium 04: Building Combination Prevention Trials
SY04.01
SY04.03
Building Combination Prevention: Trial Designs
The Role of Mathematical Modeling to Design,
Conduct, Interpret and Replicate HIV Prevention
Trials
Helen Weiss1
London School of Hygiene & Tropical Medicine, MRC Tropical
Epidemiology Group, London, United Kingdom
1
Marie-Claude Boily1
Imperial College London, Infectious Diseases Epidemiology, London,
United Kingdom
1
Combination prevention trials are based on synergies of multiple
prevention and treatment components, including interventions that
aim to change structures and environments. Such interventions are by
nature community-based rather than individually-based, and thus lend
themselves to community-randomized trials rather than individualrandomized trials.The impact of the intervention is then also assessed
at population-level. The ‘gold standard’ method for such evaluations
is the parallel-arm cluster randomised controlled trial (RCT), which is
logistically challenging, time-consuming, and expensive to implement.
We will discuss challenges of the cluster RCT, including the choice of
control arm in light of evolving standards of care for HIV treatment and
prevention, minimizing contamination between clusters, and methods of
randomisation to minimise imbalance between arms.We will also discuss
alternative designs and show examples of trials which have evaluated
the impact of community-level interventions including stepped-wedge
designs and adaptive designs. Methodological challenges in evaluating
combination prevention interventions will also be discussed, including
time-series models for surveillance data, hierarchical models to allow for
clustering in the design, and the use of causal inference methods which
are particularly useful for non-randomized designs
SY04.04
SY04.02
Scalable Adherence Interventions
Combination Prevention: From Trials to
Implementation
Sinead Delany-Moretlwe
1
University of the Witwatersrand, Wits Reproductive Health & HIV
Institute, Johannesburg, South Africa
1
NON-ABSTRACT DRIVEN
Combination HIV prevention is the application of several evidencebased interventions — including biomedical, behavioural and structural
interventions — to maximise population-level reductions in HIV incidence
in a specific setting. While strong evidence exists now for several
single approaches, evidence for how best to combine these different
interventions for maximal impact on HIV in different populations and
settings is scarce. The success of combination HIV prevention rests
on the identification of populations or places with highest rates of
infection, adaptation of interventions that are cost-effective and likely
to have highest impact on HIV in that population, and delivery of that
combination of interventions with sufficient scale, uptake and sustained
use. In this talk, we discuss how using a public health approach can
guide the pragmatic, localised application of evidence-based combination
approaches for particular populations. We highlight how advances in fields
as diverse as HIV phylogenetics and molecular epidemiology, market
research and implementation science may refine our understanding of
local transmission dynamics and enhance our ability to deliver feasible
and acceptable interventions successfully and with maximum impact. In
addition to monitoring HIV incidence trends, the importance of monitoring
the potential synergistic or antagonistic effects of these interventions in
particular populations, and the need for responsive programming in order
to minimise unintended consequences, will also be addressed.
22
Current prevention trials often aim to test combinations of different
prevention and treatment components introduced simultaneously or
sequentially in different risk populations. Choosing the best combination
package and deciding in which population it should be tested can be
challenging. Designing and interpreting the results of costly and large
community-based randomized trials (C-RCT) to evaluate complex HIV
prevention strategies can be equally challenging. Mathematical models
are widely used to predict the population-level impact of interventions
and inform policy decisions. In this talk, we will discuss why and how
mathematical models are also increasingly and innovatively being used
at the different stages of the clinical trial process: to inform product
development, design the intervention package, to help design and
conduct clinical trials, and to interpret and help generalize results. In
so doing, we will also discuss how mathematical models can be used
to address some of the challenges encountered in community-based
randomized trials.
Terrence F. Blaschke1,2
Bill and Melinda Gates Foundation, Discovery and Translational
Sciences, Seattle, WA, United States, 2Stanford University School of
Medicine, Internal Medicine, Stanford, CA, United States
1
Recognizing that suboptimal adherence is the major cause of lack
of effectiveness of ARVs for PrEP, scalable interventions to improve
adherence are needed. Many interventions have been proposed, but to
date none have been scalable and sustainable (Medication Adherence
Interventions, Evidence Report No. 208. AHRQ Publication No.12E010-EF, 2012). A limitation of most studies is the absence of reliable
measures of adherence before and after an intervention. Moreover, the
duration of benefit in those studies showing some increase in adherence
dissipates over several months. What is needed is a scalable approach
to identifying suboptimal adherence, monitoring those more likely
to continue or become poorly adherent due to known predisposing
factors, then focusing interventions on that cohort of patients. Due
to the correspondence of drug exposure to protection from infection,
sparse sampling of dosing information is insufficient, and detailed
dosing information itself, shared with the patient and the provider, can
significantly improve adherence. Modern technology allows detailed
dosing histories to be obtained unobtrusively and collected centrally
at a point in time when interventions can be applied. Combined with
approaches such as Managed Problem Solving (Gross et al., JAMA
Intern Med. 2013; 173:300-306) and Lifetime HIV Antiretroviral
Therapy Adherence Intervention: Timing Is Everything (Bangsberg and
Haberer, JAMA Intern Med. 2013; 173:306-7), progress towards scaling
interventions to large high-risk populations is now within reach.
Symposium 05: The Role of Structure Based Design in Vaccine Development and
Immunoprophylaxis
SY05.01
SY05.03
Structure-Based Design of Improved Antibodies
for HIV-1 Treatment or Prevention
Structure-Based Stabilization of the Prefusion
Closed HIV-1 Env Trimer
Louise Scharf1, Stuart A. Sievers1, Anthony P. West, Jr.1, Ron
Diskin1,2, Michel C. Nussenzweig3, Pamela J. Bjorkman1
Peter Dak Pin Kwong1
Over 30 years after the emergence of HIV-1, there is no effective
vaccine, and AIDS remains an important threat to global public health.
Following infection by HIV-1, the host immune response is unable to
clear the virus due to a variety of factors, including rapid viral mutation
and the establishment of latent reservoirs. The only target of neutralizing
antibodies is the trimeric envelope spike complex, but HIV-1 can usually
evade anti-spike antibodies due to rapid mutation of its two spike
glycoproteins. We are using structure-based protein design methods to
engineer antibodies that can resist some of the common routes of HIV-1
mutation, with the hope that the designed antibodies could be used in
passive immunotherapy methods for HIV-1 treatment and/or prevention.
SY05.02
The Stucture of the Glycan Shield of HIV
Max Crispin1
University of Oxford, Biochemistry, Oxford, United Kingdom
1
National Institutes of Health, Vaccine Research Center, Bethesda, MD,
United States
1
The HIV-1 envelope spike (Env) assumes diverse conformations to facilitate
dual roles in viral entry and immune evasion. An antigenically specific
immunogen — which only binds to broadly neutralizing antibodies and
not to non-neutralizing or poorly neutralizing antibodies — might lead
to increased elicitation of effective HIV-1-neutralizing antibodies. Here,
we report the mature unliganded crystal structure at 3.7 Å resolution,
for one of the most antigenically specific Env immunogens developed to
date, the SOSIP.664 variant of the BG505 strain of HIV-1. The unliganded
structure closely resembled recently determined antibody-bound structures
of BG505 SOSIP.664 and appeared to be in the prefusion closed state.
Because SOSIP.664 retains the ability to transition to an open CD4-bound
state, which is recognized by poorly neutralizing antibodies like the CD4induced antibody 17b as well as antibodies against the immunodominant
V3 region like 447-52D, we used structure-based design to identify mutants
with reduced binding to 17b and 447-52D. Select variants with introduced
disulfide bonds or proline mutations no longer bound CD4-induced or
V3-directed antibodies, even in the presence of CD4. Unexpectedly, these
variants did retain the ability to bind to CD4, with electron microscopy
reconstructions indicating them to remain in a closed state, even when
bound by CD4. Structure-based design can thus be used to improve the
antigenic specificity of HIV-1 Env, with conformationally fixed variants
allowing for detailed mechanistic dissection of ligand-induced transitions.
SY05.04
Reductionist Vaccine Design to Induce Broadly
Neutralizing Antibodies against HIV
William Schief1,2,3
Immunology and Microbial Science, The Scripps Research Institute, La
Jolla, CA, United States, 2IAVI Neutralizing Antibody Center at The Scripps
Research Institute, La Jolla, CA, United States, 3Ragon Institute of MGH,
MIT and Harvard, Boston, MA, United States
1
One of the major thrusts of our work on HIV vaccine development has
been to define immunogens and regimens to induce VRC01-class broadly
neutralizing antibodies. In this work we have defined a “reductionist
approach” to vaccine design that could be applied to other epitopes on
HIV and other pathogens. We will review this line of research, and weigh
opportunities and challenges to induce broadly neutralizing antibodies
against the VRC01 epitope and other conserved HIV epitopes, in light of
recently published structures of the HIV trimeric spike.
www.hivr4p.org
23
NON-ABSTRACT DRIVEN
California Institute of Technology, Division of Biology and Biological
Engineering, Pasadena, CA, United States, 2(present address) Weizmann
Institute of Science, Department of Structural Biology, Rehovot, Israel,
3
Rockefeller University, Laboratory of Molecular Immunology, New
York, NY, United States
1
Symposium 06: Harnessing Antibody Effector Functions
SY06.01
SY06.03
Evaluating HIV-1 Vaccine Induced Antibody
Effector Function to Advance Vaccine
Candidates
Human Non-neutralizing HIV-1 Envelope
Monoclonal Antibodies Limit the Number of
Founder Viruses during SHIV Mucosal Infection
in Rhesus Macaques
Georgia Tomaras1
Duke University, Duke Human Vaccine Institute, Departments of
Surgery, Molecular Genetics and Microbiology, and Immunology,
Durham, NC, United States
1
This presentation will discuss how to utilize the diverse array of humoral
measurements to evaluate and inform decisions for advancing HIV1 vaccine candidates. Antigen specific antibody responses, some of
which are Fc-mediated, correlate with decreased risk of HIV-1 infection.
Vaccine induced polyclonal antibody responses and antibody effector
function differ profoundly across vaccine regimens. Knowledge of
protective and potentially detrimental immune responses can be
garnered by systematically examining the correlations between such
responses and risk of HIV-1 infection after exposure. Currently, the
HIV-1 vaccine field has an array of humoral immune correlates of risk
obtained from both human efficacy studies, as well as from nonhuman
primate studies. Analyses that delineate the interplay among immune
responses (innate, cellular, and humoral), host genetics (e.g., FcR
expression, HLA), and HIV-1 virus diversity (i.e., within the populations/
geographic regions enrolled in vaccine efficacy studies) are likely to
be transformative for the next phase of HIV-1 vaccine design and
evaluation. New data for understanding and evaluating vaccine
induced antibody breadth and effector function will be presented.
SY06.02
A Systems Serology Approach to Vaccination
Margie E. Ackerman1, Chris Bailey-Kellogg2, Galit Alter3
Dartmouth College, Thayer School of Engineering, Hanover, NH, United
States, 2Dartmouth College, Computer Science, Hanover, NH, United
States, 3Massachusetts General Hospital, Ragon Institute of MGH, MIT,
and Harvard, Cambridge, MA, United States
1
NON-ABSTRACT DRIVEN
In vivo, antibody responses to vaccination or natural infection are highly
polyclonal, with multiple somatic variants directed to multiple epitopes
on multiple antigens. This diversity of variable domain recognition
characteristics is further complemented by diversity in the constant
domain´s ability to interact with innate immune receptors such as FcgR.
Antibody-based protection is thus derived from the sum of specificities
and activities of this polyclonal humoral milieu. We demonstrate a highthroughput, high-content platform for the biophysical interrogation of
the innate immune recruiting capacity of polyclonal, antigen-specific
antibodies capable of parsing the complex humoral milieu into components
that can be associated with relevant clinical, genetic, or functional
characteristics. Thousands of measurements of the humoral immune
response can be determined, providing a comprehensive landscape of
antibody activity and better understanding of the characteristics and
development of both protective and pathological humoral immunity that
may be critical to understanding vaccine-mediated protection. When
applied to chronically HIV infected or vaccinated subjects, dramatically
different antibody profiles are observed, and these profiles can be used
to develop computational models of antibody activity that provide insight
into mechanisms of vaccination.
24
Sampa Santra1
Center for Virology and Vaccine Research, Boston, MA, United States
1
SY06.04
Combating Cell-Cell Transmission of HIV with
Antibodies - Slaying the Trojan Horse?
Stephen Kent1
University of Melbourne, Department of Microbiology and
Immunology, Peter Doherty Institute, Melbourne, Australia
1
Developing vaccines with broad neutralizing function is a holy grail of
HIV research, but there is a potential problem with this approach. HIV
is readily isolated from cells within the semen of HIV-infected men, and
this cell-associated virus (dubbed “Trojan horse leukocytes”) is more
infectious than free virions within semen. Data emerging from clinical
studies in humans and studies conducted in the non-human primate
model of HIV infection provide evidence for a role of cell-associated
virus in HIV transmission. The extent to which neutralizing antibodies
or MHC-restricted CTLs can stop cell-cell virus transmission in unclear.
ADCC antibodies can however readily recognize foreign infected cells
and may therefore assist in preventing cell-cell transmission of HIV. We
hypothesize ADCC against infected cells may be one mechanism for the
partial efficacy of the RV144 trial.
Symposium 07: Novel Formulations and Sustained Delivery of Antiretrovirals
SY07.01
SY07.03
Injectable and Implantable Antiretroviral
Strategies for HIV Prevention
Novel On-Demand Vaginal Microbicide
Formulations: Tablets and Films
Ian McGowan1
Jill Schwartz1
University of Pittsburgh School of Medicine, Department of Medicine,
Pittsburgh, PA, United States
1
1
A fundamental challenge associated with the use of antiretroviral preexposure prophylaxis (PrEP) is that individuals must be willing to selfadminister such products daily or in a peri-coital fashion for the strategy
to be effective. Multiple Phase 2B/3 prevention trials have demonstrated
that adherence to these regimens can be sub-optimal and consequently
PrEP interventions have had highly variable effectiveness depending
on the population studied and their degree of product adherence. In
trials where PrEP adherence has been high, the intervention has been
highly effective. Long acting injectable or implantable products are
currently used for contraception and this strategy is being evaluated
for delivery of antiretroviral drugs. This approach has the potential to
provide antiretroviral PrEP for up to 3 months with a single injection and
to circumvent adherence problems associated with daily or per-coital
administration of oral/topical PrEP agents. TMC278 LA (rilpivirine) and
GSK 744 (cabotegravir) have both completed Phase 1 evaluation and
are moving into later stage development. The purpose of this talk is to
review these two products as well as the potential for the development
of long acting single antiretroviral and combination antiretroviral/
contraceptive implantable products for HIV prevention.
SY07.02
Intravaginal Rings for HIV Prevention in Women
Thesla Palanee-Phillips1
Wits Reproductive Health and HIV Institute, Johannesburg,
South Africa
1
CONRAD Eastern Virginia Medical School, Arlington, VA,
United States
SY07.04
Nanotechnology for Drug Delivery
Kim A. Woodrow1
University of Washington, Bioengineering, Seattle, WA,
United States
1
The delivery of drug combinations is a paradigm for treatment of HIV/
AIDS, cance and drug resistant bacterial infections. My laboratory is
interested in the application of engineered nanomaterials to control
the spatial and temporal delivery of a combination of agents (small
molecules, biologics, and conjugates). Strategies to combine chemically
incompatible agents may facilitate the discovery of unique drug-drug
activities, particularly unexplored combination drug synergy. In this
presentation, I will summarize our efforts to develop polymeric delivery
systems for the combination delivery of antiretroviral (ARV) drugs for HIV
prevention and treatment. ARV drug combinations have the potential
to enhance the efficacy of current prevention strategies by overcoming
low user adherence, and harnessing drug combinations with synergistic
activity and breadth of coverage against the global diversity of HIV
variants. We have developed polymeric particulate and fiber carrier
systems for delivering ARV drug combinations. The flexibility to design
the nanoarchitecture of these polymeric carriers, combined with the
versatility of drugs that can be encapsulated for controlled release,
motivate the use of these systems for topical, injectable or oral delivery
of combination agents in the fight against HIV.
NON-ABSTRACT DRIVEN
Of the more than 35 million people living with HIV, half are women. Most
women acquire HIV through heterosexual intercourse. In fact, women
are twice as likely as men to acquire HIV during vaginal sex, due in part
to biological factors that make them more susceptible. Young women
are especially vulnerable. Efforts to promote abstinence, monogamy and
the use of male condoms have not been enough to stop the epidemic,
nor are these approaches practical in many settings. In southern Africa,
young women are up to five times more likely to become infected with
HIV than young men. Microbicides are products applied inside the vagina
or rectum to protect against HIV though sex. Although microbicides are
not yet available for widespread use, researchers are making significant
strides in the development and clinical evaluation of both vaginal and
rectal microbicide products. Microbicides that incorporate antiretroviral
(ARV) drugs are showing particular promise. A vaginal microbicide could
potentially give women the means to protect themselves against HIV.
Vaginal microbicides are being designed in many forms, including gels,
films and rings, which release an active ingredient gradually over time.
Two products are currently in Phase III trials.
Vaginal rings are products designed to allow for the slow delivery of a
drug or multiple drugs to cells inside the vagina over a period of weeks
or months. One is testing a vaginal gel containing the ARV tenofovir
used before and after sex, while two trials are evaluating a vaginal ring
containing dapivirine that women use for a month at a time. I will focus
on the use of intravaginal rings in HIV prevention research.
www.hivr4p.org
25
Roundtable 01: Safer Sex In 2014: Has the Paradigm Shifted?
RT01.01
RT01.03
What Is Safer Sex in 2014? Understanding the
Biology of HIV Prevention
Missing the Future is not an Option
James A. McIntyre1,2
AIDS Foundation of Chicago, Prevention Advocacy/IRMA, Chicago, IL,
United States
1
Anova Health Institute, Johannesburg, South Africa, University of
Cape Town, School of Public Health & Family Medicine, Cape Town,
South Africa
1
Jim Pickett1
2
RT01.02
Communicating a New Safer Sex Paradigm: The
Science of Communication
Lebogang Ramafoko1
Soul City, South Africa
1
It´s not an opinion, it´s a fact. The paradigm of what “safer sex” means
has changed. Because we now have successful, scientifically proven
biomedical prevention strategies, “safer sex = condoms” has evolved to
“safer sex” with a drop-down menu. In the United States, not everyone
is comfortable with this expansion of prevention choices. Speaking to an
assembly in Frankfurt, Germany in 1963, John F. Kennedy said, “Change
is the law of life. And those who look only to the past or the present are
certain to miss the future.” So yes, change is the law. It´s good that the
law is also to buckle up, because the evolution of “safer sex” has been
a bumpy ride so far, and continues to be. What is this all about? Can we
ride it out? What needs to be done to smooth our way now and better
prepare for the new biomedical tools we will certainly have? Staying
stuck in the past and mired in the present isn´t going to get us anywhere
close to zero.
RT01.04
Policy and Program: Perspectives on Safer Sex
Nduku Kilonzo1,
LUCT, Kenya
1
NON-ABSTRACT DRIVEN
26
Roundtable 02: Prevention of Mother-to-Child Transmission Revisited
RT02.01
RT02.03
The Role of PMTCT and Very Early Treatment in
the Elimination of Pediatric HIV Infection
Prevention of Mother-to-Child Transmission
Revisited
Sarah W Read1
Louise Kuhn1
NIAID, Division of AIDS, Rockville, MD, United States
Columbia University, New York, NY, United States
1
1
RT02.02
In light of recent data on the challenges of curing HIV infected babies,
this session will review current approaches in PMTCT and how these
may need to be modified. Conceptual distinctions between prevention
and treatment in the maternal-infant context will be made and the
special issues related to each discussed.
The Case for Early Antiretroviral Treatment
towards HIV Cure in Perinatal Infection
Deborah Persaud1
Johns Hopkins University School of Medicine, Baltimore, MD, United
States
1
RT02.04
PMTCT: Mind the Gaps!
Lee Fairlie1
Paediatrics, Wits RHI, Johannesburg, South Africa
1
Although huge gains have been made in PMTCT with significant
reductions in the numbers of HIV-infected infants, there are a few areas
that still need attention to improve care for women and their infants to
ensure that current gains continue. This discussion will include where
the current programmatic and research gaps are and highlight where
there may be failures, particularly related to high-risk populations.
NON-ABSTRACT DRIVEN
The reservoir for HIV in resting memory CD4+ T cells precludes cure
with current antiretroviral drugs. Substantially limiting the size of the
latent reservoir is the first step towards drug free remission and cure. In
perinatal HIV infection, early antiretroviral treatment significantly alters
the size of latent viral reservoirs. The unique aspects of the neonatal
immune system coupled with the potential to initiate very early
treatment therefore provide a unique opportunity to assess very early
treatment and drug-free remission in neonates. This is exemplified in the
case of the ‘Mississippi Child” who experienced 28 months of drug free
remission following 18-months of antiretroviral treatment starting at 30
hours of life. The rationale for clinical trials aimed at achieving drug free
remission and cure in perinatal HIV infection will be discussed.
www.hivr4p.org
27
Roundtable 03: Diffusion of Innovation: Accelerating Along the Research to Rollout Continuum
RT03.01
RT03.04
A Case Study of Circumcision
Diffusion of Innovation: A South African
Perspective
Agnes Binagwaho1
Ministry of Health, Rwanda
1
Yogan Pillay1
Department of Health, South Africa
1
RT03.02
Male Circumcision and Pre-Exposure
Prophylaxis (PreP) Roll Out in Kenya: A Tale of
Two Innovations
Peter Cherutich1
Ministry of Health, Nairobi, Kenya
1
RT03.05
From Research Result to Public Health Impact:
What Have we Learned and How Can We Do it
Better and Faster
Mitchell J. Warren1
AVAC, New York, NY, United States
1
RT03.03
HIV Prevention: Moving from Evidence to
Demonstration Projects to Policy and Programs
Connie Celum1
University of Washington, Department of Global Health & Medicine,
Seattle, WA, United States
1
NON-ABSTRACT DRIVEN
Recent oral PrEP trials demonstrate that biomedical prevention
interventions have strong behavioral components in terms of uptake
and adherence. It is important to understand user perspectives about
antiretroviral-based prevention which impact product use; relevant
disciplines for this research are user-centered design and mental models.
When products are demonstrated efficacious, diffusion of innovation
research and social marketing can be used to design demand creation
and delivery models. Provider perspectives need to be understood
and factored in designing models for delivery of new technologies,
training needs, and policies. There are important opportunities to learn
from implementation of novel HIV prevention technologies as they
are shown to be efficacious. Demonstration projects are important to
evaluate communication strategies about efficacy and decision tools to
help potential users decide about product use, acceptable and feasible
strategies to support adherence, and cost-effectiveness of delivery.
These demonstration projects should be conducted in parallel with
clinical development of sustained release and less user-dependent
formulations, so that a mix of HIV prevention methods can be offered
to persons at risk.
28
The presentation will focus on historical experiences along the researchto-rollout continuum in HIV prevention and related fields to outline
specific lessons that could be applied going forward with upcoming HIV
prevention research results. These lessons will be used to explore what
should be done in future product introduction, who should be doing it,
and when different aspects of rollout should take place.
Roundtable 04: Research Where and With Whom it Matters
RT04.01
RT04.03
MSM: Long Road to Trial Participation in Africa
HIV Prevention Trials in Developing Countries:
Challenges and Opportunities
KEMRI-Wellcome Trust Research Programme, Kilifi, Kenya, 2University
of Oxford, Headington, United Kingdom, 3Amsterdam Institute for
Global Health and Development, Amsterdam, Netherlands
1
RT04.02
We are not Guinea Pigs: HIV Clinical Research
for Smarties
Udom Likhitwonnawut1
AVAC, Chiang Mai, Thailand
1
HIV-related stigma makes HIV research on marginalized populations in
Thailand challenging, particularly in studies involving populations with a
long-standing history of persecution and ill-treatment such as drug users
and sex workers. Despite the low HIV prevalence in the general Thai
population, HIV prevalence and incidence among injecting drug users
and MSM are consistently higher than other groups. For this reason, these
groups have become favorite subjects of various HIV prevention studies.
Involving criminalized, persecuted populations in clinical trials can lead
to disputes and opposition from various stakeholders. In Thailand, there
are some studies involving these marginalized populations that did not
invest sufficient resources and time working with other stakeholders. As
the result, the studies encountered opposition from stakeholders. The
opposition often led to spending more time and resources to enroll trial
participants, discrediting the results of the study, and a feeling of distrust
between the stakeholders. Respect and adherence to ethical research
principles can help researchers to overcome the resistance and disputes
caused by the study. How and where a study is conducted and staffed can
significantly affect informed consent process and the ability of potential
trial participants to make free decisions. The principle of beneficence
and “do no harm” can guide researchers on how to provide the best
HIV prevention methods to trial participants. Applying other HIV ethical
guidelines such as Good Participatory Practice (GPP) can also strengthen
community engagement by involving community members in HIV trials
and help create meaningful community participation in HIV research, as
well as reducing stigma, and increasing knowledge and understanding
about HIV clinical research. Lastly trial sponsors and funders have a
responsibility in site selection to ensure that the sites have the capacity
and capability for meaningful stakeholder engagement.
Fareed Abdullah1
South African National AIDS Council, South Africa
1
RT04.04
Engaging Adolescents and Young Adults in HIV
Prevention Trials
Sybil Hosek1
Stroger Hospital of Cook County, Psychiatry, Chicago, IL, United States
1
RT04.05
African Couples Testing Matters: Most
Transmissions Are in Marriage, Most
Pregnancies Are Not Immaculate Conceptions,
and Women Aren´t Just Incubators
Susan Allen1, William Kilembe1, Mubiana Inambao1, Amanda
Tichacek1, Eric Hunter2, Rwanda Zambia HIV Research Group
Rwanda Zambia HIV Research Group-Emory University, Pathology &
Laboratory Medicine, School of Medicine, Atlanta, GA, United States,
2
Emory University, Pathology & Laboratory Medicine, School of
Medicine, Atlanta, GA, United States
1
More than 120 million Africans have been tested for HIV. Despite clear
evidence that the majority of transmissions in Africa occur within stable
heterosexual couples—often during pregnancy from husbands not
known to be HIV positive—fewer than 5% of couples have been tested
and counseled together.
In this context, and because two people are required for HIV transmission,
we argue that all prevention efforts and therapy should start with Couples’
Voluntary HIV Counseling and Testing (CVCT). CVCT is effective and
affordable. CVCT is endorsed by the WHO and training materials are
available from the CDC in several languages.
The US government has recently funded large-scale trials of treatmentas-prevention (TasP), yet compared to CVCT which prevents one HIV
infection for less than $500, TasP would cost $7000 per year. Most
African countries have per capita incomes of less than $1500 and annual
per capita health expenditures less than $100. In this setting, CVCT
should be a priority to achieve cost effective HIV prevention. Moreover,
in spite of CVCT effectiveness and affordability, some ‘population TasP’
trials deliberately withhold CVCT from African couples. This behavior
is of questionable ethics. CVCT should be provided to all couples as
standard of care. The impact and cost-effectiveness of CVCT and TasP
should be assessed separately.
www.hivr4p.org
29
NON-ABSTRACT DRIVEN
Eduard J. Sanders1,2,3
Symposium 08: Mucosal Barriers to Infection
SY08.01
SY08.03
Upper Female Genital Tract and Defining Safety
Mechanisms of HIV-1 Restriction and
Dissemination Mediated by Intestinal Mucosal
Myeloid Cells
Ruth M. Greenblatt1
University of California San Francisco, Clinical Pharmacy, Medicine,
Epidemiology and Biostatistics, San Francisco, CA, United States
1
Gabriella Scarlatti1, Mariangela Cavarelli1
San Raffaele Scientific Institute, Milan, Italy
1
Multiple tissue sites of susceptibility to HIV infection are present
in the female genital tract. The specific portal of entry of virus may
depend on a range of circumstances including concurrent STIs, trauma,
hormonal exposures, male factors and the influence of topically applied
substances. The upper FGT, including the endocervix and endometrium,
are potential sites of vulnerability and should be considered in the
evaluation of prevention modalities. Additionally, many products,
including commonly used topical vehicles and hygiene aids, underwent
regulatory evaluation prior to the current era of sophisticated mucosal
immunologic research. These products and substances may need
reappraisal using modern methods, since adverse effects may occur.
SY08.02
Behavioral and Biological Factors Affecting HIV
Acquisition in Women
The infection of specific cellular niches by HIV-1 adopting mechanisms of
restriction of replication are a challenge to develop effective prophylactic
and therapeutic interventions. Specifically, the mucosal surfaces are the
predominant sites involved in the first steps of viral uptake and entry,
and CD4+ T cell depletion. There is increasing evidence that dendritic
cells (DC) and macrophages residing in the lamina propria of the mucosa
may be the first cellular targets mediating transmission. We have recently
provided the proof of principle of the active involvement of intestinal
lamina propria resident CD11c+ DCs in the uptake of HIV. Indeed, gut
DCs migrate towards and extend processes between the tight junctions
of columnar epithelium through a CCR5-dependent mechanism, take up
R5 but not X4 virus, and then transfer infection to T cells. These data
raise the question on which DCs subsets are mediating infection and, in
turn may affect the fate of the virus and the immuneresponse.
SY08.04
Douglas Kwon
1
Ragon Institute of MGH, MIT and Harvard, Massachusetts General
Hospital and Harvard Medical School, Cambridge, MA, United States
1
Rectal Microbicide Development: An Update
Ross D. Cranston1
University of Pittsburgh, Pittsburgh, PA, United States
1
NON-ABSTRACT DRIVEN
30
Symposium 09: Sustaining Durability of Responses
SY09.01
SY09.03
Clinical Translation of HIV Protein-Expressing
Cytomegalovirus Vectors
Adjuvants and Durability of Vaccine-Induced
Immunity
Louis Picker1
Marguerite Koutsoukos1
1
Oregon Health & Science University, Vaccine and Gene Therapy
Institute, Beaverton, OR, United States
1
SY09.02
SY09.04
Antibody Persistence and T Cell Balance:
Two Key Factors Confronting HIV Vaccine
Development
Integrase Defective Lentiviral Vectors for
Induction of Persistent and Functional Immune
Responses
George K. Lewis1, Tony L. Devico1, Robert C. Gallo1
Andrea Cara1
Medicine, Division of Basic Science and Vaccine Research, Baltimore,
MD, United States
1
The quest for a prophylactic AIDS vaccine is ongoing but it is now clear
that the successful vaccine must elicit protective antibody responses.
Accordingly, intense efforts are underway to identify immunogens
that elicit these responses. Regardless of the mechanism of antibodymediated protection, be it neutralization, Fc-mediated effector function,
or both, antibody persistence and appropriate T cell help are significant
problems confronting the development of a successful AIDS vaccine.
Evidence will be presented illustrating the poor persistence of antibody
responses to Env, the envelope glycoprotein of HIV-1, and the related
problem of CD4+ T cell responses that compromise vaccine efficacy by
creating excess cellular targets of HIV-1 infection. Finally, solutions to
both problems will be proposed that are applicable to all Env-based AIDS
vaccines regardless of the mechanism of antibody-mediated protection.
Istituto Superiore di Sanità, Rome, Italy
The development of an HIV-1 vaccine that elicits durable and broadlyreactive functional antibodies remains challenging but the goal of a
vaccine strategy. Integrase-defective lentiviral vectors (IDLV) represent
a new and promising delivery system for immunization purposes,
endowed with peculiar characteristics, setting them apart from the
parental integration-competent lentiviral vectors. Current data suggest
that IDLV are able to induce long-lasting and protective immune
responses in mice after a single immunization. We recently started a
study with the aim to demonstrate in monkeys that immunization with
IDLV delivering HIV Envelope protein induces sustained and functional
anti-Env antibodies (Abs) and T cell responses. The status of this work
will be discussed along with new immunization strategies that derive
from this work.
NON-ABSTRACT DRIVEN
1
GlaxoSmithKline, Vaccines Discovery & Development, Rixensart,
Belgium
www.hivr4p.org
31
Friday, 31 October
Roundtable 05: Multipurpose Technologies
RT05.01
RT05.03
MPTs: What Are they, and Why Are they
Needed?
Regulatory Considerations with Developing
Combination Products
Judy M. Manning1, Cara J. Chrisman1
Charu Mullick1
US Agency for International Development, Office of Population &
Reproductive Health, Washington, DC, United States
1
1
One of the fastest growing fields in women’s sexual and reproductive
health (SRH) is multipurpose prevention technologies (MPTs) that
simultaneously prevent unintended pregnancy, HIV, and other sexually
transmitted infections (STIs). MPTs have the potential to: 1) meet
women’s multiple SRH prevention needs in a single product; 2) achieve
efficiencies in cost of delivery of prevention products and services; and
3) leverage existing delivery channels to achieve higher levels of SRH
prevention product uptake and demand.
This presentation will provide an overview of the public health rationale
for MPTs, and the creation of the MPT Initiative, summarize recent efforts
to harmonize and prioritize the product development pipeline, and
describe high priority MPT products currently in late stage development.
Food and Drug Administration, Silver Spring, MD, United States
RT05.04
Study Designs for Evaluating the Effectiveness of
Multipurpose Technologies
Elizabeth R Brown1,2
Fred Hutchinson Cancer Research Center, Seattle, WA, United States,
University of Washington, Biostatistics, Seattle, WA, United States
1
2
RT05.02
RT05.05
State of the Art: Multipurpose Prevention
Technology Science and Product Development
Multipurpose Prevention Technology
Joseph Romano1
NWJ Group, LLC, Wayne, PA, United States
1
The development of multipurpose prevention technologies (MPT)
products involves complex and diverse scientific disciplines. Product
design and development, clinical evaluation and regulatory approval
all present challenges to the field and addressing these challenges
will require diverse expertise and innovation. Although there is active
development of early stage MPT products, there are also efforts to
develop next generation technologies that can address the technical
and market-based challenges confronted by current MPT products. This
presentation will discuss the status of current MPT development efforts,
the development and regulatory challenges these products face, and the
ongoing research and development that will improve and enable future
MPT products.
NON-ABSTRACT DRIVEN
32
Elizabeth Bukusi1
KEMRI, Nairobi, Kenya
1
Multipurpose Prevention Technologies (MPTs) are products under
development that are capable of simultaneously addressing women´s
sexual and reproductive health (SRH) including unwanted pregnancies,
STDs such as HIV and other reproductive tract infections. Women’s SRH
needs are interrelated. Unplanned pregnancies account for nearly half of
all pregnancies worldwide and lead to almost 100,000 maternal deaths
per year as a result of unsafe abortions and complications of pregnancy
and delivery. Women have inadequate access to efficacious contraception
especially in areas with high prevalence of HIV. Transmission of HIV and
other STIs such as HSV-2 and HPV are highly prevalent among women,
especially in some specific locations and in particular pockets of the
developing world. The available MPTs, male and female condoms, may
not be a realistic option for many women: condom use is inconsistent
and in addition issues of power and trust make it difficult for women to
negotiate its use. The female condom has not been widely accessible
nor affordable. Safe, acceptable and affordable technologies that
address these simultaneous RH needs in a more holistic way are
urgently needed. MPTs address these shortcomings and build on years
of contraceptive and HIV research to develop more comprehensive and
effective prevention methods. MPTs additionally address several health
priorities identified in the MDGs including: improving maternal health
(MDG 5) through access to technologies and interventions that meet the
complex range of women’s reproductive health needs; and combating
HIV/AIDS and other diseases (MDG 6) through technologies that target
STIs and inadvertently improving child health (MDG 4) by improving
their mothers’ health. Potential MPTs, both coitally-dependent and long
acting are under development. The presentation will update participants
on the status of current and future research of technologies and products
that are focused on reproductive needs of women
Friday, 31 October
Symposium 10: Challenges of Biomedical HIV Prevention Trials
SY10.01
SY10.03
HIV Prevention Trials: Their Successes and
Failures
Enrolling Adolescents in HIV Vaccine Trials:
Will We Be Ready?
Jeanne Marrazzo1
Ann E. Strode1, Catherine M Slack2
In the last several years, the HIV prevention field has experienced
unprecedented success with new approaches to biomedical prevention,
most notably with pre-exposure prophylaxis (PrEP) and medical male
circumcision. Tenofovir and emtricitabine, logical candidates for PrEP
because of their potent inhibition of the HIV-1 reverse transcriptase,
generally good safety and tolerability profiles in HIV-infected persons,
high barrier to HIV-1 resistance, and favorable pharmacokinetics, have
been approved in combination for PrEP and were recently recommended
by the WHO for persons at risk for HIV infection, especially men who
have sex with men (MSM). Despite these advances, however, the
prevention field has been buffeted by several disappointing findings
and realizations, mainly the complex interactions between adherence to
study product, perception of HIV risk, the social stigma attached to HIV
and antiretroviral medication, and the social fabric in which clinical trials
are conducted. Both VOICE and Fem PrEP trials successfully enrolled
young African women at high risk for HIV, yet adherence was low in
both. Moreover, in VOICE, participants with the highest risk profiles
were the least likely to adhere to the study drugs. Thus, more accurate
adherence measures are critical to estimate healthy participants’ product
use during HIV-1 prevention trials. Products that require minimal daily
adherence, including sustained antiretroviral delivery from vaginal rings
or injections, may be more suitable for some women. Alternatively,
successful adoption of daily PrEP may require different social and
operational strategies to support daily use. Indeed, the high rates of
retention we observed suggest that participants valued the study and its
contribution to their health. Understanding perceived motivation for and
value of study participation and investigational products, and leveraging
this when conducting biomedical HIV-1 prevention research, is critical.
SY10.02
Modifiers of PrEP Efficacy: Co-Infections, Sex,
Hormones and Microbiome
Betsy Herold1
Albert Einstein College of Medicine, Bronx, NY, United States
1
University of KwaZulu-Natal, School of Law and HIV/AIDS Vaccines
Ethics Group, Pietermaritzburg, South Africa, 2School of Laboratory
Medicine and Medical Sciences, University of KwaZulu-Natal, Applied
Health Sciences, HIV/AIDS Vaccines Ethics Group, Pietermaritzburg,
South Africa
1
Adolescents in sub-Saharan Africa are at high risk of HIV infection and
the testing of new HIV prevention technologies, such as candidate HIV
vaccines, are urgently required as part of a combination prevention
approach for this sub-group. Should an experimental HIV vaccine
show sufficient efficacy in adults, it is critical that stakeholders are fully
prepared for the enrollment of adolescents in efficacy studies. Much has
been written on the public health need to enroll adolescents into HIV
prevention studies to obtain relevant data that could be used to license
new prevention products specifically for this group. Less work has been
done to explore children´s rights to benefit from scientific progress
and whether this includes a right, if any, to participate in such studies.
This paper assumes it is both ethically and legally justifiable, in certain
circumstances, to include adolescents in HIV prevention trials. However,
their enrollment into such trials will be complex because consent,
privacy, illegal sexual activity and mandatory reporting obligations
(among others) will need to be addressed. This paper, drawing on work
funded in part by the NIH (award number 1RO1 A1094586) sets out
key norms for reflecting on these complexities. It asks how adequately
prepared countries will be to host adolescent trials, reviews progress
towards adolescent enrollment, and discusses a framework that could
be used to assess preparedness in South Africa, and even other Southern
African countries. It concludes that children, like adults, have a right
to benefit from scientific progress. Their age should not be a reason
to undermine their rights to access new HIV prevention technologies.
However, to fully realise such rights, concerted and sustained attention
must be given to promoting their welfare as trial participants.
SY10.04
Why Context Matters in Understanding the
Challenges to Clinical Trials
Jonathan Stadler1
Wits Reproductive Health & HIV Institute, Johannesburg,
South Africa
1
Recently, the uneven results of microbicide trials have drawn attention
to the political economy and social and cultural context in which trials
are conducted and the significance this has for enrollment, retention,
and adherence to study products. Drawing on data from two microbicide
trials, my talk presents an anthropological analysis of two recent
microbicide / PrEP trials and argues that a holistic perspective of the
trial as a social phenomenon may enhance our understanding of the
challenges to biomedical trials. The presentation also suggests ways of
incorporating social, scientific and ethnographic methods into clinical
trials in the endeavor to promote nuanced and thoughtful analysis.
www.hivr4p.org
33
NON-ABSTRACT DRIVEN
University of Washington, Seattle, WA, United States
1
Friday, 31 October
Symposium 11: Emerging Areas in Immunity
SY11.01
SY11.03
Systems Vaccinology: Enabling Rational Vaccine
Design with Systems Biology
Universal, MHC-E Restricted CD8+ T Cells
Participate in RhCMV Vaccine Vector-Induced
Protection Against SIV
Bali Pulendran1
Emory Vaccine Center, Atlanta, GA, United States
1
Despite their great success, we understand little about how effective
vaccines stimulate protective immune responses. Two recent
developments promise to yield such understanding: the appreciation of
the crucial role of the innate immune system in sensing microorganisms
and tuning immune responses, and advances in systems biology. In
this presentation, I will discuss how these developments are yielding
insights into the mechanism of some of the most successful vaccines
ever developed. Furthermore, such developments promise to address
a major challenge in vaccinology: that the efficacy of a vaccine can
only be ascertained retrospectively, upon infection. The identification of
molecular signatures induced rapidly after vaccination, which correlate
with and predict the later development of protective immune responses,
would represent a strategy to prospectively determine vaccine efficacy.
Such a strategy would be particularly useful when evaluating the
efficacy or immunogenicity of untested vaccines, or in identifying
individuals with sub-optimal responses amongst high risk populations,
such as infants or the elderly. We have used a systems biology approach
to identify early gene signatures that correlate with, and predict the later
immune responses in humans vaccinated with the live attenuated yellow
fever vaccine YF-17D, the seasonal and pandemic influenza vaccines,
meningococcal vaccines, pneumococcal vaccines, shingles vaccines and
the RTS,S malaria vaccine. I will review these studies, and discuss their
broader implications for vaccinology, and highlight some of the critical
insights that are emerging.
SY11.02
Resident Memory CD8 T Cells: Quantity,
Location, and Function
Kathryn Fraser1, Jason Schenkel1, Ashley Haase1, David Masopust1
University of Minnesota, Minneapolis, MN, United States
1
NON-ABSTRACT DRIVEN
Memory CD8 T cell quantity, location, and function relate to protective
efficacy. This presentation will summarize two series of investigations,
1) the characterization of resident memory CD8 T cell distribution,
differentiation, and function in mice, and 2) the use of heterologous
prime-boost-boost vaccination strategies to establish preternaturally
robust, polyfunctional, and broadly distributed SIV-specific memory CD8
T cells in rhesus macaques. New functions for resident mucosal memory
CD8 T cells will be described; as local sensors of previously encountered
antigens that broadcast innate-like alarm signals that activate local
humoral, cell-mediated, and innate immunity and confer an antiviral
state that can confer near sterilizing immunity against a viral challenge
within the mouse female reproductive tract. Additional data will be
presented that support the hypothesis that establishing sufficiently
abundant and functional memory CD8 T cells within barrier tissues may
achieve rapid protective responses at the site of pathogen exposure in
the rhesus macaque model of stringent vaginal SIV challenge.
34
Helen Wu1, Scott G. Hansen1, Katherine B. Hammond1, Collette M.
Hughes1, Jason Reed1, Abigail B. Ventura1, Reesab Pathak1, Alfred
W. Legasse2, Benjamin J. Burwitz1, Michael K. Axthelm1, Klaus
Früh1, Louis J. Picker1,2, Jonah B. Sacha1,2
Oregon Health & Science University, Vaccine & Gene Therapy Institute,
Beaverton, OR, United States, 2Oregon Health & Science University,
Oregon National Primate Research Center, Beaverton, OR, United States
1
Background: Vaccination with rhesus cytomegalovirus (RhCMV)
expressing SIV proteins results in unparalleled protection from
pathogenic SIV replication. The RhCMV-induced, SIV-specific CD8 T cells
are characterized by wide breadth, high promiscuity (responses present
in all vaccinees), and non-canonical epitope targeting. We hypothesized
that the breadth and promiscuity of the RhCMV-induced CD8 T cell
response result from unique patterns of Major Histocompatibility
Complex (MHC) restriction.
Methods: RhCMV/gag-vaccinated rhesus macaques (RMs) were MHCtyped by deep sequencing and single MHC-I transferents produced by
transfection of the allele of interest into MHC-I-null cell lines. MHC-I
transferents were pulsed with a single 15-mer peptide and used as
antigen-presenting cells in ICS with PBMC. T cell recognition of SIVinfected targets was assessed by ICS.
Results: RhCMV-induced, MHC-I-restricted CD8 T cell responses are
restricted by non-classical MHC-E molecules, including universal
“supertope” responses elicited in every vaccinee. MHC-E-restricted CD8
T cells are able to recognize both autologous and heterologous SIVinfected CD4 T cells in vitro. MHC-E is upregulated on the surface of
SIV-infected CD4 T cells, in contrast to decreased levels of classical
MHC-I molecules.
Conclusions: RhCMV elicits high frequency MHC-E-restricted responses
that explain the wide breadth, promiscuity, and novel epitope targeting
of the induced CD8 T cell response. As these MHC-E-restricted CD8 T
cells can recognize naturally processed antigen on SIV-infected cells,
they may confer protection from SIV replication that is observed in
~50% of RhCMV-vaccinated RMs. The MHC-E-restricted CD8 T cell
response may be particularly effective due to increased levels of MHC-E
on the surface of infected cells. RhCMV-induced, MHC-E-restricted CD8 T
cell responses represent a novel immune response against retroviruses.
Friday, 31 October
Symposium 11: Emerging Areas in Immunity
SY11.04
Cooperativity of HIV-specific Cytolytic CD4
T-cells and CD8 T-cells in Control of HIV Viremia
Susan Johnson1,2, Michael A. Eller1,2, Bruce T. Schultz1,2, Richard
Lu1,2, Alexander F. Oster1,2, Damien Z. Soghoian3, Agnes-Laurence
Chenine1,2, Galit Alter3, Ulf Dittmer4, Mary A. Marovich1,2, Merlin
Robb1,2, Nelson L. Michael1,2, Diane L. Bolton1,2, Hendrik Streeck1,2
US Military HIV Research Program, Silver Spring, MD, United States,
Henry M. Jackson Foundation for the Advancement of Military
Medicine, Silver Spring, MD, United States, 3Ragon Institute of MGH,
MIT and Harvard, Boston, MA, United States, 4Institute of Virology,
University Hospital Essen, University of Duisburg-Essen, Essen,
Germany
1
2
NON-ABSTRACT DRIVEN
Background: HIV-specific CD4 and CD8 T cell responses have been
shown to be critical in the control of HIV replication, though it is
unknown whether HIV-specific CD4 and CD8 T cells can act in concert.
We therefore defined the functional and transcriptional profile of HIVspecific cytolytic CD4 T cells and addressed the cooperativity between
both cellular lineages.
Methods: We used multicolor flow cytometry and Biomark Fluidigm
technology to compare transcriptional, phenotypic and functional profiles
of HIV-specific cytolytic CD4, CD4 Th1 and CD8 T cells. Additionally,
we assessed the ability of these cells to inhibit viral replication alone
or together with HIV-specific CD8 T cells and how this interaction is
complicated by HIV infection.
Results: HIV-specific CD4 cytolytic T cells were characterized by a
distinct functional and phenotypic profile in comparison to HIV-specific
Th1 CD4 T cells. The expression of CD57, KLRG-1, GrzB and Perforin
was significantly (p< 0.05) higher on HIV-specific cytolytic CD4 T cells
than Th1 cells, with T-box transcription factors eomesodermin and tbet
expressed on HIV-specific cytolytic CD4 T cells. Overall HIV-specific
cytolytic CD4 T cells showed transcriptional and phenotypic similarities
to HIV-specific CD8 T cells. Interestingly, cytolytic CD4 T cells were able
to inhibit HIV replication by an average of 40%, compared to an average
of 60% for CD8 T cells at day 3, but became infected by day 7. CD4
and CD8 T cells were also able to control viral replication synergistically,
suggesting a model where cytolytic CD4 T cells are involved in the
containment of viral replication, but dependent on the overall control of
HIV replication by HIV-specific CD8 T cells.
Conclusions: Our data suggests that HIV-specific CD4 cytolytic T cells
play a role in HIV infection, acting in concert with CD8 T cells to aid in
viral control. Overall, our results provide new insights on CD4 cytolytic T
cells functional properties and their involvement in HIV control, valuable
for vaccine development.
www.hivr4p.org
35
Friday, 31 October
Symposium 12: Differential Use of Antibodies in Prevention
SY12.01
SY12.03
Update on Clinical Development of VRC01 and
Second Generation Neutralizing CD4 Binding
Site-Specific Monoclonal Antibodies
HIV Cell-To-Cell Spread and Host
Countermeasures
Barney S. Graham
1
Vaccine Research Center, National Institute of Allergy and Infectious
Diseases, National Institutes of Health, Bethesda, MD, United States
1
VRC01 is an HIV-1 CD4 binding site-specific monoclonal antibody (mAb)
with potent and broad neutralizing activity. Phase I investigation was
initiated in uninfected persons for potential preventive (VRC 602) uses
and in HIV-infected subjects for potential therapeutic (VRC 601) uses.
The studies evaluate route and dose and the objectives include safety,
tolerability and pharmacokinetics (PK). VRC01 is administered on day 0
and at week 4 intravenously (IV) over a dose range of 1 to 40 mg/kg or
subcutaneously (SC) at 5 mg/kg.
As of 7/15/14, 40 subjects have received 74 doses. VRC01 was well
tolerated by both routes with no serious adverse events or doselimiting toxicity. At 20 mg/kg (n=5) or 40 mg/kg (n=4), the mean (±
SD) maximum serum concentrations were, respectively, 870±197 and
1611±258 mg/L after the first IV dose. In healthy adults at 20 mg/kg
(n=3) and 40 mg/kg (n=4), mean 28 day trough serum concentrations
were approximately 32.7±9.1 and 60.6±20.1 mg/L, after the first IV dose
and 40.6±10.7 and 102.6±48.8 mg/L after the second, respectively.
The clearance in healthy adults is estimated to be 466±118 mL per day
after IV administration over the 5 to 40 mg/kg (n=11) dose range. The
elimination half-life was similar in healthy and HIV-infected adults with
an overall mean value of 14.6±4.8 days.
Future development goals include additional studies of VRC01
evaluating dose, route and schedule, and testing efficacy to prevent
infection in high-risk infants and adults. The therapeutic potential of
VRC01 to diminish viremia during acute infection, prevent viral rebound
after treatment interruption, and reduce the viral reservoir during chronic
infection will also be evaluated. Second generation CD4 binding sitespecific mAbs with higher potency and breadth and extended half-life
are being explored for use as single agents or in combination with other
mAbs and treatment options.
SY12.02
Engineering Bispecific Antibodies for HIV
Prevention
David Ho1, Neal Padte1
NON-ABSTRACT DRIVEN
1
Aaron Diamond AIDS Research Center, New York, NY,
United States
36
Olivier Schwartz1
Institut Pasteur, Paris, France
1
Tuesday, 28 October
OA01.01
OA01.02
Structure-based Design of Trimeric V1V2
Antigens
A Native Linked Soluble Trimer of the HIV1 Spike Displaying Antigenic and Structural
Mimetic Properties
Jason Gorman1, Yongping Yang1, Aliaksandr Druz1, Ulrich Baxa2,
Peter D. Kwong1
NIH/NIAID/VRC, Structural Biology Section, Bethesda, MD, United
States, 2NIH/NCI, Cancer Research Technology Program, Frederick, MD,
United States
1
Background: Broadly neutralizing antibodies (bNAbs) targeted against
V1V2 of HIV-1 Env are among the most prevalent elicited by natural
infection and require less somatic mutation than those targeting other
sites, making antibodies of this class attractive targets of elicitation for
an HIV-1 vaccine. The quaternary specificity exhibited by V1V2 bNAbs
necessitates the development immunogens that effectively mimic the
trimeric cap of the functional native spike.
Methods: Recent structural work has defined the trimeric orientation
of the V1V2 domains at the apex of the viral spike, the V1V2 cap. This
V1V2 trimeric supersite was transplanted onto heterologous trimeric
scaffolds and screened for binding to quaternary specific antibodies.
Results: Here we present a trimeric V1V2 cap which recapitulates
the quaternary alignment of the three V1V2 domains present in the
native spike, as assessed by binding to quaternary specific V1V2directed bNAbs. High throughput screening of over a hundred constructs
revealed a single trimerization domain capable of presenting an
appropriate mimic of the V1V2 cap. Antibody binding was observed to
multiple strains transplanted onto the trimeric scaffold while gp120s of
the same strain did not bind. The transplanted V1V2 construct was able
to bind V1V2-bNAbs from multiple donors and importantly this trimeric
construct bound only to a single fab as observed with V1V2-directed
antibodies on the HIV-1 Spike.
Conclusions: Recent structural studies of HIV-1 trimers have provided
data to allow rational design of quaternary supersite transplants onto
scaffolds. High throughput screening with V1V2-directed antibodies
identified a construct which presents the V1V2 cap in an antigenically
similar manner to that observed with the BG505.SOSIP. The scaffolded
protein provides a simplified, well behaved alternative for use as a probe
and a candidate immunogen to focus the immune response on this
specific epitope.
Shailendra K. Sharma1, Natalia de Val2, Shridhar Bale3, Javier
Guenaga1, Andrew B. Ward2, Richard T. Wyatt1
IAVI Neutralizing Antibody Center at The Scripps Research Institute,
Department of Immunology and Microbial Science, La Jolla, CA, United
States, 2The Scripps Research Institute, Department of Integrative
Structural and Computational Biology, La Jolla, CA, United States, 3The
Scripps Research Institute, Department of Immunology and Microbial
Science, La Jolla, CA, United States
1
Background: Soluble mimetics of the HIV Env spike are of interest for
structure/function studies and immunogenic analysis. The BG505 SOSIP
trimers are the new structural gold standard, displaying an excellent
antigenic profile, molecular homogeneity and stability, allowing 5-6 Å
structural resolution. These trimers require furin co-expression, contain
unnatural cysteines and are purified by 2G12-affinity chromatography.
Many uncleaved gp140 trimers that maintain gp120-gp41 covalent
linkage by mutation of the cleavage site are suboptimal in their structural
organization, resulting in recognition by non-neutralizing mAbs.
Methods: Here, we describe covalent peptide linkage of gp120
to gp41 to create uncleaved but well-ordered trimers. The design
rationale was to provide flexibility at the cleavage site to allow native
rearrangement of gp120 and gp41 trimeric subunits mimicking the
endogenous furin cleavage event. We began this process by using a
flexible linker using one to three repeats of G4S linking JRFL gp120 to
gp41. Env deletion at residue 664 increased expression and an E168K
mutation was added to restore PG9/16 recognition. The native flexiblelinked (NFL) trimers were expressed in 293F cells and screened by a
panel of trimer-preferring bNAbs.
Results: Little trimer-associated recognition was observed, but
introduction of a I-to-P change in gp41 resulted in increased recognition
by trimer-preferring bNAbs, especially with two repeats of the flexible
linker. These trimers were purified by lectin-affinity and size-exclusion
chromatography, followed by negative selection, to isolate well-ordered
JRFL NFL trimers as confirmed by negative stain EM. Octet binding
analysis revealed recognition by most bNAbs but low recognition by
non-broad mAbs that was confirmed by EM 3D reconstructions of very
well-ordered trimers.
Conclusions: “Uncleaved” JRFL gp140-NFL2P trimers mimic cleaved
trimers in their antigenic properties and represent an exciting new
soluble spike design potentially applicable to other Envs.
www.hivr4p.org
37
ORAL ABSTRACT SESSIONS
Oral Abstract Session 01: B Cell Immunogen Design
Oral Abstract Sessions
OA01.03
OA01.04
A Recombinant HIV Envelope Trimer Selects
for Quaternary Dependent Antibodies
Targeting the Trimer Apex
An Efficiently Cleaved HIV-1 Subtype C Env
that Is Selectively Recognized by Neutralizing
Antibodies: A Platform for Immunogen
Design
Devin Sok1, Marit J. van Gils2, Matthias Pauthner1, Karen L. SayeFrancisco1, Jessica Hsueh1, Bryan Briney1, Jean-Philippe Julien1,
Peter S. Lee1, Yuanzi Hua1, John P. Moore3, Ian A. Wilson1, Rogier
W. Sanders2, Dennis R. Burton4
1
Scripps Research Institute, La Jolla, CA, United States, 2University of
Amsterdam, Academic Medical Center, Amsterdam, Netherlands, 3Weill
Medical College of Cornell University, Department of Microbiology and
Immunology, New York, CA, United States, 4Scripps Research Institute,
Department of Immunology and Microbial Science, La Jolla, CA, United
States
Background: Broadly neutralizing antibodies (bnAbs) targeting the
trimer apex of HIV envelope (Env) are favored candidates for vaccine
design and immunotherapy because of their high neutralization breadth
and potency. Methods to isolate bnAbs against this site, however, have
been limited due to the quaternary nature of the epitope region.
Methods: We report the use of a recombinant HIV envelope trimer,
BG505 SOSIP.664 gp140, as an affinity reagent to isolate quaternarydependent bnAbs from the peripheral blood of a chronically infected
donor. We chose to isolate antibodies from donor 84 from IAVI Protocol
G, from whom the antibodies PGT141-145 were isolated.
Results: We describe novel somatic variants of the PGT141-145 bnAb
family with a range of neutralization breadth and potency. Interestingly,
the new variant are highly divergent from the PGT141-145 somatic
variants being only 50-60% similar in amino acid sequence identity, but
deriving from the same gene family and sharing similar long CDRH3s.
Strikingly, variants with limited neutralization breadth were isolated
that share similar overall antibody features as the broadly neutralizing
variants, suggesting that differing evolutionary pathways can occur
within a single donor to yield differing neutralization profiles.
Conclusions: Overall, our results report for the first time the ability of
a soluble protein bait to isolate quaternary-dependent antibodies. Thus,
BG505 SOSIP.664 gp140 offers a useful tool for the isolation of trimerapex glycan-dependent antibodies and reveal a mosaic of antibody
responses against the trimer apex within a clonal family.
38
Saikat Boliar1, Supratik Das1, Manish Bansal1, Brihaspati Narayan
Shukla1, Shilpa Patil1, Tripti Shrivastava1, Sandeep Goswami1, C
Richter King2, Jayanta Bhattacharya1, Bimal K. Chakrabarti1
Translational Health Science and Technology Institute, THSTI-IAVI HIV
Vaccine Design Program, Gurgaon, India, 2IAVI, IAVI Vaccine Design,
New York, NY, United States
1
Background: The HIV vaccine field continues to study the elicitation
of broadly neutralizing antibodies (bnAbs) targeting the Envelope
glycoprotein (Env). A cleaved native HIV-1 Env, which selectively binds
to bnAbs, would be an ideal immunogen. This approach, however, has
eluded researchers because of the difficulty in maintaining the structural
integrity of cleaved Envs. Soluble BG505.664 SOSIP trimeric Envs (clade
A) display a nearly native antigenic profile, but require mutated cleavagesite, cysteine-linkages and co-expression of furin. Priming with plasmid
DNA expressing cleaved, membrane bound native Env followed by boost
with soluble Env could be a feasible solution. However, JRFL (clade B) Env
is the only known cleaved Env when expressed on the cell surface. Thus it
is important to screen other Envs that show efficient cleavage.
Methods: A total of 38 Indian subtype C env clones were screened for
cleavage. Comprehensive FACS-based binding and biochemical assays
were used to confirm cleavage and characterize the Env.
Results: We have identified 4-2.J41Env, which is efficiently cleaved
and displays similar antigenic profile to JRFL on the cell surface. A
comparative analysis of both Envs show that JRFL and 4-2.J41 Envs
bind to a cleavage dependent bnAb, PGT151 and the cleavage-defective
forms do not bind to PGT151. However, when the cytoplasmic tail (CT)
is truncated, 4-2.J41delCT Env binds to both neutralizing and nonneutralizing antibodies suggesting that the cytoplasmic domain of 4-2.
J41 Env plays an important role in maintaining its native conformation.
Codon-optimization of the envs enhances expression but, unlike JRFL,
4-2.J41 Env demonstrates similar properties to its non-codon optimized
counterpart indicating that it retains its native conformation.
Conclusions: Here, we report the identification of 4-2.J41Env, an
efficiently cleaved subtype C Env. As almost half of the global HIV-1
infections are mediated by clade C virus, this Env would advance the
design and development of HIV-1 vaccine.
Tuesday, 28 October
OA01.05
OA01.06
Minimizing Undesirable Epitope
Immunodominance on HIV-1 Env
Immunogens through Rational Immunogen
Modification
Uncleaved Soluble gp140 Constructs with
Retained Native-like Trimer Conformation
and Antigenicity
Andrew T. McGuire1, Anita M. Dreyer1, Sara Carbonetti1, Jolene A.
Glenn1, Johannes F. Scheid2, Hugo Mouquet2, Leo Stamatatos1
Seattle Biomed, Seattle, WA, United States, 2The Rockefeller University,
1
Background: Immunization with recombinant HIV-1 Env elicits
antibodies targeting epitopes on variable regions of Env that display
narrow breadth of neutralization (nNAbs), but fails to elicit antibodies
targeting conserved epitopes that display broad neutralizing activity
(bnAbs). This is thought to be in part due to the immunodominance of
the variable regions of Env, as well as the inability of recombinant Env
to activate naive B cells that give rise to bNAbs. We recently engineered
an Env capable of activating B cells expressing germline-reverted (gl)
BCRs of VRC01-class bNAbs and herein compare the ability of this Env to
stimulate B cells expressing glVRC01-class BCRs and B cells expressing
glnNAb BCRs
Methods: We developed novel assays to monitor the concurrent
activation of B cells expressing glVRC01 class BCRs and glnNAb BCRs
in response to various Envs, and assays to assess Env uptake by these
B cell lines.
Results: Unlike B cells expressing glVRC01 class BCRs, several Envs
bound to, activated, and were internalized by B cells expressing glnNAb
BCRs targeting both the CD4-BS and V3, indicating that nNAb eptiopes
are more readily presented on Env than bNAb epitopes. A direct
comparison of the magnitude of B cell activation responses, showed
that an Env specifically engineered to engage glVRC01 class B cells,
preferentially activates B cells expressing glnNAb BCRs, and that the
latter internalize this Env more efficiently than glVRC01 class B cells.
Importantly, we demonstrate that the removal of V1 V2 and V3 from
Env improves B cell activation and antigen uptake through glVRC01
class BCRs relative to CD4-BS and V3 directed nNAb BCRs.
Conclusions: Our results provide experimental evidence that explains
why immunization with recombinant Env results in the production of
nNAbs instead of VRC01 class bNAbs. However, we demonstrate that
it is possible to engineer an Env-based immunogen that eliminates
or reduces the immunodominance of nNAb epitopes and favors the
maturation process of VRC01 class bNAbs.
Ivelin Georgiev1, M. Gordon Joyce1, Yongping Yang1, Marie
Pancera1, Jason Gorman1, Priyamvada Acharya1, Guillaume
Stewart-Jones1, Aliaksandr Druz1, Cheng Cheng1, John R.
Mascola1, Peter D. Kwong1
National Institutes of Health, Vaccine Research Center, Bethesda, MD,
United States
1
Background: Soluble gp140 constructs that mimic native-like HIV-1
Env trimers are prime immunogen candidates. gp140 cleavage into
gp120 and gp41, however, is thought to be essential for mimicry of
native-like conformation and antigenicity, as evidenced by binding to
quaternary-specific broadly neutralizing antibodies (qbNAbs) but not to
non-neutralizing antibodies (NnAbs).
Methods: We employed structure-based design strategies to generate
modified uncleaved soluble gp140 trimers. The designed constructs
were characterized using a high-throughput ELISA assay against a panel
of qbNAbs and NnAbs, as well as bio-layer interferometry, gel filtration
and negative-stain EM.
Results: The designed uncleaved gp140 constructs were initially tested
in the context of the clade A HIV-1 strain BG505 and showed good
binding to qbNAbs (including highly quaternary-specific antibodies
such as PGT145 and VRC26.09) but not to NnAbs (such as F105). The
constructs exhibited a significant trimer fraction by gel filtration and
native-like conformation by negative-stain EM. The antigenicity and
conformational properties of these constructs resembled those of the
current state-of-the-art soluble gp140, BG505.SOSIP, which in contrast
requires full cleavage.
Conclusions: Uncleaved soluble gp140 constructs that mimic nativelike trimers have the advantage of not requiring cleavage, rendering
them useful candidates as protein- as well as DNA-based immunogens.
Uncleaved analogs in diverse HIV-1 strains are currently being evaluated
and immunogenicity studies are ongoing.
www.hivr4p.org
39
Oral Abstract Session 02: Microbicides: Male Partner Engagement and Sexual Behaviors
OA02.01
OA02.02
Engaging Male Partners in Women’s
Microbicide Use: Evidence from Clinical Trials
and Implications for Future Research and
Microbicide Introduction
Strategies to Improve Male Involvement and
Partner Support in the ASPIRE Trial: The
Hillbrow Experience
Michele Lanham1, Rose Wilcher2, Elizabeth T. Montgomery3,
Robert Pool4, Sidney Schuler5, Rachel Lenzi5, Barbara Friedland6,
Betty Njoroge7, Elizabeth Bukusi7, Robyn Dayton2
FHI 360, Social and Behavioral Health Sciences, Durham, NC, United
States, 2FHI 360, Research Utilization, Durham, NC, United States, 3RTI
International, Women’s Global Health Imperative, San Francisco, CA,
United States, 4University of Amsterdam, Centre for Social Science
and Global Health, Amsterdam, Netherlands, 5FHI 360, Social and
Behavioral Health Sciences, Washington, DC, United States, 6Population
Council, HIV and AIDS Program, New York, NY, United States, 7Kenya
Medical Research Institute, Research Care and Training Program, Centre
for Microbiology Research, Nairobi, Kenya
1
Background: Constructively engaging male partners in womencentered health programs such as family planning and PMTCT has
resulted in both improved health and relationship outcomes. Concerted
efforts to engage men in women’s microbicide use for HIV prevention
could make it easier for women to access and use microbicides, if an
effective product is identified.
Methods: We conducted primary and secondary analyses of male
engagement data from six qualitative studies implemented in
conjunction with microbicide trials in South Africa, Kenya, and Tanzania.
The analyses included 535 interviews and 107 focus groups with trial
participants, male partners, and community members. We synthesized
the findings across the studies and developed recommendations for
future research and microbicide introduction.
Results: The majority of women in steady partnerships wanted their
partner’s agreement to use microbicides. Women whose male partners
were resistant to microbicide use used a number of strategies to
obtain their approval. Among men who were aware of their partner’s
microbicide use, involvement ranged from opposition to agreement/
non-interference to active support. Both men and women expressed a
desire for men to have access to information about microbicides. Some
women and men said that it would be helpful if male partners could talk
with a health provider about microbicides; however, men were hesitant
to go to the clinic during the trials because of their work schedules, fear
of HIV testing, and stigma.
Conclusions: We recommend counselling women on whether and how
to involve their partners, providing couples’ counselling on microbicides,
and targeting men with community education and mass media to increase
their awareness and acceptance of microbicides. These activities should
be tested in microbicide trials, open-label studies, and demonstration
projects to identify effective male engagement approaches to include in
eventual microbicide introduction. Efforts to engage men must take care
not to diminish women’s agency.
40
Krishnaveni Reddy1, Pranitha Ramchuran1, Nombulelo Maseko1,
Amukelani Vuma1, Lizzy Gama1, Sylvia Sibeko1, Nosipho Duba1,
Helen Rees1, Thesla Palanee1
Wits Reproductive Health & HIV Institute, School of Clinical Medicine,
1
Background: In Southern Africa, men play a key role socially and
economically in womens’ lives and are often more dominant in
relationships. In terms of sexual health, males are less likely to exhibit
health seeking behavior or access HIV/STI testing and treatment as
frequently as women. This has major implications sexually and socially
for women involved in reproductive health research and hence proactive
male partner involvement and support in this area is crucial
Methods: At Wits RHI, strategies have been implemented to enhance
male involvement in the ASPIRE trial (an HIV prevention study
investigating the use of intravaginal Dapivirine ring). These strategies
include couple counseling, offering of facilitated trial participation
disclosure sessions, hosting of male health education weekend
workshops and partner invitation to study retention events. These
activities, aimed at increasing reproductive/sexual health knowledge,
includes information on contraception, STIs and HIV as well as ASPIRE
purpose and objectives, research misconceptions and importance of
partner support. Community Advisory Board members are invited to
these events to facilitate discussion as needed.
Results: Male partner attendance has been minimal due to work
related reasons, however male partner involvement and sharing of
sexual health information where possible is still encouraged at these
events. Participant feedback reveals that about 50% of participants have
disclosed study participation to partners with a favorable response in
most cases. Partners appear supportive in terms of reminding participants
of visits and product adherence with some partners being more open to
HIV/STI testing and treatment.
Conclusions: Male partner involvement in ASPIRE appears to increase
inter-partner communication and sexual health awareness with potential
to impact on study retention, product adherence and ultimately study
outcomes. Therefore at Wits RHI, male involvement strategies undergo
continuous evaluation and revision to optimize this aspect.
Tuesday, 28 October
OA02.03
OA02.04
Gel Use Disclosure in an Open-label 1%
Tenofovir Gel Trial: Perspectives from
Participants, Partners and Community Men in
KwaZulu-Natal, South Africa
Exploring Post-coital Intravaginal Cleansing
Practices among Women Enrolled in the
Microbicides Development Programme MDP
301 Clinical Trial
Kathleen M. MacQueen1, Sarah Dlamini2, Brian Perry1, Alesha
Majors1, Chitra Singh2, Diantha Pillay2, Eunice Okumu1, Sharon
Watson1
Mitzy Gafos1, Angela Crook1, Robert Pool2, Misiwe Adelaide
Mzimela3, Neetha Morar4, Andrew Vallely5, Andrew Abaasa6,
Thesla Palanee7, Oliver Mweemba8, Gita Ramjee4, Suzanna
Francis5, Anatoli Kamali6, Helen Rees7, Maureen Chisembele8,
Sheena Mccormack1, on behalf of the MDP Study Team
1
FHI 360, Durham, NC, United States, 2CAPRISA/University of Kwa Zulu
Natal, Durban, South Africa
MRC Clinical Trials Unit at UCL, London, United Kingdom, 2University
of Barcelona, Barcelona, Spain, 3Zululand University, KwaDlangezwa,
South Africa, 4HIV Prevention Research Unit, Medical Research Council,
Durban, South Africa, 5Mwanza Intervention Trials Unit, National
Institute for Medical Research, Mwanza, Tanzania, United Republic of,
6
MRC/UVRI Uganda Research Unit on AIDS, Kampala, Uganda, 7Wits
Reproductive Health and HIV Institute (WRHI), Johannesburg, South
Africa, 8University of Zambia, Lusaka, Zambia
1
Background: Disclosure of product use has been shown to influence
adherence in microbicide trials. We explored disclosure in the context of
CAPRISA 008, an ongoing open-label follow-on trial of 1% tenofovir gel
enrolling uninfected CAPRISA 004 trial participants.
Methods: In-depth interviews (n=63) and focus groups (n=8) were held
with CAPRISA 008 participants; male partners of 13 women who fully
disclosed trial participation and gel use were also interviewed. Four
focus groups were held with community men who were not partners of
CAPRISA 008 participants.
Results: Most CAPRISA 008 participants interviewed told their partner at
least some details about being in the trial or use of the gel. Motivating
factors for disclosure included perceived difficulty in hiding gel from
a partner and a desire to inform others about the benefits of the gel.
Women disclosing gel use to a partner perceived it easier to adhere
to the coitally-related regimen. Male partners of disclosers described
feelings of initial fear and apprehension of the gel followed by gradual
acceptance after being more fully informed. Non-disclosing women
described the ease of hiding gel use from their partners. Barriers to
disclosure included fear that the partner would not understand her
motivations for using the gel for HIV prevention or that he would react
negatively to her use of an intravaginal product. Community men saw
disclosure as indicative of the quality and seriousness of a relationship:
women in casual relationships could use gel without disclosing while
those in stable relationships should discuss gel use because nondisclosure inferred lack of trust. Community men felt that counselors
and pharmacists could assist women in explaining the purpose and use
of the gel to their partner.
Conclusions: Disclosure of 1% tenofovir gel use is perceived as beneficial
by women and men when it is negotiated and socially supported. Nondisclosure is a practical and acceptable option in some relationships.
Background: Post-coital intravaginal cleansing (IVC) could impact
the use of vaginal microbicide gels and limit the protective effect. In
MDP301 participants were counselled against IVC less than 1 hour after
sex and here we report their post-coital IVC practices.
Methods: We enrolled 9385 women at 13 clinics in 6 research centres:
Africa Centre (AC), Durban (DU), Johannesburg (JB) in South Africa;
Mwanza (MW) Tanzania, Masaka (MS) Uganda, and Mazabuka (MZ)
Zambia. Data on 9096 women with follow up data are included in this
analysis. Data on post-coital IVC were collected for each sex act in the
last week/4 weeks, at weeks 4, 24, 40 and 52 after enrolment.
Results: Despite counselling to the contrary, 42% (3,838/9096) of
women reported IVC within an hour after sex at least once during the
trial, mainly water or finger cleansing. IVC differed significantly across
centres: 26% in JB, 29% in AC, 31% in MW, 36% in MS, 59% in DU,
and 69% in MZ (p=0.001). In multivariable pooled analysis controlling
for centre, IVC was associated with age < =24 (AOR 1.11; CI 1.01,1.22),
< =primary education (AOR 1.16; CI 1.03,1.30), and reporting other
IV practices (AOR 1.70; CI 1.25,2,32). In multivariable centre specific
analysis, IVC was associated with younger age (JB, AC), unemployment
(MS), inconsistent gel use (MW), randomisation to 0.5% PRO2000 or
placebo gel groups (MS, MZ), larger household size (AC), Islamic religion
(MS), partner knowledge of gel (DU), other IV practices (MW) and clinic
of enrolment (DU, JB, AC). Among women who reported IVC, cleansing
was practiced after 43% of sex acts (on average per woman) differing by
centre at 33% in JB MW, 35% in MS, 43% in AC, and 48% in DU MZ.
IVC decreased from 33% in the first half to 24% in the second half of
the trial, decreasing most in JB and least in MZ.
Conclusions: Post-coital IVC is a common hygiene practice that may be
amenable to change. We need to further understand the impact of IVC
on microbicide efficacy and consider strategies to influence post-coital
IVC practices.
www.hivr4p.org
41
OA02.05
OA02.06
Language, Terminology and Understanding
of Anal Sex amongst VOICE Participants in
Uganda, Zimbabwe and South Africa
Application of a Body Map Tool to Enhance
Discussion of Sexual Behaviour in Women:
Experiences from MTN 003D
Zoe Duby1,2, Miriam Hartmann3, Imelda Mahaka4, Elizabeth T.
Montgomery3, Christopher J. Colvin1, Barbara Mensch5, Ariane
van der Straten3
Sarita Naidoo1, Kubashni Woeber1, Otilia Munaiwa2, Juliane
Etima3, Zoe Duby4, Miriam Hartmann5, Elizabeth Montgomery5,
Barbara Mensch6, Ariane van der Straten5, and the MTN 003D
Team
University of Cape Town, School of Public Health, Cape Town, South
Africa, 2Desmond Tutu HIV Foundation, Cape Town, South Africa, 3RTI
International, San Francisco, CA, United States, 4UZ-UCSF Collaborative
Research Programme, Harare, Zimbabwe, 5Population Council, New
1
Background: Despite efforts to use comprehensible terms, whether
research participants accurately interpret questions on sexual behaviour
is not well understood. MTN-003D, a qualitative ancillary study to
the VOICE HIV prevention trial in Sub-Saharan Africa, explored local
vernacular for sexual behaviours, understanding of terms for penileanal intercourse (AI) in VOICE ACASI questionnaires, and participant
narratives of AI behaviour compared to ACASI reports.
Methods: In-depth interviews (IDIs) were conducted with 88 women
(22 Ugandans, 26 Zimbabweans and 40 South Africans) in their
language of preference (Zulu, Luganda, Shona or English). A diagram of
a nude female figure was used as a visual aid to facilitate discussion of
participant interpretations of terms for AI.
Results: Findings suggest potential misreporting of AI due to ambiguity
and misinterpretation of terms in VOICE ACASI questionnaires. After
terms were clarified in IDIs, many participants claimed they had not
understood the ACASI AI question, or that they had interpreted it to
refer to vaginal sex from behind. There was also ambiguity in vernacular
terms participants themselves used. Of 88 participants, 35 had reported
AI in ACASI, but only 10 of these also disclosed AI during IDIs. Another
7 reported AI during the IDIs but had not done so in ACASI. Of the
remaining 25 reporting AI in ACASI but not in the IDI, a third needed the
term for AI to be clarified during the IDI.
Conclusions: Findings highlight challenges in developing terms for sexual
behaviour that are neither ambiguous nor open to misinterpretation.
In efforts to balance social appropriateness with non-ambiguity across
languages, terminology used in research is not always accurate or
explicit. These findings suggest that some participants misunderstood
the AI question in VOICE, which may have led to misreporting. The
challenges in accurately reporting sexual behaviour have implications
not only for clinical trials, but also for clinical practice and assessing
HIV/STI risk.
42
South African Medical Research Council, HIV Prevention Research Unit,
Durban, South Africa, 2UZ-UCSF Collaborative Research Programme,
Harare, Zimbabwe, 3MU-JHU Research Collaboration, Kampala,
Uganda, 4Desmond Tutu HIV Foundation, IIDMM University of Cape
Town, Cape Town, South Africa, 5RTI International, San Francisco, CA,
United States, 6Population Council, New York, NY, United States
1
Background: Body maps are used in sexual and reproductive health
studies to explore sex and sexuality, risks and pleasures, and to
understand individual’s perceptions of their bodies. MTN-003D, an
exploratory study of potential sources of efficacy dilution in the VOICE
trial, used a body map tool, together with in-depth interviews (IDIs), to
encourage frank discussion about sexual behaviour.
Methods: IDIs were conducted with 88 former VOICE participants in
Zimbabwe, Uganda and South Africa from December 2012 to March
2013. Body map tools, which showed the front and back outline of a
nude female figure, were used during IDIs to initiate and aid discussion
of sex. Interviewers asked women to identify and discuss genitalia and
other body parts associated with sexual behaviour, pain and pleasure;
and their responses (key words or short sentences) were noted on
the map. Analysis of annotated body maps was done together with
corresponding transcript data to ascertain the success of this tool.
Results: Body maps helped interviewers probe issues around sexual
behaviour. Women could point to body parts without having to verbalise
potentially embarrassing anatomical terms. Women were amused, shy
or embarrassed when shown the map. Most women readily labelled
maps; however some needed clarity or encouragement. Body maps
were used by most women to discuss pleasure, however only half of
the women used the tool to identify areas associated with pain. Only 6
women were reluctant to discuss their sexual behaviour (2 refused due
to religious beliefs; 1 only labelled biological functions of the body; 2
avoided looking at map; 1 uncomfortable to discuss anal sex).
Conclusions: In this study, body maps used in combination with IDIs
were feasible and effective tools to enhance discussions around sexual
behaviour, specifically anal sex. Body maps provided women with a
non-intimidating way of discussing and disclosing their sexual practises
and minimized miscommunication of anatomical terminology.
Tuesday, 28 October
OA03.01
OA03.02 LB
Preclinical Evaluation of TMC-278 LA, a
Long-acting Formulation of Rilpivirine,
Demonstrates Significant Protection from
Vaginal HIV Infection
HIV PrEP Dose Rationale for Cabotegravir
(GSK1265744) Long-acting Injectable
Nanosuspension
Olivia Snyder1, Heather Vincent1, Sophie Lachau-Durant2, Guenter
Kraus2, Peter Williams2, J. Victor Garcia1
UNC Chapel Hill, Chapel Hill, NC, United States, Janssen Research and
Development, Beerse, Belgium
1
2
Background: Vaginal HIV transmission accounts for the majority of
new infections worldwide. Prevention efforts have demonstrated mixed
success due to lack of adherence to drug regimens. New efforts to
prevent HIV transmission have focused on long−acting (LA) antiretroviral
drug formulations to circumvent this issue.
Methods: We first established critical pharmacokinetic parameters of
intramuscular (IM) injection of TMC−278 LA in mice and demonstrated
sustained drug release for 4 weeks. Humanized BLT mice were then
vaginally challenged with HIV−1 transmitted founder viruses (T/F) after
IM administration of TMC−278 LA and the extent of protection from
HIV acquisition was determined. In the 1st experiment, challenges with
the T/F virus CHO40 were performed 1 week after drug administration
(600mg/kg). In a 2nd experiment, BLT mice were challenged 1 week
after drug administration with 1 of 3 different viruses (CHO40, RHPA,
or JR-CSF). These mice were exposed to a second challenge 3 weeks
after drug administration with a different T/F virus (THRO). Infection was
determined using viral load assay and PCR analysis for vDNA in tissues.
Identity of the infecting viruses was confirmed by DNA sequencing.
Results: In the first experiment, a single IM dose of TMC−278 LA
(600 mg/kg) one week before vaginal challenge provided significant
protection from CHO40 infection (6/6 mice protected) (p=0.0047).
In contrast, 3/3 animals that received saline became infected. In the
second experiment, 6/7 BLT mice that received saline before exposure
became infected. Whereas 6/8 BLT mice that received TMC-278 LA
before the first viral challenge were protected from infection (p=0.026).
All mice exposed 3 weeks post−drug administration to a 2nd challenge
with THRO became infected.
Conclusions: TMC-278 LA offers significant protection from vaginal HIV
infection against T/F viruses. Although a wane in protection over time
was observed. These results demonstrate the potential of long−acting
antiretroviral formulations for HIV prevention.
Bill Spreen1, Alex Rinehart2, Kimberly Smith2, David Margolis1,
Susan Ford3, Steve Piscitelli1
GlaxoSmithKline R&D, Infectious Diseases, Research Triangle Park,
NC, United States, 2ViiV Healthcare, R&D, Research Triangle Park, NC,
United States, 3GlaxoSmithKline R&D, Clinical Pharmacology Modeling
and Simulation, Research Triangle Park, NC, United States
1
Background: Long acting (LA) drugs such as the integrase inhibitor
cabotegravir (CAB, GSK1265744) may bolster PrEP adherence and
efficacy. PrEP dose selection is challenging given lack of validated
surrogate markers and low rates of HIV seroconversion.
Methods: CAB PrEP dose selection was based on in vitro virology,
human antiviral activity, healthy volunteer (HV) single and repeat dose
safety and PK studies and non-human primate (NHP) PK and SHIV
rectal/vaginal challenge studies. Data were integrated in populationbased PK (PPK) analyses/dose simulations to identify a dose predicted to
be efficacious and clinically practical.
Results: In a 10-day monotherapy study in ART-naïve patients CAB 5mg/
day p.o. (n=7) yielded geomean blood plasma (BP) Ctau of 0.57 ug/mL,
>3-fold above protein-adjusted (PA)-IC90 of 0.166 ug/mL, and produced
a 2.14 log median decrease in HIV RNA, providing a PK/PD correlate.
Single injection CAB LA (SC or IM) in 58 HV evaluated ascending doses
for safety and PK. CAB LA 800mg IM BP levels exceeded Ctau of oral
5mg/day for 16 weeks. A repeat dose HV (n=40) study confirmed safety
and PK of 800mg IM dose; geomean Ctau (week 12) was 1.11 ug/mL or
7x PA-IC90. NHP PK studies targeting CAB concentration-time profiles
in BP approximating human values were followed by SHIV162p3
rectal challenges (50xTCID50) to establish POC and evaluate threshold
of protection. CAB BP ≥1xPAIC90 correlated with ≥ 97% protection.
SHIV vaginal challenge showed protection in 6/8 rhesus macaques
(300xTCID50) and 8/8 pigtail macaques (50xTCID50). PPK based
simulation of dosing regimens achieving target BP levels with quarterly
injections enabled selection of 800mg IM q12week dose.
Conclusions: CAB LA PrEP dose selection was guided by human safety,
PK and antiviral activity studies and NHP challenge studies. CAB LA
800 mg IM q12weeks is predicted to yield drug levels correlated with
robust anti-HIV activity and protection against SHIV rectal and vaginal
challenge. This dose is under study in phase 2 safety and PK trials.
www.hivr4p.org
43
Oral Abstract Session 03: Animal Model Studies of Microbicides and Injectables
OA03.03
OA03.04
Tenofovir Reservoir Intravaginal Rings
Provide Superior Pharmacokinetics and
Higher Sustained Drug Levels than Tenofovir
Matrix Rings
A Combination Vaginal Ring Releasing
Dapivirine and Darunavir
Meredith Clark1, Lara Pereira2, Justin Clark3, Todd Johnson3,
Chou-Pong Pau4, David Friend1, James Smith4, Patrick Kiser5
CONRAD Eastern Virginia Medical School, Arlington, VA, United States,
2
LifeSource Biomedical LLC, Moffett Field, CA, United States, 3University
of Utah, Salt Lake City, UT, United States, 4Centers for Disease Control
and Prevention, Division of HIV and AIDS Prevention, Atlanta, GA,
United States, 5Northwestern University, Evanston, IL, United States
1
Background: Sub-protective topical antiretroviral (ARV) levels can lead
to HIV transmission events. Therefore maintaining drug levels in tissues
is a primary goal of long acting ARV delivery systems. Matrix intravaginal
rings (IVR) show attenuating release rates over time; reservoir IVR can
deliver precise drug levels over several months. Both IVR types are being
investigated in the topical PrEP field. Here we present the in vitro and in
vivo evaluation of TFV release and pharmacokinetics (PK) of matrix and
reservoir polyurethane IVR in pigtail macaques.
Methods: Macaque-sized IVR contained 360mg TFV in a solid
hydrophilic polyurethane (HPU) matrix or 550mg TFV in a hollow-core
HPU reservoir. IVR were characterized for in vitro release. IVR were
inserted vaginally into pigtail macaques (N=4-6) for 28d; blood plasma,
vaginal secretions (VS), rectal secretions (RS), vaginal tissue (VT) and
rectal tissue (RT) biopsies were collected at various time points. TFV PK
was assessed by LC/MS-MS.
Results: Matrix IVR demonstrated time-dependent TFV release and PK
profiles, with ~1-log reductions observed for both in vitro release rate
(32 to 4 mg/d) and median TFV concentrations in VS (3-6x106 to 3-4x105
ng/mL), VT (0.9-3x104 to 0.6-1x103 ng/g), and RT (497 to 76.5 ng/g)
over the 28d treatment period. Median TFV RS levels were sustained at
< IC50 levels (186-349 ng/ml). Reservoir IVR showed time-independent
TFV release and PK profiles over the same duration. In vitro release was
~6mg/d TFV, and median TFV levels were sustained at 2-7x106 ng/g
(VS), 0.4-1.7x105 ng/g (VT), and 0.2-1.1x104 ng/g (RS).
Conclusions: TFV reservoir IVR provide superior PK control and higher
sustained vaginal TFV levels compared to TFV matrix IVR in macaques.
Previous non-human primate studies that evaluated TFV 1% gel and
tenofovir disoproxil fumarate IVR suggest these local concentrations are
likely to be protective from vaginal challenge in the macaque model.
44
Karl Malcolm1, Diarmaid Murphy1, Delphine Desjardins2,3,
Nathalie Dereuddre-Bosquet2,3, Patricia Brochard2,3, Ludivine
Perrot2,3, Alain Pruvost4, Roger Le Grand2,3, Ole Lagatie5, Leen
Vanhooren5, Maxim Feyaerts5, Jens Van Roey5
Queen’s University Belfast, School of Pharmacy, Belfast, United
Kingdom, 2Commissariat à l’Energie Atomique (CEA), Division of
Immuno-Virology, DSV/iMETI, IDMIT Center, Fontenay-aux-Roses,
France, 3Paris Sud University-11, UMR-E1, Orsay, France, 4Commissariat
à l’Energie Atomique (CEA), iBiTecS, SPI, Laboratoire d’Etude du
Métabolisme des Médicaments, F-91191, Gif-sur-Yvette, France,
5
Janssen Diagnostics, Turnhoutseweg, Belgium
1
Background: Combination microbicide vaginal rings, containing two
or more antiretrovirals targeting different steps in the HIV replicative
process, may be more effective than single microbicide products at
preventing sexual transmission of HIV. Here, we report the preclinical
development, including in vitro release and macaque pharmacokinetics,
of matrix-type silicone elastomer rings containing dapivirine (DPV;
an experimental non-nucleoside reverse transcriptase inhibitor) and
darunavir (DRV; a marketed protease inhibitor).
Methods: Macaque rings containing 25 mg DPV, 300 mg DRV and
100 mg DPV, and 300 mg DRV were manufactured and characterised
by differential scanning calorimetry. In vitro release was assessed into
isopropanol/water and simulated vaginal fluid. Macaque vaginal fluid
and blood serum concentrations for both antiretrovirals were measured
during 28-day ring use. Tissue levels were measured on day 28. Ex vivo
challenge studies were performed on vaginal fluid samples and IC50
values calculated.
Results: DRV caused a concentration-dependent reduction in the DPV
melting temperature in both solid drug mixes and in the combination
ring. In vitro release from rings was dependent on drug loading, the
number of drugs present, and the release medium. In macaques, serum
concentrations of both microbicides were maintained between 101-102
pg/mL. Vaginal fluid levels ranged between 103-104 ng/g and 104-105
ng/g for DPV and DRV, respectively. Vaginal tissue concentrations
decreased in rank order: vagina (1.8×103-3.8×103 ng/g) > cervix
(9.4×101-3.9×102 ng/g) > uterus (0-108 ng/g) > rectum (0-40 ng/g).
Measured IC50 values (HIV-1 BaL) determined from macaque vaginal
fluid samples were < 2 ng/mL for both compounds.
Conclusions: Based on these results, and in light of the ongoing clinical
progress of the 25mg DPV ring, a combination vaginal ring containing
DPV and DRV is a viable second-generation HIV microbicide candidate.
Tuesday, 28 October
OA03.05
OA03.06 LB
A Novel Intravaginal Ring (IVR) Protects
Macaques against SHIV-RT Infection and
Reduces HSV-2 Shedding after Repeated
SHIV-RT/HSV-2 Co-challenge
Rectal Specific Gels Containing Maraviroc
and/or Tenofovir Protect against Rectal SHIV
Transmission in a Macaque Model
Thomas M. Zydowsky1, Jessica Kenney1, Meropi Aravantinou1,
Shweta Ugaonkar1, Nina Derby1, Larisa Kizima1, Shimin Zhang1,
Olga Mizenina1, Jose Fernández-Romero1, Melissa Robbiani1
Population Council, HIV & AIDS Program, New York, NY, United States
1
Background: MZC gel (MIV-150, zinc acetate [ZA], carrageenan [CG])
significantly reduces vaginal SHIV-RT and HSV-2 infection in macaques
when given 8h before challenge. CG gels significantly reduce HPV
infection in mice when dosed 24h before challenge. We aim to develop
a 90d IVR releasing MZC and levonorgestrel (LNG) to prevent HIV, HSV2, HPV, and conception.
Methods: IVRs contained 3mg MIV-150±0.6mg LNG in the matrix and
30mg ZA/70mg CG in the core (open to fluids via a matrix pore). We
measured CG, MIV-150, and LNG in macaque blood and/or vaginal
swabs during and after 28d of IVR insertion. For efficacy, we exchanged
IVRs every 21d, challenging macaques with 200 TCID50 SHIV-RT/107 pfu
HSV-2 on d7, 10, 14 and 17 post IVR insertion (20 challenges total); n=4
placebo IVR, n=4 LNG IVR, n=12 MZC IVR, n=12 MZCL IVR.
Results: MIV-150 was detected in swabs and plasma 1h post IVR
insertion, peaked at d1 in swabs and d1-3 in plasma, declined from d7d28, and was undetectable 24h after IVR removal. LNG was detected in
serum within 4h, plateaued by d7, and was undetectable 24h after IVR
removal. CG was detected in swabs of most animals from d3 until the
IVRs were removed. Preliminary data show MZCL IVRs protected (92%)
macaques against SHIV-RT (1/12 vs. 4/4 infected in LNG IVR controls;
p=0.003); 2/12 MZC and 2/4 placebo IVR animals became SHIV-RT
infected (67% protection). MZC and MZCL IVRs significantly protected
compared to control IVRs (3/24 vs. 6/8 infected, p=0.002). Initial data
suggest that ~30% fewer animals became infected with HSV-2 after
treatment with MZC and MZCL IVRs (17/24 vs. 8/8 infected controls); the
frequency (18/30 vs. 23/95 time points positive; p< 0.0006) and levels
(67/180 vs. 31/570 replicates positive; p< 0.0001) of HSV-2 shedding
were significantly lower than in the controls.
Conclusions: We developed an IVR that releases APIs targeting HIV,
HSV-2, HPV, and conception. The IVR significantly reduces SHIV-RT
infection and HSV-2 shedding in macaques.
Charles Dobard1, Andrew Taylor1, Sunita Sharma1, Dinh Chuong1,
Chou-Pong Pau1, Lisa Rohan2, Ian McGowan2, Walid Heneine1
Centers for Disease Control and Prevention, Atlanta, GA, United States,
Magee Womens Research Institute, University of Pittsburgh, Pittsburgh,
PA, United States
1
2
Background: Rectal transmission of HIV is an important driver of the
HIV epidemic in several populations. Rectal microbicides containing
antiretroviral drugs are under development to prevent rectal acquisition
of HIV. Maraviroc (MVC) and tenofovir (TFV) are candidate drugs for
rectal-specific gel formulations. Previous work showed that gels
formulated for vaginal application were not optimal for rectal use. Here
we evaluated the efficacy of optimally formulated rectal gels containing
1% MVC, 1% TFV, or in combination against repeated rectal SHIV
exposures in rhesus macaques.
Methods: MVC and TFV were formulated in a rectal-specific iso-osmolar
hydrogel based formulation at neutral pH. Macaques were administered
4 ml of 1% MVC (n=6), 1% TFV (n=6), 1%MVC/1%TFV (n=6) or a
placebo (n=7) gel 30 minutes before each rectal SHIVSF162P3 challenge
(500 TCID50). Challenges were repeated twice-weekly for 5 weeks
(10 challenges). Plasma drug levels were measured 30 min after gel
application using LC/MS/MS. Infection was monitored by serology and
PCR of SHIV in plasma. Infected macaques continued to receive gel for
an additional 6-weeks to monitor impact on systemic viremia.
Results: All 7 controls were infected after a median of 4 challenges.
In contrast, 4 of 6 macaques in each group receiving either 1% MVC,
1% TFV, or MVC/TFV gel remained protected after 10 challenges
demonstrating an 83% efficacy (p=0.02). Low levels of MVC (median=4
ng/ml) and TFV (median=19 ng/ml) were detected in plasma 30 minutes
after rectal dosing, suggesting rapid absorption. There was no difference
in drug plasma concentrations between protected and breakthrough
infections. Similar plasma viremia in controls and breakthroughs was
observed reflecting the low systemic drug exposure.
Conclusions: Here we demonstrated in vivo drug release by 3 gel
formulations and showed that all provided high efficacy against repeated
rectal SHIV exposures. Our findings support the clinical development of
these gels for rectal prophylaxis against HIV acquisition.
www.hivr4p.org
45
Oral Abstract Session 04: Innate Immunity
OA04.01
OA04.02
SIV- and Vaccine-elicited NK Cell Memory in
Rhesus Macaques
Innate Lymphoid Cells are Depleted in HIV
Infection
R. Keith Reeves1,2, Haiying Li1,2, Eryn Blass1, Hualin Li1, Stephanie
Jost3, Marcus Altfeld3, Dan Barouch1
Henrik N. Kløverpris1,2, Aslam Noorbhai3, Warren Kuhn4, Marisa
Yadon1, Duran Ramsuran1, Sheperd Nhamoyebonde1, Victoria
Kasprowicz1, Bruce Walker5, Thumbi Ndung’u1, Philip Goulder6,
Abdool S. Karim7, Jenny Mjösberg8, Alasdair Leslie1
Boston, MA, United States, 2New England Primate Research Center,
Southborough, MA, United States, 3Ragon Institute of MIT, MGH and
Harvard, Boston, MA, United States
1
Background: Natural killer (NK) cells provide rapid responses to viral
infections and are typically considered to be nonspecific components of
innate immunity. However, recent studies have shown that NK cells can
also mediate antigen-specific memory in mice, but it remains unclear
whether this phenomenon also exists in primates.
Methods: In this study we evaluated NK cells from a cohort of 8 rhesus
macaques chronically infected with SIVmac251, 6 naïve controls, and
9 macaques vaccinated with replication-incompetent Ad26 vectors
expressing either HIV-1 Env or SIVmac239 Gag. Using a novel flow
cytometric assay we evaluated antigen-specific killing of autologous
dendritic cells (DCs) by highly purified hepatic and splenic NK cells.
Fluorochrome-labeled DCs were pulsed with intact SIV Gag or HIV-1 Env
and non-pulsed DCs served as intra-well controls. Purified NK cells were
co-cultured with DCs at multiple E:T ratios and specific lysis was used as
a functional determination of antigen-specificity.
Results: Splenic NK cells from SIV-infected animals were highly reactive
to Gag-pulsed DCs at a 10:1 NK:target ratio with a median specific lysis
of 40% compared to 1% in naive controls, indicating the presence
of antigen-specific NK cells in chronic infection. In our vaccinated,
unchallenged macaques, at both 10:1 and 5:1 NK:target ratios, splenic
and hepatic NK cells lysed antigen-matched targets at higher frequencies
than they did antigen-mismatched targets (P = 0.021, liver; P = 0.040,
spleen). Responses in peripheral blood were marginal, suggesting that
memory NK cells likely reside in tissues.
Conclusions: Taken together, our data demonstrate the first evidence of
NK cell memory in a primate species. Furthermore, antigen-specific NK
cell responses to SIV antigens are induced in primates following both
infection and vaccination. The longevity, functionality, and specificity of
memory NK responses in primates suggest their functional relevance in
developing vaccines against multiple pathogens, including HIV.
46
KwaZulu-Natal Research Institute for TB&HIV, K-RITH, Durban, South
Africa, 2University of Copenhagen, Dept. of Int. Health, Immunology
and Microbiology, Copenhagen, Denmark, 3University of KwaZuluNatal, Dept of General Surgery, King Edward Hospital, Durban, South
Africa, 4University of KwaZulu-Natal, Dept of ENT, King Edward
Hospital, Durban, South Africa, 5Ragon Institute, Massachusetts Institute
of Technology and Harvard, Cambridge, Boston, MA, United States,
6
Oxford University, Paediatrics, Oxford, United Kingdom, 7Center for the
AIDS Programme of Research in South Africa, CAPRISA, Durban, South
Africa, 8Karolinska Institutet, Center for Infectious Medicine, Stockholm,
Sweden
1
Background: Innate Lymphoid Cells (ILCs) lack rearranged antigen
receptors but share functional and developmental characteristics with
lymphocytes and can be subdivided into three main groups according
to their production of Th1, -Th2 and -Th17 cell-associated cytokines
named ILC1, ILC2 and ILC3s, respectively. ILCs play a crucial role in
tissue homeostasis and repair in both non-infectious and infectious
diseases. HIV-1 pathology involves lymphoid tissue destruction of the gut
mucosa associated with microbial translocation, immune activation and
disease progression in both ARV-treated and untreated individuals. We
hypothesized that HIV modulates the ILC population in blood and tissue.
Methods: We used phenotypic and transcriptional characterization of
human ILCs from a total of n=83 chronic HIV-infected and n=64 HIV
uninfected individuals recruited from 4 different cohorts in Durban,
South Africa.
Results: We show that the Th2 cytokine producing ILC2 population in
human peripheral blood is severely depleted in viremic HIV-1 infected
individuals (P< 0.0001) but preserved in those with natural immune
control of HIV-1 (P=0.003), and correlates negatively with plasma
viral load (r=-0.62, P=0.0004). This ILC2 population is depleted from
peripheral blood within 20 days of HIV-1 transmission (P=0.04) and
only partially restored upon successful antiretroviral therapy in a subset
of individuals. Similar depletion of ILC1 and ILC3 populations were
observed in chronic infection (P< 0.001) to similar kinetics as the ILC2
population, but distinct from the observed NK cell expansion dynamics.
Preliminary data from gut and tonsil tissue resident cells shows overall
reduction of ILCs skewed towards an NCR3+ ILC3 population.
Conclusions: These data demonstrate that HIV-1 infection rapidly and
profoundly modulates the tissue resident and circulating ILC population,
with great potential significance to the pathology of this disease.
Tuesday, 28 October
OA04.03
OA04.04
Impact of Systemic Immune Activation (IA)
and Inflammation on the HIV Susceptibility
of HIV- individuals with HIV Concordant or
Discordant Partners
Do CD16+ NKG2A+ NK Cells Recruited to the
Gut Combined with Passively Administered
SIV Specific Antibodies Prevent SIVmac251
Acquisition in Macaques?
Shameem Z. Jaumdally1,2, Pamela P. Gumbi1, Hoyam
Gamieldien1, Lindi Masson1, Heather B. Jaspan3,4, Caroline
Tiemessen5,6, Anabela Picton5,6, Anna-Lise Williamson1,7, David
Coetzee8, Francesca Little9, Jo-Ann S. Passmore1,7
Namal P.M. Liyanage1, Melvin N. Doster1, Francesca Caccuri1,
Poonam Pegu1, Shari N. Gordon1, Robyn Washington Parks1,
Cynthia Pise-Masison1, Luis Barcena2, Jeff Scheider2, Howard Y.
Lakouga2, Patrick Kiser2, Stephen Whitney3, Luca Schifanella1,
Monica Vaccari1, Thomas J. Hope2, Genoveffa Franchini1
University of Cape Town, Medical Virology, Cape Town, South
Africa, 2University of Cape Town, Department of Public Health and
Family Medicine, Cape Town, South Africa, 3University of Cape
Town, Immunology, Cape Town, South Africa, 4Seattle Biomedical
Research Institute, Seattle, WA, United States, 5National Institute
for Communicable Diseases of the NHLS, Johannesburg, South
Africa, 6Faculty of Health Sciences, University of the Witwatersrand,
Johannesburg, South Africa, 7National Health Laboratory Service,
Groote Schuur Hospital, Cape Town, South Africa, 8Department of Public
Health and Family Medicine, Cape Town, South Africa, 9University of
Cape Town, Department of Statistical Science, University of Cape Town,
1
Background: Studies of individuals who appear to resist HIV infection,
such as HIV- partners in HIV discordant relationships, are important for
identifying host responses or characteristics associated with protection
against HIV infection. Previously, immune quiescence has been
associated with HIV resistance. The aim of this study was to compare
the level of IA and systemic inflammation in HIV- individuals from
South Africa (SA) who were either in relationships with HIV-infected
or uninfected stable partners, to evaluate markers of HIV exposure or
resistance.
Methods: A heterosexual couples cohort of 103 HIV- individuals with
long-term stable HIV- concordant partners (HIV- unexposed) and 113
HIV- individuals with HIV+ discordant partners (HIV- exposed) were
included in this study. T cell activation and proliferation (CD38, HLADR, CCR5, Ki67) in blood was assessed by flow cytometry. Cytokines in
plasma were evaluated by Luminex.
Results: HIV- exposed individuals had lower frequencies of CD4+ T-cells
in blood expressing the CCR5 [alone (p=0.05) or in combination with
Ki67 (p=0.05) or CD38 (p=0.05)] than HIV-unexposed individuals.
Similarly, HIV- exposed individuals had significantly lower frequencies
of CD8+ T-cells in blood expressing CCR5 [alone (p=0.05) or in
combination with Ki67 (p=0.01) and CD38 (p=0.05)] and HLA-DR
(p=0.01) than their HIV- unexposed counterparts. Plasma concentrations
of IL-2 (p=0.02), IFN-γ (p=0.05) and GM-CSF (p=0.006, stayed sig. after
adjustment for multiple comparisons) were significantly lower in HIVexposed compared to HIV- unexposed individuals.
Conclusions: This study suggests that HIV- exposed individuals from
SA have an immune quiescent phenotype, with lower frequencies of
activated CCR5-expressing T-cells and CCR5 density per T cell, than their
HIV- unexposed counterparts. Since CCR5 expressing T-cells, especially
activated ones, are the preferred targets for HIV infection, this study in
HIV- discordant couples suggests that CCR5 agonists may be useful to
block HIV infection.
National Cancer Institute, NIH, Animal Models & Retroviral Vaccine
Section, Bethesda, MD, United States, 2Northwestern UniversityFeinberg School of Medicine, Robert H Lurie Medical Research
Center 303 E Superior, Chicago, IL, United States, 3Advanced
BioScience Laboratories, Inc., Rockville, MD, United States
1
Background: Growing evidence suggests that NK cells and antibodies
that mediate ADCC can control HIV infection. ADCC was shown to have
an inverse correlation with risk of HIV infection in the RV144 vaccine
trial that showed 31% protection from HIV acquisition. We recently
showed the recruitment of CD16+ NKG2A+ NK cells to the gut during
ALVAC/SIV/gp120 vaccine regimen in macaques. Here we investigated
whether NKG2A+ NK cells together with vaccine induced anti-envelop
IgG at mucosa can protect from SIVmac251 acquisition.
Methods: IgG was purified from sera following 8 additional
immunizations with ALVAC-SIV/gp120/Alum of 8 macaques previously
exposed to SIVmac251 remaining uninfected. To demonstrate that
systemically given IgG can localize to the gut, CY5 labeled antihuman IgG was subcutaneously administered to macaques and antihuman antibodies were measured in serum and rectum by ELISA and
immunohistochemistry (IHC) at 6h, 24h and weekly for 4 weeks. Two
macaque cohorts (A and B) were vaccinated with ALVAC-SIV (SIV766
Gag-Pro gp120TM) at 0, 4, 12 and 24 weeks. ALUM was administered
together with the vaccine at 12 and 24 weeks. Purified IgG will be given
to group A (n=8) subcutaneously (24h) and intra-rectally (IR) (2h) prior to
the repeated low dose challenge with SIVmac251 IR in July 2014. Group B
(n=8) and unvaccinated group (n=12) will be challenged simultaneously.
Results: Purified IgG from 8 protected animals showed 1.87% of gp120
specific activity by ELISA. Anti-human IgG was detected in serum
and rectal biopsies by ELISA and IHC up to 3 weeks post antibody
administration. Cytometric analysis of rectal biopsies showed recruitment
of NKG2A+NK (p=0.03) cells to the gut of vaccinees.
Conclusions: Our current data shows the recruitment of NKG2A+ NK
cells to the gut and successful delivery of antibodies to the gut mucosa.
Protection after SIV challenge in the end of July 2014, will confirm our
hypotheses that the vaccine mediated recruitment of NK cell together
with antibodies can prevent SIV acquisition.
www.hivr4p.org
47
OA04.05 LB
OA04.06
Integrated Systems Biology Analysis Reveals
Contrasting Role for Innate Immune Response
Genes in Conferring Risk of Infection in RV144
Trial
Loss of Viral Control in a Subset of HIVInfected Long-term Non-progressors Is
Associated with a Decline of an Array of
Antiviral Responses
Ali Filali-Mouhim1, Slim Fourati1, Francois Lefebvre1, Rasmi
Thomas2, Raphael Gottardo3, Nicole Frahm3, Stephen De
Rosa3, Peter Wilkinson1, Petra Stafova1, Amit Sabnis1, Jaranit
Kaewkungwal4, Punnee Pitisuttithum4, Sorachai Nitayanphan5,
Supachai Rerks-Ngarm6, Nelson Michael7, Jerome Kim2, Merlin
L. Robb2, Robert J. O’Connell8, M. Juliana McElrath3, Mark
Cameron1, Rafick Sekaly9
Mohamed El-Far1, Pascale Kouassi1, Odalis Asin-Milan1, Annie
Chamberland1, Mohamed Sylla1, Jean-Philippe Goulet1, Petronela
Ancuta1, Jean-Pierre Routy2, Danielle Rouleau1, Marianne Harris3,
Nicole Bernard2, Cécile Tremblay1
Vaccine & Gene Therapy Institute of Florida, Port Saint-Lucie, FL,
United States, 2U.S. Military HIV Research Program (MHRP), Walter
Reed Army Institute of Research, Bethesda, MD, United States, 3Vaccine
and Infectious Disease Division, Fred Hutchinson Cancer Research
Center, Seattle, WA, United States, 4Faculty of Tropical Medicine,
Mahidol University, Bangkok, Thailand, 5Royal Thai Army, Armed
Forces Research Institute of Medical Sciences, Bangkok, Thailand,
6
Department of Disease Control, Ministry of Public Health, Nonthaburi,
Thailand, 7U.S. Military HIV Research Program, Walter Reed Army
Institute of Research, Bethesda, MD, United States, 8U.S. Army Medical
Component, Armed Forces Research Institute of Medical Sciences,
Bangkok, Thailand, 9Department of Pathology Case Western Reserve
University, Cleveland, OH, United States
1
Centre de Recherche du CHUM, Montreal, QC, Canada, 2McGill
University, Montreal, QC, Canada, 3BC Centre for Excellence in HIV/
AIDS, Vancouver, BC, Canada
1
Background: The RV144 trial showed promise towards the development
of an effective HIV vaccine. Antibodies to HIV Env V1V2 loop, Envspecific IgA and HLA class II DQB1*06 allele with high Env IgA were all
significantly associated with risk of infection.
Methods: We used an integrated systems biology approach to identify
novel correlates of immunogenicity and/or protection. Env-specific
stimulated peripheral blood mononuclear cells from RV144 participants
were used to test the hypothesis that innate pro-inflammatory responses
would demarcate RV144 case-control groups.
Results: A case/control gene expression analysis using 133 controls and
27 cases yielded few significantly differentially expressed genes between
cases and controls. To adjust for the heterogeneity and the case/control
class imbalance, we employed a stratification strategy using a treebased classification method with IgA, V1V2 and DQB1*06 as predictor
variables and the infection status as the outcome. The obtained tree
model showed a balanced accuracy of 61% and identified two different
risk groups. Cases in the first group had low IgA titers and did not express
the DQB1*06 allele. Transcriptional profiling showed downregulation
of the type II interferon induced genes (Fisher enrichment test p value
=0.04) in these cases compared to controls. Cases in the second Group
had high IgA and Low V1V2 titers. Transcriptional profiling showed,
interestingly, up-regulation of several type I interferon genes including
antiviral genes (Fisher enrichment test p value =10-6) in these cases
compared to controls.
Conclusions: Herein we have identified a gene expression signature,
segregating case and control donors and which, counterintuitively,
shows contrasting roles for innate immune response genes in conferring
vaccine induced protection. These results have important implications
for the development of more effective HIV vaccine strategies.
48
Background: Understanding molecular mechanisms of natural disease
control in the absence of treatment in a minor group of HIV-infected
subjects, identified as slow progressors (SP), represents a logical
approach towards a potential functional cure.
Methods: In the current study we aimed to identify molecular signatures
associated with viral control in our Canadian cohort of HIV-infected SP
(Study # CTN 247; subjects maintaining CD4 counts over 500/microliter
for over 7 years, in the absence of treatment). To this end, we have
identified 5 subjects that have experienced a sudden loss of viral control
(average increase of viral load: 5 to 79 fold) and a decline in the absolute
CD4 counts (average loss: 211 cells/microliter). We used the Illumina
technology to study genome-wide transcriptional profiles in peripheral
blood mononucleated cells (PBMCs) isolated from these subjects before
(Visit 1) and after (Visit 2) the loss of virological control.
Results: Our analysis identified 1,381 probe sets corresponding to
1,268 genes that were differentially expressed between V1 and V2
(nominal p-value < 5%). Among these genes, 864 were downregulated
and 517 were upregulated at V2 versus V1. Of note, among the downregulated genes, several members of both innate and adaptive anti-viral
responses were identified such as APOBEC3G, IL-32, the T cell receptor
CD96 (known to associated with the superior quality of CD8 T cells in
HIV-infected Elite controllers), Lck, LAT, JAK1, ITK and the IL-7 receptor
(IL-7R). Our validation assays are currently in progress to validate the
expression of these genes at the protein level.
Conclusions: Our current study takes advantage of the power of the
whole genome transcriptional analysis on privileged samples from HIVinfected SP subjects before and after the loss of virological control to
identify new targets for therapeutic interventions and also to identify
early biomarkers to predict disease progression.
Tuesday, 28 October
OA05.01
OA05.02
Durable Suppression of Established
Transmitted Founder Replication in
Infected BLT Humanized Mice by Vectored
ImmunoTherapy
An HIV DNA Vaccine Delivered by
Electroporation and Boosted by rVSV HIV-1
Gag Is Safe and Immunogenic in Healthy HIVuninfected Adults
Cailin Deal1, Yong Ouyang2, Christin M. Hong1, Dong Sung An3,
David Baltimore2, Alejandro B. Balazs1
Christine M. Hay1, Gregory J. Wilson2, Marnie Elizaga3, Sue Li4,
Nidhi Kochar4, Mary Allen5, Magdalena Sobieszczyk6, Ian Frank7,
Nicole Frahm3, Georgia D. Tomaras8, Michael Egan9, Michael
Pensiero5, John Eldridge9, NIAID HIV Vaccine Trials Network
Harvard University, The Ragon Institute of MGH, MIT and Harvard,
Cambridge, MA, United States, 2California Institute of Technology,
Biology, Pasadena, CA, United States, 3University of California at Los
Angeles, Department of Translational Sciences, Los Angeles, CA, United
States
1
Background: Recent reports in humanized mice and monkeys have
found that broadly neutralizing antibodies (bNAbs) can suppress
the replication of laboratory strains of HIV and SHIV while bNAb
concentration remains high. Vectored ImmunoProphylaxis (VIP) results
in long-lived bNAb expression following a single intramuscular (IM)
injection of a specialized viral vector, and this approach has been
demonstrated as a means of durably suppressing viral load. However,
previous reports of VIP-delivered bNAbs for HIV therapy required prior
antiretroviral drug therapy to reduce viral load to prevent escape.
Methods: Humanized BLT mice were infected IV with the REJO.c
transmitted molecular founder strain of HIV. A low dose of combination
antiretroviral therapy (ART) was administered to these animals for 5
weeks, followed by a single IM injection of VIP expressing VRC07 or
luciferase. Mouse plasma was analyzed by ELISA to determine antibody
concentration and by qPCR to determine viral load. Cellular fractions
were analyzed by flow cytometry to quantify human CD4 cells over
time. After sacrifice, plasma was subjected to a clinically validated
ultrasensitive PCR-based viral load assay.
Results: We detected viral loads of 105 copies/mL in infected mice prior
to low-dose ART treatment, which resulted in a transient reduction and
rebound to pre-therapy loads. Following VIP administration, we observed
a rapid increase in the blood concentration of VRC07. Mice expressing
VRC07 exhibited a sharp decline in viral load to undetectable levels and
an increase in CD4 cells over four weeks and this effect was sustained
for the remaining 8 weeks of the study. In contrast, mice expressing
luciferase exhibited increasing viral loads with concomitant decreases in
CD4 cells throughout the study.
Conclusions: Our results demonstrate that VIP expressing VRC07 is
sufficient to suppress actively replicating transmitted founder virus at
high viral load and support efforts to move Vectored ImmunoTherapy
into clinical trials with infected patients.
University of Rochester Medical Center, Rochester, NY, United States,
Vanderbilt University, Nashville, TN, United States, 3Fred Hutchinson
Cancer Research Center, Seattle, WA, United States, 4SCHARP - FHCRC,
Seattle, WA, United States, 5DAIDS/NIAID/NIH, Rockville, MD, United
States, 6Columbia University, New York, NY, United States, 7University
of Pennsylvania, Philadelphia, NY, United States, 8Duke University,
Durham, NC, United States, 9Profectus Biosciences, Tarrytown, NY,
United States
1
2
Background: Eliciting HIV-specific immune responses through vaccination
remains an important goal in preventing HIV. Here we present safety,
tolerability, and immunogenicity data from a phase Ia trial of a novel HIV1 multi-antigen (MAG) DNA vaccine delivered by electroporation (EP) with
DNA IL-12 adjuvant and boosted with an rVSV HIV-1 Gag vaccine.
Methods: HVTN 087 enrolled 100 healthy adults in a multicenter,
randomized, double-blinded, placebo-controlled study. Participants
received 3,000 mcg HIV-MAG (gag/pol, env, nef/tat/vif) DNA vaccine coadministered with IL-12 DNA at 0, 250, 1000, or 1500 mcg (N=22/group)
or placebo (N=3/group) intramuscularly by EP at 0, 1 and 3 months
boosted by rVSV Gag vaccine or placebo at 6 months. Participants were
assessed for reactogenicity, tolerability, and adverse events. CD4+ and
CD8+ T-cell responses to HIV potential T-cell epitope (PTE) peptides were
measured by intracellular cytokine staining (ICS) 2 weeks after 3rd and
4th vaccinations.
Results: EP was generally well tolerated. Local and systemic reactogenicity
symptoms were generally mild to moderate. After the 4th vaccination
some subjects experienced moderate to severe systemic symptoms and
several experienced transient lymphopenia. After DNA prime, CD4+ T-cell
responses to any PTE were detected in 77% of subjects and CD8+ T-cell
responses in 40%. Gag-specific CD4+ T-cell response rates after DNA
prime increased significantly following rVSV Gag boost, from 15% to
86%. Gag-specific CD8+ T-cell responses also increased significantly from
6% after DNA prime to 26% after rVSV Gag boost. IL-12 DNA 1500 mcg
increased the magnitude of CD8+ T-cell responses compared to no IL-12
DNA (p=0.02).
Conclusions: HIV-1 DNA vaccination given by EP with IL-12 and boosted
with HIV-rVSV is safe and immunogenic. DNA/EP prime dramatically
improves T-cell responses after a single rVSV boost compared with rVSV
homologous prime-boost (HVTN 090). IL-12 DNA does not increase
response rates but increases the magnitude of CD8+ T-cell responses.
www.hivr4p.org
49
Oral Abstract Session 05: Vaccine, Viral Latency and Cure
OA05.03
OA05.04
Elicitation of Immune Responses by a DNA/
MVA Vaccine in ART Treated Patients in a
Treatment Interruption Trial
Early Initiation of ART in Acute HIV Infection
(Fiebig I to III) Does Not Preclude the
Development of HIV-specific Cellular Immune
Responses
Harriet L. Robinson1, Melanie Thompson2, Sonya Heath3,
Stephen J. Brown4, Bentley Sweeton2, Kathy Williams2, Pamela
Cunningham3, Miles Viljanen4, Rahul Basu1, Yongxian Xu5,
Suefen Kwa1
GeoVax, Inc., Smyrna, GA, United States, 2AIDS Research Consortium
of Atlanta, Atlanta, GA, United States, 3University of Alabama at
Birmingham, Department of Medicine, Birmingham, AL, United States,
4
AIDS Research Alliance, Los Angeles, CA, United States, 5The Hope
Clinic of the Emory Vaccine Center, Decatur, GA, United States
1
Background: GV-TH-01, a Phase 1 open-label trial of GOVX-B11, a
DNA/MVA prime-boost regimen, in HIV infected patients on ART was
undertaken to evaluate safety and vaccine-elicited T cell responses, and
to explore viral rebound during analytical treatment interruption (TI).
Methods: Patients who began ART within 18 months of seroconversion
and had sustained plasma HIV-1 RNA < 50 c/mL for at least 6 months
were enrolled. Patients received a total of 4 inoculations at intervals of
8 weeks. 2 of pGA2/JS7 DNA (3mg) followed by 2 of MVA/HIV62B (108
TCID50). At 8 weeks after the last immunization, plus an efavirenz washout if needed, participants entered a TI phase of 12 weeks, after which
ART was reinstituted. T cell responses were scored for IFNg or IL2 by
flow cytometry following stimulation with Gag, Env and Pol peptides.
Responses were considered positive if ≥2-fold higher than pre-vaccination.
Results: 8 of 9 men completed all vaccinations. For the 8, median age
was 37.5 yrs, baseline CD4 count was 691/µL (501-1612/µl) and all had
HIV-1 RNA < 50 c/mL. Median viral load prior to ART was 5.1 log10
c/mL (2.6-7.2 log10 c/mL). No serious adverse events occurred. After
the 1st or 2nd MVA/HIV62B immunization, Gag-specific CD8 T cells were
boosted over pre-vaccination levels in 7 out of 8 (P< 0.05) whereas
Gag-specific CD4 T cells were boosted in 5 of 8 patients (P=0.2). 6 of 8
patients elicited previously undetectable CD8 responses whereas 5 of 8
elicited previously undetectable CD4 responses to Gag epitopes. Gp120
or gp41-specific antibody responses were boosted in 3 of 8 patient and
2 of 8 patients respectively. Excluding one acute seroconverter, the
median reduction in HIV-1 RNA at weeks 2, 6, and 12 compared to preART levels was -2.2, -1.3 and -0.8 log10 c/mL.
Conclusions: This trial demonstrates the potential for GOVX-B11 to
boost both T cell and antibody responses in a therapeutic setting. A
placebo-controlled trial will be required to further assess the therapeutic
benefit of the vaccine.
50
Alexandra Schuetz1,2, Yuwadee Phuang-Ngern1, Rapee
Trichavaroj1, Nittaya Phanuphak3,4, Rungsun Rerknimitr5,
Suchada Sukhumvittaya1, Surat Jongrakthaitae1, Silvia RattoKim2,6, James Fletscher3, Eugene Kroon1,3, Nitaya Chomchey3,4,
Robert J. O’Connell1, Viseth Ngauy1, Praphan Phanuphak3,4,5,
Nelson L. Michael6, Jerome H. Kim6, Mark S. De Souza1,3,4,
Jintanat Ananworanich2,3,6, RV254/SEARCH 010 Study Group
1
Armed Forces Research Institute of Medical Sciences – United States
Component, Retrovirology, Bangkok, Thailand, 2Henry M. Jackson
Foundation for the Advancement of Military Medicine, Bethesda,
MD, United States, 3SEARCH, Bangkok, Thailand, 4The Thai Red Cross
AIDS Research Centre, Bangkok, Thailand, 5Chulalongkorn University,
Department of Medicine, Bangkok, Thailand, 6United States Military
HIV Research Program (MHRP), Walter Reed Army Institute of
Research, Silver Spring, MD, United States
Background: High viral load (VL) and rapid loss of CD4+ T cells are a
hallmark of acute HIV infection. The association between the subsequent
emergence of HIV-specific CD8+ T cells and the decrease in VL support
that CD8+ T cells are crucial in the initial control of viral replication.
However, little is known about the kinetics of emerging T cell responses
during early acute infection (Fiebig [F] I to III) and the effect of early
initiation of antiretroviral treatment (ART) on the development of HIVspecific T cells.
Methods: 28 patients with acute HIV infection (11 FI/II, 17 FIII) were
enrolled in the RV254/Search 010 study and immediately received ART.
HIV-specific immune responses against Gag and Env peptide pools were
determined in PBMC using IFN-γ intracellular cytokine staining prior to
ART initiation and 6 and 24 months post-ART.
Results: At time of diagnosis, no HIV-specific T cell responses were
detected in FI/II, while in FIII predominantly Gag-specific CD4+
responses were observed in 2/17 (12%) and CD8+ responses in 4/17
(24%) patients. Median plasma VL in FI/II was 5.5 log10 copies/ml vs.
6.7 log10 copies/ml in FIII (p=0.001). At 6 and 24 months post-ART
all patients had undetectable VL. However, 6 months post-ART there
was no significant difference observed in the frequency of Gag-specific
CD4+ and CD8+ responses between patients treated in FI/II (2/11 [18%]
and 6/11 [55%], respectively), and FIII (4/17 [23%], and 8/17 [47%],
respectively). The frequency of Gag-specific CD8+ T cell responses was
maintained 24 months post-ART, primarily in patients that initiated ART
in FIII (FI/II CD4+: 0/11, CD8+: 4/11 [36%]; FIII CD4+: 5/17 (29%), CD8+:
9/17 [53%] p>0.05).
Conclusions: HIV-specific T cell responses, primarily CD8+ mediated
against Gag, were detected at FIII but not FI/II. However, after 6 months
of ART, both groups had similar numbers of responders maybe due to
viremia during the initial post-ART period or ongoing viral replication in
privileged sites.
Tuesday, 28 October
OA05.05
OA05.06 LB
Engineered Gag-specific T-cell Receptors
Redirect Polyclonal CD8+ T-cells to Clear HIV1-infected CD4+ T-cells from ART-treated
Patients
Structure of HIV-1 gp120 V1V2 in Complex
with Human mAb 830A Reveals a 5-Stranded
Beta Barrel Conformation and Integrinbinding Site
Hongbing Yang1, Sandrine Buisson2, Giovanna Bossi2, Gemma
Hancock1, Rebecca Ashfield2, Annelise Vuidepot2, Tara Mahon2,
Peter Molloy2, Joanne Oates2, Zoe Wallace1, Namir Hassan2, Bent
K. Jakobsen2, Lucy Dorrell1
Ruimin Pan1, Miroslaw K. Gorny2, Susan Zolla-Pazner2,3, XiangPeng Kong1
University of Oxford, Nuffield Department of Medicine, Oxford, United
Kingdom, 2Immunocore Ltd, Abingdon, Oxfordshire, United Kingdom
1
Background: HIV-1 establishes a stable latent reservoir in resting CD4+
T cells that is not eliminated by ART. New immunotherapeutics are
needed to deliver a ´shock and kill´ in order to reduce viral reservoirs.
We evaluated the antiviral efficacy of engineered high affinity immunemobilising monoclonal T cell receptors (ImmTACs) that redirect CD8+ T
cells to kill HIV-1-infected CD4+ T cells.
Methods: HIV-1 ImmTACs comprised engineered high affinity T cell
receptors (TCRs) specific for the HIV-1 gag epitope, SLYNTVATL (SL9)
and its common escape variants, fused to a humanized CD3-specific
single chain variable fragment. Their antiviral potency was evaluated
in viral inhibition and killing assays with ex vivo CD4+ and CD8+ T cells
from HLA-A*0201-positive ART-treated patients. Fluorescent-labelled
TCRs were also used to quantify target epitope density on infected cells.
Results: HIV-1 ImmTACs significantly enhanced ex vivo CD8+ T
cell-mediated inhibition of endogenous HIV-1 replication at low
concentrations (1-10 nM) and CD8+/CD4+ ratios of 1:1 and 1:10 (mean,
SD: 72%, 14%, p=0.008; 55%, 15%, p=0.004 respectively) despite
low epitope expression on the cell surface (< 50/cell). More potent
inhibition was achieved when CD8+ T cells from healthy HLA-A*0201+
donors were used (mean, SD: 85%, 8%, p< 0.0001). Killing of infected
cells was confirmed by lack of viral recrudescence after wash-out of the
ImmTAC from the culture and was maximal within 48 hours of ImmTAC
exposure. Of note, resting infected CD4+ T cells were eliminated almost
as efficiently as activated cells (mean 85% vs. 95%). The potency of the
ImmTACs was highly correlated with the level of intracellular HIV-1 gag
expression (r2=0.48, p< 0.0001).
Conclusions: ImmTACs can efficiently redirect polyclonal CD8+ T cells to
kill activated and resting HIV-1-infected CD4+ T cells from HIV-1 positive
patients, at low effector:target ratios and low epitope densities. Our
data support further evaluation of ImmTACs as a component of HIV-1
eradication strategies.
NYU School of Medicine, Department of Biochemistry and Molecular
Pharmacology, New York, NY, United States, 2NYU School of Medicine,
Department of Pathology, New York, NY, United States, 3Veterans
Affairs New York Harbor Healthcare System, New York, NY, United
States
1
Background: The first and second variable regions (V1V2) of gp120
play vital roles in the function of HIV-1 envelope (Env). V1V2, which
harbors multiple glycans and is highly sequence diverse, is located at
the Env apex and stabilizes the trimer. It shields V3 and the co-receptor
binding site in the pre-fusion state and exposes them upon CD4 binding.
Recent structural data, including that of the scaffolded V1V2 in complex
with PG9/PG16 and the trimeric SOSIPs, suggested that V1V2 forms
a Greek key motif with 4 beta strands (named strands A-D). However,
regions of V1V2 including the integrin-binding site were missing from
those structural visualizations, and our understanding of the V1V2
structure-function is thus not yet complete.
Methods: We have determined a crystal structure of the V1V2 scaffold
(V1V2ZM109-1FD6) in complex with 830A, an anti-V1V2 human mAb with
an epitope known to overlap the integrin-binding site.
Results: Our complex structure revealed that V1V2 has a 5-stranded
beta barrel structure with the region of the integrin-binding site forming
a kink (AAs 178-180) followed by an additional beta strand (strand C’)
before strand D. The complete barrel structure naturally presents the
glycans on its outer surface and packs conserved hydrophobic residues,
including the RV144 sieve residue Ile181, in its core,. The two lengthvarying regions in V1 and V2 form two extended loops towards one end
of the barrel. The epitope of 830A is discontinuous with three segments:
it centers at Thr175, Tyr177, Leu179 and Asp180 at the kink overlapping the
integrin-binding site (AAs 179-181) and also includes residues Arg153 and
Val154 in V1 and Leu193, Ile194 and Ser195 at the C-terminus of V2.
Conclusions: The V1V2 region forms a 5-stranded beta barrel, a unique
structure that can function as an independent module.
www.hivr4p.org
51
Oral Abstract Session 06: B Cell Repertoires for Protection
OA06.01
OA06.02 LB
Evolution of Antigen-specific B-cell Receptor
Repertoires in Early SIV Infection
Role of Intestinal Microbiota in Shaping the
B Cell Repertoire in HIV Infection and Env
Vaccination
Eva J. Archer1,2,3, Rachael Bashford-Rogers2, Brenna Hill1, Daniel
C. Douek1, Paul Kellam2, Richard A. Koup1
Vaccine Research Center, NIAID, NIH, Bethesda, MD, United States,
Wellcome Trust Sanger Institute, Hinxton, United Kingdom, 3University
of Cambridge, Cambridge, United Kingdom
1
2
Background: Broadly neutralizing antibodies to HIV arise only in a
fraction of cases after years of infection, while early antibody responses
to HIV infection typically do not neutralize circulating virus or provide
protection from infection. Detailed understanding of the early antibody
response can inform vaccine and immunogen design. We use nextgeneration sequencing to track the evolution of the antibody repertoire
generated in response to SIV infection in macaques.
Methods: Naive, memory, and antigen-specific B cells were sorted from
blood, lymph nodes, and bone marrow of rhesus macaques infected
with SIVmac251 at 4, 10, and 24 weeks post infection. B cell receptor
heavy chain sequences were amplified using Ig-specific primers and
sequenced using MiSeq 2x300bp paired end reads to obtain full
length VDJ sequences. Sequences were analysed using a bioinformatics
platform to track V, D, and J gene usage; mutation from germline;
CDR3 sequence and length; and clonality and diversity of the sample
populations over time.
Results: Antigen-specific B cells were first detected in the bone marrow,
blood, and lymph nodes at 4 weeks post infection using a SIV gp140
trimeric probe. The fraction of SIVgp140 positive memory B cells
increased throughout infection, up to 1.5% of total B cells in lymph nodes
at 24 weeks post infection. Antigen specific B cells show accumulation
of mutations in the VH gene compared to nonspecific memory B cells,
and organize naturally into clusters of clonally related sequences from
acute and early chronic infection.
Conclusions: This study is a high-resolution analysis of memory and
antigen-specific B cells in different spatial compartments showing the
dynamics of the humoral immune response from acute to early chronic
SIV infection.
52
Larry (Huaxin) Liao1, A.M. Trama1, W.B. Williams1, M.A. Moody1,
Nathan Vandergrift1, G.D. Tomaras1, D.J. Marshall1, T. Gurley1,
J. Whitesides1, J. Eudailey1, A. Foulger1, R. Parks1, C. Stolarchuk1,
K.E. Lloyd1, K. Soderberg1, J.R. Mascola2, R. Koup2, L. Corey3,
G.B. Nabel2, P. Gilber4, C. Morgan3, J. Maenza3, M. Keefer5, S.
Hammer6, G. Churchyard7, D.C. Montefior1, B.S Graham2, L.R.
Baden8, T.B. Kepler9, B.F. Haynes1
Duke University Medical Center, Human Vaccine Institute, Durham,
NC, United States, 2National Institute of Allergy and Infectious Diseases,
Vaccine Research Center, Bethesda, MD, United States, 3University of
Washington, Fred Hutchinson Cancer Research Center, Seattle, WA,
United States, 4University of Washington, SCHARP, Fred Hutchinson
Cancer Research Center, Seattle, WA, United States, 5University of
Rochester Medical Center, Division of Infectious Disease, Rochester,
NY, United States, 6Columbia University Medical Center, New York,
NY, United States, 7The Aurum Institute, Johannesburg, South Africa,
8
Brigham and Women’s Hospital, Boston, MA, United States, 9Boston
University, Boston, MA, United States
1
Background: The memory B cells in intestine contain a subset of cells
reactive with commensal bacteria. In acute HIV-1 infection (AHI), virus
replication is prominent in the gastrointestinal tract with early depletion
of CD4+ T cells and destruction of germinal centers. Antibodies in
serum and mucosal fluid in AHI are targeted to HIV-1 Env gp41, are
cross-reactive, non-neutralizing and do not select viral escape. Thus we
hypothesize that pre-transmission commensal bacteria antibodies that
cross-react with HIV-1 antigens may shape the B cell response to HIV-1.
Methods: Variable region gene segments of Ig heavy- and light-chain
(VH and VL) were isolated from single plasma cells and memory B cells
sorted from individuals with AHI and early HIV-1 infection, and from
vaccinees who received the prime-boost vaccination in HVTN Phase
II trials with VRC DNA and rAd5 vaccine containing clades A, B and
C env gp140 genes. The Isolated VH and VL genes were expressed as
recombinant mAbs for characterization.
Results: The predominant Env antibody response in blood in early HIV1 infection is to gp41. Of 412 mAbs isolated from terminal ileum plasma
cells, 19 (5%) were HIV-1-reactive, 14 of which were gp41-reactive,
and of these, 11 (79%) were cross-reactive with gut flora. One gut flora
antigen that reacted with gp41 antibodies was identified as E.coli RNA
polymerase. From the vaccinees with VRC DNA prime and rAd5 boost
(HVTN204 and HVTN082), 90.2% of 254 antibodies derived from
HIV-1 Env-specific memory B cell were gp41-reactive. Vaccine-induced
gp41 mAbs failed to neutralize HIV-1 transmitted/founder viruses,
and remarkably, 82% of the gp41 antibodies were cross-reactive with
intestinal commensal -bacterial antigens.
Conclusions: Acute HIV-1 infection and Env gp140 vaccination may
have induced dominant HIV-1 gp41 antibody responses resulted in part
from expansion of preexisting cross-reactive, mutated, memory B clones
that were triggered pre-HIV infection by commensal bacterial antigens.
Tuesday, 28 October
OA06.03
OA06.04
Plasmablast Phenotype and Mucosal
Antibodies to V2 in Vaccine-induced
Protection Against SIVmac251
Use of Enzyme-digested Virus-like Particles
as Probes for Flow Cytometric Sorting of HIVspecific Neutralizing Ab-producing B-cells
Luca Schifanella1,2, Nicolo` Binello1, Monica Vaccari1, Shari N.
Gordon1, Francesca Caccuri1, Matthew Blackburn1, Melvin Doster1,
Namal Liynage1, Poonam Pegu1, Xiaoying Shen3, Georgia D.
Tomaras3, David Venzon4, Don Stablien5, Susan W. Barnett6, Dan
Barouch7, Sanjay Phogat8, Genoveffa Franchini1
Evan M. Cale1, Nicole A. Doria-Rose1, Tommy Tong2,3, Ema T.
Crooks2,3, Richard Nguyen1, David R. Ambrozak1, Stephen P.
Perfetto1, Mario Roederer1, James M. Binley2,3, John R. Mascola1
1
NIH/NCI, Vaccine Branch, Bethesda, MD, United States, 2Università
degli Studi di Milano, Department of Biomedical and Clinical
Sciences - Section of Infectious Diseases and Immunopathology
‘L. Sacco’, Milano, Italy, 3Duke Human Vaccine Institute, Durham,
NC, United States, 4NIH/NCI, Biostatistics and Data Management
Section, Bethesda, MD, United States, 5The EMMES Corporation,
Rockville, MD, United States, 6Novartis Vaccines and Diagnostics
Inc., Cambridge, MA, United States, 7Harvard Medical School, Beth
Israel Deaconess Medical Center, Boston, MA, United States, 8Sanofi
Pasteur, Swiftwater, PA, United States
Background: We have recently recapitulated the RV144 vaccine efficacy
in a SIVmac251 model. In our study, rectal anti-cyclic V2 IgG antibodies
correlated with a decrease risk of SIVmac251 acquisition (p=0.0063).
Analysis of the homing markers on plasmablast (PB) resulted in a higher
frequency of alpha4beta7+ PBs in animals with higher levels of IgG and
IgA to cyclic V2 in rectal mucosal.
Methods: We investigated the homing potential in 5 different vaccine
strategies by measuring alpha4beta7 and CXCR3 as markers for gut
mucosa and inflammatory sites, respectively in a total of 118 macaques.
The immunoglobulin (Ig) expression on PB and the mucosal antibody
responses were assessed in all groups. We studied a cohort of macaques
immunized 4 times with ALVAC-SIV and twice with gp120/alum or
gp120/MF59. A second cohort was immunized twice with DNA-SIV/
gp120/alum or once Ad26-SIV, and boosted with ALVAC-SIV/gp120/
alum twice. Finally, an additional group was immunized 4 times with
NYVAC-SIV boosted twice with gp120/alum.
Results: Both ALVAC-SIV/gp120/alum and DNA-SIV/gp120/alum
immunized animals were protected from SIVmac251 mucosal acquisition
(p=0.029 and p=0.014, respectively). However, no protection was
observed with ALVAC-SIV/gp120/MF59, NYVAC-SIV/gp120/alum and
Ad26 primed regimens. DNA-SIV/gp120/alum and ALVAC-SIV/gp120/
alum immunizations increased the frequency of alpha4beta7 plasmablasts
(p=< 0.0001 and p=0.015) while ALVAC-SIV/gp120/MF59, NYVACSIV/gp120/alum increased the frequency of CXCR3+ plasmablasts
(p=0.0001 and p=0.004 respectively). We are completing Ad26-SIV/
gp120 and DNA-SIV/gp120/alum studies as well as the analysis of Ig
expression and correlations with mucosal antibody responses.
Conclusions: Preliminary results suggest that different vaccine
modalities are able to alter the plasmablast homing and the effector
functions of the antibody response at mucosal sites that may correlate
with protection.
NIAID, National Institutes of Health, Vaccine Research Center,
Bethesda, MD, United States, 2Torrey Pines Institute for Molecular
Studies, San Diego, CA, United States, 3San Diego Biomedical Research
Institute, San Diego, CA, United States
1
Background: Isolating new broadly neutralizing antibodies (bnAbs)
provides fresh insights for rational HIV-1 vaccine design. Several sites
of vulnerability to bnAbs are now well defined, and it is likely that
additional targets exist. Virus-like particles (VLPs) that express Env
trimers display all of the natural epitopes and are therefore attractive
candidates for use as probes. However, VLP surfaces are also decorated
with nonfunctional forms of HIV Env. Exposing VLPs to proteases clears
nonfunctional Env, leaving native trimeric Env intact. The resulting
“trimer-VLPs” preferentially bind to nAbs compared to non-neutralizing
antibodies and therefore may have gained sufficient selective power for
use as B cell probes to identify new bnAbs.
Methods: PBMC from HIV-infected patients were stained with the
enzyme-digested trimer-VLPs, and VLP-positive memory B cells were
sorted by flow cytometry. Multiplex RT-PCR was performed on cell lysates
to amplify kappa, lambda, and heavy chain genes. Cloned antibodies
were assessed for neutralization breadth and potency against a panel of
Env-pseudoviruses using the TZM-bl neutralization assay.
Results: VLPs were used to sort B cells from the donor of the broad
CD4bs antibody VRC13. The sort yielded relatives of VRC13, as well
as unrelated sequences that exhibited high sequence divergence from
germline VH genes and/or long CDR3 regions, which are characteristics
of known bnAbs. Cloning and characterization of these Abs is ongoing.
Conclusions: Unlike many previously used B cell probes, VLPs express
multiple epitopes including those for trimer specific antibodies which
allows for simultaneous sorting of B-cells of different specificities.
Additionally, these probes are devoid of non-functional forms of Env
and therefore preferentially bind to neutralizing Ab-producing B cells.
VLPs therefore hold great promise for isolating novel HIV-specific bnAbs
that can better inform vaccine design.
www.hivr4p.org
53
OA06.05
OA06.06
Viral Escape Pathways from Broadly
Neutralising Antibodies Targeting the HIV
Envelope Cleavage Site Enhance MPER
Mediated Neutralisation
Maturation Pathways for the Broad and
Potent Antibody 10E8 using a Combination of
Next Generation Sequencing, Bioinformatics
and Functional Analysis
Constantinos Kurt Wibmer1,2, Daniel J. Sheward3, Jinal N.
Bhiman1,2, Nonkululeko Ndabambi3, Debra H. Elliot4, Julie
Rouelle4, Ashley Smira4, Salim S. Abdool Karim5, James E.
Robinson4, Lynn Morris1,2,5, Carolyn Williamson3,5, Penny L.
Moore1,2,5
Cinque S. Soto1, Gilad Ofek1, M. Gordon Joyce1, Baoshan Zhang1,
Krisha McKee1, John R. Mascola1, Peter D. Kwong1
Centre for HIV & STIs, National Institute for Communicable Diseases,
NHLS, Johannesburg, South Africa, 2Faculty of Health Sciences,
University of the Witwatersrand, Johannesburg, South Africa, 3Institute
of Infectious Disease and Molecular Medicine (IIDMM) and Division
of Medical Virology, University of Cape Town and NHLS, Cape
Town, South Africa, 4Tulane University Medical Center, Department
of Pediatrics, New Orleans, LA, United States, 5Centre for the AIDS
Programme of Research in South Africa (CAPRISA), Nelson R Mandela
School of Medicine, University of KwaZulu-Natal, Durban, South Africa
1
Background: A preventative HIV-1 vaccine will likely elicit broadly
neutralising antibodies (bNAbs). The diverse modes of neutralisation by
bNAbs leads to variable escape pathways in each epitope. Documenting
escape and identifying common elements for each site of vulnerability
will inform immunogen design. Here, we map a newly described bNAb
specificity at the gp120-gp41 interface, and define viral escape in
CAPRISA donor CAP248.
Methods: Escape mutations were identified from longitudinal singlegenome envelope sequences, and confirmed by mutagenesis and
neutralisation assays. CAP248 plasma responses were mapped
longitudinally to determine the kinetics of bNAb maturation in response
to neutralisation escape. A monoclonal antibody (mAb), CAP248-30.2B,
was isolated by B-cell culture and characterised.
Results: The CAP248-30.2B mAb recapitulated the plasma breadth, but
was significantly less potent due to an unusual neutralisation plateau.
The CAP248-30.2B mAb was sensitive to deletion of the N611 glycan in
most heterologous viruses, but this mutation had a less significant effect
on CAP248 plasma, suggesting a swarm of cleavage site directed bNAbs
in CAP248 with differential glycan dependence. Viral escape mutations
from CAP248 bNAbs accumulated in the gp160 cleavage sites (positions
500, 502, 505, and 507-509) as well as proximal sites in gp41. These
changes collectively resulted in 100-fold enhanced neutralisation by the
MPER bNAbs 4E10 and 10E8. The binding of 4E10 or 10E8 to captured
virus enhanced CAP248-30.2B binding.
Conclusions: New bNAbs targeting the gp120-gp41 interface, such as
those in CAP248, have provided an additional target for HIV-1 vaccines.
The enhancement of MPER mediated neutralisation by CAP248 escape
mutations in the cleavage site suggests a synergistic relationship
between cleavage site and MPER bnAbs, similar to that between V1V2
and CD4 binding site bNAbs. Delineation of CAP248 virus-antibody coevolution may thus provide a blueprint for the co-induction of these two
antibody classes.
54
NIAID, NIH, Vaccine Research Center, Bethesda, MD, United States
1
Background: Antibodies that target the membrane-proximal external
region (MPER) of the HIV-1 gp41 subunit represent one of the few
categories of antibodies that effectively neutralize HIV-1. Such antibodies
provide potential templates to guide vaccine strategies, but details of
their generation and maturation have been unclear.
Methods: From the PBMCs of a single time point of donor N152, we used
454 Pyrosequencing to obtain the sequences of the variable domains of
heavy and light antibodies from B cell transcripts. Bioinformatics sieving
of gene families and structure-based sequence signatures delineated
potential transcripts from the maturation pathway of the effective MPERdirected antibody 10E8. We calculated maturation intermediates along
these pathways, synthesized heavy and light chains, and used transient
transfection to express antibodies, which we assessed for neutralization
and for MPER-peptide affinity.
Results: We identified 235 heavy chain sequences with V, D and J
segments matching the mature 10E8 antibody and 147,691 light chain
sequences with V and J segments matching the mature 10E8 antibody.
These sequences were further sieved with structural signatures. Pathways
- based on maximal intermediates, minimal sequence reversions,
and minimal N-nucleotide additions - for heavy and light chain were
identified, and we paired heavy and light chain sequences for the
unmutated common ancestor (UCA) along with three inferred heavy and
light chain sequences. Unexpectedly, the CDR H3 of the UCA differed
substantially from the mature sequence. Nonetheless, all of the paired
intermediates, including the UCA, showed binding to MPER peptide,
while only the two most mature intermediates showed neutralization.
Binding to MPER in the context of lipid micelles, meanwhile, mirrored
that of neutralization.
Conclusions: The combination of cross-sectional NGS data, bioinformatics,
and functional analysis can be used to define the maturation pathways for
an antibody, even with cross-sectional data.
Tuesday, 28 October
OA07.01
OA07.02
Characteristics of Young Black Men who Have
Sex with Men Enrolled in HPTN 061
Through the Rabbit Hole: Considering the
Situational Experience of Risk among Men
who Have Sex with Men in the Context of HIV
Prevention
Christopher Chauncey Watson1, Jonathan Paul Lucas2, Darrell P.
Wheeler3
The George Washington University, Public Health Research Clinic,
Washington, DC, United States, 2FHI 360, Science Facilitation, Durham,
NC, United States, 3Loyola University Chicago, School of Social Work,
Chicago, IL, United States
1
Background: Men who have sex with Men (MSM) comprise the largest
proportion of new HIV diagnosis in the United States (CDC, 2012).
The prevalence among Black MSM is higher than that of Latino or
White MSM (Jeffries, 2012). Most studies documenting HIV incidence
among Black MSM have been based on cross-sectional studies of HIV
surveillance. Very few studies have estimated incidence among young
Black MSM, who continue to be disproportionately infected at higher
rates than other racial/ethnic groups.
Methods: HPTN 061 (The BROTHERS Study) was a six US city study
designed to determine the feasibility and acceptability of a multicomponent HIV prevention intervention for Black MSM. Men were
recruited from 7/09 to 10/10 through a multi-pronged approached.
Once found eligible, men participated in 3 visits (baseline, 6 mo, 12 mo)
visits which included ACASI interviews, demographic collections, HIV/
STI testing and counseling, social/sexual network questionnaire and the
opportunity to work with a Peer Health Navigator.
Results: Of the 1553 Men recruited in HPTN 061, 519 were < 30.
Overall HIV incidence for HPTN 061 was 3% but was 5.9% among < 30
y.o. Younger men had more STIs, no usual place for healthcare and more
identified barriers to receiving health care. A multivariate analyses found
HIV incidence was associated with younger age (HR 3.4, CI: 1.4-8.3)
and unprotected receptive anal intercourse with HIV positive partners or
unknown status partner (HR 4.1, 1.9-9.1). Over 50% of the individuals
reported no work and no health coverage.
Conclusions: In the largest longitudinal cohort of Black MSM to date
in the US, HIV incidence was highest among Young BMSM. We found
significant drivers for HIV acquisition among Young Black MSM to
include; access to health care services, higher sexual risk behaviors, and
occurrence of STIs. Additional research is needed to focus primarily on
this population to develop effective prevention strategies.
Martina Brostrom1, Nicola Desmond2,3
UNAIDS, the Joint United Nations Programme on HIV/AIDS, Geneva,
Switzerland, 2Malawi-Liverpool-Wellcome Trust, Blantyre, Malawi,
3
Liverpool School of Tropical Medicine, Department of Clinical Sciences,
Liverpool, United Kingdom
1
Background: After 3 decades of research and programming for MSM,
in 2014, HIV remains a major public health challenge in this population.
Even in countries where MSM have access to HIV services, civil
liberties and community structures, prevalence rates are consistently
high. This warrants development and scale up of bespoke, innovative
and meaningful HIV combination prevention programmes that better
respond to the diverse social and health needs of MSM. A better insight
into the experiences of risk and the impact on sexual practices among
MSM is required to inform such approaches.
This paper provides a metasynthesis of the linkages between the
situational experience of risk for MSM and unprotected sexual
intercourse and explores the implications for HIV prevention.
Methods: A systematic search of nine electronic databases was
conducted and complemented by a free text search on Google scholar
on the name and publications of key authors and an inspection of the
references of identified articles. Six qualitative studies that investigated
the situational experience of risk among MSM were identified. A
metaethnographic synthesis was undertaken to describe, analyse and
interpret the studies.
Results: The synthesis resulted in the identification of 6 themes: losing
control; relationship building; fatalism, homophobia and stigma of HIV;
prevention literacy and confusion; risk management strategies and the
biography of individuals.
Conclusions: MSM are well aware of the risks of contracting HIV through
unprotected anal intercourse and deploy prevention strategies to stay
safe and healthy, but what they conceive as safe and risk is different
from that of medical professionals. Frequently MSM are ambiguous
about risk and this is reflected in their sexual practices. Those working
in HIV prevention must be aware of the lay risk management strategies
deployed by MSM and engage with them.
www.hivr4p.org
55
Oral Abstract Session 07: Risk and Prevention for MSM
OA07.03
OA07.04
Sexual Behaviour Profile of Gay and Other
Men who Have Sex with Men Enrolled in the
PROUD Pre-exposure Prophylaxis Open-label
Pilot Study in England
Structural Stigma Affects Access to Pre- and
Post-exposure Prophylaxis and HIV Risk
among Men who Have Sex with Men (MSM) in
the United States
Mitzy Gafos1, David Dolling2, Monica Desai2, Gemma Wood2,
David Dunn2, John Saunders3, Nicola E. Mackie4, Alexandra
Meijer5, Amanda Clarke6, Christine Bowman7, Charles Lacey8,
Lisa Southon9, Clare Oakland10, Chris Higgs11, David White12, Iain
Reeves13, Michael Brady14, Julie Fox15, Anthony Nardone16, on
behalf of the PROUD Study Team
Catherine Oldenburg1, Amaya Perez-Brumer2, Mark
Hatzenbuehler2, Douglas Krakower3, David Novak4, Matthew
Mimiaga1,5,6, Kenneth Mayer3,5,7
1
MRC Clinical Trials Unit at UCL, Institute of Clinical Trials & Methodology,
London, United Kingdom, 2MRC Clinical Trials Unit at UCL, London,
United Kingdom, 3Ambrose King Centre/Barts Sexual Health Centre,
London, United Kingdom, 4Jefferiss Wing, St Mary’s Hospital, London,
United Kingdom, 556 Dean Street Clinic, London, United Kingdom, 6Elton
John Centre, Brighton & Sussex University Hospital Trust Sussex House,
Brighton, United Kingdom, 7Royal Hallamshire Hospital, Dept of GU
Medicine, Sheffield, United Kingdom, 8Monkgate Health Centre, Dept
of GU Medicine, York, United Kingdom, 9Manchester Centre for Sexual
Health, Manchester, United Kingdom, 10Centre for Sexual Health & HIV
Research, Mortimer Market Centre, London, United Kingdom, 11John
Hunter Clinic, St. Stephen’s Centre, Chelsea & Westminster Hospital,
London, United Kingdom, 12Medical Innovation Development Research
Unit (MIDRU), Birmingham Heartlands Hospital, Birmingham, United
Kingdom, 13Homerton Sexual Health Services, London, United Kingdom,
14
Caldecot Centre, London, United Kingdom, 15St Thomas’ Hospital,
London, United Kingdom, 16Public Health England, London, United
Kingdom
Background: The public health benefit of PrEP will depend on effective
targeting of higher risk individuals. We report the baseline sexual
behaviour of men enrolled in England’s PROUD pilot study.
Methods: In PROUD, eligible HIV negative gay/MSM, aged 18+, who
report recent and intended condomless anal sex, are randomized to
receive Truvada as PrEP immediately or after 12 months.
Results: The PROUD cohort of 470 men are a median age of 35 (IQR 34-36),
79% white, 58% UK born, 60% university educated, 82% employed, 47%
in a relationship and 96% self-identify as gay. Anal sex was reported with a
median of 10 (IQR 5-20) partners total, and 7 (IQR 2-15) new partners in the
last 90 days. Condoms were not used with a median of 2 (IQR 1-5) partners
for receptive sex and 3 (IQR 1-6) for insertive. Reasons for no condom at last
condomless sex were: 66% less pleasure, 51% don’t like condoms, 33%
partner didn’t like condoms, 27% condoms not discussed, 23% and 22%
under influence of drugs or alcohol (68% consumed alcohol weekly and
73% used recreational drugs in the last 90 days). At last condomless sex,
45% assumed partner to be HIV-, 27% HIV+ on ART, 23% HIV unknown,
5% HIV+ not on/unknown ART. When having condomless anal sex,
29% felt at little or no risk, 47% somewhat at risk, 23% at high risk. Risk
reduction strategies reported were: 39% use condoms, 26% ask partner
to use condom, 38% chose HIV- partners, 29% use strategic positioning,
23% seek partners on ART; only 16% reported no strategy. In the last year
39% had used PEP (19% >=2 times); 87% had visited a clinic >=2 times
for HIV and 82% for STI testing. 37% reported a bacterial rectal STI, which
was associated with higher numbers of condomless sex partners (OR 1.09
receptive p< 0.001, 1.04 insertive p=0.011).
Conclusions: PROUD is enrolling gay men who are selective condom users
with high rates of STIs, who despite regular clinic attendance and use of risk
reduction strategies, including sero-sorting and PEP, are likely to be at high
risk of HIV acquisition.
56
Harvard School of Public Health, Department of Epidemiology, Boston,
MA, United States, 2Columbia University Mailman School of Public
Health, Department of Sociomedical Sciences, New York, NY, United
States, 3Beth Israel Deaconess Medical Center, Boston, MA, United
States, 4OLB Research Institute, Cambridge, MA, United States, 5The
Fenway Institute, Boston, MA, United States, 6Massachusetts General
Hospital, Department of Psychiatry, Boston, MA, United States, 7Harvard
School of Public Health, Department of Global Health and Population,
Boston, MA, United States
1
Background: Stigmatizing social environments (structural stigma) are
hypothesized to affect HIV risk through several pathways, including
decreased access to HIV prevention services.
Methods: In August 2013, members of the largest MSM social networking
site in the US completed a survey about current HIV prevention practices.
State-level structural stigma was based on a previously validated
composite index: 1) density of same-sex couples; 2) proportion of
Gay-Straight alliances per public high school; 3) 4 state laws related
to sexual orientation; and 4) public opinion toward homosexuality.
This information was linked to survey responses via participants’ state
of residence. Multivariable logistic generalized estimating equations
were used to assess the relationship between structural stigma and 3
outcomes: (1) unprotected anal sex (UAS) in the previous 3 months; (2)
having taken or heard of HIV pre-exposure prophylaxis (PrEP) and postexposure prophylaxis (PEP); and (3) comfort discussing male-male sexual
behavior and HIV prevention with primary care providers, adjusting for
demographic and socioeconomic covariates.
Results: Among 5,321 HIV-uninfected MSM, lower levels of structural
stigma were associated with decreased odds of UAS (aOR 0.97, 95% CI
0.94-0.99), greater odds of awareness of (aOR 1.05, 95%CI 1.02-1.09)
and taking PEP (aOR 1.10, 95% CI 1.01 to 1.19), awareness of (aOR 1.04,
95% CI 1.01-1.08) and taking PrEP (aOR 1.13, 95% CI 1.04-1.23), and
comfort discussing male-male sex (aOR 1.08, 95% CI 1.05-1.11) and
HIV prevention strategies (aOR 1.05, 95% CI 1.02-1.08) with primary
care providers.
Conclusions: MSM living in highly stigmatizing environments report
greater individual-level risk behaviors and decreased knowledge
and access to biomedical HIV prevention strategies, placing them
at heightened vulnerability for HIV. Legal reforms to protect sexual
minorities are needed to reduce HIV risk and support existing HIV
prevention efforts among MSM.
Tuesday, 28 October
OA07.05
OA07.06 LB
Multiple HIV Counselling Sessions and Safe
Sex Practice Helped Bangkok Men who Have
Sex with Men Stay HIV-uninfected: Thailand
2006-2014
Project PrEPARE: High Levels of Medication
Adherence with Continued Condomless Sex
in U.S. Men who Have Sex with Men in an Oral
PrEP Adherence Trial
Wipas Wimonsate1, Sarika Pattanasin1, Anuwat Sriporn1,
Pikunchai Luechai1, Kesinee Satumay1, Narongritt Tippanonth1,
Nutthawoot Promda1, Anchalee Varangrat1, Anupong
Chitwarakorn2, Timothy H. Holtz1,3
Kenneth H. Mayer1,2,3, Steven Safren1,4,5, Jessica Haberer3,6, Steven
Elsesser1, William Clarke7, Craig W. Hendrix7, Mark Marzinke7,
Matthew J. Mimiaga1,4,8
Thailand-MoPH U.S. CDC Collaboration, Nonthaburi, Thailand, Ministry
of Public Health, Department of Diseases Control, Nonthaburi, Thailand,
3
Division of HIV/AIDS Prevention, U.S. Centers for Disease Control and
Prevention, Atlanta, GA, United States
2
1
2
Background: In prior surveys, Thai men who have sex with men (MSM)
have had a low level of HIV awareness and prior history of testing, and
significant levels of unsafe sex. We hypothesized that reinforcement
of HIV risk-reduction strategies through multiple counselling sessions
would be associated with remaining HIV uninfected.
Methods: We enrolled Thai MSM from the Bangkok metropolitan area
into a five-year cohort study with four-monthly visits (maximum of 16
visits). At every visit, MSM received a comprehensive counselling session:
HIV transmission information; risk-reduction strategies; HIV testing;
and provision of condoms and lubricants. Logistic regression was used
to investigate if number of visits was associated with remaining HIV
uninfected.
Results: From April 5, 2006 to March 24, 2014, 1,260 HIV-uninfected
MSM, age ≥18 years, were enrolled, and followed and tested for HIV
every four months. The mean number of follow-up visits during the
study period was 11 (Standard Deviation, SD, 5). We detected 239
MSM with incident HIV infection. Logistic regression showed the odds of
remaining HIV uninfected were 1.3 times as high for each subsequent
visit (Adjusted Odds Ratio, AOR, 1.29, 95% Confidence Interval, CI, 1.241.34), and associated with pre-study practice of only insertive sex (AOR
2.53, 95% CI 1.53- 4.16), protected anal sex (AOR 1.75, 95% CI 1.242.45), and never joining in group sex (AOR 1.73, 95% CI 1.24-2.42),
when adjusted for pre-study age group, education level, employment,
living arrangement, recreational drug use, coerced sex, history of STI
testing, history of HIV testing, and HIV knowledge.
Conclusions: Regardless of other pre-study demographic and behavioral
factors, more visits and pre-study safe sex practices were associated
with a higher odds of remaining HIV uninfected. Safe sex messages and
retention in comprehensive HIV testing services should be strengthened
among Thai MSM.
Fenway Health, The Fenway Institute, Boston, MA, United States,
Beth Israel Deaconess Medical Center, Division of Infectious Diseases,
Boston, MA, United States, 3Harvard Medical School, Medicine, Boston,
MA, United States, 4Massachusetts General Hospital, Psychiatry, Boston,
MA, United States, 5Harvard Medical School, Psychiatry, Boston, MA,
United States, 6Massachusetts General Hospital, Medicine, Boston,
MA, United States, 7Johns Hopkins University School of Medicine,
Pathology, Baltimore, MD, United States, 8Harvard School of Public
Health, Epidemiology, Boston, MA, United States
1
Background: Pre-exposure prophylaxis (PrEP) has been shown to
decrease HIV incidence in MSM, but optimal protection is dependent
on medication adherence. The data presented here are from a pilot
study, designed to assess whether an intervention based on Life-Steps,
an effective intervention to enhance HAART adherence in HIV-infected
patients, could improve PrEP adherence among men who have sex with
men (MSM). who reported recent condomless anal sex.
Methods: Between 11/2012 and 12/2013, 55 Boston-area HIV- MSM
who were interested in initiating PrEP were enrolled in Project PrEPARE,
and randomized to Life-Steps which included 4 weekly sessions that
addressed barriers and facilitators of PrEP adherence, or to a control
condition that provided supportive counseling. Adherence was monitored
by Wisepill, and unprotected sex was assessed via daily text messaging.
Results: Participants were primarily White (94%) and well-educated
(64% had completed college), and sociodemographic factors were
similar in both randomized groups. Over the 6 months’ observation,
90% of the MSM in each group took at least 80% of their daily pills
on a weekly basis, as measured by Wisepill and drug levels, and this
did not differ by study condition. Condomless sex rates did not change
significantly in either group over time. About 90% of the time during
the study, the men in both groups either had drug levels consistent with
taking most of their PrEP doses and/or used condoms during sex. No
incident HIV infections occurred during the study.
Conclusions: Most MSM in this study demonstrated levels of PrEP
adherence consistent with protective benefit, whether they received
Life-Steps or supportive counseling. Given the high rates of adherence
in both groups, the data suggest that MSM who elect to receive
open-label PrEP can be highly adherent and self-protective. New PreP
adherence interventions should focus on at risk persons who anticipate
or demonstrate difficulties with routine medication adherence.
www.hivr4p.org
57
Oral Abstract Session 08: Correlates of Protection and Exposure
OA08.01
OA08.02
Evaluation of Mucosal Tissue Explants as ex
vivo Surrogates of in vivo Vaccination of Nonhuman Primates (NHPs) and Humans
Evolutionary Analysis Identifies an MX2
Haplotype Associated with Natural Resistance
to HIV-1 Infection
Carolina Herrera1, Ronald Veazey2, Alexandra Schuetz3, Natalia
Olejniczak1, Agnès-Laurence Chenine4, Sorachai Nitayaphan3,
Jaranit Kaewkungwal5, Punnee Pitisuttithum5, Supachai RerksNgarm6, Robert J. O’Connell4, Jean-Louis Excler4, Jerome H. Kim4,
Robin Shattock1
Irma Saulle1, Mara Biasin1, Federica Gnudi1, Salomè Ibba1,
Micaela Garziano1, Manuela Sironi2, Daria Trabattoni1, Sergio Lo
Caputo3, Francesco Mazzotta3, Antonio Caruz4, Luca De Gioia5,
Mario Salvatore Clerici6
Imperial College, Infectious Diseases, London, United Kingdom,
2
Tulane National Primate Research Center, Tulane, LA, United States,
3
Armed Forces Research Institute of Medical Sciences, Bangkok,
Thailand, 4Walter Reed Army Institute of Research, Rockville, MD,
United States, 5Mahidol University, Bangkok, Thailand, 6Ministry of
Public Health, Bangkok, Thailand
2
1
Background: HIV vaccine trials have revealed the need to establish
better correlations between NHP and human studies. This project aimed
to use ex vivo cervicovaginal and colorectal explants from NHPs and
from participants of the ongoing trial RV305 to assess B-cell responses
elicited upon vaccination with ALVAC/AIDSVAX B/E. Having shown that
in vivo vaccination elicits specific anti-gp120 HIV-1CM244 IgG and IgA
systemically and mucosally, we assessed the ex vivo infectibility of
NHP and human explants and the neutralization profile of tissue culture
supernatants
Methods: Six Rhesus macaques were immunized with ALVAC/AIDSVAX
B/E following RV144 regime. Colorectal and cervicovaginal biopsies were
obtained 4 weeks pre-vaccination, and 2 weeks after each immunization.
Serum was collected at the same time points. Vaginal and colorectal
biopsies were obtained at week 26 from participants enrolled in RV305
having been vaccinated at weeks 0 and 24 with ALVAC/AIDSVAX B/E,
AIDSVAX B/E, or ALVAC, or placebo in each group. Neutralization profile
of non-infected explant supernatants against two CRF01_AE isolates was
assessed in TZM-bl cells. Explants were challenged ex vivo with SHIVBaL
or SHIVSF162P3 or with HIV-1 CM235-LucRT2A and cultured for 15
days. Infectibility was determined by measurement of p27/p24 viral
antigen in culture supernatants. Limited unblinding of RV305 samples
was performed allowing correlation of samples to groups
Results: Moderate neutralization was observed in all NHP and human
explants and in NHP serum. Furthermore, limited ex vivo infectibility
was detected in NHP mucosal explants after vaccination. Variable levels
of infection were observed in human samples corresponding potentially
to individuals having received vaccine or placebo within each group
Conclusions: While awaiting for further unblinding, this study supports
our hypothesis that ex-vivo mucosal tissue models could represent
a new tool to assess mucosal B-cell responses to vaccination in NHP
studies and during clinical trials
58
University of Milan, Biomedical and Clinical Sciences, Milan, Italy,
Scientific Institute IRCCS MEDEA, Bosisio Parini, Milan, Jamaica,
3
S. Maria Annunziata Hospital, Florence, Italy, 4University of Jaen,
Immunogenetic Unit, Jaen, Spain, 5University of Milan-Bicocca,
Biotechnology and Biosciences, Milan, Italy, 6Don Gnocchi Foundation,
Milan, Italy
1
Background: The human and macaque MX2 (myxovirus resistance 2)
proteins efficiently restrict HIV-1 and other simian immunodeficiency
viruses, but have a modest effect against retroviruses that infect nonprimate species. This observation points to species-specific virus-host
interactions that may result from evolutionary arms races. This research
project, thus, aimed to demonstrate the relevance of MX2 variants to
HIV-1 infection susceptibility
Methods: We analysed:
1) evolutionary history of MX2 in placental mammals (PAML analysis);
2) structural models of MX2;
3) genotyping analysis of rs2074560 in the MX2 gene in 3 independent
case-control cohorts of HIV-1 exposed seronegative individuals (HESN);
and
4) in vitro HIV-1 replication plus IFNα-stimulated MX2 mRNA expression
in peripheral blood mononuclear cells (PBMC) from healthy controls (HC)
grouped according to their MX2 genotype.
Results: MX2 evolved adaptively in mammals with distinct sites
representing selection targets in rodents and primates; pervasive
selection mainly involves residues in loop 4, previously shown to
carry antiviral determinants. We also demonstrated that recent distinct
selective events have driven the frequency increase of two haplotypes
in human populations of Asian and European ancestry. The Asian
haplotype carries a susceptibility allele for melanoma; the European
haplotype is tagged by rs2074560, an intronic variant. By analysis of
three independent HESN cohorts with different geographic origin and
exposure route we verified that the ancestral (G) allele of rs2074560
protects from HIV-1 infection with a recessive effect (combined p value
of 1.55x10-4). In line with these findings, the G allele is associated with
lower in vitro HIV-1 replication and increased MX2 expression.
Conclusions: Results herein establish a role for MX2 as a central element
of antiviral response in mammalian species and a possible target for
therapeutic intervention in HIV-1 treatment and prevention
Tuesday, 28 October
OA08.03
OA08.04
Systematic Analysis of HIV-1 Env Epitopes
of Two HLA Class I Alleles Associate with
Different Rates of HIV Infection in the
Pumwani Sex Worker Cohort
Comprehensive Sieve Analysis of
Breakthrough HIV-1 Sequences in the RV144
Vaccine Efficacy Trial
Meika EI Richmond1,2, Christina A. Daniuk1, Rupert E. Capina1,
Joshua Kimani3, Charles Wachihi3, Makubo Kimani3, Thomas
Bielawny1, Mark G. Mendoza1, T. Blake Ball1,2,3, Francis A.
Plummer2,3,4, Ma Luo1,2
Public Health Agency of Canada, National Lab for HIV Immunology,
Winnipeg, MB, Canada, 2University of Manitoba, Medical Microbiology,
Winnipeg, MB, Canada, 3University of Nairobi, Nairobi, Kenya, 4Public
Health Agency of Canada, National Microbiology Lab, Winnipeg, MB,
Canada
1
Background: HIV exposed seronegative (HESN) individuals provide a
unique opportunity to study natural immunity to HIV-1. Our previous
studies on HESN women from the Pumwani sex worker cohort showed
that A*01:01 is associated with slower seroconversion while B*07:02 is
associated with rapid seroconversion. Understanding why these alleles
associate with different infection rates and their epitope characteristics
may provide clues for design of an effective HIV vaccine.
Methods: We screened 1820 peptides (9mer overlapping by 8aa) of
HIV-1 clade A/D Env for A*01:01 and B*07:02 epitopes using iTopia
Epitope Discovery System. Binding eptiopes were analyzed for affinity,
off-rates and validated by IFNγ ELISpot. Selected tetramers were used
(A*0101:VI/VK, B*0702:SL/KL) for flow cytometry analysis of CD8 T cell
memory (CCR7/CD45Ra) and differentiation (CD27/CD28) phenotypes.
Results: A*01:01 bound 20 epitopes, while B*0702 bound 64. No
differences where seen in peptide binding affinity or off-rate between the
two alleles. ELISpot responses were stronger in A*01:01+ patients (p<
0.0001) and a higher proportion of A*01:01 epitopes is in the constant
regions. Tetramer specific CD8s had distinct memory and differentiation
profiles than non-specific CD8s (p=0.0002). Additionally, CD8 T cells
specific for A*01:01 epitope YI were more likely to have an effector
memory (CCR7-CD45RA-) phenotype than B*07:02 specific-CD8 T cells
(p=0.0002). Moreover, CD8 T cells specific for both A*01:01 epitopes
(YI/VK) were highly antigen experienced (CD27-CD28-) compared to
those specific for B*07:02 epitopes (p< 0.0001).
Conclusions: These data are consistent with our previous study of HIV-1
Gag epitopes of these alleles. A*01:01 recognizes fewer epitopes than
B*07:02, suggesting narrow immune responses targeting conserved
regions may be better than broad immune responses. Additionally,
A*01:01 associates with higher effector memory and higher antigen
experience, suggesting high cytolytic activity and potentially more
effective responses.
Paul T. Edlefsen1, Morgane Rolland2, Tomer Hertz1, Sodsai
Tovanabutra2, Andrew J. Gartland1, Allan C. deCamp1, Craig A.
Magaret1, Hasan Ahmed1, Raphael Gottardo1, Michal Juraska1,
Connor McCoy3, Brendan B. Larsen4, Eric Sanders-Buell2, Chris
Carrico5,6, Sergey Menis5,6, Meera Bose2, Miguel A. Arroyo7, Robert
J. O’Connell8, Mark S. deSouza8, Sorachai Nitayaphan7, Punnee
Pitisuttithum9, Jaranit Kaewkungwal7, Supachai Rerks-Ngarm10,
Merlin L. Robb2, Jason S. McLellan11, Ivelin S. Georgiev11, Tatsiana
Kirys11, Peter D. Kwong11, Jonathan M. Carlson12, Nelson L. Michael2,
William R. Schief5,6,13, James I. Mullins4, Jerome H. Kim2, Peter B.
Gilbert1, RV144 Sequencing Team
1
Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease
Division, Seattle, WA, United States, 2US Military HIV Research Program,
Silver Spring, MD, United States, 3Fred Hutchinson Cancer Research Center,
Public Health Sciences Division, Seattle, WA, United States, 4University
of Washington, Department of Microbiology, Seattle, WA, United States,
5
University of Washington, Department of Biochemistry, Seattle, WA, United
States, 6The Scripps Research Institute, IAVI Neutralizing Antibody Center
and Department of Immunology and Microbial Sciences, La Jolla, CA,
United States, 7AFRIMS, Royal Thai Army Component, Bangkok, Thailand,
8
AFRIMS, US Army Component, Bangkok, Thailand, 9Mahidol University,
Faculty of Tropical Medicine, Bangkok, Thailand, 10Thai Ministry of Public
Health, Nonthaburi, Thailand, 11Vaccine Research Center, NIAID, NIH,
Bethesda, MD, United States, 12Microsoft Research, Redmond, WA, United
States, 13Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United
States
Background: The RV144 clinical trial showed the partial efficacy of
a prime-boost pox-protein vaccine regimen with an estimated vaccine
efficacy (VE) of 31% for protecting low-risk Thai volunteers against
acquisition of HIV-1. The impact of vaccine-induced immune responses
can be investigated through sieve analysis of HIV-1 breakthrough
infections (infected vaccine and placebo recipients).
A V1/V2-targeted comparison of the genomes of HIV-1 breakthrough
viruses from vaccine and placebo recipients identified two V2 amino acid
loci that differed between the vaccine and placebo groups, corroborating
the finding that V2-specific antibodies in vaccine recipients were
associated with a reduced risk of HIV-1 infection.
Methods: Here we extended the V1/V2 sieve analysis to the entire HIV1 genome using an array of sieve analysis methods based on individual
sites, k-mers and genes/proteins.
Results: Across the HIV-1 proteome, we identified 56 amino acid sites or
“signatures” and 119 k-mers that differed between the vaccine and placebo
groups; among these, 19 signature sites and 38 k-mers were located in Envgp120, Gag, and Pro that constituted the RV144 vaccine. The nine signature
sites in Env-gp120 were significantly more likely to be known antibodyassociated sites than non-signature sites (p = 0.0021). In particular, one
signature in V3 (317) overlapped with a hotspot of antibody recognition,
and sites 369 and 424 are linked to CD4 binding site neutralization.
Conclusions: Since signature sites were not preferentially restricted to
the vaccine immunogens and because most of the statistical significance
did not withstand multiplicity adjustment, we predict that few of the
56 genetic signatures found across the HIV-1 proteome are strongly
linked to the RV144 vaccine-induced immune pressure. The collection
of statistical methods and tools we employed constitutes an analysis
platform applicable to sieve analysis of breakthrough infection genomes
in general vaccine efficacy trials for diverse pathogens.
www.hivr4p.org
59
OA08.05
OA08.06 LB
SIVsmE660 Envelope Variants from
Breakthrough Infections Following DNA/
MVA Vaccination of Rhesus Macaques are
Susceptible to Neutralization
Microbicide-vaccine Combination Provides
Significant Protection against Vaginal SHIV162P3 Challenge in Cynomolgous Monkeys
Stacey A. Smith1, Samantha Burton1, Sharmila Reddy1, Katie
Kilgore1, Eric Hunter1, Harriet Robinson2, Rama Amara1, Cynthia
Derdeyn1
Emory University, Atlanta, GA, United States, 2GeoVax, Inc., Atlanta,
GA, United States
1
Background: SIVmac239-based DNA prime/MVA boost vaccine
regimens including GM-CSF or CD40L adjuvants were tested in rhesus
macaques, along with an MVA only regimen. Significant protection was
observed against a low dose repeat intra-rectal SIVsmE660 challenge.
We investigated whether vaccine-induced antibodies resulted in
breakthrough infection by neutralization resistant variants following
challenge.
Methods: We identified the SIVsmE660-derived transmitted/
founder envelope (T/F Env) sequence for 14 vaccinated and 4
control monkeys using SGA. T/F Env expression vectors were used to
generate pseudovirus, which was tested for neutralization sensitivity to
heterologous pooled serum from chronically SIV-infected monkeys, and
autologous vaccinated monkey serum collected at 13 weeks post-MVA
boost (3-4 weeks prior to challenge).
Results: All T/F Envs were susceptible to neutralization by the chronic
SIV-infected serum pools to varying degrees, but there was no significant
difference between Envs from vaccinated vs. control monkeys, or
between vaccine groups. In fact, the most neutralization resistant Env
variants were found in two control monkeys who had no pre-challenge
anti-SIV immunity. All T/F Envs from vaccinated monkeys were also
susceptible to neutralization by the week 13 autologous serum at IC50s
of up to 1:3,000. Finally, a recently described signature in the Env
gp120 C1 region, 45A/47K, did not determine resistance of the T/F Envs
against heterologous or autologous neutralization.
Conclusions: These results demonstrate that antibodies capable
of neutralizing the autologous T/F Envs at titers exceeding those
estimated to be necessary for protection against mucosal transmission
were generated by these vaccination regimens. Even though the
breakthrough T/F Envs were sensitive to neutralization, these animals
were not protected. Thus, increasing the magnitude, breadth, and/
or local mucosal production of these antibodies may be required to
enhance their protective potential.
60
Roger Le Grand1, Nathalie Nathalie Bosquet1, Stefania
Dispinseri2, Leslie Gosse1, Delphine Des Jardins1, Shen Shen3,
Georgia Tomaras3, Nicola Hopewell4, Susan Barnett5, Hela
Saidi1, Rodolphe Thiebaut6, Gabriella Scarlatti2, Alethea Cope4,
Robin J. Shattock4
Commissariat à l’Energie Atomique et aux Energies Alternatives,
Division of Immuno-Virology, IDMIT Center, Paris, France, 2San
Raffaele Scientific Institute, Milan, Italy, 3Duke Human Vaccine
Institute, Durham, NC, United States, 4Imperial College, London, United
Kingdom, 5Novartis Vaccines, Cambridge, MA, United States, 6Institut
de Santé Publique, d’Epidémiologie et de Développement – ISPED,
Bordeaux, France
1
Background: Although a number of new biomedical prevention tools
have demonstrated variable success against HIV infection in clinical
trials including a partially effective microbicide (tenofovir gel) and
a modestly protective vaccine (the Thai RV-144 trial), their combined
introduction could provide more potent protection. This study directly
explores potential positive interactions.
Methods: We used a nonhuman primate model to determine whether
combining a partially effective microbicide (1% tenofovir gel) with
an envelope based vaccine could provide enhanced efficacy against
repeat intravaginal challenge with SHIV-162P3. Vaccinated animals
received three nasal priming doses containing a combination of gp140
TV1 (Clade C) and SF162 (clade B) constructs administered with R848
(TLR7/8 agonist) followed by two intramuscular boost immunizations
administered with MF59.
Results: Tenofovir gel provided a 46% reduction in infection following
6 consecutive low dose intravaginal challenges (P=0.04), where
tenofovir gel in vaccinated animals provided 81% reduction (P=0.02)
and the vaccine alone failed to protect against SHIV-162P3 infection
(P=0.85). Following 12 consecutive challenges the the combination
group maintained a sustained protection of 63% (P=0.0006) while the
microbicide group only provided 14% reduction in infection (P=0.02).
In a second phase, protected animals were challenged a further 12
times in the absence of microbicide. Protected animals in the vaccine
group initially receiving tenofovir gel when challenged in the absence
of gel maintained protection (p=0.01) with a total risk reduction over 22
exposures of 38%, while animals initially receiving tenofovir gel were
fully susceptible to infection.
Conclusions: These important findings offer the possibility that
combined implementation of new biomedical prevention strategies may
provide significant reduction in HIV incidence and argues for accelerated
assessment of potentially beneficial combinations through randomised
controlled clinical trials.
Tuesday, 28 October
OA09.01
OA09.02
Why Women at High Risk for HIV-1 Infection
Did Not Join the VOICE Study in Uganda: A
Qualitative Community Study
Community Engagement in a Volatile
Community Post-marikana for a Phase III
Microbicide Ring Trial
Teopista Nakyanzi1, Samuel Kabwigu1, Doreen Kemigisha1,
Sophie C. Nanziri1, Patrick Ndawula1, Stella Nanyonga1, Juliane
Etima1, Flavia M. Kiweewa1, Rhonda White2, Lisa Noguchi3,
Clemensia Nakabito1
Cheryl Emily Louw1, Ntswaki Rose Masilo1, Marthie de Villiers1,
Annalene M. Nel2, Michelle Isaacs2
Makerere University–Johns Hopkins University Research Collaboration,
Kampala, Uganda, 2FHI 360, Community Program, Durham, NC, United
States, 3Microbicide Trials Network, Washington, DC, United States
Madibeng Centre for Research, Brits, South Africa, 2International
Partnership for Microbicides, Paarl, South Africa
1
1
Background: HIV-1 prevalence in Uganda remains among the highest in
the world, and recent surveillance data have not seen a significant drop
in HIV-1 incidence. The VOICE study evaluated safety and effectiveness
of daily use of female-controlled ARV-based oral and topical vaginal
HIV prophylaxis. Despite being at risk of HIV-1 acquisition, and having
limited prevention tools, many women declined to join the study. We
describe the underlying factors that hindered at-risk women from
joining VOICE.
Methods: Women aged 18-45 in high-risk communities identified
by the Uganda HIV/AIDS sero-Behavioral Survey 2004-05 were
sensitized about the VOICE study. Prospective participants’ questions
and concerns were addressed, and those willing to be screened on
site were systematically pre-screened using an IRB-approved checklist
with multiple risk questions. Presumptively eligible women were given
appointments and reminded by telephone calls.
Results: From November 2009 to December 2011, among the 3,217
women sensitized in the community, only 25% (n=820) were interested
and pre-screened. Of those expected to turn up, only 43% (n=356)
showed for appointments. Among those who did not turn up (n=464),
78% (n=362) did not know or were not sure of their partners HIV status
and feared finding out their HIV status; 58% (n=269) were not living
with their partner(s) and could not choose from which partner to obtain
permission; and 30% (n=139) had no income and feared losing financial
support from their partners. Other factors included fear of side effects,
myths and misconceptions about clinical research. Others needed more
time to think about participation, and the rest promised to come for
screening but later changed their minds.
Conclusions: Women are challenged by the fear of knowing their HIV
status, exacerbated by lack of independence to make decisions about
study participation. There is more need to sensitize communities about
HIV prevention research, participant challenges and involving male
partners in research.
Background: When Madibeng Centre for Research (MCR) began its
work on a Vaginal Microbicide Ring Phase III trial the surrounding
community was peaceful. Wage negotiations in the surrounding mines
became tense leading up to the Marikana massacre on 16 August 2012,
just more than 4 months into the trial. The situation in the area remained
tense and also led to male partner aggression towards their female
partners. Partners viewed the trial and the ring with suspicion resulting
in some participants being forced to stop using the ring and to withdraw
from the trial.
Methods: With participant consent, MCR strategized on ways to reach
out to the affected participants’ partners and to engage the local
community:
•
Partners were visited personally and given appropriate
information on the trial
•
Partners were invited to attend the research centre (RC) to
view procedures and interview the Principal Investigator
•
Upscaled community engagement and education occurred in
the communities close to Marikana
•
Participants were educated and equipped to deal with partners
who did not wish them to use the ring, or who wanted them
to withdraw against their will
Results: Partners who accepted a visit from the Community Liaison
Officer were in favour of the trial and their female partners’ participation
on the trial. Some partners visited the RC to observe all procedures
before accepting that their female partners could continue on the trial.
Women felt more equipped to deal with partner objections to the trial.
Early trial withdrawal for partner related issues decreased substantially
after the intervention.
Conclusions: The situation around a research centre could change
dramatically overnight. Unrelated incidents could have a ripple effect
on the clinical trials in the area. Partners may transfer their frustration
and anger onto their female partners, affecting their compliance to the
protocol. With appropriate intervention using a multi-pronged approach
it is possible to turn such a situation around.
www.hivr4p.org
61
Oral Abstract Session 09: Engaging, Recruiting and Retaining Trial Participants
OA09.03
OA09.04
Employing a Youth-centered Approach to
Investigate HIV Risk among Adolescents
and Young Adults in an HIV Hyper-endemic
Setting
WhatsApp!: Use of Mobile Technology in
Optimizing ASPIRE Study Retention and
Adherence at the Wits Reproductive Health
and HIV Institute, Johannesburg
Janan Dietrich1, Laura Cotton2, Stefanie Hornschuh1, Martin van
der Watt1, Cari L. Miller2, Glenda Gray1, Mark Brockman2, Angela
Kaida2, on behalf of the AYAZAZI Study Team
Krishnaveni Reddy1, Helen Rees1, Thesla Palanee1
1
Perinatal HIV Research Unit, Soweto, South Africa, 2Simon Fraser
University, Faculty of Health Sciences, Burnaby, BC, Canada
1
Background: Adolescents and young adults (AYA) (10-24 years)
are significantly at risk of HIV acquisition across Southern Africa.
The continued burden of HIV transmission among AYA reflects poor
prioritization of youth-focused and engaged research yielding an
inadequate understanding of intersecting factors necessary to implement
accessible and effective HIV prevention programs. Use of youthcentred approaches within HIV research and programming presents a
critical opportunity to engage youth, build capacity, and support youth
leadership in the HIV response.
Methods: AYAZAZI (‘Knowing Themselves’ in Zulu) is a youth-centred,
inter-disciplinary cohort study that aims to link socio-behavioural,
structural, clinical, and biomedical data to understand HIV acquisition
risk among AYA aged 16-24 years living in Soweto, South Africa. We
aim to enroll a cohort of 400 AYA (HIV-negative or HIV status unknown)
followed biannually for three years.
Results: AYAZAZI uses a youth-centred approach to engage AYA
throughout the research process. A Soweto-based Adolescent Community
Advisory Board provides overall study guidance and oversight. We
engaged the expertise and lived experiences of community members
in the development of the survey and clinical protocol to ensure youthappropriate language and priorities. AYA research assistants from Soweto
were hired and trained to support recruitment and to administer surveys.
We engaged with several youth-focused community partners to conduct
outreach and raise awareness of AYAZAZI. AYA research assistants will
receive ongoing training to support skill development and participants
will have access to regular knowledge exchange forums to support
youth leadership around HIV knowledge in the community.
Conclusions: Youth-centred, inter-disciplinary approaches that engage
young people throughout the research process are critical to support,
empower, and build local capacity required to prevent HIV infection and
reverse the high HIV risk environment among youth in South Africa.
62
Background: Mobile technology is an evolving communication
medium within clinical trials for participant/staff interaction and
represents a personalized discrete contact platform that is available 24
hours a day. Previously, short message service (SMS) communication
between staff and participants positively impacted study retention but
was primarily unidirectional due to associated costs. The development
of low cost instant messaging (IM) programs opens new opportunities
for bidirectional communication between participants and staff.
Methods: At Wits RHI, during locator information collection, most
participants indicated having access to smartphones and the WhatsApp
IM program. Staff then embarked on an array of strategies to utilize
this communication method for retention and adherence optimization.
Community health workers (CHWs) were provided with smartphones
to allow continuous communication with allocated participants for visit
reminders, on study related matters such as engagement activities/
events, clinic closures and the occasional check-ins. The Investigator of
Record (IoR), study coordinator (SC) and clinicians also use WhatsApp
to communicate with participants on clinical and study related issues
discretely.
Results: Participants respond to staff in real time on WhatsApp due
to low costs, ease of covert use and ability to raise sensitive issues.
Consequently, reporting of adverse events and adherence issues occurs
and is documented and followed up as needed. In addition, participants
are able to timeously communicate their challenges in attending
scheduled visits so they can be re-scheduled accordingly which then
impacts retention.
Conclusions: The ease of use and cost efficiency of IM contributes to
optimizing retention and adherence as well as adverse event reporting;
factors essential in determining safety and effectiveness of the study
product by allowing more frequent communication between staff and
participants.
Tuesday, 28 October
OA09.05
OA09.06
Turning around Poor Retention: A Cape Town
Experience
Reasons Boston MSM Enrolled in Placebo
Controlled PrEP Trials Did Not Continue Using
PrEP when it Was Available in an Open Label
Study
Karen Dominguez1, Katherine Gill1, Leader N. Kanyiki1, Ben
Brown1, Linda-Gail Bekker2
1
Desmond Tutu HIV Foundation, Cape Town, South Africa, 2Desmond
Tutu HIV Foundation, IIDMM University of Cape Town, Cape Town,
South Africa
Background: Retention in prevention trials is critical as it underpins
adherence and safety monitoring. Retention requires motivation and
action on behalf of participants and staff. As a site participating in a
Phase III tenofovir 1% vaginal gel study, we experienced poor participant
retention shortly after initiation in October 2011. Based in a township in
Cape Town, a severe increase in missed visits began when participants
travelled out of the province for the December holidays.
Methods: We calculated a site specific monthly retention rate using
number of visits attended divided by number of visits expected. Novel
ideas were urgently implemented to get participants already enrolled to
return for follow-up visits. Ideas included reminder and birthday SMSes,
recognition rewards for prompt and regular visits, transport for visits, as
well as Saturday clinics. The enrolment process was made more stringent
by instituting a pre-screen and discussion group prior to screening in
an effort to ensure participant commitment. A rewards points system
was designed as an incentive to come for participant visits and more
points could be earned for visits on scheduled dates. The points could
be accrued and redeemed for various prizes.
Results: The site enrolled 153 female participants, ages 18-30. Retention
reached its low point with a rate of 45 %. Following and with continued
use of the interventions, an average of 81% retention rate was achieved
for the remainder of the year . Methods that have been more popular
with the participants include the points system incentive, recognition,
and vehicle transport for visits.
Conclusions: Migration is a challenge for retention. Many trial sites
in South Africa face this challenge. Our team has become pro-active
in making retention a priority to prepare for migration and retain
participants throughout the year. Through careful participant selection
and use of methods to increase motivation and reduce “hassle” factors,
retention was enhanced in this cohort.
Christopher Chianese1, Kenneth Mayer1, Marcy Gelman1, Lori
Panther1, Robert Grant2,3
Fenway Health, Boston, MA, United States, 2Gladstone Institute of
Virology and Immunology, University of California, San Francisco, CA,
United States, 3University of California, San Francisco, CA, United States
1
Background: Boston participants who enrolled in the CDC’s PrEP safety
study and the iPrEX PrEP efficacy trial were offered open label access
to tenofovir-emtricitabine (TDF/FTC) for PrEP in the iPrEX Open Label
Extension (OLE). We identified participants who enrolled and did not
take TDF/FTC at the Boston study site and evaluated study records to
characterize reasons for not being administered TDF/FTC.
Methods: Participants who were not dispensed TDF/FTC were identified.
Visit note source documentation for each participant was analyzed
to determine why participants were ineligible for drug dispensation.
Reasons for drug dispensation ineligibility for each participant were then
coded and sub-divided into common themes.
Results: Of 79 participants who enrolled in the CDC Safety study and
87 who enrolled in iPrEx, 56% enrolled in iPrEx OLE at the Boston site
between 7/7/2011 and 3/29/2012. Median age was 44; 65% white,
23% Black, 12% other. Of 93 participants enrolled, 37% were not
dispensed TDF/FTC. Median age of participants not dispensed TDF/FTC
was 47; 59% white, 35% Black, 6% other. Of 34 participants, 38% did
not perceive they had high enough risk for HIV to justify PrEP; 18%
were concerned about side effects; 15% had active clinically significant
medical problems (including cardiac disease, pulmonary disease, DM
requiring hyperglycemic medication); 11% were taking contraindicated
medications; 6% had seroconverted after completing the CDC Safety
study, 6% felt pill-taking would be too burdensome; 6% indicated it was
because of personal choice but did not provide reasons.
Conclusions: Because PrEP study participants changed clinically and
behaviorally over time, these findings demonstrate that ongoing clinical
examination and behavioral assessment of PrEP users is important,
and PrEP delivery should not be automatic. Additional exploration into
contextual factors that influence an individual’s decision to take PrEP
is needed as PrEP becomes one of several HIV prevention modalities.
www.hivr4p.org
63
Oral Abstract Session 10: Bacterial Vaginosis and HSV-2: Impact on Genital Immunity
OA10.01
OA10.02
The Effects of Hormones and Vaginal
Microflora on the Content of MUC1, MUC4,
MUC5AC and MUC7 in the Cervicovaginal
Fluid (CVF)
HSV-2-driven Changes in α4β7 Expression
Correlate with Increased Susceptibility to
SHIV ex vivo and in vivo
Bernard J. Moncla1,2, Catherine Chappell1, Brian M. Debo2, Ingrid
S. Macio3, Katherine E. Bunge1, Sharon L. Hillier1,2
University of Pittsburgh, Ob/Gyn and Reproductive Sciences,
Pittsburgh, PA, United States, 2Magee-Womens Research Institute,
Microbiology, Pittsburgh, PA, United States, 3Magee-Womens Research
Institute, Clinical Research, Pittsburgh, PA, United States
1
Background: Mucins are glycoproteins that protect and lubricate
epithelial surfaces. In humans, they can be secreted (MUC5AC, MUC7)
or membrane bound (MUC1, MUC4, MUC16). Mucins can trap HIV but
little is known about the impact of the vaginal microbiota and hormonal
status on the mucin content in vaginal and cervical fluid. The objective of
this study was to measure the quantity of 5 MUCs in the CVF of women
under different hormonal conditions, stratifying for vaginal microflora.
Methods: CVF was collected via catamenial cup from 165 healthy
asymptomatic reproductive aged women in the follicular (n=27) or
proliferative (n=26) phase, women using levonogestrol (LNG) IUDs
(n=28), DMPA (n=29) or combined oral contraceptives (n=27) and
29 post-menopausal women. Vaginal smears were evaluated using
the Nugent criteria for bacterial vaginosis (BV) among reproductive
age women. The MUC content of the samples was evaluated using
ELISA. Student’s t-test and one-way analysis of variance with post-hoc
comparisons made using Bonferroni’s multiple comparisons procedure
were used to assess statistical significance.
Results: MUCs 1, 4, 5AC and 7 were detected at significantly higher
concentrations in CVF with increasing Nugent score (P< 0.05), while
MUC16 was not linked to BV status. By contrast, MUC content was
minimally impacted by hormonal status. There was no statistically
significant difference in MUC1 and MUC5AC among women in the
different hormonal groups. MUC4 was significantly increased among
post-menopausal women (P=0.005, comparing post-menopausal
women to all others), and women using a LNG IUD had lower levels
of MUC7 in the CVFs compared to other reproductive age women
(P=0.006).
Conclusions: In this study, vaginal microflora had a greater impact
on MUC content of CVF than hormonal status. Women with BV have
increased levels of secreted and membrane bound mucins, indicating
that microbiota induce changes in the expression of cervical mucins
which could contribute to an increased risk of HIV.
64
Diana Goode1, Rosaline Truong1, Meropi Aravantinou1, James
Blanchard2, Agegnehu Gettie3, Melissa Robbiani1, Elena Martinelli1
Population Council, New York, NY, United States, 2Tulane National
Primate Research Center, Tulane, LA, United States, 3Aaron Diamond
AIDS Research Center, New York, NY, United States
1
Background: The availability of highly susceptible HIV target cells
that can rapidly reach the mucosal lymphoid tissues may increase the
chances of an otherwise rare transmission event to occur. α4β7high CD4+
T cells are highly susceptible to HIV infection, they home specifically to
the GALT and HIV-gp120 interacts with α4β7. We hypothesized that HSV2, which augments the risk of HIV acquisition, modulates the expression
of α4β7 and other homing molecules in the vaginal tissue increasing its
susceptibility to HIV and SIV.
Methods: To test this hypothesis we used an in vivo rhesus macaque
(RM) model of HSV-2 vaginal infection and we developed a new ex vivo
model of macaque vaginal explants.
Results: In vivo, we found that HSV-2 chronically infected RMs were
more susceptible to vaginal SHIVSF162P3 infection. Ex vivo HSV-2 infection
increased the susceptibility of vaginal tissue to SHIV. Notably, in vivo
HSV-2 infection decreased expression of α4β7 on blood α4β7high CD4+ T
cells. In uninfected RMs the frequency of these cells inversely correlated
in blood and vaginal tissue. Ex vivo HSV-2 infection of vaginal explants
increased the frequency of α4β7high CD4+ T cells and α4β7high CD80+ DCs
in the mucosa and this directly correlated with HSV-2 replication. HSV2 also increased the frequency of CD103+ CD4+ T cells, however, this
negatively correlated with HSV-2 replication. The expression of α4β7
on migratory CD4+ T cells and the frequency of CD62L+ CD4+ T cells
also correlated with HSV-2 replication. Importantly, the HSV-2-driven
increase in the frequency of α4β7high CD4 T directly correlated with SHIV
replication in the HSV-2 infected tissues.
Conclusions: Our results suggest that HSV-2 infection modulates the
expression of homing molecules. In particular the HSV-2-driven increase
in the availability of α4β7high CD4+ T cells and DCs correlates with increased
susceptibility to SHIV infection. Thus, blocking a4b7 may decrease the
risk of HIV vaginal acquisition in HSV-2 infected and potentially HSV-2
uninfected women.
Tuesday, 28 October
OA10.03
OA10.04
Incident Herpes Simplex Virus Type 2
Associated with HIV Infection in Phambili
Mucosal Integrity Factors Are Perturbed
during Bacterial Vaginosis: A Proteomic
Analysis
James G. Kublin1, Zoe Moodie2, Barbara Metch2, Linda-Gail
Bekker3, Gavin Churchyard4, Maphoshane Nchabeleng5, Koleka
Mlisana6, Mary Allen7, Larry Corey8, Glenda Gray9, on behalf of
the HVTN 503 Study Team
HIV Vaccine Trials Network, Seattle, WA, United States, 2FHCRC,
SCHARP, Seattle, WA, United States, 3University of Cape Town, The
Desmond Tutu HIV Centre, Cape Town, South Africa, 4Aurum Institute
for Health Research, Klerksdorp, South Africa, 5Medunsa HIV Research
Unit, Medunsa, South Africa, 6University of KwaZulu Natal, Durban,
South Africa, 7DAIDS, NIAID, NIH, Vaccine Clinical Research Branch,
Vaccine Research Program, Rockville, MD, United States, 8FHCRC,
HVTN, Seattle, WA, United States, 9Perinatal HIV Research Unit, Chris
Hani Baragwanath Hospital, Soweto, South Africa
1
Background: In the Phambili preventative HIV-1 vaccine efficacy trial
conducted in South Africa, herpes simplex virus type 2 (HSV2) infection
at enrollment increased the risk of HIV-1 acquisition in men but not
in women. We monitored for incident HSV2 and potential associations
with HIV infection.
Methods: HSV2 prevalence at enrollment and incidence at 24 months
on study were determined by validated western blot assay. HIV-1
testing occurred at specified intervals and RNA PCR was used to confirm
HIV-1 infection. Multivariate logistic regression modeling was used to
investigate associations.
Results: HSV2 prevalence at enrollment was 31% (248/798; 95%
CI 28-34%); prevalence of HSV2 was significantly higher in women
(49%) than in men (16%); p< 0.001. Prevalence increased with age for
both genders, with 45% of men and 75% of women 27 years or older
infected. Prevalence varied by site, but not by circumcision status after
adjustment for age. HSV2 incidence at 24 months was 11% (43/384;
95% CI 8-14%); acquisition of HSV2 was significantly higher in women
(20%) than in men (7%); p< 0.001. Incident HSV2 infection among
men was positively associated with age (OR=1.2 per 1 year increase)
and negatively associated with smoking dagga (OR=0.2) and being
apart regularly from a main partner (OR=0.2). Among women, having a
casual/anonymous partner within 6 months prior to screening was the
only significant factor associated with incident HSV2 infection (OR=3.6).
Sixty HIV-1 infections occurred within participants’ first 24 months in
study, of whom 30 were HSV2 positive at enrollment. Among the other
30, 11 were diagnosed as HSV2 positive prior to or concurrent with
HIV infection diagnosis; 19 were HSV2 negative at the time of HIV1 diagnosis. By 24 months, HIV infection was associated with HSV2
infection, adjusted for age, both for men (OR=5.6) and for women
(OR=3.7).
Conclusions: HSV2 acquisition is markedly associated with HIV-1
infection in both heterosexual men and women in South Africa.
Kelly Arnold1, Kenzie Birse2, Lyle Mckinnon3, Jiriam Lingappa4,
Rick Novak5, Garrett Westmacott6, T. Blake Ball6, Doug
Lauffenburger1, Adam Burgener6
MIT, Boston, MA, United States, 2University of Manitoba, Winnipeg,
MB, Canada, 3CAPRISA/University of KwaZulu Natal, Durban,
South Africa, 4University of Washington, Seattle, WA, United States,
5
University of Illinois, Chicago, IL, United States, 6Public Health Agency
of Canada, Winnipeg, MB, Canada
1
Background: Mucosal inflammation, such as is found during bacterial
vaginosis (BV), has been associated with increased risk of HIV infection
in women. However, the mechanisms and/or drivers of HIV-susceptibility
are not well understood. Here we utilized a global proteomics approach,
coupled to multivariate computational modeling, to examine impact of
BV on the female genital tract mucosa in two distinct populations.
Methods: Cervicovaginal lavage samples collected from North American
individuals (Group A: BV+, n=8; BV-, n=26) and Kenya (Group B: BV+,
n=10; BV-, n=10) were analyzed by tandem mass spectrometry. Data
was analyzed using a combination of hierarchical clustering, pathway
analysis, and multivariate (Lasso) modeling.
Results: Approximately 650 host (human) and 100 bacterial protein
factors were identified in mucosal samples. BV+ individuals showed
significantly affected host protein expression including 52 in group A
and 92 in group B (p< 0.05). Microbiome proteins were also affected,
including significant reduction of Lactobacillus sp. factors (8, p< 0.05) and
elevation of numerous (>20 species) anaerobic bacterial proteins (50)
in BV (p< 0.05). Hierarchical clustering revealed two distinct branches
of upregulated/downregulated protein groups. Functional enrichment
analysis of downregulated clusters revealed that cornified envelope
(Grp A:1.2E-3; B: 6.6E-6) and keratinization (A: 1.8 E-4; B: 1.1E-4) tissue
integrity factors were common to both datasets. Lasso multivariate
modeling further identified 14 specific biomarkers (distinguishing BV+
individuals with 90% accuracy) that associated with increased host
metabolism, membrane integrity, and toll-like receptor signaling.
Conclusions: These data suggest that BV-associated inflammation
affects tissue integrity factors in the vaginal compartment, and may help
to explain BV-related HIV infection.
www.hivr4p.org
65
OA10.05
OA10.06 LB
Effect of Bacterial Vaginosis on Markers of
Genital Tract Inflammation and Mucosal
Immunity: Mechanisms for Susceptibility to
HIV Infection
Association of Tenofovir (TFV) Detection with
Reduced Risk of Herpes Simplex virus Type-2
(HSV-2) Acquisition in the VOICE (MTN 003)
Study
Andrea Ries Thurman1, Thomas D. Kimble1, Tina D.
Cunningham2, Betsy C. Herold3, Pedro Mesquita3, Ashley Huber3,
Raina N. Fichorova4, Hassan Y. Dawood4, Titilayo Fashemi4,
Neelima Chandra1, Jill L. Schwartz1, Gustavo F. Doncel1
Jeanne Marrazzo1, Lorna Rabe2, Cliff Kelly3, Edward Livant2, Z.
Mike Chirenje4, Barbra Richardson3, James Dai3, Jeanna Piper5,
Sharon Hillier2, the VOICE Study Team
CONRAD, Eastern Virginia Medical School, Norfolk, VA, United States,
Eastern Virginia Medical School, School of Public Health, Norfolk, VA,
United States, 3Albert Einstein College of Medicine, Infectious Diseases
and Pediatrics, Bronx, NY, United States, 4Brigham and Women’s
Hospital, Harvard Medical School, Laboratory of Genital Tract Biology,
1
2
Background: Bacterial vaginosis (BV), a highly prevalent alteration of
vaginal flora, has been linked to an increased risk of HIV acquisition
and transmission in observational studies, but the underlying biologic
mechanisms are not known. We measured markers of subclinical
vaginal inflammation, endogenous antimicrobial activity, and vaginal
flora in women with BV and repeated sampling 1 week and 1 month
after completion of metronidazole therapy.
Methods: Longitudinal, open label study (ClinicalTrials.gov
#NCT01347632) of 33 women with a Nugent score of 4 or higher.
All women had genital swabs, cervicovaginal (CV) fluid lavage (CVL)
and vaginal biopsies obtained at enrollment (active BV) and received
7 days of metronidazole treatment. Repeat sampling was performed
approximately 1 week and 1 month after completion of therapy.
Results: The CVL from women with resolved BV (Nugent 0 - 3) had
significantly higher anti-HIV activity compared to women with abnormal
microbiota (Nugent 4 - 10). The mean anti-HIV activity of women with
BV was < 0, indicating that their CVL enhanced HIV infection of TZMbl
cells in vitro. Genital tract interleukins IL-1β and ICAM-1 levels were
significantly higher (p < 0.05) and SLPI levels were lower
(p = 0.02) in the presence of BV compared to normal microbiota. AntiHIV activity of CVL was significantly associated with the Nugent score
and genital IL-1β and IL-8 levels in a multivariate model controlling for
participant race and visit. Although women with BV had significantly
lower numbers of CD45+, CD3+ and CD8+ vaginal immune cells
compared to racially matched women without BV, they had significantly
greater numbers of CCR5+ cells and increased numbers CD4+ cells and
CD4/CD8 ratios.
Conclusions: These data support that BV is associated with changes
in select soluble immune mediators, an increase in mucosal HIV target
cells and a reduction in endogenous CV antiviral activity, which may
contribute to the increased risk of HIV acquisition.
66
1
University of Washington, Seattle, WA, United States, 2Magee Women
Research Institute, Pittsburgh, PA, United States, 3SCHARP - FHCRC,
Seattle, WA, United States, 4University of California San Francisco
Research Programme, Harare, Zimbabwe, 5DAIDS, NIAID, NIH,
Bethesda, MD, United States
Background: In sub-Saharan Africa, HSV-2 infection is common, and
increases risk of HIV transmission and acquisition. TFV gel applied before
and after vaginal intercourse provided partial protection from HSV-2
acquisition in CAPRISA 004. We assessed efficacy of 1% vaginal TFV gel
in preventing HSV-2 acquisition in women in the VOICE Study, a 5-arm,
randomized, double-blind, placebo-controlled trial that studied daily use
of oral and vaginal tenofovir for HIV-1 pre-exposure prophylaxis.
Methods: From September 2009-June 2011, 5,029 women were
enrolled in VOICE in South Africa, Uganda, and Zimbabwe. 1,004
women were randomized to receive TFV 1% gel. Testing for HSV-2 typespecific antibody (Focus HerpeSelect EIA) was performed on plasma
from enrollment and study exit (positive result index value >3.5). TFV
measured in plasma (first quarterly visit) or vaginal swab (6 month
visit) was used as a measure of gel use. Using Poisson regression, we
analyzed the association between TFV detection in these samples and
HSV-2 acquisition.
Results: Of women in the TFV gel arm, 566 were HSV-2 uninfected
at enrollment (56%); of these, 531 (94%) had first quarter plasma TFV
and end-of-study HSV-2 serology available. Over follow-up of 511 PY,
92 incident HSV-2 cases occurred: 77 in women with no plasma TFV
detected and 15 in those with plasma TFV (incidence 20.2 (95% CI
15.9-25.2) vs 11.6 (6.5-19.1), respectively). Plasma TFV detection was
associated with reduced risk of HSV-2 seroconversion (unadjusted
incidence rate ratio (IRR) 0.57 (0.33-1.00; P=0.049); IRR adjusted for
country, age, marital status, ≥ 2 sex partners, hormonal contraception
type, anal sex, and HIV status 0.54 (0.31-0.97; P=0.039)). With TFV
detection in vaginal swabs, adjusted IRR was 0.71 (0.45-1.1; P=0.122).
Conclusions: Detection of plasma TFV among TFV gel users was
associated with reduced risk of HSV-2 acquisition after controlling for
potential risk modifiers, including sexual behavior and HIV-1 acquisition.
Oral Abstract Session 11: Vaccine Development: Emerging Insights
OA11.01
OA11.02
African Early Infection Cohort as a Platform
for Vaccine Discovery: The IAVI Protocol C
Experience
Intradermal HIV-DNA Given with or without
Intradermal Electroporation Is Safe and
Highly Immunogenic in Healthy Swedish HIV1 DNA/MVA Vaccinees
IAVI Human Immunology Lab, London, United Kingdom, 2MRC/
UVRI, Uganda Research Unit on AIDS, Entebbe, Uganda, 3Project San
Francisco, Kigali, Rwanda, 4ZEHRP, Lusaka, Zambia, 5CGMRC-KEMRI,
Kilifi, Kenya, 6KAVI-Institute of Clinical Research, University of Nairobi,
Nairobi, Kenya, 7Emory University, Atlanta, GA, United States, 8Aurum
Institute, Rustenburg, South Africa, 9IAVI, New York, NY, United States,
10
UAB, Birmingham, AL, United States, 11La Jolla Institute for Allergy
and Immunology, La Jolla, CA, United States, 12Duke Human Vaccine
Institute, Durham, NC, United States, 13University of Oxford, Oxford,
United Kingdom, 14Blood Systems Research Institute, San Francisco, CA,
United States
1
Background: Early HIV host-virus interactions provide important clues
to vaccine discovery and conducting acute infection studies in Africa
with multiple clades is vital to development of a global vaccine.
Methods: We followed cohorts at high risk of HIV acquisition at 9
research centers in East and Southern Africa to identify volunteers with
incident HIV infection. Infection date was estimated from serial antibody,
p24 and/or PCR results. Volunteers were invited to bring a sexual partner
for 1 visit. Follow up was monthly for the first 3 months post-infection,
quarterly through 2 years, and every 6 months thereafter. At every visit,
volunteers had a symptom-directed exam including medications, and
blood for viral load, CD4+ T cell counts and PBMCs.
Results: From 2006-2011, 614 seroconverters and 406 sexual partners
were enrolled. Follow up is ongoing; to date over 2,500 person years of
study time has provided a valuable platform for characterizing the HIV
genetic bottleneck at transmission and founder virus characteristics. T
cell responses in the acute phase have been shown to be associated with
virus control and viral replicative capacity with both set point viral load
and CD4+ T cell decline. Disease progression varied by sub-type, with
subtype-C and D-infected volunteers progressing faster than A, and HLA
alleles associated with outcomes have been characterized, such as B*44,
B*45, B*57 and B*81. The development and isolation of neutralizing
antibodies 2-3 years post infection is being described and the level of
circulating memory T follicular helper cells has been associated with the
development of such antibodies. Sample panels have provided valuable
to the development of pseudovirions and validating assays to detect
recent infection.
Conclusions: Prospective data and samples from early infection cohorts
are increasingly valuable for vaccine discovery work. Identifying host
and viral factors associated with acquisition and early viral control
across clades can inform the design of a global HIV vaccine.
Charlotta Nilsson1,2,3, Bo Hejdeman4, Karina Godoy-Ramirez1,
Teghesti Tecleab1, Gabriella Scarlatti5, Andreas Bråve1,2, Patricia L.
Earl6, Richard R. Stout7, Merlin L. Robb8, Robin Shattock9, Gunnel
Biberfeld1,2, Eric Sandström4, Britta Wahren2
The Public Health Agency of Sweden, Solna, Sweden, 2Karolinska
Institutet, Department of Microbiology, Tumor and Cell Biology,
Stockholm, Sweden, 3Karolinska Institutet, Department of Laboratory
Medicine, Huddinge, Sweden, 4Södersjukhuset and Karolinska
Institutet, Venhälsan and Department of Education and Clinical
Research, Stockholm, Sweden, 5IRCCS San Raffaele Scientific Institute,
Milan, Italy, 6National Institutes of Health, National Institute of Allergy
and Infectious Diseases, Bethesda, MD, United States, 7Bioject Inc.,
Tualatin, OR, United States, 8Walter Reed Army Institute of Research,
Department of Retrovirology, Rockville, MD, United States, 9Imperial
College, Department of Infectious Diseases, Division of Medicine,
London, United Kingdom
1
Background: DNA-based vaccines have been shown to be safe
but weakly immunogenic in humans. We compared safety and
immunogenicity of intradermal (id) vaccination with and without
electroporation (EP) in a phase I randomized placebo-controlled trial of
a HIV-DNA prime HIV-MVA boost vaccine in healthy Swedish volunteers
Methods: Twenty-five volunteers were randomized to receive HIV-DNA
plasmid vaccine 0.6 mg id using a needle-free device (Zetajet) with (n=16)
or without id EP (Dermavax) (n=9). An additional five volunteers were
placebo recipients. HIV-DNA plasmids encoding Env gp160 subtypes A,
B, and C; rev B; Gag A and B and RTmut B were given at weeks 0, 6 and
12. Boosting with recombinant MVA-CMDR expressing HIV-1 Env, Gag
and Pol of CRF01_AE (HIV-MVA) or placebo was performed at weeks 24
and 40. Nine of the vaccinees received a subtype C CN54 gp140 protein
boost in combination with HIV-MVA.
Results: The intradermal EP delivery was very well tolerated. After three
HIV-DNA immunizations, the IFN-γ ELISpot response rate to Gag was
slightly higher in HIV-DNA EP recipients (5/15, 33%) than in HIV-DNA id
recipients (1/7, 14%, p=0.6158). The first HIV-MVA or HIV-MVA+gp140
vaccination increased the IFN-γ ELISpot response rate to an overall
18/19 (95%). CD4+ and/or CD8+ T cell responses to Gag or Env were
demonstrable in 94% of vaccinees. A balanced CD4+ and CD8+ T cell
response was noted, with 78% and 71% responders, respectively. IFN-γ
and IL-2 production dominated the CD4+T cell response to Gag and Env.
The CD8+ response to Gag was broader with expression of IFN-γ, IL-2,
MIP1β and CD107. No differences were seen between groups. Binding
antibodies were induced after the second HIV-MVA+/-gp140 in 93%
of vaccinees to subtype C Env. The highest titers were seen among EP/
gp140 recipients.
Conclusions: Intradermal electroporation of HIV-DNA was well tolerated.
Strong cell-mediated and antibody-mediated immune responses were
elicited by the HIV-DNA priming and HIV-MVA boosting regimen,
irrespective of id EP use.
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67
Jill Gilmour1, Anatoli Kamali2, Etienne Karita3, William Kilembe4,
Eduard J. Sanders5, Omu Anzala6, Susan Allen7, Vinodh Edward8,
Fran Priddy9, Matt A. Price9, Gladys Macharia5, Joshua Baalwa10,
Shane Crotty11, Thomas Denny12, Elise Landais11, Persephone
Borrow13, Jianming Tang10, Michael Busch14, Jessica Prince7, Dan
Claiborne7, Pascal Poignard11, Pat Fast9, Eric Hunter7
OA11.03
OA11.04
VLP-expressing DNA/MVA Vaccines: The
Effect of Schedule and Regimen on Antibody
Magnitude and Avidity
Vaccine Enhancement Confirmed among Men
in HVTN 503-S, Final Results from a Recall
Study of Phambili Participants
Susan P. Buchbinder1,2, Christine Hay3, Nicole Grunenberg4, Paul
Goepfert5, Tomaras Georgia6, Kelly Seaton6, Alicia Sato7, Marnie
Elizaga4, Xuesong Yu7, Peter Gilbert7, Bernard Moss8, Harriet
Robinson9
Zoe Moodie1, Barbara Metch1, Mary Allen2, Linda-Gail Bekker3,
Gavin Churchyard4, Maphoshane Nchabeleng5, Koleka Mlisana6,
James Kublin1, Glenda E. Gray7,8
San Francisco Department of Public Health, Bridge HIV, San Francisco,
CA, United States, 2University of California at San Francisco, Medicine,
Epidemiology and Biostatistics, San Francisco, CA, United States,
3
University of Rochester Medical Center, Rochester, NY, United States,
4
5
University of Alabama-Birmingham, Birmingham, AL, United States,
6
Duke Human Vaccine Institute, Durham, NC, United States, 7SCHARPFHCRC, Seattle, WA, United States, 8National Institute of Allergy and
Infectious Diseases, National Institutes of Health, Bethesda, MD, United
States, 9GeoVax, Inc., Smyrna, GA, United States
Institutes of Health, Vaccine Research Program, Division of AIDS,
Bethesda, MD, United States, 3University of Cape Town, Desmond
Tutu HIV Foundation, Cape Town, South Africa, 4Aurum Institute for
Health Research, Johannesburg, South Africa, 5University of Limpopo,
Medunsa, MEDUNSA HIV Clinical Research Unit, Mankweng-E, South
Africa, 6University of KwaZulu-Natal, Centre for AIDS Programme
for Research in South Africa, Durban, South Africa, 7University of the
Witwatersrand, Perinatal HIV Research Unit, Johannesburg, South
Africa, 8South African Medical Research Council, Cape Town, South
Africa
Background: HIV-1 vaccines ideally induce high magnitude and
avidity antibody (Ab) responses to the envelope glycoprotein (Env). In
particular, the highly conserved immunodominant region (IDR) of gp41
is an important target for virion capture and ADCC. We compared Ab
responses between different regimens and schedules in HIV negative
healthy participants (pts) to evaluate the addition of a 3rd MVA dose,
spacing between MVA doses, and the inclusion of co-expressed GM-CSF
on Ab magnitude and avidity.
Methods: The JS7 (D) and GEO-D03 DNA (Dg) and HIV62 MVA (M)
vaccines produce virus-like particles displaying trimeric, membranebound Env; the GEO-D03 DNA co-expresses GM-CSF. HVTN 205
enrolled 150 pts in DDMM (0,2,4,6 mo.) and 75 pts in an MM_M
(0,2,6 mo.) regimens ; HVTN 094 enrolled 15 pt each in DgDgMM_M
(0,2,4,6,10 mo) and DgDgM_M (0,2,4,8 mo.) regimens.
Results: All regimens were safe and well tolerated. Two MVA doses
generated IgG to IDR gp41 in 73%-100% of participants. The 3rd MVA
dose significantly increased the magnitude (HVTN 205) and avidity
(HVTN 205 and 094) of this IgG. This response was durable, with 58%
of pts still positive 6 months after the 3rd MVA dose in HVTN 205, and
the median magnitude among persistent responders declining only
½ log. Increased spacing between the 1st and 2nd MVA inoculations
showed significantly enhanced avidity but no difference in magnitude
or response rates. Including co-expressed GM-CSF in the DNA prime did
not enhance the magnitude or avidity of the Env-specific Ab. At the end
of the immunization regimen in HVTN 094, median avidity indices were
34 and 23 for the 3- and 2-dose MVA arms.
Conclusions: The addition of a spaced 3rd MVA boost to a DDMM
regimen enhances the magnitude and avidity of the Env-specific Ab
response, and appears to be durable 6 months after immunization
without additional boosting. ADCC and virion capture assays are being
run to evaluate the unique immune profile and promise of this regimen
for protection against clade B infection.
Background: Final analysis of the Phambili study, assessing the MRK
Ad5 gag/pol/nef subtype B preventive HIV-1 vaccine in South Africa,
showed a higher rate of HIV-1 infection in the vaccine group compared
to placebo after 42 months of follow-up, with a statistically significant
difference among men, irrespective of circumcision or Ad5 serostatus,
but not amongst women. As most follow-up occurred after participant
unblinding, results may have been influenced by ascertainment bias
related to dropout. Consequently, HVTN 503-S was initiated in June
2013 to recall all uninfected participants for additional HIV testing.
Methods: Phambili participants who were HIV uninfected at last
Phambili contact were contacted for a follow-up visit which included HIV
testing, risk prevention counseling, behavioral risk assessment and, for
men, circumcision assessment. Cox proportional hazards models were
used to estimate the vaccine:placebo hazard ratio (HR) from time of
enrollment in Phambili.
Results: Overall, 464 (67%) of the 695 uninfected Phambili participants
were enrolled in 503-S, with no differences in contact disposition
between treatment or gender groups. Sites were unable to contact
95 (14%). Among the 187 participants who dropped out of Phambili,
37% enrolled. The median time from last Phambili HIV test to 503-S
testing was 33 months (range 27-77). For women, 8 new infections were
diagnosed among vaccinees (n=99) and 14 among placebos (n=106).
Among men, 11 new infections were diagnosed in the vaccine group
(n=130) and 3 among placebos (n=129). The Phambili HR was 2.46
(95% CI 1.22-4.93) among men, adjusted for baseline HSV-2; with the
additional 503-S follow-up, it was 2.75 (1.49, 5.06). 11 new infections
were diagnosed among Phambili dropouts.
Conclusions: Sites successfully located and scheduled participants
despite more than two years since participants’ last Phambili visit. HIV
testing results supported the conclusion of vaccine enhancement of HIV1 acquisition among men.
1
68
1
2
OA11.05
OA11.06 LB
HIV-specific Antibody in Rectal Secretions
Following Late Boosts in RV144 Participants
(RV305)
HVTN 097: Evaluation of the RV144 Vaccine
Regimen in HIV Uninfected South African Adults
1
1
Armed Forces Research Institute of Medical Sciences, Department of
Retrovirology, Bangkok, Thailand, 2Henry M Jackson Foundation for
the Advancement of Military Medicine, Bethesda, MD, United States,
3
Walter Reed Army Institute of Research, U.S. Military HIV Research
Program, Silver Spring, MD, United States, 4Mahidol University, Faculty
of Tropical Medicine, Bangkok, Thailand, 5Ministry of Public Health,
Department of Disease Control, Nonthaburi, Thailand, 6Cooper Human
Systems, Nashua, NH, United States, 7National Institute of Infectious
Diseases, AIDS Research Center, Tokyo, Japan
1
Background: Rectal mucosa is the primary site of HIV acquisition
during anal intercourse. Anti-HIV envelope antibodies were identified
as a correlate of risk in the RV144 efficacy trial but the presence of
vaccine-induced antibodies in rectal secretions has not been previously
assessed.
Methods: HIV-specific antibodies in rectal secretions were measured.
HIV uninfected RV144 male vaccinees (n=26) were randomized to
receive late boosts (RV305) consisting of 2 injections of ALVAC-HIV
and/or AIDSVAX B/E or placebo at 0 and 6 months. Rectal secretions
collected with Merocelâ sponges. Total and HIV envelope gp120
A244gD specific IgG and IgA antibodies were measured by ELISA at
study entry and 2-week post first and second vaccinations.
Results: Total IgG and IgA were detected in 97% (0.2-484.8 µg/mL) and
86% of samples (2.0-3677.4 µg/mL), respectively, throughout the three
times points. Anti-A244gD IgG responses were undetectable in ALVACHIV/AIDSVAX B/E and AIDSVAX B/E groups, but one sample in ALVACHIV group had a low level of anti-A244gD IgG at baseline. Two weeks
post first injection, anti-A244gD IgG was present in 70% and 57% (0.711.6 and 1.0-13.4 µg/mL) of ALVAC-HIV/AIDSVAX B/E and AIDSVAX B/E
groups, respectively. Post second injection, response rates decreased to
33% in the ALVAC-HIV/AIDSVAX B/E group (0.6-2.4 µg/mL) and to 29%
in the AIDSVAX B/E group (2.6-9.3 µg/mL). No responses were detected
in the ALVAC-HIV group. IgA antibodies to A244gD were not detected in
any of the tested samples.
Conclusions: IgG antibodies to A244gD were detectable in rectal
secretions in the majority of RV144 vaccine recipients who received
late boosts with ALVAC-HIV/AIDSVAX B/E or AIDSVAX B/E implying a
possible mechanism for vaccine protection following anal HIV exposure.
IgA responses to HIV gp120 were absent from all vaccination groups
despite total IgA being higher than total IgG. Merocelâ sponges are
effective in collecting rectal secretions for antibody measurement.
Glenda E. Gray1,2, Erica Andersen-Nissen3, Nicole Grunenberg4,
Ying Huang5, Surita Roux6, Fatima Laher7, Craig Innes8, Niya
Gu9, Carlos DiazGranados9, Sanjay Phogat9, Carter Lee10, Edith
Swann11, Jerome Kim12, Robert O’Connell12, Nelson Michael12,
Britta Flach3, Steve DeRosa13, Nicole Frahm13, Lynn Morris14, David
Montefiori15, Peter Gilbert5, Georgia Tomaras15, Julie McElrath3,13,
Lawrence Corey4, HVTN 097
South African Medical Research Council, Cape Town, South Africa,
Perinatal and HIV Research Unit, Soweto, South Africa, 3Cape Town HVTN
Immunology Laboratory/Hutchinson Centre Research Institute, Cape Town,
South Africa, 4Fred Hutchinson Cancer Research Center, HVTN, Seattle, WA,
United States, 5Fred Hutchinson Cancer Research Center, SCHARP, Seattle,
WA, United States, 6Desmond Tutu HIV Foundation, IIDMM University of
Cape Town, Cape Town, South Africa, 7Perinatal HIV Research Unit,Faculty
of Health Sciences, University of the Witwatersrand, Soweto, South Africa,
8
Aurum Institute for Health Research, Klerksdorp HVTN CRS, Klerksdorp,
South Africa, 9Sanofi Pasteur, Swiftwater, PA, United States, 10Global
Solutions for Infectious Diseases, South San Francisco, CA, United States,
11
Division of AIDS, National Institute of Allergy and Infectious Diseases,
National Institutes of Health, Bethesda, MD, United States, 12U.S. Military
HIV Research Program, Walter Reed Army Institute of Research, Silver
Spring, MD, United States, 13Fred Hutchinson Cancer Research Center,
HVTN Laboratory Program, Seattle, WA, United States, 14National Institute
for Communicable Diseases of the NHLS, Johannesburg, South Africa,
15
Duke University Human Vaccine Institute and the Center for HIV/AIDS
Vaccine Immunology, Duke University School of Medicine, Durham, NC,
United States
1
2
Background: Following the RV144 trial demonstration of 31% vaccine
efficacy in preventing HIV infection in Thailand, HVTN 097 was
designed to evaluate the same regimen in South Africans to ascertain
whether their immune response profiles were similar to Thais. This
study was conducted in preparation for evaluating a similar clade C
HIV vaccine regimen in South Africa. Our study was critical as previous
studies have demonstrated that age, gender and BMI impact vaccineinduced immune responses.
Methods: ALVAC-HIV (vCP1521) expressing HIV-1 Env (clade E, gp120
from strain 92TH023 and clade B TM gp41 from strain HIVLai, and Gag
and protease (clade B) was administered at baseline then 1, 3 and 6
months later with AIDSVAX® B/E, bivalent HIV gp120 subtypes B (MN)
and E (A244) adsorbed to alum at the last 2 vaccinations. Immune
responses were measured 2 weeks after the last immunization.
Intracellular Cytokine Staining identified response rates and frequencies
of HIV-specific T cells expressing IFN-γ and/or IL-2. Binding antibodies
to HIV-1 gp120 and V1V2, IgG subclass, and antibody functions
(ADCC, avidity index, nAb, and virion capture) are ongoing. Immune
responses were stratified by age, gender and BMI.
Results: In 68 participants, overall peak response rates of Env-specific
CD4+ T cells expressing IFN-γ and/or IL-2 was 70.6% (95% CI 58.9%,
80.1%), similar to or greater than responses in RV144. Although not
significant, participants < 25 years old had higher response rates
observed than those ≥26 (76.0% vs. 55.6%, p=0.108). Response rates
in females (75%; 95%CI 56.6%, 87.3%) were similar to males (67.5%;
95%CI 52.0%, 79.9%). Response rates stratified by BMI categories of
< 25, 25-30, >30 were 63.6%, 82.4% and 85.7%, respectively (p<
0.1 for BMI ≥25).
Conclusions: Response rates and magnitudes of Env-specific CD4+
T cells in South Africans induced by the same vaccine regimen used
in RV144 were at least comparable to or better than those induced in
RV144. Age/gender or BMI did not affect CD4+ T-cell response rates.
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69
Siriwat Akapirat , Chitraporn Karnasuta , Sirinan Madnote ,
Hathairat Savadsuk1, Jiraporn Puangkaew1, Surawach
Rittiroongrad1, Sandhya Vasan1, Viseth Ngauy1, Merlin L. Robb2,3,
Jean-Louis Excler2,3, Punnee Pitisutthithum4, Supachai RerksNgarm5, Nelson L. Michael3, Mark S. de Souza6,7, Jerome H. Kim3,
Robert J. O’Connell1, Nicos Karasavvas1, on behalf of the RV305
Study Group
1
Oral Abstract Session 12: Towards Broadly Neutralizing Antibody Induction
OA12.01
OA12.02
Maturation of Broadly Neutralizing V1V2directed Antibodies in the Context of
Autologous Viral Escape
Development of a V1/V2-targeting
Quaternary-specific Broadly Neutralizing
Lineage
Jinal N. Bhiman1,2, Nicole A. Doria-Rose3, Constantinos Kurt
Wibmer1,2, Daniel J. Sheward4, Carolyn Williamson4,5, Salim
S. Abdool Karim5, Peter D. Kwong3, John R. Mascola3, Lynn
Morris1,2,5, Penny L. Moore1,2,5
Elise Landais1, Bryan S. Briney2, Sergei L. Kosakovsky-Pond3,
Daniel T. MacLeod1, Yolanda Lie4, Paul Algate5, Dennis R. Burton2,
Terri Wrin4, Po-Ying Chan-Hui5, Pascal Poignard1,2, The IAVI
Protocol C Investigators & The IAVI African HIV Research Network
Centre for HIV and STIs, National Institute for Communicable Diseases
of the National Health Laboratory Services, Johannesburg, South Africa,
2
School of Pathology, Faculty of Health Sciences, University of the
Witwatersrand, Johannesburg, South Africa, 3Vaccine Research Center,
National Institute of Allergy and Infectious Diseases, National Institutes
of Health, Bethesda, South Africa, 4Institute of Infectious Diseases
and Molecular Medicine, Division of Medical Virology, University
of Cape Town and NHLS, Cape Town, South Africa, 5Centre for the
AIDS Programme of Research in South Africa (CAPRISA), University of
KwaZulu Natal, Durban, South Africa
1
1
Background: A family of 12 anti-V1V2 monoclonal antibodies (mAbs)
was previously isolated from an HIV-1 superinfected individual,
CAP256. This lineage emerged 30-34 weeks post-infection (w.p.i.) with
individual mAbs isolated at different time-points showing varying levels
of maturation and neutralization activity. Mutations R166S or K169E
became fixed at 176 w.p.i. and resulted in complete viral escape from all
mAbs. Here we investigated the effect of earlier mutations at positions
166 and 169 on the maturation of the CAP256-VRC26 lineage.
Methods: Potential escape mutations were identified using single
genome envelope amplification at 11 time-points from 6-94 w.p.i.
Mutations were introduced into the sensitive superinfecting virus and
tested in neutralization assays against the 12 mAbs to assess their role
in escape.
Results: By 42 w.p.i., just prior to the development of breadth, the
wildtype 169K was replaced with either a 169I/R/Q/T in all sequences,
indicating early immune pressure at this site. Introduction of K169I/Q
mutations into the superinfecting virus abrogated neutralization by
CAP256-VRC26.01 and 12, the least broad members of the family,
with less effect on other lineage members. Similarly, at position 166,
we observed a K166R mutation in 14/15 sequences at 94 w.p.i. that
abrogated neutralization by all but the four broadest mAbs, CAP256VRC26.03, 04, 08 and 09. As 166R and 166K are the two most prevalent
immunotypes at this position (in 70% and 14% of all global viruses,
respectively), the increased breadth of these four mAbs is consistent with
their ability to tolerate both residues.
Conclusions: Mutations that arose in the viral population soon after the
expansion and subsequent maturation of the CAP256-VRC26 family
exposed later lineage members to multiple immunotypes, thereby
increasing their neutralization breadth. These data illustrate how a
gradual process of autologous viral escape shaped the maturation of
this mAb response and provides insights for future sequential vaccine
regimens.
70
The International AIDS Vaccine Initiative, Neutralizing Antibody Center,
La Jolla, CA, United States, 2The Scripps Research Institute, Department
of Immunology and Microbial Sciences, La Jolla, CA, United States,
3
University of California San Diego, Department of Medicine, La Jolla,
CA, United States, 4Monogram Biosciences Inc, San Francisco, CA,
United States, 5Theraclone Sciences Inc., Seattle, WA, United States
Background: Designing a vaccine capable of eliciting broadly neutralizing
antibodies (bNAbs) to HIV requires a better understanding of these Ab
maturation pathways as they occur by interplay with HIV envelope
glycoprotein evolution. IAVI Protocol C is a large longitudinal cohort
of primary HIV-1 infection in sub-Saharan Africa. The development of
bNAb responses was evaluated in 385 Protocol C donors and individuals
with bNAb specificities targeting diverse highly conserved epitopes were
selected for mAb isolation and maturation studies.
Methods: Donor PC64 serum neutralizing activity was mapped to
a N160-glycan-dependent, kifunensine-sensitive V1V2 quarternary
epitope. The PG9-like neutralizing activity was detectable as early as
18 months post infection (mpi), peaked at 36 mpi and waned slightly
during 2 years of additional follow-up. Envelope glycoprotein genes
from PC64 were cloned at 10 different time points between 1 and 48
mpi. High throughput B-cell activation and functional screening were
used to isolate mAbs with PG9-like neutralizing activity at 6 time points
post infection. Deep sequencing of memory B-cells from longitudinal
PBMCs samples was performed using the MiSeq (Illumina) platform.
Results: Ten related mAbs with PG9-like neutralizing activity were
isolated from the month-36 sample. The Abs use the VH3-15*01 and
VK3-20*01 genes and possess long CDRH3s (25 amino acids) that
contain tyrosine residues. Deep sequencing of 12 longitudinal samples
spanning 4 years of infection suggested the emergence of the lineage
around 9 mpi, identifying a sequence with 99.66% nucleotide identity
to germline. Analysis of Env sequences showed evidence of escape
mutations and an increase in diversity after 12 mpi, likely a consequence
of selection pressure from the PG9-like lineage.
Conclusions: Additional Ab isolated from different time points are
currently being characterized. The study will provide important insight
regarding the maturation pathways of bNAbs and critical information for
vaccine design.
OA12.03
OA12.04
Development of Broadly Neutralizing AntiHIV-1 Antibodies during Natural Infection
through Early Epitope Acquisition and
Subsequent Maturation
Investigating Epitope Exposure on Native
Trimers
Sergey Menis1, Chris Carrico2, Dan Kulp1, William Schief1,3
The Scripps Research Institute, Department of Immunology and
Microbial Sciences, La Jolla, CA, United States, 2Buck Institute for
Research on Aging, Novato, CA, United States, 3Ragon Institute of MGH,
MIT and Harvard, Boston, MA, United States
1
D. Noah Sather1, Sara Carbonetti1, Delphine Malherbe2, Franco
Pissani3, Andrew B. Stuart1, Ann J. Hessell2, Spyros Kalams4,
Nancy L. Haigwood2, Leonidas Stamatatos1
Seattle Biomedical Research Institute, Seattle, WA, United States,
Oregon National Primate Research Center, Beaverton, OR, United
States, 3Henry M. Jackson Foundation for the Advancement of Military
Medicine, Bethesda, MD, United States, 4Vanderbilt University,
Nashville, TN, United States
2
Background: Approximately 20-30% of HIV-1 infected subjects
develop broadly neutralizing antibodies (bNAbs), which are prototypes
for the types of antibodies that will likely need to be elicited by a
successful HIV-1 vaccine. Delineating the key early events that lead
to the development of bNAbs during infection may help guide the
development of immunogens and vaccine regimens to prevent HIV-1
infection.
Methods: We followed two HIV-1 positive subjects from before they
developed bNAb activity until several years after breadth was detected
in the plasma. We studies viral Envelope evolution over the course of
infection and studied its relationship to the development of bNAbs in
early infection.
Results: Both subjects developed bNAb activity one year post infection,
which ultimately mapped to the membrane proximal external region
(MPER) and the CD4 receptor binding site. For one subject, we were
able to identify anti-MPER activity in the earliest plasma sample that
exhibited no bNAb activity, indicating that this epitope specificity was
acquired very early on, but that these antibodies initially were not able to
mediate neutralization. Escape mutations within the bNAb epitopes did
not arise in the circulating envelopes until bNAb activity was detectable
in the plasma, indicating that this early response was not sufficient to
drive viral escape. Finally, we found that global anti-Envelope binding
avidity significantly increased at the time bNAbs developed, indicating
that antibody maturation was a key step.
Conclusions: We report one potential mechanism by which bNAbs
develop during natural infection in which an epitope target is acquired
very early on during the course of infection. After sufficient time
and antibody maturation, the response matures to acquire broadly
neutralizing activity, forcing the virus to escape. These findings reinforce
a key role for antibody maturation in the development of bNAbs, and
highlight the necessity of developing vaccine regimens that are able to
drive somatic hypermutation.
Background: HIV infection typically generates autologous but not
broadly neutralizing antibodies, presumably due to immunodominance
of strain-specific epitopes on the native spike. Consistent with this view,
it has been reported that rabbit immunization with the BG505 SOSIP
soluble trimer, a faithful mimic of the native spike, induces potent
BG505-neutralizing but not broadly neutralizing responses. We sought
to employ the known structure of the BG505 SOSIP trimer to investigate
the exposure of variable and conserved epitopes on the native spike.
Methods: We computationally built a large ensemble of glycosylated
BG505 SOSIP trimers with a variety of backbone and glycan
conformations to approximate the motions of the native trimer. We then
identified potential antibody epitopes in each member of the ensemble
in an unbiased and exhaustive manner. Finally, we computed the
frequency at which each position in the trimer is observed in an epitope.
This frequency represents a quantitative estimate of the exposure to
antibody for any particular position.
Results: Antibodies with common CDR lengths access epitopes in the
variable regions (V1, V2, V4, V5) considerably more frequently (220
times more frequently) than in the conserved CD4-binding-site (CD4bs).
Glycosylation is the primary culprit: in the absence of glycans, the
variable regions are accessed only 7 times more frequently than the
CD4bs. Dominant V5-directed responses may further reduce responses
to conserved CD4bs epitopes, by competition. Increased CDRH3 length
allows antibody access to hidden epitopes but also increases access to
variable epitopes.
Conclusions: Variable epitopes in V1, V2, V4, V5 are predicted to be
immunodominant on native spikes, purely on the basis of exposure.
Antibodies against these epitopes likely inhibit induction of broadly
neutralizing antibodies by trimer immunogens. Our analysis may
guide design of trimers and immunization regimens for more favorable
immune responses.
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71
1
OA12.05
OA12.06 LB
An Inflammatory Profile that Predicts the
Development of Neutralizing Antibody
Breadth
Induction of Antibodies with Long Variable
Heavy Third Complementarity Determining
Regions by Repetitive Boosting with
AIDSVAX® B/E in RV144 Vaccinees
Anne Sophie Dugast1, Kelly Arnold2, Michelle Hoffner1,
Florencia Pereyra1, Michael Seaman3, Bruce Walker1, Douglas
Lauffenburger2, Galit Alter1
States, 2MIT, Department of Biological Engineering, Cambridge, MA,
United States, 3Center for Virology and Vaccine Research, Boston, MA,
United States
1
Background: Over the past decade, there has been an exponential
increase in the discovery of new broadly neutralizing Abs (bNAbs);
however the mechanism by which these Abs are elicited remains unclear.
Interestingly, compelling evidence from multiple studies suggests that
high viremia and associated immune activation are potential drivers for
the development of bNAbs. Here we sought to dissect the inflammatory
signals associated with bNAb evolution in a large cohort of subjects
that spontaneously control viral replication (Controllers) and to define
whether viremia and inflammation can be unlinked as predictors of the
induction of bNAbs.
Methods: Neutralizing activity and 19-plex cytokine analyses were
performed in 103 Controllers and 7 acutely infected individuals who
went on to develop bNAbs versus 8 who did not. Acute subjects were
followed longitudinally over 12 months. Partial least square regression
and a decision tree analysis were used to define multivariate cytokine
signatures associated with the evolution of bNAbs across cohort groups.
Results: We observed that high plasma levels of CXCL13, sCD40L and
IP10 were enriched in Controllers that evolved bNAbs compared to those
that did not (p=0.001). While none of the cytokines independently
predicted bNAb evolution, we found that a specific pattern of cytokine
production was associated with the evolution of bNAbs, consisting of
high plasma levels of IP10 in combination with CXCL13 or sCD40L. This
profile was validated in a cohort of chronically infected patients that
evolved bNAbs, and predicted the evolution of neutralizing breadth in a
cohort of acutely infected patients that went on to develop bNAbs.
Conclusions: Overall, this unique inflammatory signature points to
a very specific mechanism for the evolution of neutralizing antibody
breadth, that is induced as early as a few weeks post infection, pointing
to a role of specific adjuvants or inflammatory cues that if co-delivered
during immunization, may promote more effective affinity maturation.
72
Michael Anthony Moody1, David Easterhoff1, Thaddeus C.
Gurley1, John F. Whitesides1, Dawn J. Marshall1, Andrew
Foulger1, Krissey E. Lloyd1, Robert Parks1, Justin Pollara1, Ryan
Duffy1, Shaunna Shen1, Jerome H. Kim2, Nelson L. Michael2,
Merlin L. Robb2, Robert J. O’Connell3, Sandhya Vasan3, JeanLouis Excler2, Supachai Rerks-Ngarm4, Jaranit Kaewkungwal5,
Punnee Pitisuttithum5, Sorachai Nitayaphan3, Faruk Sinangil6,
Donald Francis6, Carter Lee6, Thomas B. Kepler7, S. Munir Alam1,
Guido Ferrari1, David C. Montefiori1, Hua-Xin Liao1, Georgia D.
Tomaras1, Barton F. Haynes1
Duke University Medical Center, Duke Human Vaccine Institute,
Durham, NC, United States, 2US Military HIV Research Program,
Rockville, MD, United States, 3AFRIMS, Bangkok, Thailand, 4Thai
Ministry of Public Health, Nonthaburi, Thailand, 5Mahidol University,
Bangkok, Thailand, 6Global Solutions for Infectious Diseases, South San
Francisco, CA, United States, 7Boston University, Boston, MA, United
States
1
Background: The Env gp120AE.A244 used for boosting in the ALVAC/
AIDSVAX® B/E RV144 trial expressed a dominant V2 linear epitope
centered on lysine (K) at position 169, and a subdominant peptideglycan epitope recognized by V1V2 broad neutralizing antibodies
(bnAbs) CH01 and PG9 and the unmutated ancestor antibody (Ab) of
CH01. V2-specific Abs isolated from RV144 vaccinees displayed tier-1
strain-specific neutralization (AE.92TH023) and bound K169-containing
linear epitopes. The RV305 trial recruited 90 RV144 participants to
return after 6 years for two boosts with either ALVAC+AIDSVAX® B/E
or AIDSVAX® B/E alone to evaluate the effect of this boost on vaccineinduced immunity.
Methods: B-cell repertoires of 4 RV305 vaccinees with the greatest
breadth of serum neutralization (A3R5 assay) were studied by antigenspecific memory B-cell sorting and recombinant Ab generation. Two
vaccinees received ALVAC+AIDSVAX® B/E and two were boosted with
AIDSVAX® B/E protein in alum. PCR-amplified monoclonal Abs (mAbs)
were characterized in binding, neutralization, and ADCC assays.
Results: The boosts increased VH mutation frequency from that seen
following the initial RV144 vaccine regimen (RV305 mean 5.50%, 258
mAbs, 4 vaccinees; RV144 mean 2.60%, 105 mAbs, 12 vaccinees)
and expanded a population of Abs with heavy third complementarity
determining regions (HCDR3s) > 22 amino acids (RV305 30/258;
RV144 1/105). Similar to V1V2 bnAbs- and other neutralizing Abs with
long HCDR3s, these mAbs principally used D2/D3 and JH6. Moreover,
35.2% of mAbs were sensitive to PNGase F native deglycosylation of
Env gp120AE.A244, including 9 mAbs with long HCDR3s. Four vaccineinduced N156QN160Q-sensitive V2 mAbs were isolated that are being
characterized for neutralization capacity.
Conclusions: Repetitive boosting of RV144 vaccinees expanded a pool
of Abs with many of the characteristics of V1V2 bnAbs. Expansion of this
subdominant group of antibodies by vaccination may represent a step
forward in the quest to induce bnAbs.
Oral Abstract Session 13: ARV Exposure & Efficacy in the Genital Tract
OA13.01
OA13.02
Defining Pharmacokinetic and
Pharmacodynamic linkages between Genital
Tissue and Lumen Compartments
Phase 1 Safety & Pharmacokinetic Trial of a
Polyurethane Tenofovir Disoproxil Fumarate
Intravaginal Ring in Healthy, Low-risk U.S.
Women
University of Pittsburgh, Pittsburgh, PA, United States, 2Magee-Womens
Research Institute, Pittsburgh, PA, United States, 3Johns Hopkins
University, Baltimore, MD, United States, 4IPM, Silver Springs, MD,
United States
1
Background: Early clinical trials of topical microbicides only defined
drug levels in tissue and luminal fluids. We are now determining if the
drug quantity is sufficient to inhibit HIV. We theorize increased drug
levels will correlate to decreased HIV infection.
Methods: In the FAME-02 clinical trial, 60 women were randomized to
use dapivirine (DPV) gel or film, placebo or active. After 7 daily doses of
product, a 10 ml cervicovaginal lavage (CVL) and two vaginal biopsies
were taken. For pharmacokinetic (PK) determination, DPV was quantified
using LC-MS/MS. For pharmacodynamic (PD) activity, CVL was tested
using an in vitro TZM-bl assay and tissue was tested using the ex
vivo challenge assay. PK/PD relationships were determined by linear
regression.
Results: Women using the films and gels had no serious adverse
events and found them to be acceptable. However, 5 women in the
DPV film arm had not placed the films correctly and were excluded
from this analysis. DPV levels in CVL ranged from 174-834 ng/ml in film
compared to 489-794 ng/ml in gel users. DPV levels in vaginal tissue
ranged from 0.06-25 ng/mg in film compared to 25-79 ng/mg in gel
users. CVL showed a strong correlation between amount of DPV and
inhibition of HIV infection from film (P < 0.0001; r2 = 0.635) and gel (P
< 0.0001; r2 = 0.684) users. Likewise, vaginal biopsies showed a strong
correlation between the DPV levels and inhibition of HIV infection in the
tissue from film (P = 0.002; r2 = 0.349) and gel (P < 0.0001; r2 = 0.530)
users. For both CVL and tissue, there was no significant difference in
drug levels or activity between film or gel users.
Conclusions: To define the potency of a topical microbicide, PK and PD
relationships are being defined. CVL had 10-100-fold higher DPV levels
than vaginal tissue which was reflected in better PK/PD correlations in
CVL as compared to tissue. While luminal concentrations were higher,
these data show DPV was effectively delivered to both lumen and tissue
compartments.
Marla J. Keller1, Lilia Espinoza1, Mark A. Marzinke2, Pedro M.
Mesquita1, Ashley M. Huber1, Lorna K. Rabe3, Bruce Frank4, Jason
McConnell4, Mark Mitchnick4, Craig W. Hendrix2, Patrick F. Kiser5,
Betsy C. Herold1
Albert Einstein College of Medicine, Bronx, NY, United States, 2Johns
Hopkins University School of Medicine, Baltimore, MD, United States,
3
Magee-Womens Research Institute, Pittsburgh, PA, United States,
4
Particle Sciences Inc., Bethlehem, PA, United States, 5Northwestern
University, Evanston, IL, United States
1
Background: The prodrug tenofovir disoproxil fumarate (TDF) is more
potent than tenofovir (TFV) against HIV in vitro & provides greater
protection against genital herpes in a murine model. Sustained TDF ring
delivery for 28 days (d) completely protected macaques from multiple
vaginal simian-HIV challenges. Protection was associated with mean TFV
levels in vaginal fluid (VF) of 1.8 x 105 ng/mL. The objectives of this trial
are to evaluate the safety & pharmacokinetics of a reservoir-type TDF &
placebo polyurethane (PU) IVR when used by women continuously for
14 d.
Methods: A randomized, single blind, placebo controlled trial of 30
women is ongoing in Bronx, NY. Clinical safety is assessed by pelvic
exam & colposcopy. Swabs (2 proximal, 1 distal to IVR) and plasma are
collected 1, 3, 7 & 14 d after ring insertion & 2 & 7 d after ring removal
for drug levels. Cervical tissue is obtained for TFV & TFV-diphosphate
levels & ex vivo HIV challenge studies. Anti-HIV & anti-HSV-2 activity of
VF as a marker of pharmacodynamics is ongoing.
Results: To date, 11 women completed the study (5 TDF, 6 placebo).
The mean age is 31 years (range 25-37). There are 4 product related
Grade 1 adverse events due to vaginal discharge (3 TDF, 1 placebo) & no
product related colposcopic findings. No subject reported ring removal
or expulsions. Bacteria were not detected by culture of fluid from the
core of 9 IVRs tested. Median VF TFV levels at 1, 3, 7 & 14 d after
IVR insertion are 3, 4, 6 & 9 x 104 ng/mL, respectively. There was no
significant difference in VF TFV levels during ring use but levels declined
significantly 2 & 7 d after removal to 3 & 0.4 x 103 ng/mL, respectively
(p=0.002).
Conclusions: TDF & placebo PU vaginal rings are well tolerated & TFV
levels exceed the clinical correlate of protection previously observed with
TFV gel dosing before & after sex in women (~103 ng/mL). Sustained TDF
IVR delivery may achieve drug concentrations needed to prevent HIV &
herpes & overcome the low adherence seen in prior microbicide trials.
www.hivr4p.org
73
Charlene S. Dezzutti1,2, Lisa C. Rohan1,2, Katherine Bunge1,
Nathan Ehrilich2, Leslie Meyn2, Philip Graebing2, Mark Marzinke3,
Craig Hendrix3, Brid Devlin4, Sharon L. Hillier1,2
OA13.03
OA13.04
Multicompartmental Pharmacokinetics of
Tenofovir 1% Gel Using the BAT 24 Regimen
Versus Daily and Single Pericoital Dosing
Population Pharmacokinetic Model of Vaginal
Tenofovir 1% Gel in the Cervicovaginal Fluid
Jill L. Schwartz , Debra Weiner , Angela Kashuba , David Archer ,
Vivian Brache4, Courtney A. Schreiber5, Beatrice A. Chen6, Alfred
Poindexter7, Andrea Thurman1, Jaim J. Lai2, Kuo H. Yang3, Craig
Sykes3, Christine Mauck1, Betsy Herold8, Charlene Dezzutti6,
Gustavo F. Doncel1
1
2
3
1
CONRAD/EVMS, Arlington, VA, United States, 2FHI-360, Durham, NC,
United States, 3University of North Carolina, Chapel Hill, NC, United
States, 4Profamilia, Santo Domingo, Dominican Republic, 5Perelman
School of Medicine, University of Pennsylvania, Philadelphia, PA,
United States, 6University of Pittsburgh, Pittsburgh, PA, United States,
7
Advances in Health, Houston, TX, United States, 8Albert Einstein
College of Medicine, Bronx, NY, United States
1
Background: This multicenter study evaluated 90 women for
multicompartment concentrations of tenofovir (TFV) and its active
metabolite (TFV diphosphate [TFV-DP]) after vaginal administration of
TFV 1% gel regimens (BAT24, pericoital and daily).
Methods: TFV concentrations were measured in blood plasma,
cervicovaginal fluid (CVF), & tissue, and TFV-DP was measured in tissue,
using LC-MS/MS, after a single study-related sex act (SA) and after two
weeks of twice weekly study-related sex (MA). For the SA, participants
were randomized to BAT24, precoital or postcoital and to sample
collection at 4, 12 or 72h after study-related sex. For the MA, they were
re-randomized to BAT24, precoital, postcoital or daily (sample collection
at 4, 12 or 72h and again at 5, 10 or 14 days after last study-related
sex). Anti-HIV activity was measured in cervicovaginal lavages (CVL) by
single-round infection assay.
Results: Plasma TFV median concentrations were low (≤8ng/mL).
Generally BAT24 produced the highest TFV in CVF and tissue, and the
highest TFV-DP in tissue. Population composite median profiles of TFV
and TFV-DP exposure in the tissues of BAT24 were 1-2x and 2-5x higher,
respectively, compared to other arms. All regimens provided mean TFVDP ≥1800 fmol/mg, which persisted for up to 72 hours. The terminal
half-lives of TFV and TFV-DP in the tissues were approximately 47h and
58h, respectively. Mean % HIV inhibition of CVL was ≥95% at 4h and
≥82% at 12h, but decreased to 52% by 72h, pooling all regimens.
Conclusions: Although BAT24 generally produced higher TFV and TFVDP, all dosing regimens yielded mean TFV-DP concentrations that were
above a threshold associated with protection in nonhuman primates
(~103 fmol). Decrease in % HIV inhibition in CVL at 72h was likely due
to dilution. Persistently high TFV-DP concentrations in cervicovaginal
HIV target tissues provide the basis for a forgiving dosing regimen.
Based on these concentrations, the potential of a single pericoital dose
should be explored.
74
Kuo-Hsiung Yang1, Jill L. Schwartz2, Craig Sykes1, Gustavo F.
Doncel2, Angela D.M. Kashuba1
UNC Eshelman School of Pharmacy, Chapel Hill, NC, United States,
CONRAD/EVMS, Arlington, VA, United States
1
2
Background: Tenofovir (TFV) gel effectiveness trials have measured
TFV concentrations in cervicovaginal fluid (CVF) from trial participants
to determine product adherence and to assess the relationship between
adherence and effectiveness. Herein we present a model which describes
TFV gel CVF pharmacokinetics (PK).
Methods: We evaluated data (N=111) from 2 PK studies. TFV
concentrations were measured in CVF after a single dose (CONRAD095),
1 study-related sex act & 2 weeks of twice weekly study-related sex
acts (CONRAD 113). Nonlinear mixed effects population PK modeling
was performed with NONMEM 7.2. Inter-individual variability (random
effects) was evaluated with an exponential error model while residual
variability was assessed with additive & proportional error model. 3
Monte-Carlo Simulation runs were performed to generate prediction
intervals for BAT24, peri-coital, and daily dosing (stead-state) regimens.
Results: A 3 compartment, bolus input, linear kinetic model with
a vaginal gel depot compartment, 1st-order absorption (Ka) into CVF
(Vb), clearing into Vc with distributional clearance (CLd), and 1st-order
elimination (CLt) best described the data. Fixed effects for Ka, CLt, CLd,
Vb, Vc were: 1.54 (1/h), 9.56 & 0.04 x 10-4 (L/h), 106 & 2.56 x 10-4
L. The inter-individual variability (CV%) for the parameters were: 23,
82, 221, 45, and 218 with a full covariance matrix on CLt, CLd, Vc,
and Vb (residual additive error: 0.65; proportional: 0.32). Monte-Carlo
Simulations from the model predicted that 95% of the population would
have CVF TFV concentrations falling below 2ng/mL (LLQ) by week 6
after last gel dose for peri-coital & BAT24 regimens, and week 7 for daily
dosing(median terminal T1/2= 43h).
Conclusions: This model highlights the very long cervicovaginal decay
of TFV gel after single or multiple applications. Furthermore, it can be
used to predict CVF TFV concentrations, and is therefore useful for
objective assessment of clinical trial adherence & product development
benchmarks.
OA13.05 LB
OA13.06
Sex Matters: MTN-011 Phase 1 Study
on the Impact of Sex on Tenofovir
Gel Pharmacokinetics (PK) and
Pharmacodynamics (PD)
Population Pharmacokinetic Modelling of
Dapivirine
Betsy C. Herold1, Cliff Kelly2, Beatrice Chen3, Robert Salata4, Mark
Marzinke5, Charlene Dezzutti3, Lisa Levy6, Beth Galaska Burzuk7,
Jeanna Piper8, Ian McGowan7, Craig W. Hendrix5, MTN 011
1
Albert Einstein College of Medicine, Bronx, NY, United States,
SCHARP (Statistical Center for HIV/AIDS Research and Prevention)
3
Magee Womens Research Institute, Pittsburgh, PA, United States, 4Case
Western Reserve University, School of Dental Medicine, Cleveland, OH,
United States, 5Johns Hopkins University School of Medicine, Baltimore,
MD, United States, 6Family Health International, Washington, DC,
United States, 7Magee Women Research Institute, Pittsburgh, PA, United
States, 8National Institute of Allergy and Infectious Diseases, National
Institutes of Health, Bethesda, MD, United States
1
Neliette Van Niekerk1, Jeremy Nuttall2, Stefan Zeiser3, Annalene
Nel1
International Partnership for Microbicides, Clinical Affairs, Paarl,
South Africa, 2International Partnership for Microbicides, Product
Development, Silver Spring, MD, United States, 3Kinesis Pharma BV,
PK/PD Modelling, Breda, Netherlands
Background: Tenofovir (TFV) gel reduced HIV acquisition when applied
pericoitally in CAPRISA 004, but not with daily dosing in VOICE,
attributed largely to poor adherence. The timing of gel dosing relative
to sex also may have contributed to trial outcomes as sex/semen may
modify local TFV concentration. To identify optimal gel dose timing,
MTN conducted a phase 1 PK/PD study of TFV 1% gel applied 1 hour
before (-1h), 24 hours before (-24h), or 1 hour before and 1 hour after
(BAT) sex compared to dosing without coitus.
Methods: The trial enrolled 24 evaluable couples from 2 US sites.
Women completed 2 baseline no gel visits (no sex, sex) and 5 gel visits
(-1 h + sex, -1 h no sex, -24 h + sex, -24 h no sex, and BAT + sex).
Cervicovaginal lavage (CVL), vaginal and cervical tissues, and plasma
were collected ~2 h after sex with matching collection times at no sex
visits and assayed for drug (PK) and CVL anti-HIV activity in vitro (PD).
Results: BAT dosing achieved the highest (CVL: 3.5×105 ng/mL; cervical:
129 ng/mg; vaginal: 258 ng/mg) and -24 h + sex the lowest (CVL:
2.9×103 ng/mL; cervical: 1.46 ng/mg; vaginal: 5.3 ng/mg) TFV levels.
Compared to dosing without sex, mean TFV levels after sex decreased
42% and 78% (1.33x105ng/mL, p=0.005 and 8.53x103 ng/mL, p<
0.001) in CVL and decreased 74% and 55% (13.92 ng/mg, p=0.04 and
2.64 ng/mg, p< 0.001) in cervical tissue with -1 h and -24 h dosing,
respectively. Vaginal tissue decreases were even greater. In contrast,
mean plasma TFV was 128% higher (1.61 ng/mL, p< 0.01) following
sex with -1 h dosing, presumably reflecting greater absorption.
Postcoital CVL anti-HIV activity increased significantly from a median
[IQR] baseline of 55[54]% in the absence of gel to 99[7], 77[57], and
100[0.4] with -1, -24, or BAT dosing, respectively. The antiviral activity of
CVL correlated significantly with drug level.
Conclusions: Timing of dosing relative to sex impacts TFV gel PK/
PD. Pericoital dosing or sustained delivery may be optimal for PrEP,
particularly with poor adherence.
Background: A population pharmacokinetic (PK) model has been
developed for dapivirine delivered from the Dapivirine Vaginal Ring-004
(25 mg dapivirine). The model describes the uptake, distribution and
elimination of dapivirine, and allows the simulation of scenarios that
have not been tested, allowing informed decision-making without the
need for additional clinical trials.
Methods: The population PK model was developed to describe
dapivirine levels in vaginal fluid (CVF) at the cervix and introïtus, as well
as in plasma, based on data from two Phase I trials, IPM 013 and IPM
024, which evaluated the local and systemic PK of dapivirine from Ring004 in healthy, HIV-negative women. The model was validated with
results from the dapivirine-only arm in IPM 028, a drug-drug interaction
study with miconazole nitrate.
Results: Different types of model structures were evaluated. The
observed dapivirine plasma and CVF concentrations were best described
using a model with direct transition from the fluid to the plasma
compartment, including a peripheral compartment. Visual predictive
checks were good, and fixed and random effects parameters could
be estimated with high precision. Although the initial purpose of the
model was to predict dapivirine CVF levels in cases of ring loss/removal,
it also proved very useful in assessing non-adherence to ring use in the
ongoing IPM 027 Phase III trial. The effect of different ring use/removal
scenarios on dapivirine plasma and CVF concentrations was simulated
and compared to expected levels associated with perfect ring use over
28 days. This assisted in defining a range of dapivirine plasma and CVF
levels that may be indicative of non-adherence to ring use over 28 days.
Conclusions: The population PK model for dapivirine allows the
simulation of ring use scenarios not tested in clinical trials (effect on
drug levels of ring removal, prediction of accumulation for successive
ring insertions), and is a very useful tool to support non-adherence
evaluations in Phase III.
www.hivr4p.org
75
2
Oral Abstract Session 14: Host Factors: Injury, Acquisition and Infection
OA14.01
OA14.02
Protective HLA Alleles Reduce Markers of
Gut Damage and Microbial Translocation
and Preserve the Cellular Immune Response
during Acute HIV-1 Infection
Combined Effect of HLA-C*04:01 and
KIR2DS4 on Increased HIV Viral Loads
Daniel Claiborne1, Jessica Prince1, Eileen Scully2, Jakob
Kopycinski3, Gladys Macharia3, Krystelle Nganou-Makamdop4,
Jianming Tang5, Paul Goepfert5, Tianwei Yu1, Shabir Lakhi6,
William Kilembe6, Daniel Douek4, Jill Gilmour3, Matthew Price3,7,
Marcus Altfeld8, Susan Allen1,6, Eric Hunter1
Emory University, Atlanta, GA, United States, 2Ragon Institute of MGH,
MIT and Harvard, Boston, MA, United States, 3IAVI, London, United
Kingdom, 4NIAID, NIH, Bethesda, MD, United States, 5University of
Alabama at Birmingham, Birmingham, AL, United States, 6ZEHRP,
Lusaka, Zambia, 7UCSF, Epidemiology & Biostatistics, San Francisco,
CA, United States, 8Heinrich Pette Institute, Leibniz Institute for
Experimental Virology, Hamburg, Germany
Alex Olvera1, Susana Pérez-Alvarez1,2, Carmina Ganoza3, Javier
Lama3, Nicole Bernard4, Jorge Sanchez3, Christian Brander1,2,5
IrsiCaixa AIDS Research Institute - HIVACAT, Badalona, Spain,
Universitat Autònoma de Barcelona, Barcelona, Spain, 3Asociación
Civil IMPACTA Salud y Educacion, Lima, Peru, 4Research Institute of
the McGill University Health Centre, Montreal, QC, Canada, 5Institució
Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain
1
2
1
Background: Efforts to elucidate protective host factors in HIV-1 infection
often rely on cross-sectional data to determine factors influencing
pathogenesis. Here, we study a cohort of 127 acutely infected Zambians
with longitudinal CD4 counts up to 5 years post infection to identify
novel HLA alleles associated with disease progression.
Methods: Cox proportional hazard models were used to elucidate
protective and deleterious HLA alleles. Plasma cytokines were measured
at seroconversion (median 45 days post infection), 3, and 6 months post
infection using the Luminex platform. Multicolor flow cytometry was
used to assess cellular activation.
Results: In a multivariable Cox model, HLA-B*1401, B*57, B*5801, B*81,
DQB1*02, and DRB1*15 were found to provide significant protection
from CD4 T cell decline. Protective HLA class I alleles were associated
with significantly lower plasma lipopolysaccharide (LPS) levels and fewer
activated (CD38+, PD-1+) CD8+ T cells early in acute infection, a time
point before these protective alleles significantly altered plasma viral
load (VL). Furthermore, protective HLA class I alleles were associated
with a decrease in IL-10 levels over time and lower levels of iFABP in
plasma at 6 months post infection. This data suggests that protective
cellular immune responses operate early in acute infection before
control of plasma VL is detected.
Conclusions: This study of a well-characterized subtype C Zambian
cohort of acutely infected individuals provides an opportunity to
elucidate host immunogenetic factors contributing to HIV-1 pathogenesis
and to investigate the underlying immunological mechanisms. This
data suggests that protective HLA alleles can influence disease course
early in acute infection by maintaining gut integrity, limiting microbial
translocation, and ultimately preserving functional cellular immune
responses by reducing inflammation.
76
Background: It is well known that relative viral control can be achieved
by some HIV infected individuals, especially those expressing particular
HLA class I and KIR genes. Since KIR genes use HLA alleles as ligands,
HLA and KIR polymorphisms may both be related to relative HIV
control. However, specific HLA-KIR binding combinations are largely
unknown and the effect of these combinations on HIV viral control is
just beginning to emerge.
Methods: We assessed potential associations between HLA alleles
and KIR genes with HIV viral load and CD4 counts in a cohort of 235
untreated individuals with HIV clade B infection in Lima (Peru). All
individuals were HLA typed at a 4-digit resolution, 170 of them were
also typed for KIR genes. For the KIR2DS4 locus, full (f) and deleted (d)
alleles were differentiated. Differences in viral load and CD4 counts were
tested by Mann-Whitney test and multiple comparisons effects were
controlled applying False Discovery Rate (FDR) and q-value calculation.
Results: Nineteen HLA alleles and 3 KIR genes were associated with
differences in viral load and/or CD4 counts (based on uncorrected p <
0.05). Of these, only the association between HLA-C*04:01 with high
viral load remained statistically significant after multiple comparison
correction (p=0.0001, q=0.0097). Interestingly, the 3 KIR genes showing
differences with p< 0.05 were all putative HLA-C*04:01 ligands. For this
reason we evaluated the combined effect of C*04:01 with the KIR2L1,
KIR2DS1 and KIR2DS4. Our results show that the association between
HLA-C*04:01 and elevated viral load was due to the co-expression of
KIR2DS4f, but not KIR2DS4d.
Conclusions: Our data indicate that the association between
HLA-C*04:01 expression and high viral load and reduced CD4 count is
due to a NK cell component that has not been appreciated before. This
observation, together with other reports on HLA-KIR allele combination
and variable HIV control may help shed further light on immune
mechanisms of HIV control and guide HIV immunogen design.
OA14.03
OA14.04 LB
Immune Correlates Identified in the
RV144 Vaccine Efficacy Trial Impact HIV-1
Acquisition Only in the Presence of Certain
HLA Class II Genes
Transmitted Escape Mutations Lead to
Accelerated HIV-1 Disease Progression and
Largely Define the Relative Contribution of
HLA Alleles to Control
Heather Prentice1,2, Daniel Geraghty3, Georgia D. Tomaras4,
Youyi Fong5, Wyatt Nelson3, Gustavo Kijak1,2, Susan ZollaPazner6, Sorachai Nitayaphan7, Supachai Rerks-Ngarm8, Jaranit
Kaewkungwal9, Punnee Pitisuttithum9, Nelson L. Michael1, Peter
Gilbert5, Jerome H. Kim1, Rasmi Thomas1,2
Jonathan M. Carlson1, Malinda Schaefer2, Chanson Brumme3,
Nico Pfeifer1, Roger Shapiro4, Thumbi Ndung’u5, John Frater6,
Simon Mallal7, Mina John7, David Heckerman1, Philip Goulder6,
Zabrina Brumme8, Eric Hunter2
U.S. Military HIV Research Program (MHRP), Walter Reed Army
Institute of Research, Silver Spring, MD, United States, 2Henry M.
Jackson Foundation for the Advancement of Military Medicine,
Bethesda, MD, United States, 3Clinical Research Division, Fred
Hutchinson Cancer Research Center, Seattle, WA, United States, 4Duke
University Human Vaccine Institute and the Center for HIV/AIDS
Vaccine Immunology, Duke University School of Medicine, Durham,
NC, United States, 5Statistical Center for HIV/AIDS Research and
Prevention, Vaccine and Infectious Disease Division, Fred Hutchinson
Cancer Research Center, Seattle, WA, United States, 6Veterans Affairs
New York Harbor Healthcare System and the Department of Pathology,
New York University School of Medicine, New York, MD, United States,
7
Department of Retrovirology, US Army Medical Component, AFRIMS,
Bangkok, Thailand, 8Department of Disease Control, Ministry of Public
Health, Nonthaburi, Thailand, 9Center of Excellence for Biomedical
and Public Health Informatics BIOPHICS, Mahidol University, Bangkok,
Thailand
1
Background: Two immune correlates were identified as predictors of
risk of infection in the RV144 HIV-1 vaccine clinical trial. Binding of
plasma IgA antibodies (IgA) to HIV-1 envelope (Env) correlated directly
with acquisition whereas binding of IgG antibodies to an antigen
containing the variable regions 1 and 2 of Env (IgG) correlated inversely
with acquisition. We hypothesized that HLA class II molecules expressed
on antigen presenting cells modulate CD4 T cell stimulation of antibody
production by B cells involved in vaccine-induced responses.
Methods: HLA class II genes were genotyped in the case-control group
of 205 uninfected and 41 infected RV144 vaccinated volunteers. The
interaction of HLA-DPB1, DQB1 and DRB1 alleles with IgA and IgG
immune responses was tested for an effect on acquisition by logistic
regression.
Results: DQB1*06 (p=0.002, q=0.06), DPB1*05 (p=0.03, q=0.19),
and DPB1*13 (p=0.04, q=0.20) had a significant interaction with IgA
antibody responses on acquisition. Increased IgA levels associated
with increased risk for infection only in the presence of DQB1*06. In
contrast, IgA levels did not associate with increased risk for infection
in the presence of DPB1*05 and *13. Further, high IgG antibody levels
directly correlated with the presence of DPB1*13 allele. IgG V1V2
antibodies were associated with reduced acquisition risk only in the
presence of DPB1*13 (OR=0.3, p=0.007) with no association observed
in the absence of DPB1*13. The observed interactions for DQB1*06 with
IgA and DPB1*13 with IgG remained significant in a multivariable model
when testing other primary variables including IgG avidity, antibodydependent cellular cytotoxicity, neutralizing antibodies, and Env-specific
CD4+ T cells.
Conclusions: These associations suggest that certain HLA class II genes
modulated the effect of antibody responses to the RV144 vaccine and
impacted HIV-1 acquisition. Understanding this HLA class II mechanism
will enable improved HIV vaccine design.
1
Microsoft Research, Redmond, WA, United States, 2Emory Vaccine
Center, Atlanta, GA, United States, 3UBC Centre for Excellence in
HIV/AIDS, Vancouver, BC, Canada, 4Beth Deaconess Medical Center,
Cambridge, MA, United States, 5University of KwaZulu Natal, Durban,
South Africa, 6University of Oxford, Oxford, United Kingdom, 7Murdoch
University, Murdoch, Australia, 8Simon Fraser University, Burnaby, BC,
Canada
Background: The extent of intra- and inter-host adaptation of HIV
to HLA-mediated immune responses remains a central challenge for
vaccine design, as the presence of escape mutations in circulating HIV
sequences may compromise vaccine-induced immunity and reduce the
protective effects of certain HLA alleles. One strategy is to vaccinate
against escaped epitopes, though it is unclear whether such epitopes
can elicit effective immune responses.
Methods: We developed a probabilistic model of HIV sequence
evolution, trained on >4,000 clade B and C sequences with matched
HLA types, that yields a natural metric of the extent of HLA-specific
adaptation of each sequence. HIV adaptation to HLA was strongly
associated with VL and CD4 counts in cross-sectional and longitudinal
early infection cohorts, validating our models and confirming the role
autologous adaptation plays in disease progression.
Results: Within transmission pairs, the extent to which the donors’ Gag,
Pol and Nef sequences were “pre-adapted” to the linked recipients’ HLA
alleles predicted recipient VL 24 mo post infection (Spearman r =0.39,
p=0.0005, N=81) and time to CD4< 250 (HR = 5.97, p = 0.03, N=48).
In cross-sectional chronic cohorts, individuals for whom the regional
circulating viral sequences were well adapted had higher viral loads.
Furthermore, the extent to which circulating HIV strains were adapted
to individual HLA alleles predicted the VL associated with those alleles
(Spearman r=0.77, p=0.009).
Conclusions: Transmission of HIV pre-adapted to host HLA leads to
accelerated disease progression and largely explains the extent to
which an HLA is relatively protective or hazardous. This observation
argues that the immune system is unable to mount effective responses
against these adapted epitopes, even when they represent the initial
challenge variant, and suggests that vaccination strategies that target
such adapted epitopes must achieve a level of response that is not
observed in natural infection.
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OA14.05
OA14.06
Inhibiting the Host Exonuclease TREX1
Induces a Localized and Protective Host
Interferon Response against Acute HIV
Infection in vivo
Candidate Loci Associated with AIDS Virus
Replication Identified by Whole Genome
Sequencing of SIV-Infected Macaques
Lee Adam Wheeler1, Judy Lieberman1
Harvard Medical School, Boston, MA, United States
1
Adam Ericsen1, Gabriel Starrett1, Justin Greene1, Michael
Lauck1, Mariel Mohns1, Brian Cain1, Ngoc Pham1, Muthuswamy
Raveendran2, David Rio Deiros2, Jason Weinfurter2, Adam Bailey1,
Julie Karl1, Roger Wiseman1, Donna Muzny2, Thomas Friedrich1,
Richard Gibbs2, Jeffrey Rogers2, David H. O’Connor3
University of Wisconsin - Madison, Madison, WI, United States,
Baylor College of Medicine, Houston, TX, United States, 3University of
Wisconsin - Madison, Pathology and Laboratory Medicine, Madison,
WI, United States
1
Background: Viral infections typically trigger innate immune sensors
to produce type I interferons (IFNs). HIV infection however, does not
activate these pathways. We previously showed that by inhibiting the
cytosolic exonuclease TREX1 in vitro, HIV DNA accumulated in the
cytoplasm and up-regulated IFNs that reduced HIV replication and
subsequent spreading.
Methods: Here, we use RNA interference to investigate the effect of
knocking down trex1 expression in CD4+ cells on HIV transmission in
human cervicovaginal explants, and in vivo, using a small animal model
of HIV.
Results: In the absence of TREX1, IFNs and interferon response
elements (IREs) are induced in vaginal tissue exposed to HIV and
effectively suppress viral replication, thereby blocking HIV transmission
to human cervicovaginal tissue and delayed transmission to humanized
mice for ~3-4 weeks relative to mock-treated controls. Protection
from HIV transmission was coupled with a marked increase in type I
IFN expression. Expression of inflammatory cytokines (IL-1b, IL-6, IL8), which facilitate HIV transmission, is significantly reduced relative
to mock- and LPS-treated controls. Direct treatment of vaginal tissue
with recombinant IFNβ also decreased viral replication (albeit less
effectively than inhibiting trex1), though it did not block the induction
of inflammatory cytokines. This suggests that the accumulation of viral
DNA in the cytosol of infected cells, which TREX1 is known to degrade,
is an important trigger to the induction of these genes both in human
tissues and in vivo.
Conclusions: From these data, we conclude that, in the absence
of TREX1 expression, the host interferon response can effectively
control viral replication and block dissemination for several weeks.
Further characterization of TREX1 function in vivo will allow for a
better understanding of the mechanism by which HIV evades the host
antiviral response and could provide new targets for drug and/or vaccine
development to prevent HIV transmission.
78
2
Background: A small percentage of HIV-infected people and SIVinfected macaques spontaneously control chronic-phase viral replication.
This control is associated with certain major histocompatibility complex
(MHC) class I alleles. However, only a minority of individuals with these
protective MHC class I alleles suppress viral replication, suggesting
additional loci act in concert with MHC genetics to mediate viral control.
Methods: We sequenced whole genomes of 12 SIV-infected Mauritian
cynomolgus macaques (6 controllers and 6 progressors), all of whom
share the control-associated MHC haplotype termed M1, and identified
genetic variation that differentiated controllers from progressors. We
assembled a second cohort to prospectively test the role of one such
region on SIV control.
Results: We found the highest densities of control-associated variation
on chromosomes 2, 3, 7, and 9. One of these regions is near (within
~400 kilobases) a cluster of genes involved in antigen presentation
and CD8(+) effector cell function, including the gene encoding the
cytotoxic granule protein granzyme B (GzmB). The inheritance of a
single granzyme B allele was associated with chronic-phase control
of SIVmac239 (P=0.0001) and higher expression of GzmB by CD3(-)
CD8(+) NK cells during early infection. We then assembled a cohort
of MHC-identical M1(+) animals to prospectively test whether M1(+)
animals with this “protective” GzmB allele control SIV infection; however,
only 1 of 4 animals in this cohort controlled SIV.
Conclusions: Although we did not confirm a causal relationship between
GzmB genetics and MHC-associated control of SIV, we established
a framework for identifying candidate control-associated loci and
prospectively testing causality. We are currently assessing whether any
of these regions differentiate a larger cohort of Indian-origin rhesus
macaque controllers (n=20) from the Indian-origin rhesus population in
general (n=144).
Oral Abstract Session 15: PrEP and Microbicide Adherence in Women
OA15.01
OA15.02
Hidden Heterogeneity: Uncovering Patterns
of Adherence in Microbicide Trials
To Give or Not to Give PK Results: An Ethical
Dilemma for Researchers & Regulators in
Uganda
Lori Miller1, Richard Hayes1
London School of Hygiene & Tropical Medicine, London, United
Kingdom
Juliane Etima1, Clemensia Nakabiito1, Carolyne A. Akello1, Samuel
Kabwigu1, Teopista Nakyanzi1, Josephine Nabukeera1, Miriam
Hartmann2, Elizabeth Montgomery2, Barbara Mensch3, Ariane
Van der Straten2,4
Background: Understanding participant adherence is critical to interpret
results from HIV prevention microbicide trials. Typically trials report
microbicide adherence based on the average of the number of selfreported sex acts which are covered by gel during the trial. Past trial
estimates indicate high adherence, often around 80-95%. In reality, the
population of participants may be composed of subgroups of women
who have different patterns of gel use over time. Thus, a longitudinal
approach to analyse adherence data may provide a better understanding
of how participants use study products over follow-up. This study sought
to identify subgroups of women with different adherence trajectories
over the course of several completed microbicide trials.
Methods: Longitudinal latent class analysis (LLCA) was used to identify
groups of participants with different adherence trajectories over trial
follow-up. Self-reported use of gel at sex acts was analysed from four
microbicide trials: MDP 301, HPTN 035, Cellulose Sulfate CONRAD, and
Carraguard. LLCA models were selected for each trial based on best fit
and interpretably, and compared across trials.
Results: Preliminary results identified several subgroups of women
with different adherence patterns over time, demonstrating that not
all participants had consistently high adherence. The different groups
included very high adherers throughout the trial, those with diminishing
adherence over time, and those with mid to high adherence. Interestingly,
groups with initially low adherence which increased over time were not
identified.
Conclusions: LLCA revealed that trials are actually composed of a
heterogeneous mixture of adherence patterns among trial participants.
LLCA provided a more nuanced understanding of self-reported data.
Results from trajectory analysis can be used to identify participant
characteristics associated with different adherence patterns, and this
information may be helpful for recruitment or adherence counselling for
future microbicide trials.
Collaboration, Kampala, Uganda, 2Women’s Global Health Imperative,
RTI International, San Francisco, CA, United States, 3Population Council,
New York, NY, United States, 4Center for AIDS Prevention Studies
(CAPS), UCSF, Dept. of Medicine, San Francisco, CA, United States
1
Background: During the VOICE trial, self-reports and monthly product
counts were used to measure adherence. Despite high reported
adherence, drug pharmacokinetic (PK) levels indicated widespread nonuse. The second stage of MTN 003D, a qualitative ancillary study to
VOICE, retrospectively provided participants with PK results in hopes
of encouraging candid discussion of low adherence. As the first PrEP
study to provide such results directly to participants, researchers and
regulators in Uganda were challenged with several ethical concerns.
Methods: After weighing risks and benefits of disclosing individual PK
results during in depth interviews and prior to focus group discussions,
tools were developed to ensure these were presented in a clear, yet
neutral manner using pictures of a teapot with varying amounts of
liquid. Nonetheless, regulators were concerned about the lack of health
benefit and possible victimization including blame and/or ill treatment
of participants for non-use. Study teams underwent training on the
meaning of results, how to provide them in a non-judgmental manner;
and how to handle participants’ reactions. Regulators’ concerns were
allayed during a face-to-face meeting with members of the study team
who described how results would be provided and how participants,
who might be upset by the results, would be counseled.
Results: In Uganda, 51participants received PK results; reactions
ranged from disbelief to joy. No social harms were noted. Use of the
tools facilitated discussions, even after initial denial of results. Indeed,
while only 3/41 “low adherers” initially accepted their results, during the
course of the interview many went on to provide reasons for non-use.
Conclusions: Despite ethical concerns, provision of PK results can be
done in a safe and ethical manner, which reduces the possibility of
victimization. The provision of these results can provide useful insights
into participants’ adherence challenges and future design of trials.
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79
1
OA15.03
OA15.04
The Effect of Presentation of Pharmacokinetic
(PK) Drug Results on Self-reported Study
Product Adherence among VOICE Participants
in Zimbabwe
Disclosure of Pharmacokinetic (PK) Drug
Results Promotes Open Discourse on Nonadherence among Women in VOICE
Petina Musara1, Otillia Munaiwa1, Imelda Mahaka2, Nyaradzo M.
Mgodi1, Miriam Hartmann3, Lisa Levy4, Elizabeth T. Montgomery3,
Cynthia I. Grossmann5, Zvavahera Mike Chirenje1, Ariane van der
Straten3, Barbara Mensch6
University of Zimbabwe-University of California San Francisco
Collaborative Research Programme, Harare, Zimbabwe, 2Zimbabwe
AIDS Prevention Project, University of Zimbabwe, Harare, Zimbabwe,
3
Women’s Global Health Imperative; RTI International, San Francisco,
CA, United States, 4FHI-360, Durham, NC, United States, 5DAIDS/NIH/
NIMH, Bethesda, MD, United States, 6Population Council, New York,
NY, United States
1
Background: Honest reporting of participant product use is critical
to understanding challenges to adherence in HIV PrEP trials. Previous
studies, like VOICE, have shown a huge gap between self reported
adherence and actual PK data. MTN-003D study was conducted after
VOICE trial to elicit truthful reports about product non-use in VOICE.
The study was conducted in 2 stages, the first without PK drug results
provided to participants, the second with.
Methods: We conducted in-depth interviews and focus group
discussions. Guides included questions assessing factors affecting
women’s use of study product. We enrolled 74 women at 2 Zimbabwe
sites (26 in stage 1; 48 in stage 2.) Nine participated in both stages,
and constitute the sample for this analysis. We analyzed changes in
responses to adherence questions among the women in both stages to
gain insight into their adherence challenges. Data from both stages were
coded, summarised and compared using NVivo.
Results: Five out of the 9 women reported high adherence in stage
1, yet admitted to non-use in stage 2. PK drug results showed that 2
were low adherers and 3 were inconsistent users. Zero of the 5 reported
any major challenges to product use in stage 1, but rather described
facilitators such as supportive sexual partner / family, perceived high
HIV risk and regular use of reminders. In stage 2, participants divulged
several reasons for product non-use, including joining the study for
health services, partner disapproval, side effects, decreased sexual
pleasure, negative influence, perceived reduced HIV risk, and difficulty
swallowing tablets. One participant reported high adherence in stage
1, which she maintained in stage 2, despite PK results to the contrary.
Three participants had consistent responses in stages 1 and 2. Their PK
drug results indicated that they were high adherers.
Conclusions: Presentation of drug PK results to study participants
may facilitate more honest reporting of product use and challenges to
adherence.
80
Ariane van der Straten1,2, Petina Musara3, Juliane Etima4,
Kubashni Woeber5, Elizabeth T. Montgomery1, Miriam
Hartmann1, Lisa Levy6, Thola Benny7, Helen Cheng1, Jeanna
Piper8, Cynthia I. Grossman9, Barbara Mensch10, MTN-003D
Study Team
Women’s Global Health Imperative; RTI International, San Francisco,
CA, United States, 2University of California San Francisco, CAPS,
Department of Medicine, San Francisco, CA, United States, 3UZ-UCSF
Collaborative Research Programme, Harare, Zimbabwe, 4Makerere
University –Johns Hopkins University Research Collaboration, Kampala,
Uganda, 5Medical Research Council, Durban, South Africa, 6FHI 360,
Durham, NC, United States, 7Desmond Tutu HIV Foundation, Cape
Town, South Africa, 8DAIDS/NIAID/NIH, Bethesda, MD, United States,
9
NIMH/NIH, Bethesda, MD, United States, 10Population Council, New
1
Background: In VOICE, ≥50% of women assigned to daily active oral
tablets or vaginal gel had undetectable drug in all plasma samples
tested. MTN-003D is an ancillary study using in-depth interviews (IDI)
and focus group discussions (FGD), together with disclosure of PK
results, to explore participants’ adherence challenges during VOICE.
Methods: We systematically recruited VOICE participants, with plasma
samples tested (median=6 samples), and PK results defined as low
(0%, N=85), inconsistent (>0% to< 75%, N=26) or high (≥75%; N=20)
based on frequency of drug detection. The PK results were disclosed
one-on-one and initial reactions captured by interviewers. 72 IDI and
12 FGD were conducted and summarized in debriefing reports for rapid
review of key findings. IDI participants also ranked 20 theme cards
listing adherence challenges.
Results: There were 131 participants (32 South Africa, 48 Zimbabwe, 51
Uganda). The most common reaction to disclosed PK results, by group
was: surprise (40%; low), acceptance (42%; inconsistent) and happiness
(65%; high). While the majority accepted their PK results in Zimbabwe
and South Africa, the opposite happened in Uganda. Nevertheless, even
those disagreeing with results discussed adherence challenges. Those
in the low/inconsistent groups described how they disposed of unused
products. Preliminary findings indicate that most reported initiating use;
however, low adherers later discontinued or poorly executed product
use. Experienced/worried about side effects and products may be harmful
were among the 3 top ranked adherence challenges in both the low and
high PK groups. Not receiving enough support was ranked third in the
low group; but much lower (11th) in the high group.
Conclusions: Provision of PK results seemingly promoted candid
discussions around poor adherence and experience with products
in VOICE. Detailed analyses of transcripts should clarify the meaning
behind site differences in PK results’ reactions, and adherence challenges
reported by each PK groups.
OA15.05
OA15.06 LB
Bounding the Seasons of Pre-exposure
Prophylaxis: When and why Women at
Higher Risk of HIV Might Suspend Use
Proportion Days Covered as a Measure of
Adherence in US PrEP Subjects
Keith Rawlings1, David Magnuson2, Robertino Mera Giler2
Emily E. Namey , Amy Corneli , Kawango Agot , Khatija Ahmed ,
Jacob Odhiambo2, Joseph Skhosana3
1
1
2
3
FHI 360, Social & Behavioral Health Sciences, Durham, NC, United
States, 2Impact Research and Development Organization, Kisumu,
Kenya, 3Setshaba Research Centre, Pretoria, South Africa
Gilead Sciences, Medical Affairs, Foster City, CA, United States, 2Gilead
Sciences, Drug Sfty & Public Health, Foster City, CA, United States
1
Background: Now that Truvada as pre-exposure prophylaxis (PrEP) has
been shown to be effective, critical questions about implementation
center on when individuals should take PrEP and for how long. Periods
of PrEP use, suggested to correspond with periods of increased HIV
risk, have been termed seasons of PrEP. Understanding when and why
women would suspend PrEP use is essential for effective counseling on
initiating and stopping PrEP. We explored how African women at higher
risk of HIV might define the end of a season of PrEP, to help inform
counseling guidelines.
Methods: As part of a larger study exploring PrEP and risk compensation
among women at higher risk of HIV in Bondo, Kenya, and Soshanguve,
South Africa, we conducted qualitative semi-structured interviews with
30 women from each site. Participants created timelines of their sexual
contacts over the past six months and were asked to imagine they had
taken PrEP during this period. Referencing their timelines, the women
discussed whether and why they would have taken a break from PrEP,
or would do so in the future.
Results: A third of the women reported they would have suspended their
use of PrEP during the past six months. Nearly half foresaw a reason to
suspend PrEP use in the future, citing reasons related to partnership
issues, phases of life, and life events. Partnership issues ranged from no
current partner to concepts of trust and stability in relationships. Phases
of life included pregnancy and aging out of frequent sex. Life events
focused on extended travel or festive seasons that could disrupt PrEP
use. In some of these contexts, suspension of PrEP was related to a
perceived reduction in HIV risk. In others, the potential for risk stayed
the same or increased.
Conclusions: Women recognize circumstances, separate from daily
adherence issues, in which they will stop PrEP. These may or may not
correspond to lower risk contexts. Counseling is needed when women
transition off PrEP to discuss risk and strategies for maintaining sexual
health.
Background: In 2014 the CDC and the WHO recommended the use
of once daily Truvada® (TVD, tenofovir/emtricitabine) for pre-exposure
prophylaxis (PrEP) in combination with other strategies to reduce the risk
of sexually acquired HIV-1 in adults at high risk. A recent analysis of
PrEP subjects showed that four doses per week significantly reduced HIV
acquisition. The objective here is to evaluate PrEP adherence in a large
US administrative database.
Methods: The proportion of days covered (PDC) is defined as the percent
of days TVD was supplied during the first 180 days of therapy. Only
subjects with ≥2 Rx were included in the analysis. We used national
electronic data from ~ 55% of all US retail pharmacies between January
2012 and March 2014. A previously described algorithm (Mera et al.
ICAAC 2013) was used to identify TVD for PrEP. Quantile (median)
regression was used to estimate the adjusted medians of several risk
factors.
Results: 18358 prescriptions were analyzed from 3253 unique
individuals. A total of 1107 individuals met the inclusion criteria.
The median PDC was 64.3%, 95% CI 61.4 - 67.3%. The multivariate
quantile regression showed that gender was significantly associated
with median adherence higher in males (77%, 95% CI 73-80%) than
females (34%, 95% CI 29-40%; p< 0.01). Adherence also significantly
increased by calendar year, with individuals prescribed TVD in 2013
having higher medians (70%, 95% CI 65-74%) than those in 2012
(50%, 95% CI 46-56%). There was a significant relationship between
age and adherence, with older subjects having a higher median PDC
(4% more for 10 years difference).
Conclusions: A recent analysis suggests that drug levels consistent with
≥4 TVD pills/wk resulted in a HIV risk reduction of 86-100%. Overall
adherence to TVD for PrEP in this retail data base population is consistent
with levels that have shown significant reduction in HIV transmission in
clinical trials. This is the first assessment of adherence to PrEP among
individuals outside of a clinical trial setting.
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81
1
Oral Abstract Session 16: Novel Vaccine Concepts
OA16.01
OA16.02
Live Adenovirus Type 4 Recombinants Induce
Durable Neutralizing Antibody Responses
to Influenza H5 with High Levels of Somatic
Hypermutation in Humans
Non-HIV-derived (Poly)peptides as Primary
Immunogens Followed by Envelope Boost
Elicit Cross-clade Neutralizing HIV-1
Antibodies
Jinghe Huang1, Byong Kang1, Adam Wheatley2, Sandeep
Narpala2, Adrian Mcdermott2, April Poole1, Robert Bailer2, Richard
Koup2, Stephen Migueles1, Marc Gurwith3, Mark Connors1
Mei-Yun Zhang1, Zheng Yang1, Jingjing Li1, Qingsheng Liu1,
Tingting Yuan1, Yanyu Zhang1, Li-Qing Chen2, Qi Lou2,
Huazhong Yin2, Dimiter S. Dimitrov3
1
NIAID/LIR, HIV-Specific Immunity Section, Bethesda, MD, United
States, 2NIAID, NIH, Vaccine Research Center, Bethesda, MD, United
States, 3PaxVax Inc., La Jolla, CA, United States
1
University of Hong Kong, Hong Kong, China, 2Zhejiang Academy of
Medical Sciences, Hangzhou, China, 3National Cancer Institute, NIH,
Frederick, MD, United States
Background: Induction of durable antibody responses capable of
neutralizing diverse HIV-1 isolates is the most important goal in HIV
vaccinology. Broad and potent HIV Env-specific antibodies isolated from
humans are characterized by binding to conserved sites on Env and
by high levels of hypermutation. Live virus recombinant vectors may
provide prolonged exposure to transgene- encoded antigen and may
more potently drive affinity maturation than non-replicating vectors. We
report results of a small Phase 1 trial of mucosal immunization with live
adenovirus type 4 encoding influenza H5 HA (Ad4-H5-Vtn) as a model
antigen. We explored its ability to induce durable, broad neutralizing
antibody responses through upper respiratory tract application.
Methods: Live Ad4-H5Vtn (103-108 viral particles (VP)) was administered
by tonsillar swab or by nasal spray in a dose escalation study. Controls
received (1010 VP) as enteric-coated capsules. Neutralizing antibodies
were measured by a pseudotype HA lentiviral vector system. B cells
were isolated using fluorescent HA probes. Immunoglobulin genes were
cloned for mutation analysis, and re-expressed to measure neutralization
potency and breadth.
Results: The vaccine was well tolerated and symptoms were consistent
with mild URI. Tonsillar and nasal immunizations were the most
immunogenic routes inducing neutralizing antibody responses at day
60 (ID50 = 809 and 666 respectively)which rose during the first 6
months. B cells that expanded early were specific for both H1 and H5
and were directed to the conserved stalk, with H5 HA-monospecific cells
expanding by 6 months. The median level of hypermutation was 3.6%
compared to germline at week 4 and 7.4% at weeks 34-52. However,
some antibodies contained 13-34% mutation and neutralized group 1
and 2 viruses.
Conclusions: A single dose of live recombinant Ad4-H5 can induce
highly mutated antibodies, suggesting that mucosal administration
is a promising platform for the induction of HIV-specific neutralizing
antibodies.
Background: A fundamental challenge to develop an effective HIV-1
vaccine is to identify vaccine immunogens that can initiate and maintain
immune responses leading to elicitation of broadly neutralizing HIV1 antibodies (bnAbs) through complex maturation pathways. We
have previously found that HIV-1 envelope glycoproteins (Env) lack
measurable binding to putative germline predecessors of known bnAbs
and proposed to search for non-HIV-derived immunogens that could
initiate their somatic maturation.
Methods: Using b12 as a model bnAb and yeast display technology, we
identified five non-HIV-derived (poly)peptides that bound to its putative
germline and intermediate antibody predecessors.
Results: Rabbit immunization with the (poly)peptides alone induced
high titers of antibodies cross-reactive with Envs, and serum IgGs
neutralized SF162 and JRFL. Priming rabbits with the (poly)peptides
followed by boosts with an gp140SF162 trimer and a resurfaced Env
(RSC3) induced antibodies that competed with mature b12 for binding
to gp140SF162 trimer and neutralized clade B, C and E isolates. In contrast,
rabbits immunized only with gp140SF162 and RSC3 did not compete
with mature b12 for binding to gp140SF162 trimer and neutralized only
clade B isolates. Following the priming with the (poly)peptides, the
boosting (secondary) immunogens and the order of different boosting
immunogens in rabbit immunization significantly affected the profile
and neutralization potency of the induced rabbit antibodies.
Conclusions: Our study is the first to provide an evidence that appears
to support the concept that non-HIV-derived (poly)peptides may serve as
primary immunogens to initiate immune responses leading to elicitation
of cross-clade neutralizing antibodies.
82
OA16.03
OA16.04
Mechanisms of Protection Observed with
Varicella Zoster Virus as a Vaccine Vector in
the SIV Macaque Model
Genetic Assessment of B-cell Responses
Following Immunization with HIV Env Protein
and Adjuvants or SHIV (AD8) Infection in
Nonhuman Primates
University of Toronto, Department of Immunology, Toronto, ON,
Canada, 2University of Toronto, Department of Medicine, Toronto, ON,
Canada, 3Mount Sinai Hospital, Department of Microbiology, Toronto,
ON, Canada, 4Public Health Agency of Canada, National HIV and
Retrovirology Laboratories, Ottawa, ON, Canada
1
Background: Among the HIV-vaccine strategies tested to date, those
involving recombinant viral vectors with ongoing replication such as
those from the Herpesvirus family show the most promise. We have
taken advantage of a well‑studied virus, Varicella Zoster Virus (VZV),
as a vector for HIV antigens and anticipate the induction of a durable
and multifaceted immune response against HIV when adopting this
approach. VZV establishes a latent infection with cycles of sub-clinical
reactivation that can potentially boost the immune response against HIV
antigens throughout life; an advantage over prime-boosting schemes
against HIV.
Methods: We have recently utilized VZV as an SIV vaccine vector
in the cynomolgus macaque model of HIV and demonstrated that
approximately 33% of the animals vaccinated a single time with a
VZV-SIV construct and challenged intra-rectally with multi low-dose
SIV, controlled SIV viral load down to undetectable levels after initial
infection. In addition, the remaining vaccinees maintained their setpoint viral loads significantly lower than controls (p< 0.001). Immune
responses associated with viral control have been assessed by multiparameter flow cytometry in various tissues including blood, rectal
tissue, lymph nodes and broncho-alveolar sites.
Results: Analysis of the systemic immune response showed the presence
of a pool of TEM cells that correlated to a reduction in the set-point viral
load (p< 0.001). We also have found evidence that Th17 response in
the gut and lymph nodes may play an important role in the control
of viral load (p< 0.05). We are currently investigating the dynamics
between different T cell populations (TEM, Tregs, Th17 and Th22) as well
as immune activation longitudinally during the infection phase of this
trial with emphasis on the interplay between blood and mucosa.
Conclusions: The sustained viral clearance observed in this study
offers an exciting platform to investigate possible immune correlates of
protection against SIV.
Joseph Francica1, Zizhang Sheng2, Yoshiaki Nishimura1,
Zhenghai Zhang2, Manmohan Singh3, Derek T. O’Hagan3, Daniel
Douek1, Thomas Kepler4, Malcolm Martin1, Lawrence Shapiro2,
Robert Seder1
NIH, Bethesda, MD, United States, 2Columbia University, New York,
NY, United States, 3Novartis Vaccines and Diagnostics Inc., Cambridge,
MA, United States, 4Boston University, Boston, MA, United States
1
Background: An effective preventive vaccine for HIV-1 should generate
potent neutralizing antibody responses against the HIV envelope (Env).
However, broadly-neutralizing antibodies from HIV+ individuals often
have unique characteristics such as extensive somatic hypermutation
(SHM), long CDRH3 regions and VH gene restriction. Therefore, we
monitored the development of these characteristics in the preclinical
setting.
Methods: NHP were immunized with HIV Env clade C protein alone,
or with a panel of clinical adjuvants or were infected with SHIV(AD8)
virus. Env-specific B cells were sorted, barcoded, deep sequenced, and
analyzed using a newly curated NHP Ig reference database. In total we
analyzed VH gene usage, SHM, and CDRH3 length from over 58,000
unique Ig heavy chains.
Results: The adjuvants tested here did not significantly increase germline
divergence compared to unadjuvanted protein, nor did they give rise to
B cells with long CDRH3 regions. However, animals and individual reads
with higher SHM showed evidence of a narrowed VH gene repertoire
and were enriched in the IGHV3Q gene. By comparison, antibodies
from SHIV infected animals showed an increase in SHM in animals with
better neutralization responses. Better neutralization responses further
correlated with higher viral loads and viral Env diversity, indicating that
chronic antigen stimulation and diversity is likely a driving force for
higher SHM and the development of antibody potency and breadth.
Finally, we correlated the presence of 3 VH genes: IGHV4A, 4D and 3J
with better neutralization, indicating it may be possible to identify a VH
gene signature of neutralization.
Conclusions: This study is the first comprehensive comparison of
antibody genetics using vaccine and infection models in NHP. Our
data suggest that future vaccine regimens should be designed to mimic
chronic viral infection, utilizing heterologous Env immunogens and
prolonged innate and antigen stimulation to drive affinity maturation
and neutralization breadth.
www.hivr4p.org
83
Catia T. Perciani1, David O. Willer2,3, Kamaran Haq2,3,
Oluwadamilola H. Iwajomo2,3, Jacqueline K. Chan2,3, Richard
Pilon4, Joseph M. Antony2,3, Paul Sandstrom4, Kelly S.
Macdonald1,2,3
OA16.05
OA16.06
Expanded Epitope Recognition by Vaccination
with DNA Encoding Novel Immunogens
Comprising HIV Conserved Elements
Vaccine Induced Distinct Circulating Memory
CD4+ T-cells with T-follicular Helper Cell
Commitment in Humans
Barbara K. Felber1, Xintao Hu1, Viraj Kulkarni1, Antonio Valentin1,
Margherita Rosati1, Candido Alicea1, Morgane Rolland2, Sylvie Le
Gall3, Niranjan Y. Sardesai4, Beatriz Mothe5, Christian Brander5,6,
James I. Mullins2, George N. Pavlakis1
Antje Heit1, Frank Schmitz2, Miranda Moore3, Sarah Gerdts3, Britta
Flach3, Harlan Robins3,4, Alan Aderem2,5, Stephen de Rosa3,5, M
Juliana McElrath3,5
National Cancer Institute at Frederick, Frederick, MD, United States,
2
University of Washington, Seattle, MD, United States, 3Ragon Institute
of MGH, MIT, and Harvard, Boston, MD, United States, 4Inovio
Pharmaceuticals, Inc., Blue Bell, PA, United States, 5IrsiCaixa AIDS
Research Institute - HIVACAT, Autonomous University of Barcelona,
Barcelona, Spain, 6Institucio Catalana de Recerca i Estudis Avancats
(ICREA), Barcelona, Spain
1
Background: HIV sequence diversity and potential “decoy” epitopes
are hurdles in the development of an effective AIDS vaccine. To target
immune responses towards invariable viral regions we engineered
DNA-based immunogens encoding conserved elements (CE) of HIV-1
selected on the basis of stringent conservation, functional importance,
and association with immune control.
Methods: Macaques were immunized by IM injection followed
by electroporation with DNA plasmids expressing CE or complete
immunogens (gag, env), alone or in heterologous prime-boost regimens.
Immune responses elicited by CE from gag (HIV, SIV) and env (HIV) were
compared to those obtained upon vaccination with DNA expressing the
complete antigens.
Results: Macaques vaccinated with CE gag or CE env DNA developed
robust CE-specific cytotoxic T cells (Granzyme B+, CD107+). CE-specific
T cells were found only in ~50% of animals vaccinated with DNA
encoding the complete antigen and these responses targeted fewer
CE per animal. Importantly, boosting CE-primed macaques with DNA
expressing full-length antigen increased both magnitude and breadth
of the CE responses.
Although designed as CTL vaccine, p24CE DNA vaccine induced Ab
responses able to recognize p55gag and targeting a broad range of linear
epitopes. In contrast, antibodies induced by p55gag DNA vaccination
failed to recognize p24CE or linear CE epitopes. Interestingly, boosting
of p24CE DNA primed macaques with p55gag DNA increased Abs
recognizing HIV p24gag as well as p24CE proteins, thereby inducing
broadest immunity.
Conclusions: Combination of conserved elements and full-length
immunogen provides a novel strategy to increase the magnitude and
breadth of cellular and humoral immunity while targeting efficiently the
conserved regions of the virus. This allows for the development and
expansion of subdominant responses and greater breadth of immune
response.
84
Fred Hutchinson Cancer Research Center, VIDD, Seattle, WA, United
States, 2Seattle Biomedical Research Institute, Seattle, WA, United States,
3
4
Adaptive Biotechnologies, Seattle, WA, United States, 5University of
Washington, Seattle, WA, United States
1
Background: Peripheral blood CD4+ T follicular helper (pTfh) cells were
recently identified in humans, but it is unclear whether they resemble
a stable population of antigen-specific memory cells responsive upon
antigen re-encounter. We investigated pTfh kinetics, population diversity
and their relationship to B-cell responses in vaccine recipients.
Methods: PBMC samples, longitudinally collected from volunteers
enrolled in HVTN068 from either the DNA/Ad5 or Ad5/Ad5 arm as
well as from healthy, non-vaccinated donors were used to identify three
distinct CXCR5+ pTfh subsets by their individual expression pattern of
PD1 and ICOS. Multicolor flow cytometry, qPCR and deep sequencing
of the TCRB repertoire of individual CD4+ T cell subsets was applied to
characterize and phenotype the individual pTfh subsets.
Results: Amongst the different pTfh subsets, the vaccine-specific PD1/
ICOS double-positive memory pTfh (pTfhdp) showed a unique clonal
expansion one week after boosting. Simultaneously, vaccine-specific,
IgG expressing plasmablasts appeared in the circulation. At the same
time, proliferating pTfhdp cells up-regulated germinal center Tfh genes,
and became enriched for Th1-like cells. Strikingly, during the peak
response the T-cell receptor clonal repertoire of pTfhdp was completely
exchanged and identical T-cell clones were shared with PD1+ ICOSmemory pTfh (pTfhPD1) cells.
Conclusions: We conclude that vaccination induces memory pTfhPD1
that respond with a rapid but transient expansion and differentiation
into activated pTfhdp following antigen reencounter.
Oral Abstract Session 17: Mucosal Target and Effector Cells
OA17.01
OA17.02
Immune Activation and HIV Target Cells in the
Adolescent Female Genital Tract
Functional Dynamics of the Interaction
between HIV gp120 and α4β7
Smritee Dabee1, Heather B. Jaspan1,2, Shaun L. Barnabas1,3,
Shameem Z. Jaumdally1, Hoyam Gamieldien1, David Lewis4,5,
Thola Bennie3, Angel Phuti3, Clive M. Gray1,6, Anna-Lise
Williamson1,6, Thomas J. Hope7, Francesca Chiodi8, Robin
Shattock9, Jo-Ann S. Passmore1,6, Linda-Gail Bekker3,10, Women’s
Initiative in Sexual Health (WISH)
Fatima Nawaz1, Claudia Cicala1, Katija Jelicic1, Raffaello Cimbro2,
Donald Van Ryk1, Jocelyn Ray1, Joseph Hiatt1, Catherine
Schwing1, Danlan Wei1, Massimiliano Pascuccio1, Aftab A. Ansari3,
Anthony S. Fauci1, James Arthos1
Background: The biological mechanisms underlying HIV risk in
younger women is unclear. As HIV is primarily transmitted across the
genital mucosa and preferentially infects activated CD4+ cells, we
investigated the influence of age and sexually transmitted infections
(STIs) on CD4+ target cell immune activation from the genital tract of
adolescent females from South Africa.
Methods: As part of a longitudinal cohort study aiming to enrol 150
adolescent women between the ages of 16-22 years at the Desmond
Tutu Youth Centre, Masiphumele, Cape Town [part of the EDCTP funded
Women’s Initiative in Sexual Health (WISH)], cervical mononuclear
cells were obtained from 35 adolescents and the T-cell expression of
activation and proliferation markers (CD38, HLA-DR, Ki67) was measured
by FACS. Women were screened for bacterial vaginosis (BV) and STIs (C.
trachomatis, N. gonnorhoea, T. vaginalis, M. genitalium, HSV-1 & 2 and
HPV). For comparison, 11 HIV-negative adult women were also included.
Results: Adolescents (median 18 years; IQR 18-19) had significantly
higher frequencies of activated CD4+ T-cells (CD38+, HLADR+,
CD38+HLADR+: each p< 0.0001) than adult women. In contrast,
adolescents had significantly lower frequencies of CD4+ T-cells expressing
CCR5 (p=0.02) than adult women. When adolescents were stratified
according to age (16-17, 18-19 and 20-22 year old), the 16-17 year old
females generally had higher levels of activated and proliferating T-cells
compared to the other age groups, significantly so for the proliferating
CD4+ T-cells (Ki67+; ANOVA: p=0.048). The prevalence of STIs and BV
were high in adolescents, with 40% testing positive for C. trachomatis.
Adolescents infected with C. trachomatis tended to have higher levels of
T-cell activation compared to those without an STI, significantly so for
CD38+HLADR+ T-cells (p=0.01).
Conclusions: Heightened levels of genital immune activation found in
South African adolescent females, partly due to the presence of STIs,
could put them at higher risk of HIV infection.
2
Background: Many isolates of HIV and SIV exhibit a specific affinity
for integrin α4β7, the gut homing receptor. CD4+ T cells expressing
α4β7 represent an ideal target for productive infection during mucosal
transmission. However, many details surrounding the interaction
between HIV and α4β7 remain unresolved. We hypothesize that the
evolution of a specific interaction between HIV and α4β7 provides the
virus a means to increase the efficiency of infection in mucosal tissues.
Disrupting this interaction may provide a strategy to reduce or prevent
mucosal transmission. Here we identify key features of the α4β7-HIV
gp120 interaction that facilitate viral replication.
Methods: We evaluated the capacity of gp120, by binding to α4β7,
to induce cellular proliferation of primary CD4+ T cells using CFSE
proliferation assays. We also examined the manner in which α4β7 is
physically linked to CD4 using high-resolution confocal microscopy and
fluorescence resonance energy transfer (FRET) on primary CD4+ T cells.
Results: We find that CD4 and α4β7 are closely associated (< 10nM) on
the surface of CD4+ T cells. Both gp120 and soluble MAdCAM mediated
costimulation of cellular proliferation in an α4β7-dependent manner.
Conclusions: Although α4β7 is not an entry receptor we find that it
is closely associated with CD4 on the cell membrane. This association
suggests that α4β7 plays a role in T cell proliferation. We determined
that both gp120 and MAdCAM costimulate CD4+ T cells and facilitate
cellular proliferation and viral replication. We hypothesize that one of the
selective pressures leading to a specific interaction between gp120 and
α4β7 involves the capacity of α4β7 to promote metabolic activity that
is conducive to viral replication. In the context of mucosal transmission,
that is generally inefficient, these features of HIV interactions with α4β7provide a means to increase transmission efficiency. Future studies
will address the potential of targeting this interaction as a means of
interfering with transmission.
www.hivr4p.org
85
Institute of Infectious Disease and Molecular Medicine, University of
Cape Town, Department of Clinical Laboratory Sciences, Cape Town,
South Africa, 2Seattle Biomed, Seattle, WA, United States, 3Desmond
Tutu HIV Foundation, Cape Town, South Africa, 4Institute of Infectious
Disease and Molecular Medicine, University of Cape Town, Department
of Microbiology, Cape Town, South Africa, 5National Institute for
Communicable Diseases, Johannesburg, South Africa, 6National Health
Laboratory Services, Cape Town, South Africa, 7Feinberg School of
Medicine, Northwestern University, Department of Cell and Molecular
Biology, Chicago, IL, United States, 8Karolinska Institutet, Department
of Microbiology, Tumor and Cell Biology, Stockholm, Sweden, 9Imperial
College London, Department of Infectious Diseases, London, United
Kingdom, 10Institute of Infectious Disease and Molecular Medicine,
University of Cape Town, Cape Town, South Africa
1
NIAID, National Institutes of Health, Bethesda, MD, United States,
Johns Hopkins University School of Medicine, Baltimore, MD, United
States, 3Emory University School of Medicine, Atlanta, MD, United
States
1
OA17.03
OA17.04
Preferential HIV Susceptibility of Specific
CD4+ T Cell Subsets in the Cervix
Expression of MAIT Cells in Blood and Genital
Mucosa of HIV Infected and Uninfected
Women
Vineet R. Joag1, Lyle R. McKinnon2,3, Segen Kidane1, Mark Yudin4,
Steve Kimwaki5, Stephanie Rainwater6, Julie Overbaugh6, Omu
Anzala5,7, James Arthos8, Joshua Kimani5, Rupert Kaul1,2,5
University of Toronto, Immunology, Toronto, ON, Canada, 2University
of Toronto, Medicine, Toronto, ON, Canada, 3CAPRISA, Durban, South
Africa, 4University of Toronto, Obstetrics and Gynecology, Toronto, ON,
Canada, 5University of Nairobi, Medical Microbiology, Nairobi, Kenya,
6
University of Washington, Fred Hutchinson Cancer Research Centre,
Seattle, WA, United States, 7University of Nairobi, Kenya AIDS Vaccine
Initiative, Nairobi, Kenya, 8National Institute of Allergy and Infectious
Diseases, Bethesda, MD, United States
1
Background: A better understanding of the cellular targets of HIV in
the female genital tract may inform the development of HIV vaccines
and microbicides. Proposed correlates of cellular susceptibility include
expression of the HIV co-receptor CCR5, mucosal homing integrins, and/
or CD69, a marker of early immune activation.
Methods: We used a CCR5-tropic HIV pseudovirus to quantify HIV entry
into unstimulated cervical CD4 T cell subsets in a rapid, flow cytometrybased assay. We also assessed the metabolic state of cervical cell subsets
based on Ki67 expression, since proliferating cells are more susceptible
to productive HIV infection.
Results: HIV entry into cervical T cells was CD4 and CCR5 dependent.
Viral entry was approximately three-fold higher into cervical CD4 T cells
compared to blood (p=0.0003), although within a given participant,
virus entry was strongly correlated between the two compartments
(r=0.748, p=0.0002), suggesting shared host determinants of HIV entry.
Cervical CD4 T cells that expressed a4b7 (p=0.003), a4b1 (p=0.0002),
or CD69 (p=0.0004) were approximately two-fold more susceptible to
HIV entry; CCR5 expression was increased at least two-fold on a4b7+
(p< 0.0001) and CD69+ (p< 0.0001) CD4 T cells, but only 1.2 fold on
a4b1+ (p=0.0002) CD4 T cells. Cervical a4b7+ CD4 T cells expressed
two-fold higher levels of the proliferation marker Ki67 (p=0.003), which
was not seen in CCR5+ (p=0.95), CD69+ (p=0.46) or a4b1+ (p=0.24)
CD4 T cells.
Conclusions: Preferential HIV entry into immunologically activated and
a4b7+ cervical CD4 T cells was probably due, at least in part, to increased
CCR5 expression. Increased HIV entry into cervical a4b1+ CD4 T cells
did not correlate with CCR5 overexpression, and the mechanism of
enhanced cell entry in this cell subset is unknown. Markers of metabolic
activity and proliferation were specifically increased in cervical a4b7+
CD4 T cells suggesting that this subset may be particularly suited to
sustain productive HIV infection.
86
Anna Gibbs1, Edwin Leeansyah2, Andrea Introini1, Taha Hirbod1,
Joshua Kimani3, Terry B. Ball4,5, Kristina Broliden1, Johan K.
Sandberg2, Annelie Tjernlund1
Karolinska Institutet, Department of Medicine, Solna, Stockholm,
Sweden, 2Karolinska Institutet, Department of Medicine, Huddinge,
Stockholm, Sweden, 3University of Nairobi, Department of Medical
Microbiology, Nairobi, Kenya, 4University of Manitoba, Department of
Medical Microbiology, Winnipeg, MB, Canada, 5Public Health Agency of
Canada, National Microbiology Laboratory, Winnipeg, MB, Canada
1
Background: Mucosa-associated invariant T (MAIT) cells are a recently
described antimicrobial T cell population, which is abundant in blood
and mucosal surfaces. MAIT cells have been shown to be reduced in
blood, however preserved in mucosa of HIV infected patients. The aim
of this study is to define number, localization, and functional profile of
MAIT cells ex vivo in tissue samples from the female genital tract (FGT)
of HIV infected and uninfected women.
Methods: Cervical tissue samples were collected from Swedish women
undergoing hysterectomy (n=15), and paired PMBC and ectocervical
biopsies were collected from HIV seropositive female sex workers (n=20),
HIV seronegative female sex workers (n=17), and HIV seronegative lower
risk women (n= 21) from Kenya. An advanced multicolor flow cytometry
panel was used to enumerate MAIT cells and determine their functional
profile. In situ staining was performed to define the distribution of the
MAIT cells in the tissue samples.
Results: MAIT cells represent ∼2% out of the total amount of mucosal
T cells in the cervical and endometrial compartments of the Swedish
HIV uninfected women. The majority of the MAIT cells are CD8+T cells
and they produced high levels of IFN- γ, TNF, IL-17, Granzyme B, and
moderate levels of IL-22 after E. coli stimulation. MAIT cells were located
close to the basal membrane of ectocervix and in the submucosal
compartment of the endometrium. We will evaluate the samples from
the Kenyan women in regards to the percentage of MAIT cells, in blood
and in the mucosa, of HIV infected vs. uninfected women.
Conclusions: MAIT cells are present in the FGT of Swedish HIV
uninfected women and they display microbial reactivity by producing
pro-inflammatory cytokines and cytolytic molecules. The production
of IL-17 and IL-22 may allow MAIT cells to play a role in regulating
homeostasis and integrity of the mucosal barrier in FGT. This may be
an important mechanism to defend the mucosal genital barrier integrity
and prevent dissemination of local infections.
OA17.05
OA17.06
Rapid Loss of Th17 Cells after SIV Infection
May Underlie Mucosal Dysfunction
No Increase in Activated T Cells and Limited
Increase in Adenovirus-specific T Cells in
Colon and Rectal Mucosa Following HIV-1
DNA/rAd5 Immunization
University of Washington, Pharmaceutics and Washington National
Primate Research Center, Seattle, WA, United States, 2University of
Washington, Department of Comparative Medicine, HIC/Comparative
Pathology Program, Seattle, WA, United States, 3Emory University,
Emory Vaccine Center, Yerkes National Primate Research Center,
Atlanta, GA, United States, 4Harvard University, Beth Israel Deaconess
Medical Center, Boston, MA, United States, 5Frederick National
Laboratory for Cancer Research, AIDS and Cancer Virus Program, SAICFrederick, Inc., Frederick, MD, United States
1
Background: During chronic HIV/SIV infection, CD4+IL-17-producing T
cells (TH17) are significantly depleted from mucosal tissues, and their
absence is highly associated with GI dysfunction. However, the kinetics
of immune dysfunction, in sequence with microbial translocation and
inflammation remain unknown. Herein we elucidate the kinetics of acute
host/pathogen interactions with the goal of identifying early events after
HIV infection that reveal potential prevention strategies.
Methods: We collected longitudinal biopsies and blood at early acute
timepoints from six rhesus macaques following intrarectal SIV challenge
with SIVmac239x (100,000 TCID50). Rectum, colon, jejunum, lymph
nodes, and blood were collected pre-SIV (day -21) and days 3, 7, 14,
28, 42, and 63 post-challenge. Immunophenotype, functionality,
inflammation, virus kinetics, and microbial translocation were measured.
Results: Strikingly, TH17 cells were significantly depleted from all GI
tract sites by day 3 post-SIV infection (rectum: p=.005; colon: p=.032;
jejunum: p=.047). This depletion was neither correlated to generalized
CD4 depletion, nor to generalized loss of cytokine production
(exhaustion/anergy). The marked loss of TH17 cells appeared to be
unique to this particular subset, as we did not observe dramatic early
loss of IL-17+CD8+ T cells or innate lymphoid cells. Additionally, we
found that loss of TH17 cells preceded overt inflammation and microbial
translocation, indicating that this deficiency may underlie subsequent
mucosal dysfunctions observed during HIV/SIV.
Conclusions: Taken together, our data indicate that the loss of TH17 cells
from the GI tract at the earliest stages of infection uniquely precedes
microbial translocation and a systemic proinflammatory state. The
preservation of TH17 cells may, therefore, be an important factor in
preventing HIV transmission and pathogenesis. Furthermore, targeted
TH17 cell retention could serve as a novel inhibitory strategy against HIV
pathologies.
Stephen De Rosa1, Nicole Frahm1, Gabriela Diaz1, Paul Newling1,
Daryl Morris1, Cecilia Morgan1, Barney Graham2, Edith Swann3,
Georgia Tomaras4, Janine Maenza1, Ann Duerr1, M Juliana
McElrath1
Vaccine Research Center, NIH, Bethesda, MD, United States, 3DAIDS/
NIAID/NIH, Bethesda, MD, United States, 4Duke University, Durham,
NC, United States
1
2
Background: HVTN076 is an exploratory vaccine trial examining
mucosal immune responses to the NIH Vaccine Research Center
multiclade HIV-1 DNA/rAd5 vaccine, also used in the HVTN505 test-ofconcept study.
Methods: 17 healthy Ad5 seronegative HIV-uninfected adults enrolled;
16 completed all vaccinations (3xDNA, rAd5; 0, 4, 8, 24 wks). Mucosal
biopsies were obtained from sigmoid colon and rectum by flexible
sigmoidoscopy at baseline and 1 month after rAd5. Rectal biopsies
by anoscopy were obtained 2 weeks after final DNA and 1 week after
rAd5. Single-cell suspensions following collagenase digestion were
assayed directly for phenotyping and after overnight rest for intracellular
cytokine staining. HIV-specific IgG was measured in serum and mucosal
secretions.
Results: HIV-specific CD4+ and CD8+ T cells were detected in blood at
1 month post rAd5 (69 and 44% of participants for Gag, respectively),
but none were detected in colon/rectum samples at any timepoint.
Ad-specific CD4+ T cells, measured in response to the hexon coat
protein, were detected at baseline in blood (92%) and colon (87%).
Hexon-specific response rate and magnitude increased in blood after
vaccination, but this increase was less common in colon/rectum. The
proportion of CD4+ T cells expressing CCR5 or activated CD4+ T cells
(Ki-67+BcL-2lo) in the rectum was unchanged after rAd5 vaccination.
Env-specific IgG was detected in serum and mucosal secretions.
Conclusions: The vaccine regimen did not induce detectable HIV-specific
T cells in colonic/rectal mucosa, an important site for HIV acquisition.
These cells may be present, but require in vitro expansion for detection.
Pre-existing mucosal Ad-specific T cells were detected in the majority of
these Ad5 seronegatives. Activation of such cells is a potential concern
for enhanced risk of HIV infection after rAd5 vaccination; however, postvaccination increase in Ad-specific T cells was detected in few vaccinees.
Vaccine-induced activation of T cells in bulk, as measured by Ki-67/
BcL-2, was not detected.
www.hivr4p.org
87
Laura E. Richert-Spuhler1, Tiffany Hensley-Mcbain1, Jillian Gile1,
Michael Koday1, Deborah H. Fuller1, Brian Johnson2, Melon T.
Nega3, Cara Appel2, Thomas Vanderford3, R. Keith Reeves4, Jacob
D. Estes5, Brandon F. Keele5, Nichole R. Klatt1
Oral Abstract Session 18: Evaluation of Novel Interventions
OA18.01
OA18.02
HbAHP-25 Inhibits HIV-1 Entry into Cells and
Neutralizes gp120 Induced Inflammation
Bypass of Quality Control in Protein Folding
Pathways Induces Specific Misfolding of
HIV Envelope V2 Loop: Implications for
Iminosugars as Antivirals
Tahir Bashir1, Mandar Patgaonkar2, Selva Kumar3, KVR Reddy1
National Institute for Research in Reproductive Health [NIRRH],
Division of Molecular Immunology & Microbiology, Mumbai, India,
2
Tata Institute of Fundamental Research, Department of Biological
Sciences, Mumbai, India, 3Dr. D.Y. Patil University CBD Belapur,
Department of Bioinformatics, Mumbai, India
1
Background: HIV/AIDS is one of the major causes of death worldwide.
Most of the new HIV infections occur through heterosexual mode of
transmission. Hence, preventing HIV infection at this portal of entry is
pivotal in eradicating AIDS epidemic.
Methods: HbAHP-25, a peptide against CD4 binding domain of gp120,
was designed in silico by molecular docking using Z-dock software.
Binding of this peptide to gp120 was evaluated using ELISA and flow
cytometry. Activity of HbAHP-25 against various strains of HIV-1 was
evaluated on TZM-bl cells and PBMCs using Luciferase and p24 assay
in presence of seminal plasma and vaginal fluid. Competitive ELISA
was performed to determine effect of HbAHP-25 on CD4 binding to
gp120. Cell viability and epithelial monolayer integrity in presence of
HbAHP-25 was determined by MTT assay and Immunofluorescence.
Multiplex cytokine assay was performed to determine inflammatory
status of peptide and its role in gp120 mediated inflammation.
Results: HbAHP-25 exhibits significant anti-HIV activity against various
strains of HIV-1 in a dose dependent fashion. Results from ELISA and
SPR reveal direct interaction between HbAHP-25 and gp120; it binds to
a site proximal to CD4 binding site on gp120, and has partial epitope
similarity with VRC01. HbAHP-25 retains its anti-HIV potential in
presence of vaginal fluid & seminal plasma. It didn’t affect cell viability/
monolayer integrity even at concentrations 4-5 fold higher than its
IC50. HbAHP-25 didn’t affect tight junction proteins and did not show
any significant induction of pro/anti-inflammatory cytokines; instead it
reduced gp120 induced activation of pro-inflammatory cytokines viz.,
IL-1α, IL-6, IL-8 and GM-CSF.
Conclusions: HbAHP-25 has potent anti-HIV activity by blocking gp120CD4 interaction and alleviating gp120 induced inflammation in vaginal
cells, suggesting its plausibility as a potential prophylactic/microbicidal
agent. Binding site of HbAHP-25 on gp120 can also be a potential site
for vaccine development.
88
Simon Spiro1, Nicholas McCaul2, Ronald Derking3, Dominic
Alonzi1, Raymond Dwek1, Rogier Sanders3, Ineke Braakman2,
Nicole Zitzmann1
University of Oxford, Institute of Glycobiology, Oxford, United
Kingdom, 2University of Utrecht, Cellular Protein Chemistry, Utrecht,
Netherlands, 3University of Amsterdam, Department of Medical
Microbiology, Amsterdam, Netherlands
1
Background: The HIV Env glycoprotein is vital for cell targeting and
entry. During folding in the endoplasmic reticulum the sequential
trimming of glucose residues from the protein’s glycans by ER
α-glucosidases is crucial in targeting it to the chaperones calnexin and
calreticulin (CNX/CRT). Iminosugars are a class of glucose-mimetic drugs
that inhibit α-glucosidases preventing association of Envelope with CNX/
CRT. NB-DNJ is an iminosugar that is antiviral against all clades of HIV
and no escape mutants have ever been detected even after months of
treatment in vitro. We used NB-DNJ to study the role of CNX/CRT in Env
folding and to better understand its antiviral functions.
Methods: We used radioactive pulse-chase labelling to study the effects
of NB-DNJ on Env oxidative folding in the ER and then conformational
monoclonal antibody binding to examine the structure of the final Env
protein, using both gp120 and BG505 SOSIP trimers. We also used
molecular clones to study the effects of the drugs on the production of
whole viral particles and derivative mutants.
Results: In the presence of NB-DNJ nascent gp120 oxidised unusually
rapidly, after which there was a slow isomerisation of the disulphides
back into an apparently native conformation. However, antibody
mapping of the secreted protein showed the V2 loop was in an aberrant
conformation. HIV mutants that lacked the V1/V2 loop were partially
resistant to the effects of the drug. While the proportion of misfolded
protein correlated well to α-glucosidase inhibition, the antiviral effect did
not- instead full suppression of virion infectivity occurred when only a
small proportion of the proteins were misfolded.
Conclusions: This lack of correlation can be explained by the need for
HIV envelope proteins to trimerise and then cluster in order to function;
the misfolding of a single protomer could then affect the greater whole,
thus explaining the selectively antiviral effects of iminosugars. CNX/CRT
is vital for the folding of V1/V2 but not for the rest of Env.
OA18.03
OA18.04
A Role for Scavenger-like Lymphocyte
Receptor CD6 in HIV-1 Viral Infection
Testing a Fast-dissolving Tablet Containing
a Recombinant Live Biotherapeutic Product,
MucoCept-CVN, in the Non-human Primate
Model for Colonization
Immunoreceptors Group, Institut d’Investigacions Biomèdiques
August Pi i Sunyer (IDIBAPS), Centre Esther Koplovitz, Barcelona, Spain,
2
CELLEX-IDIBAPS-HIVACAT, Barcelona, Spain, 3Infectious Diseases and
AIDS Unit, Hospital Clínic de Barcelona, Barcelona, Spain, 4Hospital
Clínic de Barcelona, Department of Immunology, Barcelona, Spain,
5
Faculty of Medicine, Universitat de Barcelona, Department of Cellular
Biology, Immunology and Neuroscience,, Barcelona, Spain
1
Background: CD6 is a lymphocyte receptor that physically colocalizes with the TCR/CD3 complex at the immunological synapse
where interacts with CD166/ALCAM, an adhesion molecule of the Ig
superfamily. The extracellular region of CD6 exihibits three SRCR
(scavenger receptor cysteine-rich) domains, a highly conserved module
of the innate immune system. Accordingly, CD6 allow lymphocytes to
sense the presence of bacterial cell wall components, and soluble CD6
(sCD6) improves mouse survival following lethal bacterial-challenge. Our
objective was to analyze whether CD6 would recognize HIV-1 structures.
Methods: sCD6 neutralization assays were performed in PBMC using
recombinant NL4.3-Renilla (X4) virus. Recombinant gp120, sCD6 and
sCD4 binding was analyzed by ELISA and Western blot. Overlapping
15-mer peptides derived from HIV-1 MN strain gp120 were obtained
from the NIH and used for ELISA competition assays. CD6 and CD4
receptors colocalization assays were analyzed by confocal microscopy.
The sCD6 mediated inhibition of gp120 binding to human PBMC was
analyzed by FACS.
Results: We report that sCD6 significantly inhibits HIV-1 infectivity (90%
at a dose of 10µg/ml) (p< 0.01). Additionally, sCD6 binds to HIV-1 gp120
in a dose-dependent manner. The CD6-binding region is localized to
a linear sequence of the V3 loop important for chemokine receptor
interaction (P6283). The interaction of sCD6 with gp120 is enhanced by
prebinding of sCD4 to gp120 (p< 0.001) suggesting that sCD6 inhibitory
activity is mediated by blocking the gp120/coreceptor interaction. In
addition, we demonstrate the interaction and colocalization between
CD6 and the CD4 receptor. Finally, sCD6 inhibited the gp120 binding to
human PBMC, in a dose-dependent manner (p< 0.05).
Conclusions: Ours results suggest that sCD6 directly interacts with V3
loop of HIV-1 gp120 and CD4 receptor and likely mediates anti-HIV-1
activity playing a role in the virological synapse. Furthermore, sCD6
could prevent HIV entry being useful as a new HIV-1 treatment.
Laurel Lagenaur1,2, Peter Lee2, Thomas Parks2
NCI, Bethesda, MD, United States, 2Osel Inc., Mountain View, CA,
United States
1
Background: Osel developed a recombinant live biotherapeutic
product, MucoCept-CVN based on human vaginal Lactobacillus jensenii
expressing the HIV-1 entry inhibitor Cyanovirin-N for prevention of
vaginal transmission of HIV-1. We previously reported that macaques
colonized with a live bacterial preparation in hydroxyethylcellulose
showed a 63 % reduction of SHIVSF162P3 acquisition followed repeated
vaginal challenge. We formulated MucoCept-CVN as a fast dissolving
vaginal tablet, which would allow for discreet, coital-independent,
female controlled use. Tablets were potent (5.8 x 1011 colony forming
units (CFUs)/gram) and stable for at least one year at 4oC and 25 oC .
Macaques were dosed with 1 tablet or 5 tablets delivered over 5 days.
We examined colonization of the MucoCept-CVN bacteria in the rhesus
macaque vagina at 14 and 21 days post dosing and found that 83%
were highly colonized.
Methods: Fast dissolving tablet were formulated (~110 mg in a 1 x 1.5
cm mold). Tablet friability and dissolution were tested. Stability of the
product was monitored for 1 year at 4oC and 25oC and for 2 months at
37oC. A single tablet or multiple tablets were administered to 18 rhesus
macaques and vaginal colonization was monitored at 14 and 21 days
post dosing by culture and PCR.
Results: Tablets were potent and contained 5.8 x 1011 colony forming
units (CFUs)/gram. The tablets were stable 1 yr at 4oC and 25 oC and for
1 month at 37oC. The tablets maintained integrity when handled and
dissolved quickly. High levels of colonization ~108 CFU/vaginal swab of
colonization were achieved in 15/18 macaques by day 21 post dosing.
Two macaques had lower colonization levels ~105 CFU/swab and in one
macaque bacteria were detected by PCR only.
Conclusions: MucoCept-CVN can be formulated and delivered as a
conventional fast dissolving tablet dosage form. Colonization was
achieved in 83% of macaques. The tablets were easy to handle and
stable for >1 yr. This tablet formulation can be manufactured on large
scale for ease of use.
www.hivr4p.org
89
Esther Carrasco1, Cristina Escoda1, Carmen Alvarez-Fernández2,
Sonsoles Sanchez-Palomino2, Esther Carreras1, José Maria
Gatell2,3, Teresa Gallart4, Felipe García2,3, Núria Climent2, Francisco
Lozano1,4,5
OA18.05
OA18.06
The Role of Semen on Vaginal HIV-1
Transmission and on the Efficacy of Maraviroc
as a Topically Applied Microbicide
Post Coital Assessment of Topical Microbicide
Formulations in the Macaque Model
Dorothy L. Patton1, Yvonne Sweeney1, Lisa C. Rohan2,3
Olivia Snyder , Angela Wahl , Michael Swanson , Rae Ann
Spagnuolo1, J. Victor Garcia1
1
1
1
UNC Chapel Hill, Chapel Hill, NC, United States
1
Background: All mucosal exposures to HIV occur in the presence of
semen. Currently, there is no consensus on the effect of semen on HIV
transmission or on the potential effectiveness of topical microbicides.
Here, we use an in vivo animal model of mucosal HIV transmission
to establish the effect of semen on vaginal HIV infection and on the
efficacy of topical microbicides.
Methods: We utilized bone marrow/liver/thymus (BLT) humanized mice;
a model validated for the study of vaginal HIV transmission and HIV
prevention strategies. We first evaluated the transmission of transmitted/
founder viruses in the presence or absence of human semen. In
addition, we also evaluated the effect of semen on cell-associated
HIV transmission. Lastly, we evaluated the efficacy of topically applied
microbicides in the presence of semen using the CCR5 antagonist,
maraviroc. Log rank Mantel-Cox was used to analyze the data.
Results: To determine the effect of semen on vaginal transmission HIV1CH040, a transmitted founder (T/F) virus, was resuspended in human
semen and vaginally administered to BLT mice. Efficient transmission
was observed regardless of the presence (6/6) or absence (4/4) of
semen. No differences were noted in the levels of peripheral viral load
or CD4+ T cell decline between the two groups. When cell-associated
HIV was used for challenge in the presence of semen, 3/4 BLT mice
became infected compared to 4/4 in the control arm. When animals
were treated vaginally with maraviroc and then challenged with HIV in
semen, complete protection was observed (6/6).
Conclusions: Our results demonstrate that semen does not enhance
transmission of either cell-free or cell-associated HIV. In addition, semen
does not diminish the protective effect of maravioc from vaginal HIV
infection when applied topically. Our results establish a new paradigm
for the evaluation of HIV prevention strategies that includes human
semen in the context of cell-free and cell-associated virus.
90
University of Washington, Obstetrics & Gynecology, Seattle, WA,
United States, 2University of Pittsburgh, School of Pharmacy, Pittsburgh,
PA, United States, 3Magee Women’s Research Institute, Pittsburgh, PA,
United States
1
Background: The pigtailed macaque post-coital model provides
assessment of the safety of developing topical microbicide products with
daily use and with coitus. We have conducted studies using the postcoital safety model to establish its usefulness with gel and film product
formulations.
Methods: Study arms of four to 24 female macaques completed each
post-coital safety experiment. Formulations tested include two placebo
films (polyvinyl alcohol and cellulose based platforms; n=4 each), one
active gel (reduced glycerin tenofovir; n=8), one placebo gel (HEC;
n=24), and no product (n=24). Experiment days 1 and 3 consisted
of two vaginal exams (Time-0 and Time-30 minutes, in reference to
product application), with no opportunity for coital activity. On days 2
and 4, each female macaque was housed with a male macaque for
15minutes shortly after vaginal sampling and intravaginal product
administration. Females were sedated for post-coital exam including
colposcopy, vaginal flora, pH and smear. Days 5 and 8 each consisted of
a single vaginal exam. Safety measures include colposcopy to monitor
the mucosal integrity of the cervicovaginal tissues, assessments for
vaginal microflora fluctuations, notations of vaginal pH, and Gram stain
analysis of vaginal smears to detect inflammatory response by PMN
infiltration.
Results: Coitus increases cervicovaginal erythema, petechiae and
ecchymosis notations at colposcopy. None of the tested formulations
exacerbated these or induced other colposcopic findings. Vaginal pH
generally decreased after exposure to formulations other than HEC gel,
but increased when semen was present after coitus. Vaginal flora and
PMN presence were not altered by the formulations tested.
Conclusions: These experiments indicate that vaginal gel and film
formulations can be assessed in the macaque post-coital safety model.
Additionally, the reduced glycerin formulation of tenofovir gel is shown
to be safe by parameters assessed in this model.
Oral Abstract Session 19: Good Participatory Practices in HIV Prevention
OA19.01
OA19.02
Inclusion of Transgender and Gender Nonconforming Communities in Preventive HIV
Vaccine Research at the HIV Vaccine Trials
Network (HVTN)
Breaking Barriers: Do Research Interests Meet
the Needs of the Lesbian, Gay, Bisexual and
Transgender (LGBT) Community in Kenya?
Fred Hutchinson Cancer Research Center, Vaccine and Infectious
Disease Division, Seattle, WA, United States, 2HIV Vaccine Trials
Network, Communications and Community Engagement Units, Seattle,
WA, United States, 3HIV Vaccine Trials Network, Clinical Development
Unit, Seattle, WA, United States
1
Background: The HVTN has developed practices to increase
representation of transgender and gender non-conforming (trans*)
communities in its clinical trials since 2007. Recognizing that this
population has elevated HIV incidence, the HVTN has undertaken
ongoing efforts to engage and include this population in preventive
vaccine trials so that they contribute to finding a vaccine for use by
those who are most in need.
Methods: A multidisciplinary Transgender Working Group was formed
in 2007. The group has reviewed and revised data collection forms and
protocol template language; developed trainings to improve cultural
responsiveness; and provided a forum for development of research
proposals to better understand and serve trans* participants.
Results: This focus on the inclusion of trans* people has led to several
HVTN policy and practice changes. Demographics forms collect data on
sex at birth and gender identity as 2 independent variables. Protocols
require pregnancy testing and use of birth control only by those who are
biologically capable of bearing children. HVTN505 was HVTN’s first HIV
vaccine study to specify eligibility of male-to-female transwomen (not
grouped with MSM). Focus groups were conducted with transwomen
to collect information about barriers and facilitators to enrollment in
HIV vaccine trials. Ongoing analysis of trans* participants in HVTN trials
continues to increase understanding.
Conclusions: While sexual orientation has become a standard part of
demographics vocabulary in clinical trials, gender identity remains poorly
understood. Laboratory reference ranges and immunogenicity analyses
are often based on birth sex, thus complicating the interpretation of
laboratory data from trans* participants. Culturally responsive trial
conduct is enhanced by consultation with community stakeholders and
by attention to the needs and experiences of trans* people in clinical
and outreach settings. Collecting and sharing data regarding trans*
people in clinical HIV prevention trials is critical.
Minority Persons Empowerment Group (MPEG), Programs, Thika,
Kenya, 2International AIDS Vaccine Initiative (IAVI), External Relations,
Nairobi, Kenya, 3amfAR, GMT, New York, NY, United States,
4
International AIDS Vaccine Initiative (IAVI), Research Preparedness External Relations, Nairobi, Kenya
1
Background: LGBT community in Kenya continues to be criminalized
under the penal code. Research with LGBT individuals in the context of
dynamic human rights and heightened criminalization requires higher
scrutiny of ethical guidance and community ownership. The LGBT and
research community in Kenya aimed to develop partnership for common
research agenda and ethical guidelines.
Methods: A two-day participatory workshop with 24 LGBT leaders
with experience with research was convened to identify current ethical
challenges, solutions, HIV and Sexual Orientation and Gender Identity
(SOGI) related research priorities in Kenya.
Results: A mismatch between researchers’ priorities and community
needs has led to inadequate community consultations. A complex
informed consent process, misdirected motivation for participation (e.g.
transport reimbursement and health care) was identified as the top
ethical challenges. HIV research priorities identified by LGBT included
the need to demonstrate the link between homophobia, transphobia
and HIV; factors affecting adherence; impact of gender-based violence
on HIV infection among LGBT. Other areas included effects of drug and
substance use, investigation of causes behind the higher incidence rate
among MSM sex workers compared to female sex workers practicing
anal sex. They recommended mainstreaming of SOGI and human
rights issues in HIV research; studying the impact of the heightened
criminalization on “coming out” and on participation in research. A
national representative LGBT research advisory group was formed to
advise, guide and facilitate linkages between researchers and LGBT
community.
Conclusions: There is need for greater involvement of LGBT community
in all stages of research . Strengthening the capacity of LGBT
organizations is instrumental in ensuring ethical conduct of HIV research
in rights constrained settings. Sustainable partnership lies in matching
researchers’ agenda with community research needs and interests.
www.hivr4p.org
91
Gail B. Broder1,2, Michele P. Andrasik1,3, Shelly T. Karuna1,3,
Transgender Working Group of the NIAID-funded HIV Vaccine
Trials Network
Jonah M. Chinga1, George V. Owino2, Kent Klindera3, Prince N.
Bahati4
OA19.03
OA19.04
Words & Images Have Consequences: Gay
Men, Prevention Research and the Doubleedged Sword of the Media
The Utilization of Good Participatory
Practice (GPP) during the Planning and
Implementation of a PrEP Study among Black
MSM
Cindra Feuer1, Michael Ighodaro2, Brian Kanyemba3, Kay
Marshall1, James Pickett4, Paul Semugoma5, Maaza Seyoum6
AVAC, New York, NY, United States, IGLHRC, New York, NY,
United States, 3Desmond Tutu HIV Foundation, Cape Town,
South Africa, 4IRMA, Chicago, IL, United States, 5ANOVA Health
Institute, Health4Men Programme, Cape Town, South Africa, 6IAVI,
Johannesburg, South Africa
1
2
Background: Homophobia, stigma and criminalization hamper efforts
to include MSM in HIV research and programming in Africa when gay
men and other MSM are at high risk for HIV. Trial sites and community
advocates must consider how to best make use of media to “humanize”
MSM and support research while protecting individual confidentiality
and safety and the stability of trial sites, especially as new harsh anti-gay
laws are passed.
Methods: After a Washington Post article quoted gay Africans and
forced a Nigerian man to seek asylum in the US advocates were inspired
to develop media engagement best practices. At ICASA 2013, AVAC and
IRMA convened two sessions for MSM to collect strategies to maximize
benefits and minimize harms when engaging media. Conversations
with journalists explored strategies for reporting on MSM. We are
documenting tactics to share with researchers, advocates and media.
Results: Participants discussed courting journalists and harnessing
personal stories to gain media attention while protecting identities.
They noted the need to balance safety with visibility, and strategized
how to do both. Media was seen as critical to counteract homophobia
with public health messages for inclusion of MSM in HIV research and
services. Yet even positive stories that include names or other identifying
details can put individuals and organizations at risk. Experienced
advocates shared methods to respond to and avoid crises, including the
use of media generated by MSM. Journalists noted the importance of
educating editors.
Conclusions: Best practices for garnering media coverage that is
sensitive, safe, accurate and benefits vulnerable groups in HIV research
should be shared with advocates and researchers, particularly in
criminalized settings. Communication plans particular to MSM will
assure recruitment, retention and protection of research participants
and integrity of research sites. Education of journalists about needs of
vulnerable populations and benefits of research programs should be a
priority.
92
Jonathan Paul Lucas1, Georgette King1, Phaedrea Watkins1,
Christopher Chauncey Watson2, Craig S. Hutchinson3, Christine
Rogers1, S Wakefield4, Sheldon D. Fields5
FHI 360, Science Facilitation, Durham, NC, United States, 2The George
Washington University, Washington, DC, United States, 3UCLA, Semel
Institute for Neuroscience & Human Behavior, Los Angeles, CA, United
States, 4HIV Vaccine Trials Network, Seattle, WA, United States, 5Florida
International University, Nicole Wertheim College of Nursing & Health
Sciences, Miami, FL, United States
1
Background: Black men who have sex with men (BMSM) in the United
States (US) are disproportionately impacted by HIV despite having no
greater risk profile than their White counterparts. Deficits in AIDS/HIV
knowledge and distrust in research among BMSM have been cited as
common barriers to vital research engagement. To address potential
concerns and build community rapport, community engagement
planning for HPTN 073, a Demonstration project designed to assess
the initiation and correlates of daily pre-exposure prophylaxis (PrEP) use
by BMSM, required innovative approaches to ensure robust community
involvement during the study design and pre-implementation process.
Methods: In 2013 the HPTN 073 Community Working Group utilized
GPP guidelines to conduct three half-day community consultations
with BMSM service providers, community based organizations and
stakeholders in Washington, DC; Chapel Hill, North Carolina; and Los
Angeles, California. Consultation attendees (n=138) were provided
overviews of global PrEP research and the HIV epidemic among BMSM
in the US, as well as a description of HPTN 073. Participants provided
feedback on HPTN 073 study design and implementation plans.
Results: Feedback from the community consultations resulted in
alterations to the HPTN 073 social marketing campaign. The changes
were incorporated to support community engagement, encourage
study enrollment and increase BMSM PrEP awareness. Cultural
competency training for site staff and healthcare providers was added
and modifications were made to HPTN 073’s participant recruitment,
retention and adherence plans.
Conclusions: Engaging community members in problem-solving
solutions to issues that affect them is one of the fundamental principles
of HIV prevention research. It is a critical strategy for partnership building
to advance HIV prevention efforts, particularly with marginalized
groups. Effective utilization of GPP can aid in the development of shared
responsibility and community ownership of HIV prevention research
initiatives.
OA19.05
OA19.06
Assessing GPP in Action: How FACTS
001 Formalised the Implementation of
Stakeholder Engagement in a Large-scale
Prevention Trial
Challenges with Participant Reimbursement:
Experiences from CAPRISA 008 - A Post-trial
Access Study
Wits Reproductive Health & HIV Institute, Johannesburg, South Africa,
Perinatal HIV Research Unit, Soweto, South Africa, 3The Aurum
Institute, Johannesburg, South Africa, 4The Aurum Institute, Rustenburg,
South Africa, 5Desmond Tutu HIV Foundation, Cape Town, South
Africa, 6MatCH Research, Pietermaritzburg, South Africa, 7Medunsa
Clinical Research Unit, GaRankua, South Africa, 8Qhakaza Mbokodo
Research Clinic, Ladysmith, South Africa, 9Setshaba Research Centre,
Soshanguve, South Africa
1
2
Background: The Good Participatory Practice (GPP) Guidelines and Tools
for Biomedical HIV Prevention Trials aim to ensure effective stakeholder
engagement throughout trials. GPP Guidelines (2nd edition) have been
published, but experience with translation from principles to practice is
limited. We evaluated the formal implementation of 16 GPP principles
within the FACTS 001 trial, a phase III licensure trial of tenofovir 1% gel
conducted across 9 sites in South Africa.
Methods: Prior to trial initiation, staff and community advisory boards
(CABs) at all sites received GPP training. Sites subsequently produced
comprehensive GPP plans; these were reviewed and updated quarterly,
and sites reported monthly on the implementation of the plans. We
evaluated 44 GPP plans and 129 monthly reports produced by sites
between January 2012 and July 2013 to determine the extent to which
the GPP principles were applied and found effective.
Results: FACTS sites effectively applied all of the GPP principles relevant
to sites before and during implementation of the trial. This was achieved
through support from a full-time GPP manager based in the CORE team,
formal incorporation of GPP procedures into the Manual of Procedures,
extensive staff and CAB trainings, and regular review of site materials.
While plans were not always implemented as designed, sites developed
site-specific strategies and used new tools to meet specific needs.
Challenges included staff turn-over at sites, and the need for repeat
training in GPP planning and reporting. Balancing the requirement for
multiple tools to monitor GPP formally, without making monitoring
burdensome, was a key lesson learned.
Conclusions: The GPP guidelines provided a framework for the
development of effective stakeholder engagement in the FACTS trial.
Site-specific GPP plans provide useful over-arching guidance and
strategies needed for rapid response situations.
Centre for the Programme of AIDS Research in South Africa, HIV
Prevention Research, Durban, South Africa, 2Sandra Rotman Centre,
University Health Network, Toronto, ON, Canada, 3Dalla Lana School
of Public Health and Joint Centre for Bioethics, University of Toronto,
Toronto, ON, Canada, 4Mailman School of Public Health, Columbia
University, Department of Epidemiology, New York, NY, United States
1
Background: Reimbursement of trial participants is a controversial
issue, with post-trial access studies necessitating new strategies and
models. CAPRISA 008, a post-trial access study testing the feasibility of
using family planning services to rollout a pre-licensure HIV prevention
intervention (1% tenofovir gel), tried to balance the real life scenario of
no reimbursement for attendance at public sector clinics, with recognition
of the burden borne by participants in the preceding efficacy trial, by
offering minimal reimbursement for study visits. This reimbursement
was meant to subsidise transport to the clinic, but impacted negatively
on accrual, retention and participant morale.
Methods: A review of relevant local and international guidelines was
conducted by CAPRISA’s bioethicist, as were consultations between the
senior protocol team, participants and the community. Subsequently
the institutional policy on reimbursement was revised, as per guideline
recommendations. A revised reimbursement schedule for this study
was submitted to, and approved by the University of Kwazulu-Natal’s
Biomedical Research Ethics Committee (UKZN BREC).
Results: The review of guidelines recommended compensation for
participant time, travel and inconvenience. The retention rate for
December 2012 was 63.0% (standard 90%); UKZN BREC approval of
the revised rate occurred in February 2013, and retention in March 2013
rose to 92.2%. To date the overall retention is 94.6% (March 2014).
Conclusions: Changes in reimbursement schedules between efficacy
and post-access trials, without prior consultation with participants
and community representatives, may negatively impact accrual and
retention, with the potential to reduce sample size and impair the
validity of the findings. The question remains if community members
who did not participate in the efficacy study, should receive the same (if
any) reimbursement as the original participants, if included in post-trial
access studies.
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93
Sinazo Pato1, Deborah Baron1, Busi Nkala2, Phumeza Mzizi2,
John Mdluli3, Ireen Mosewu4, Leader Kanyiki5, Duduzile
Lembethe6, Zonke Mabude6, Ronald Moate7, Isaac Ngema8,
Ross Malamatsho9, Cynthia Dlamini1, Sinead Delany-Moretlwe1,
Glenda Gray2, Helen Rees1
Kathryn T. Mngadi1, Janet Frohlich1, Carl Montague1, Jerome
Singh1,2,3, Nelisiwe Nkomonde1, Nomzamo Mvandaba1,
Fanelesibonge Ntombela1, Londiwe Luthuli1, Quarraisha Abdool
Karim1,4, Leila Mansoor1
Oral Abstract Session 20: Reproductive Hormones and HIV Risk
OA20.01
OA20.02 LB
Medroxyprogesterone Acetate Enhances
HIV-1 Uptake and Transcytosis, but not
Replication, in Primary Human Genital
Epithelial Cells
The Contraceptive MPA, Unlike NET,
Modulates Expression of Immune Function
Genes and Increases HIV-1 Infection in
Cervical Tissue Explants and PBMCs
Victor H. Ferreira1, Aisha Nazli1, Jessica K. Kafka1, Kristen Mueller1,
Michel J. Tremblay2, Alan Cochrane3, Charu Kaushic1
Roslyn M. Ray1, Chanel Avenant1, Johnson M. Moliki1,
Janet P. Hapgood1
1
McMaster University, Hamilton, ON, Canada, 2Laval University, Laurier,
QC, Canada, 3University of Toronto, Toronto, ON, Canada
1
Background: Half of all people living with HIV worldwide are women,
yet early events in the female genital tract are poorly understood
particularly in the context of female sex hormones or hormonal
contraceptives, such as medoxyprogesterone acetate (MPA), which have
been correlated with increased HIV susceptibility and transmission.
Methods: Genital epithelial cell (GEC) cultures were prepared from
human tissues and grown in the presence or absence of physiological
concentrations of estrogen (E2), progesterone (P4) or MPA prior to HIV-1
exposure. HIV uptake was measured by p24 ELISA in disrupted GECs and
localization of virus was confirmed by electron microscopy (EM). Various
inhibitors were used to determine the pathways involved in viral uptake.
HIV infection and replication were determined by using highly sensitive
real time PCR assays. Apical recycling and transcytosis were assessed
by measuring p24 antigen in apical or basolateral GEC supernatants,
respectively, and infectiousness of the virus was determined by TZM-b1
assay.
Results: HIV uptake was significantly increased within endometrial and
cervical GECs grown in the presence of MPA. HIV-1 uptake by GECs
primarily took place via endocytosis, since treatment with Dynasore, an
endocytosis inhibitor, significantly decreased HIV uptake. EM confirmed
that HIV-1 was localized to intracellular vesicular compartments. Despite
uptake into GECs, no early or late reverse transcription products,
integrated HIV DNA or spliced RNA transcripts were measured, regardless
of hormone exposure. Nevertheless, HIV-1 transcytosis was significantly
increased among GECs grown in the presence of P4 and MPA and this
virus was infectious.
Conclusions: These results suggest that female sex hormones,
particularly MPA, regulate HIV transcytosis across the epithelium and HIV
uptake into GECs, but not replication. Ongoing studies are investigating
whether enhanced transcytosis in the absence of productive infection,
would result in increased HIV transmission to target cells.
Background: The synthetic progestins, medroxyprogesterone acetate
(MPA) and norethisterone enanthate (NET-EN), are widely used in
developing countries as injectable contraceptives, where disease burden
is high. Some studies suggest that MPA, unlike NET, increases HIV-1
acquisition in women. Whether MPA and NET differentially affect HIV-1
infection and the expression of key genes relevant to HIV-1 acquisition
via differential molecular mechanisms, is key to understanding choice of
progestin contraceptive for HIV-1 prevention.
Methods: Regulation of selected genes was investigated in cervical
tissue explants and peripheral blood mononuclear cells (PBMCs) by qRTPCR, western blotting and Luminex assays, in response to physiologically
relevant doses of progestogens. Infection assays were performed in the
absence and presence of HIV-1 using HIV-1BAL-RENILLA or HIVpNL4.3 IMCs.
The GR specific antagonist RU486 or GR siRNA knockdown were used
to determine the role of the GR in modulating ligand-specific effects.
Results: In PBMCs, MPA like dexamethasone (DEX, a GR specific agonist),
showed anti-inflammatory effects, decreasing pro-inflammatory IL6, IL8
and RANTES levels and increasing anti-inflammatory GILZ gene expression
levels, while NET and progesterone (P4) did not. In primary cervical tissue
explants, DEX and MPA repressed IL6 and IL8 and increased GILZ gene
expression levels. Differential gene expression by MPA versus NET and P4
were mediated via the GR in PBMCs. Similarly, MPA and DEX, unlike NET
and P4, increased HIV-1 replication in viable PBMCs. In primary cervical
explants, MPA, but not NET increased HIV-1 replication.
Conclusions: Collectively, the data suggest that NET, unlike MPA,
would be a safer choice of injectable progestin contraceptive in young
women in high risk areas for HIV-1 infection. The molecular basis for
this choice most likely involves differential effects of MPA as compared
to NET and P4, on transcription of immunomodulatory genes, due to
their differential actions via the ubiquitous GR.
94
The University of Cape Town, Molecular and Cell Biology, Cape Town,
South Africa
OA20.03
OA20.04
Hormonal Regulation of Early Innate Immune
Responses to HIV Infection by Dendritic Cells
from the Human Female Reproductive Tract
Injectable Contraceptive Use Correlates with
Increased HIV Target Cells at the Cervix in
Young South African Women
Marta Rodriguez-Garcia1, Zheng Shen1, John V. Fahey1, Austin W.
Boesch2, Margaret E. Ackerman2,3, Charles R. Wira1
Elizabeth H. Byrne1, Melis Anahtar1, Kathleen Doherty1,2, Gregory
Olson1, Brittany Bowman1, Nikita Padavattan3, Zaza Ndhlovu1,3,
Musie Ghebremichael1, Bruce Walker1, Thumbi Ndung’u1,3, Krista
Dong1, Douglas S. Kwon1
Background: Dendritic cells (DC) in tissues differ considerably from
classical in vitro models. DCs are key for capturing and transferring HIV
to CD4+ T cells as well as inducing adaptive immune responses. A major
gap remains in our knowledge regarding phenotype, innate responses
and regulation of DCs in the female reproductive tract (FRT), one of the
main portals of entry for HIV. Here we investigated hormonal regulation
of DC from different anatomical sites in the FRT and their early innate
responses to HIV.
Methods: Tissues from the endometrium (EM), endocervix (CX) and
ectocervix (ECX) were digested prior to DC purification using CD14 and
CD1a magnetic bead selection. Cells were analyzed by flow cytometry
or stimulated with HIV for 3 and 24 hr after estradiol (E2) treatment
(5x10-8M). Supernatants were analyzed for 9 different antimicrobials
with anti-HIV activity, using a multiplex assay developed by us.
Results: We identified distinct populations of DC/macrophages in
the FRT, based on their expression of CD11c and CD11b. Population
distribution was different between tissues, with DCs more abundant in
EM and ECX than CX. Higher expression of HLA-DR was detected in
DC from the ECX, but no differences were found in CD14 and CD103
expression between tissues. Analysis of supernatants revealed that
CD1a+ and CD14+ cells constitutively produced HNP1-3, SLIP, RANTES,
Elafin, Lactoferrin, and MIP3alpha. Treatment with E2 for 24h increased
production of HNP1-3, RANTES, MIP3alpha and SLPI. HIV stimulation
induced HNP1-3, RANTES and Elafin within 3hr, but interestingly when
DC were pretreated with E2, the stimulatory effect of HIV was undectable.
Conclusions: Different DC populations exist in the FRT that may play
different roles in HIV pathogenesis. DC produce antimicrobials shortly
after HIV challenge and these responses are modified by E2. Overall,
our results indicate hormonal regulation of innate immune responses
by DC to HIV, which may have consequences for alterations in HIV
susceptibility in the FRT.
States, 2Vanderbilt University School of Medicine, Nashville, TN, United
States, 3HIV Pathogenesis Programme, Durban, South Africa
1
Background: The use of progestin-only injectable contraception (IC) has
been epidemiologically associated with increased risk of HIV acquisition,
but it remains unclear whether this relationship is causal and, if so, the
mechanisms by which progestin-only contraceptives increase risk.
Methods: Demographic and behavioral data along with blood
and female genital tract (FGT) samples were obtained from a cohort
of 18-23 year old HIV uninfected women in Durban, South Africa.
Cervical cytobrush cells were analyzed by flow cytometry and plasma
progesterone measured by chemiluminescence. Cytokines in cervicovaginal lavage (CVL) were measured by luminex.
Results: Women using IC disproportionately accounted for new HIV
infections during the study (RR 2.55 [1.90-3.42], p=0.0018). While
31.5% of women in the cohort used IC, 72.7% of women who became
HIV positive used IC (8 of 11); 9.09% of infected women did not use
contraception. This difference was not accounted for by behavioral or
demographic differences. The use of IC was significantly associated
with a higher frequency of CD4+CCR5+ T cells of CD45+ cells in the
cervix relative to those not on hormonal contraception (1.18% [0.33,
4.53] vs. 0.29% [0.13, 0.48], respectively; p=0.036). Women with high
endogenous progesterone (>0.3ng/mL, luteal phase) in the absence
of IC use had significantly increased frequency of CD4+CCR5+ T cells
in the cervix relative to those with low progesterone (follicular phase;
1.16% [0.51, 3.76] of CD45+ cells vs 0.29% [0.13, 0.48], respectively;
p=0.042). High endogenous progesterone was also associated with
a higher frequency of cervical CD4+CCR5+HLADR+CD38+ T cells
(p=0.034). There was no observed difference based on IC use in soluble
cytokine or chemokine levels in CVL.
Conclusions: The increased frequency of HIV target cells at the site of
exposure in the context of IC and high endogenous progesterone provides
a potential biological mechanism for the increased HIV acquisition risk
for women using injectable progestin-only contraceptives.
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95
Geisel School of Medicine at Dartmouth, Physiology and
Neurobiology, Lebanon, NH, United States, 2Thayer School of
Engineering, Hanover, NH, United States, 3Geisel School of Medicine at
Dartmouth, Microbiology and Immunology, Hanover, NH, United States
1
OA20.05 LB
OA20.06
Increased Number of HIV Target Cells in
Zimbabwean vs US Women
Hormonal Contraception Use and Risk of
Female-to-Male HIV Transmission Risk in a
Zambian Cohort
Sharon L. Achilles1,2, Felix G. Mhlanga3, Allen T. Matubu3, Kevin A.
Stoner2, May A. Beamer2, Z. Mike Chirenje3, Sharon L. Hillier1,2
University of Pittsburgh, Obstetrics, Gynecology and Reproductive
Sciences, Pittsburgh, PA, United States, 2Magee-Womens Research
Institute, Pittsburgh, PA, United States, 3UZ - UCSF, Harare, Zimbabwe
1
Background: Hormonal contraception (HC), especially depot
medroxyprogesterone acetate (DMPA), has been associated with
increased HIV risk. Several investigators are evaluating the impact of
HC on immune cells and immune mediators in the female genital tract
(FGT). Understanding baseline differences in these immune factors in
different populations is critical to the evaluation of the impact of HC.
Our objective was to compare immune cells from a Sub-Saharan African
(SSA) population of women and a North American (US) population of
women using identical sampling and analytical methods.
Methods: Women in Pittsburgh, PA (US) and in Harare, Zimbabwe (SSA)
aged 18-34 who had not used hormonal or intrauterine contraception
for >30 days and DMPA for >10 months were enrolled into a study
designed to quantify systemic and FGT immune cell populations.
Enrolled women screened negative for HIV, gonorrhea, chlamydia,
trichomonas and active herpes. Immune cell populations collected from
blood and endocervical cytobrushes were quantified by flow cytometry.
All flow cytometric data were gated in a single lab.
Results: Eighty US women and 80 SSA women were enrolled. SSA
women had a higher mean number of CD4 cells 3443 vs 1674, p <
0.001), CD4CCR5+ cells (1696 vs 887, p=0.002), and activated CD4
cells (expressing CD69) (2066 vs 1233, p< 0.02) in the cervix compared
to US women. SSA women also had more CD4CCR5+ cells (3775 vs.
1547, p< 0.001) and CD4CCR6+ cells (11,053 vs. 3869, p< 0.001), and
had fewer activated CD4 cells (expressing CD69) (2085 vs. 2951, p<
0.01) in the PBMCs compared to US women.
Conclusions: There are significant differences in the numbers and
activation status of both peripheral and local CD4 cells in US and SSA
women, with SSA women having both greater mean CD4 and CCR5
expressing T-cells compared to US women. Studies evaluating the
impact of HC on FGT immune cells should consider baseline cellular
populations. HC induced changes in immune cells may be mitigated by
the number and type of cells at baseline.
96
Kristin M. Wall1,2, William Kilembe3, Htee Khu Naw3, Ilene Brill3,
Bellington Vwalika3, Elwyn Chomba3, Brent Johnson1, Lisa
Haddad1, Amanda Tichacek2, Susan Allen2
Emory University, Atlanta, GA, United States, 2Rwanda Zambia HIV
Research Group, Atlanta, GA, United States, 3Rwanda Zambia HIV
Research Group, Lusaka, Zambia
1
Background: The association between hormonal contraception (HC) use
and risk of both male-to-female and female-to-male transmission HIV
transmission is highly debated. Very little data exists related to femaleto-male transmission risk and HC use. USAID, FHI360, and the World
Health Organization have called for more evaluations of this association.
Methods: HIV discordant couples in which the man was negative and the
woman was positive (M-F+) were identified after couples’ voluntary HIV
counseling and testing from 1995-2012 in Lusaka, Zambia. Discordant
couples were followed longitudinally and demographic, behavioral,
clinical, and family planning measures were collected at baseline and
3-monthly. Men were re-tested for HIV every three months. Multivariate
Cox models evaluated time to HIV acquisition among men.
Results: Among 1654 M-F+ couples, 226 incident infections occurred
over 3366 couple-years (6.71/100 couple-years; 95%CI: 5.87-7.65).
171 (76%) infections were genetically linked to the study partner. No
interaction between genital ulceration/inflammation and contraception
was observed. Use of injectables (HR=0.7; 95%CI:0.4-1.4), OCPs
(HR=1.3; 95%CI:0.7-2.1), or implants (HR=0.8; 95%CI:0.2-2.8) in
the past three months was not associated with genetically linked HIV
transmission from women to men relative to non-HC controlling for:
woman’s age, number of previous pregnancies, woman’s log viral load
at baseline, male circumcision status, pregnancy status, sex frequency
with and without a condom, sperm on a wet mount, male and female
genital inflammation and ulceration, and time interval since enrollment.
Conclusions: Our results add to a small and inconclusive body of
literature. Over 17 years of follow-up, we found no statistically significant
association between HC use and female-to-male HIV transmission.
These findings support the continued use of HC methods for pregnancy
prevention and Prong 2 of PMTCT.
Oral Abstract Session 21: Viral Transmission Studies
OA21.01
OA21.02
Transmission of Pre-adapted Viruses
Determines the Rate of CD4 Decline in
Seroconverters from Zambia
The Sequence of the α4β7-binding Motif
on Gp120 of Transmitted/Founder Viruses
Contributes to the Dependence on the
Integrin for HIV Infection
Emory University, Atlanta, GA, United States, 2Microsoft Research,
Redmond, WA, United States, 3Zambia Emory HIV Research Project,
Lusaka, Zambia, 4University of Alabama-Birmingham, Birmingham, AL,
United States, 5IAVI, San Francisco, CA, United States, 6IAVI, London,
United Kingdom
1
Background: HIV escapes adaptive cellular immunity by selecting
mutations that are associated with the individual’s HLA-I alleles.
These mutations can be transmitted but the impact of this process on
pathogenesis is poorly understood.
Methods: In 169 transmission pairs, we studied the transmission of HIV
polymorphisms in Gag, Pol and Nef by Sanger sequencing of population
amplicons in the donor (D) and the linked-recipient (LR)
(≤3 months post-transmission). Polymorphisms statistically-linked to
HLA alleles or located in well-defined CTL epitopes were quantified
according to each LR’s HLA alleles and associated with their set-point
VL and CD4 counts.
Results: The majority of polymorphisms (83.6%) were transmitted from
the D to the LR and a significant fraction (17.3%) was already adapted
to the LR’s HLA (11.6% escape and 6.2% epitope-located). A Spearman
correlation analysis showed that transmission of Pol polymorphisms
irrelevant to the LR’s HLA was associated with a diminished set-point
VL (p=0.003). This association was lost (p=0.4) when other variables
known to determine set-point VL (gender-p=0.01; B*57-p=0.02; HLA-B
sharing-p=0.006; replicative capacity (RC)-p=0.008) were included in
a Generalized Linear Model. An in-depth analysis of survival curves
(log-rank test) for different CD4 endpoints (200-350 cells/ul) showed
that the proportion of transmitted HLA-linked polymorphisms relevant
to the LR’ HLA in Gag was consistently associated with a faster CD4
decline (p=0.0004). When other factors (gender, protective alleles, allele
sharing, RC and set-point VL) were considered in a Cox Proportional
Hazard Model, the proportion of transmitted HLA-linked polymorphisms
in Gag remained the only variable significantly associated with CD4
decline (p=0.03).
Conclusions: Because most Gag, Pol and Nef polymorphisms are
transmitted, newly infected individuals can receive a pre-adapted variant
that leads to an accelerated disease progression (faster CD4 decline)
without showing a significant effect on set-point VL.
Simone I. Richardson1,2, Elin Gray1, Nonhlanhla Mkhize1,2, Daniel
Sheward3, Bronwen Lambson1,2, Kurt Wibmer1,2, Lindi Masson3,
Lise Werner4, Nigel Garett4, Jo-Ann Passmore3, Salim AbdoolKarim4, Carolyn Williamson3, Penny Moore1,2, Lynn Morris1,2
Centre for HIV and STI’s, National Institute for Communicable
Diseases, Johannesburg, South Africa, 2School of Pathology, University
of the Witwatersrand, Johannesburg, South Africa, 3Institute of
Infectious Diseases and Molecular Medicine, University of Cape Town,
Cape Town, South Africa, 4Centre for the AIDS Programme of Research
in South Africa (CAPRISA), Nelson R Mandela School of Medicine,
University of KwaZulu-Natal, Durban, South Africa
1
Background: The integrin α4β7, which mediates the trafficking of T
lymphocytes to the gut associated lymphoid tissue (GALT), a site of rapid
HIV replication, has been described as an attachment factor for the V2
loop of the envelope protein gp120. We aimed to study the factors that
influence dependence on α4β7 for replication of transmitted/founder
viruses including cytokine levels in cervicovaginal lavage (CVL), STI
infections and the sequence of the tripeptide α4β7-binding motif.
Methods: All-trans retinoic acid-activated CD4+ T cells were incubated
with or without HP2/1 (anti-α4 antibody) or Act-1 (anti-α4β7 antibody)
prior to adding virus. Infectious virus was prepared using envelope genes
of the transmitted/founder (T/F) virus from 8 individuals in the CAPRISA
Acute Infection cohort. Replication was monitored by p24 ELISA.
Changes in viral sequence were generated by site-directed mutagenesis.
Results: T/F viruses with the highest dependence on α4β7 for replication
had P/SDI/V motifs while those with lower dependence were LDI/L.
Mutation of viruses with LDI/L motifs to P/SDI/V resulted in increased
dependence on α4β7 for replication while the reverse mutation restricted
the ability of the viruses to enter cells. T/F viruses from individuals
diagnosed with bacterial vaginosis (BV) at the time of virus isolation had
significantly higher dependence on the integrin for replication. Levels of
IL-7, a cytokine that upregulates α4β7 expression, correlated with α4β7
dependence in the CVL shortly after transmission. Both BV status and
high IL-7 levels in the CVL were associated with the P/SDI/V motifs in a
larger cohort of 28 CAPRISA 002 participants.
Conclusions: P/SDI/V motifs are more common among South African
HIV subtype C viruses accounting for 35% of variants. These data
suggest that viruses with P/SDI/V motifs favour α4β7 reactivity at
transmission influenced by the presence of BV and IL-7 cytokine levels.
These findings may lead to vaccine and therapeutic opportunities in
which α4β7 reactivity is exploited.
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97
Daniela Monaco1, Dario Dilernia1, Malinda Schaeffer1, Kristine
Dennis1, Jonathan Carlson2, Jessica Prince1, Daniel Claiborne1,
William Kilembe3, Shabir Lakhi3, James Tang4, Matt Price5,
Paul Farmer1, Richard Kaslow4, Jill Gilmour6, Susan Allen1, Paul
Goepfert4, David Heckerman2, Eric Hunter1
OA21.03
OA21.04
HIV Replicative Capacity of Transmitted
Viruses Is Associated with Early Immune
Activation, Exhaustion and Establishment of
the Viral Reservoir
In-vitro Fitness of HIV-1 Transmitted/Founder
versus Non-transmitted Full-length Genome
Infectious Molecular Clones
Jessica L. Prince1, Daniel T. Claiborne1, Gladys Macharia2, Luca
Micci1, Benton Lawson1, Eileen Scully3, Jakub Kopycinski4,
Thomas Vanderford1, Jianming Tang5, Tianwei Yu1, Shabir Lakhi6,
William Kilembe6, Guido Silvestri1, Paul Goepfert5, Matthew A.
Price2,7, Marcus Altfeld8, Mirko Paiardini1, Jill Gilmour2, Susan
Allen1,6, Eric Hunter1
Emory University, Atlanta, GA, United States, 2International AIDS
Vaccine Initiative (IAVI), London, United Kingdom, 3Ragon Institute
of MGH, MIT and Harvard, Boston, MA, United States, 4Imperial
College London, London, United Kingdom, 5University of Alabama
at Birmingham, Birmingham, AL, United States, 6Zambia Emory HIV
Research Project, Lusaka, Zambia, 7UCSF, San Francisco, CA, United
States, 8Heinrich Pette Institute, Leibniz Institute for Experimental
Virology, Hamburg, Germany
1
Background: Determining the host and viral factors that shape the
trajectory of early HIV-1 pathogenesis is key for developing rational
prevention strategies. Previously, we showed that in individuals recently
infected with HIV-1 subtype C, low viral replicative capacity (RC) as
defined by the transmitted Gag sequence, was associated with a delayed
loss of CD4 T cells independent of set point VL and host immunogenetic
factors. We hypothesize that low RC leads to a muted inflammatory
response characterized by reduced immune activation, might attenuate
infection of memory T cell subsets and preserve critical CD4 T cell
homeostasis.
Methods: Levels of plasma cytokines at seroconversion were measured
using a Luminex platform. Flow cytometry was used to assess markers
of activation (CD38+HLADR+), exhaustion (PD-1 and CD57), and
proliferation (Ki-67) on CD4 and CD8 cells. Cell associated viral DNA in
CD4 memory populations was quantified with qPCR.
Results: RC was positively correlated with levels of inflammatory
cytokines in plasma and with activation of both CD8 (p=0.01) and
central memory (CM) CD4 T cells (p=0.002). Low RC was associated
with CD8 T cells that were less exhausted (p< 0.001) and more cytotoxic
(p=0.002). RC was positively correlated with proliferation (p=0.003) and
with the level of cell associated viral DNA in CM CD4 T cells (p=0.01),
a population highlighted to be integral for the maintenance of latency
and preferentially spared in non-pathogenic SIV infection. Consistent
with previous studies, we observed that cellular immune activation,
proliferation, exhaustion, and cell associated viral DNA in CM CD4 T
cells were all associated with the rate of disease progression.
Conclusions: This study highlights the integral role that RC of
the transmitted virus plays in defining several facets of HIV-1
immunopathology. Understanding the complex interactions between
HIV and the immune system will be crucial for designing innovative
prevention strategies.
JP and DC contributed equally to this work
98
Martin J. Deymier1, Zachary Ende1, Daniel T. Claiborne1, William
Kilembe2, Susan Allen1,2, Eric Hunter1,2
Emory University, Atlanta, GA, United States, 2Zambia Emory HIV
Research Program, Lusaka, Zambia
1
Background: In ~80% of heterosexual transmissions of HIV-1, an
infected individual with a diverse viral quasispecies transmits a single
viral variant, the Transmitted/Founder (TF), to a naïve host. Evidence is
building that TF variants are enriched for certain genetic and phenotypic
characteristics that presumably enhance the efficiency of transmission.
However, the mechanisms involved are largely ambiguous, partially
because studies using full-length genomes in transmission pairs are
lacking.
Methods: We have performed HIV near full-length (NFL) single genome
amplification from six subtype C acutely infected individuals and each
of their chronically infected virologically linked partners in the ZambiaEmory HIV Research Project. Phylogenetic analysis performed on the 118
NFL genomes (mean 18/transmission pair) confirms epidemiologically
linked transmission as well as infection by a single viral variant in each
case. We have generated 5 TF & 34 non-transmitted (NT) full-length
infectious molecular clones from 5 transmission pairs and assayed for
particle infectivity by dividing the virus titer on TZM-bl cells by the RT
activity of the virus stock.
Results: The particle infectivity of the TF compared to the median of
the NT variants for all matched transmission pairs was not statistically
significant (p=0.22). However, particle infectivity correlated with the
amount of glycosylation on the Env V1-V4 region (R=0.40, p= 0.01) as
well as with replication in PBMCs for a subset of tested viruses (R=.823
p=0.01), suggesting that previous findings showing less glycosylation
on TF viruses could mean lower replicative capacities in vitro. However,
preliminary data suggests that lower replicating, less glycosylated viruses,
may preferentially productively infect monocyte-derived dendritic cells.
Conclusions: Understanding the characteristics of TF viruses that allow
for efficient transmission will aid in prophylaxis and early intervention
efforts.
OA21.05
OA21.06 LB
Genetic Footprints within the HIV-1 Envelope
Glycoprotein Associated with Transmission in
Men who Have Sex with Men
Cryptic Multiple HIV-1 Infection Revealed by
Early, Frequent, and Deep Sampling during
Acute Infection
Damien C. Tully1, Colin B. Ogilvie1, Rebecca Batorsky1, Karen
A. Power1, Hunter Bedard1, Aaron Seese1, Molly Amero1, Sue
Bazner2, Jake Tinsley3, Niall J. Lennon4, Matthew R. Henn4, Eric
Rosenberg2, Kenneth H. Mayer3, Heiko Jessen5, Marcus Altfeld1,6,
Todd M. Allen1
Gustavo Hernan Kijak1,2, Eric Sanders-Buell1,2, Agnes-Laurance
Chenine1,2, Michael Eller1,2, Nilu Goonetilleke3, Rasmi Thomas1,2,
Sivan Leviyang4, Elizabeth Harbolick1,2, Meera Bose1,2, Phuc Pham1,2,
Celina Oropeza1,2, Kultida Poltavee1,2, Anne Marie O’Sullivan1,2,
Melanie Merbah1,2, Margaret Costanzo1,2, Hui Li5, Will Fischer6, Feng
Gao7, Leigh Anne Eller1,2, Robert J. O’Connell8, Samuel Sinei9, Lucas
Maganga10, Hannah Kibuuka11, Sorachai Nitayaphan8, Morgane
Rolland1,2, Bette Korber6, Francine McCutchan12, George Shaw5,
Nelson Michael1, Merlin Robb1,2, Sodsai Tovanabutra1,2, Jerome Kim1
Background: The global spread of HIV -1 has been fueled by sexual
transmission with the epidemic disproportionately affecting men who
sex with men (MSM). As the epidemic in MSM continues unabated,
understanding the virus-host interactions responsible for transmission
may be critical for the development of an HIV vaccine and other
prevention strategies.
Methods: To elucidate the nature of the transmitted/founder (TF) virus
following rectal transmission, we developed a novel analytical strategy
utilizing deep sequencing data from a cohort of 67 acutely infected
MSM subjects.
Results: Empirical analyses revealed that deep sequencing could not
only reliably infer the TF virus but also discriminate between single
and multiple HIV infections. Using this approach we found that most
transmissions resulted from a single infection with only 16% of
individuals exhibiting evidence of multiple variant transmissions. We
extended this study to identify signature mutations that may be favored
at transmission between viruses originating from heterosexual exposure
versus those from MSM. Here, we focused on a comprehensive analysis
of Env sequences from 125 early subjects (Fiebig I-III) to discern the
genetic imprint on the underlying composition of the viral quasispecies.
A number of genetic signatures were identified in gp120 and the gp41
cytoplasmic tail. One signature pattern specifically enriched in TF viruses
from MSM was the loss of an N-linked glycosylation site at position
362 in the C3 region adjacent to the CD4 binding site. The loss of
this glycosylation motif has previously been associated with chronic
infection and implicated in increased cell-to-cell fusion activity and a
high apoptosis inducing phenotype.
Conclusions: Taken together, these findings provide unique insight into
the events of early transmission in MSM and reveal potentially important
mechanistic differences that may exist between the different routes of
sexual transmission that are not yet fully understood.
U.S. Military HIV Research Program (MHRP), Walter Reed Army Institute
of Research, Silver Spring, MD, United States, 2U.S. Military HIV Research
Program (MHRP)/ Henry M. Jackson Foundation, Silver Spring, MD,
United States, 3School of Medicine, The University of North Carolina at
Chapel Hill, Chapel Hill, NC, United States, 4Department of Mathematics
and Statistics, Georgetown University, Washington, DC, United States,
5
Perelman School of Medicine, University of Pennsylvania, Philadelphia,
PA, United States, 6Theoretical Biology, Los Alamos National Laboratory,
Los Alamos, NM, United States, 7Duke Human Vaccine Institute, Duke
University Medical Center, Durham, NC, United States, 8Armed Forces
Research Institute of Medical Sciences, Bangkok, Thailand, 9Walter Reed
Project, Kericho, Kenya, 10Mbeya Medical Research Programme, Mbeya,
Tanzania, United Republic of, 11Makerere University-Walter Reed Project,
Kampala, Uganda, 12Independent Consultant, Silver Spring, MD, United
States
1
Background: In acute HIV-1 infection (AHI) single genome sequencing
(SGS) revealed a strong bottleneck at transmission, with 60-90% of
sexual infections being established by a single transmitted/founder (T/F)
virus. We combined early and frequent sampling with targeted deep
sequencing (TDS) to study viral evolution during AHI.
Methods: We studied 7 HIV(-) at entry high-risk RV217 volunteers (2
M and 5 F, all with sexual risk) with documented HIV nucleic acid (NA)
conversion after twice-weekly testing. Starting at d2-7 (d0: first NA+
date), we studied 8-9 consecutive plasma samples (mean sampling
interval: 4.1d; peak viremia: d10-18; 1-5 samples were from pre-peak
viremia) by HIV SGS and TDS (Ion Torrent; limit of detection: 0.5%).
Results: 6/7 persons had pre-peak viremia SGS profiles consistent with
infection by a single T/F virus. However, in 4 persons, additional variants
were detected by TDS: in 3 persons at d2-7 (frequency: 0.5-4.3%),
and in one at d21. Viral populations evolved at dramatic rates, but with
different patterns. In #1,the minor variant circulated at < 5% until d17,
then increased to 57% by d31. In #2, the minor variant increased from
3.5% (d7) to 93% (d21), and then decreased to < 0.5% by d42. In #3 the
minor variant was at 0.5-1% between d7-16, then became undetectable,
but was 53% at d181. In participant #4, 2 minor variants were detected
at d21 (0.5-1.4%), increasing by d28 to 16-36%, respectively. Full length
genetic distances between cognate major and minor variants were 1.02.2%, consistent with acquisition of multiple viruses from the same donor.
Inter-variant recombinants were detected from d21 onward. During early
AHI, both major and minor variants acquired CTL-escape mutations.
Conclusions: We show that in apparent single infections minor variants
can occur at levels not detectable by SGS (i.e., cryptic multiple infection).
Furthermore these variants contribute to viral evolution, which may
have profound implications for HIV pathogenesis, cure, treatment, and
vaccine design.
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99
States, 2Massachusetts General Hospital, Infectious Disease, Boston,
MA, United States, 3The Fenway Institute, Fenway Health, Boston, MA,
United States, 4The Broad Institute of MIT and Harvard, Cambridge, MA,
United States, 5HIV Clinic Praxis, Jessen, Berlin, Germany, 6HeinrichPette-Institut, Viral Immunology, Hamburg, Germany
1
Oral Abstract Session 22: Cell and Tissue Models of ARVs for Prevention
OA22.01
OA22.02
Oral Maraviroc and Tenofovir for HIV
Prevention in Women: An Ex vivo and
Translational Approach
Pharmacodynamic Activity in Ectocervical
and Colonic Tissue of Dapivirine, Maraviroc,
and Combination Topical Gels for HIV
Prevention
Melanie R. Nicol1, Heather MA Prince2, Cindi W. Emerson1, Julie
AE Nelson2, Kristine B. Patterson2, Elizabeth J. Geller2, Myron S.
Cohen2, Angela D.M. Kashuba1,2
Charlene Dezzutti1,2, Sarah Yandura2, Lin Wang2, Brid Devlin3,
Jeremy Nuttall3, Lisa C. Rohan1,2
University of North Carolina at Chapel Hill, Eshelman School of
Pharmacy, Chapel Hill, NC, United States, 2University of North Carolina
at Chapel Hill, School of Medicine, Chapel Hill, NC, United States
University of Pittsburgh, Pittsburgh, PA, United States, 2Magee-Womens
Research Institute, Pittsburgh, PA, United States, 3IPM, Silver Springs,
MD, United States
Background: Our previous studies demonstrate the protective
concentration of tenofovir diphosphate (TFVdp) in vaginal explants is
>10-fold higher than in TZM-bl cells. Here we investigate maraviroc’s
(MVC) efficacy in cells and vaginal explants, and determine the explant’s
prediction potential of a dose-challenge study from biopsies of volunteers
given an oral dose of MVC+ tenofovir disoproxil fumurate (TDF).
Methods: TZM-bl cells (n=3) and vaginal explants (n=5 donors) were
incubated 24h in MVC 0.01-500ug/mL prior to challenge with HIV-1 JRCSF. Combination MVC+tenofovir (TFV) was also used in cells to define
the effects of drugs combined. Compared to undosed controls, efficacy
was assessed using a luciferase reporter assay in cells, and spliced
RNA 24-72h post-inoculation in explants. A dose-challenge study was
performed in 6 HIV-, pre-menopausal women administered a single
600mg MVC+600mg TDF dose. 24h post-dose, 4 vaginal+cervical
biopsies were collected for viral challenge and evaluated for infection
in the same manner as explant tissue. HIV protection was defined as
spliced RNA within one standard deviation of background.
Results: In vaginal explants, MVC protective efficacy waned after 24h.
Within 24h, MVC EC50 was 9.7ug/mL, which was >1000-fold higher
than the EC50 in TZM-bl cells (0.006 ug/mL). Additivity of MVC+TFV was
confirmed for HIV protection. The TZM-bl model and the explant model
predicted 100% and 16% efficacy, respectively, 24h after a 600mg
MVC+TDF dose. In the healthy volunteers, protection was observed in
50% (3/6) of vaginal biposies and 67% (4/6) of cervical biopsies, with
50% (3/6) of women having complete protection against HIV challenge.
Conclusions: Similar to TFVdp, cell models overestimated the efficacy
of MVC in vaginal explants. Data from TZM-bl cell monolayers over
predicted, and tissue explants under predicted, efficacy in a healthy
volunteer dose-challenge study. Tissue concentrations at 24h after
single high-dose of MVC+TDF were moderately protective against HIV
infection.
Background: Dapivirine (DPV), a non-nucleoside reverse transcriptase
inhibitor, and maraviroc (MVC), a CCR5 antagonist, were formulated
into aqueous gels to prevent mucosal HIV transmission. We hypothesize
the combination gel will have more potency against HIV infection of
mucosal tissue as compared to either single drug gel.
Methods: Dilutions of 0.05% DPV, 0.1% MVC, and 0.05% DPV/0.1%
MVC gels were evaluated on polarized ectocervical and colonic mucosal
explant cultures exposed to HIV-1BaL. After an overnight culture, the
explants were washed and medium replenished in the basolateral
compartment. Every 3 to 4 days, supernatant was collected and
replenished for up to 21 days. HIV-1 replication was monitored in culture
supernatant by p24 ELISA.
Results: Dilutions of the gels for ectocervical tissue began at 1:20 for
DPV concentrations of ~75900 nM and MVC concentrations of ~97500
nM, while dilutions for colonic tissue began at 1:2000; a 100-fold more
dilute than what was used for the ectocervical tissue. For ectocervical
tissue, 7590 nM of DPV resulted in complete tissue protection while
97500 nM of MVC was partially protective (6 of 8 explants showed no
HIV replication). The combination gel at 7590 nM of DPV/9750 nM
of MVC completely protected the tissue, while 759 nM of DPV/975
nM of MVC was partially protective (6 of 8 explants showed no HIV
replication). For colonic explants, DPV gel diluted to 759 nM completely
protected the tissue, higher dilutions showed no protection. MVC gel
diluted to 975 nM showed no substantial protection of the colonic
tissue. The combination gel diluted to 759 nM of DPV/975 nM of MVC
completely protected the colonic tissue while the 10-fold higher dilution
was partially protective (6 of 8 explants showed no HIV replication).
Conclusions: Combining both drugs in a single formulation demonstrated
modest synergy. Collectively, these data provide a rationale for further
testing of these products as dual compartment microbicides.
1
100
1
OA22.03
OA22.04
Expression, Activity, and Regulation of
Phosphorylating Enzymes in Genital and
Colorectal Tissues and Immune Cells
Transport and Transport Properties of
Tenofovir from Microbicide Gels into Vaginal
Tissue: Analysis Using Raman Spectroscopy
Minlu Hu1,2, Tian Zhou1,2, Charlene S. Dezzutti1,3, Sharon L.
Hillier1,3, Lisa C. Rohan1,2,3
Oranat Chuchuen1, Marcus H. Henderson1, Marinella G. Sandros2,
Angela D.M. Kashuba3,4, David F. Katz1,5
Magee-Womens Research Institute, Pittsburgh, PA, United States,
University of Pittsburgh School of Pharmacy, Department of
Pharmaceutical Sciences, Pittsburgh, PA, United States, 3University of
Pittsburgh, Department of Obstetrics, Gynecology, and Reproductive
Sciences, Pittsburgh, PA, United States
Duke University, Biomedical Engineering, Durham, NC, United
States, 2University of North Carolina at Greensboro, Nanoscience,
Greensboro, NC, United States, 3University of North Carolina at Chapel
Hill, Eshelman School of Pharmacy, Chapel Hill, NC, United States,
4
University of North Carolina School of Medicine, Department of
Infectious Diseases, Chapel Hill, NC, United States, 5Duke University,
Department of Obstetrics and Gynecology, Durham, NC, United States
2
Background: Studies of oral tenofovir (TFV) have revealed that TFV
and tenofovir diphosphate (TFV-DP) levels are 100× higher in colorectal
tissue than in cervical/vaginal tissue after a single oral dose. Multiple
phosphorylating enzymes (PEs) play a role in TFV activation. However,
limited data is available regarding the expression and activity of these
enzymes in the female genital and colorectal tissue relative to immune
cells.
Methods: mRNA expression for 7 PEs (AK2, AK4, NME1, NME2,
CKMT1, CKMT2, CKB) in fresh surgical human tissue samples (cervical
n=6, vaginal n=5, colorectal n=5), a vaginal epithelial cell line (VK2),
and a T cell line (PM1), was evaluated using qRT-PCR. Intracellular
TFV-DP formation was tested in VK2 and PM1 cells with or without
medroxyprogesterone acetate (MPA) and progesterone (P4) using an LCMS/MS method. Differences in TFV-DP conversion were assessed using
Student’s t-test.
Results: Vaginal, ectocervical, and colorectal tissues had similar
expression of PEs except for AK2, which was present at 15-28× higher
levels in colorectal tissue than in ectocervical or vaginal tissues (p<
0.05). The vaginal epithelial cell line was shown to have 10-10,000×
higher expression of CKB, CKMT1, CKMT2, AK2, and AK4 as compared
to levels found for the T cell line (p< 0.05). MPA treatment resulted
in a 3-fold increase in TFV-DP in the epithelial cell line (p< 0.01) and
a 30% decrease in the T cell line (p< 0.01) as compared to controls.
P4 treatment resulted in a nearly 4-fold increased TFV-DP level in the
epithelial cell line (p< 0.01) and no change in the T cell line.
Conclusions: The increased levels of AK2 in colorectal tissue suggest
that AK2 may contribute to the increased levels of TFV-DP observed in
colorectal tissues. The higher level of PEs observed in vaginal epithelial
cell line compared to T cell line, suggests that TFV-DP found in tissues
may be predominantly associated with epithelial cells. The impact of
reproductive hormones on PEs warrants further investigation.
1
Background: Common drug release assays use a liquid sink receptor
compartment. Permeability assays measure net transport through tissue
specimens of varying thickness. These do not give concentration vs.
depth in tissue, nor distinguish drug partitioning at the vehicle-tissue
interface from rate of transport in tissue. We developed a rapid, noncontact method using confocal Raman spectroscopy to measure drug
partitioning and concentration vs. depth in intact tissue layers (epithelium
vs. stroma) and to translate such data to drug diffusion coefficients. We
report here on results for Tenofovir released from its clinical gel.
Methods: Fresh porcine vaginal tissue specimens were treated with
1% Tenofovir gel in a Transwell assay for 2-8 hr at 37 °C. Gel was
applied to either epithelial or stromal tissue surfaces. Results for spatiotemporal concentration profiles were fit to a drug diffusion model to
obtain diffusion coefficients in epithelium and stroma. To determine
partition coefficients, tissue specimens were incubated by submersion in
1% Tenofovir gel and equilibration over 6 h.
Results: Tenofovir concentrations exhibited diffusion-like time- and
depth-dependent distributions in tissue. Diffusion and partition
coefficients in epithelium ranged 7x10-9 - 3x10-8 cm2/s, and 0.5 - 0.8,
respectively. Initial measurements gave ≥ 1 log increase in diffusion
coefficient in stroma. Measurements were referenced to classical
permeability data.
Conclusions: This standardizable label-free method characterizes drug
concentration distributions in tissue and gel vehicles, determining the
fundamental gel-tissue partition coefficient and diffusion coefficients
in gel, epithelium and stroma. Results suggest that the epithelium
presents a potential rate-limiting barrier to Tenofovir permeation across
vaginal mucosa. This is more incisive and pharmacologically useful
information than results of traditional methods that do not distinguish
transport across the two layers; transport parameters can be input to
computational PK models.
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101
1
OA22.05
OA22.06 LB
Mucosal Tissue Explants as Surrogates for in
vivo Efficacy of Microbicides
Predicting Effective Truvada® PrEP Dosing
Strategies With a Novel PK-PD Model
Incorporating Tissue Active Metabolites and
Endogenous Nucleotides (EN)
Carolina Herrera1, Ronald Veazey2, Angela Kashuba3, Javier
García Pérez4, José Alcamí4, Karl Malcolm5, Robin Shattock1
Tulane National Primate Research Center, Tulane, LA, United States,
3
The University of North Carolina, Chapel Hill, NC, United States,
4
Instituto de Salud Carlos III, Madrid, Spain, 5Queen’s University Belfast,
Belfast, United Kingdom
1
2
Background: Validity of the non-human primate (NHP) model is often
questioned due to the lack of correlation with clinical trials in humans.
We hypothesize that in vivo dosing of candidate microbicide conferring
protection against HIV-1/SIV transmission in mucosal sites, can be
predicted with a surrogate model of ex vivo tissue explant cultures,
through intra-tissular drug pharmacological measurements and ex vivo
infection of tissue explants
Methods: Gel-formulated nucleotide reverse transcriptase inhibitor
tenofovir (TFV), and entry inhibitor, maraviroc (MVC), alone or in
combination at fully and partially inhibitory doses were tested. Rhesus
macaque and human cervicovaginal and colorectal tissue explants were
exposed to gels for 1 h followed by addition of virus for 2 h. Wild type
isolates (BaL, YU.2, SIVmac32H and RT-SHIV) and NRTI-escape mutants
(point mutations K65R +/- M184V in YU.2 and SIVmac32H) were used.
Tissue concentrations (PK/PD parameters) of TFV, TFV diphosphorylated
(dp) and MVC were assessed at different time points during 15 days of
culture. Infection was assessed by measurement of p24/p27 in culture
supernatants
Results: In NHP and human explants high tissue drug levels and low rates
of elimination were predictive of drug antiviral efficacy with negative
linear dose-response relationships observed between explant drug
levels and p24/p27 concentrations. Greater potency with combination
gels was seen in NHP than in human tissue. Opposite drug distribution
was observed between both species with higher PK values in colorectal
than in cervicovaginal explants in NHP. Greater loss of viral replication
fitness was seen with SIV RT mutations in NHPs than in human explants
with mutant HIV-1
Conclusions: Ex vivo dose-challenge studies with human and NHP
explants confirmed robustness of the explant model and its potential
as surrogate for in vivo studies refining the prediction of candidate
microbicides efficacy in clinical trials and reducing the number of NHP
euthanized
102
Mackenzie L. Cottrell1, Kuo H. Yang1, Heather M.A. Prince1, Craig
Sykes1, Nicole White1, Stephanie Malone1, Evan S. Dellon2, Ryan
D. Madanick2, Nicholas J. Shaheen2, Julie A. Nelson3, Ronald
Swanstrom3, Kristine B. Patterson2, Angela D.M. Kashuba1
University of North Carolina Eshelman School of Pharmacy, Chapel
Hill, NC, United States, 2University of North Carolina School of Medicine,
Chapel Hill, NC, United States, 3University of North Carolina Center for
AIDS Research, Chapel Hill, NC, United States
1
Background: Failure
of
daily
tenofovir(TFV)
disoproxil
fumarate(TDF)±emtricitabine(FTC) chemoprophylaxis in women has
been attributed to poor adherence. Yet as few as 2 doses/week has been
shown effective in MSM. Here, we provide dosing strategy predictions
for female genital tract and colorectal tissue(RT) utilizing a PK-PD
model that incorporates mucosal tissue TFV diphosphate(dp) and FTC
triphosphate(tp) PK, concentrations of ENs(dATP and dCTP) and their
respective molar ratios that prevent HIV infection.
Methods: TZM-bl cells were treated with 0.03-10µg/ml TFV and 0.0330µM FTC for 24h before HIV-1JR-CSF challenge. TFVdp, FTCtp, dATP and
dCTP were quantified and infection measured by luciferase. R was used
to fit an Emax model of TFVdp:dATP or FTCtp:dCTP vs %inhibition of
infection. 48 women were given a single dose of TDF or FTC at 50,
100 or 200% of the licensed dose. TFVdp, FTCtp, dATP and dCTP were
measured in cervical(CT), vaginal(VT) and RT tissue over 48h using
LC-MS/MS. NONMEM7.3 was used to fit a population PK model and
subsequent monte-carlo simulations.
Results: From TZM-bl cells, the EC90(0.086 for TFV and 0.585 for FTC;p<
0.001) was used as the clinical target. An 8 compartment linear model
best described tissue PK. By 7 days FTC200mg daily achieved ratios
>EC90 for >85, 50 and 75% of the population in VT, CT and RT; and
TDF300mg daily achieved ratios >EC90 for < 50%, < 50% and 100% of
the population in VT, CT and RT. For RT 2 doses/week maintained >EC90
in 100% of the population. TDF+FTC maintains >EC90 for >75 and 50%
of the population in VT and CT with daily dosing and 100% in RT with
2 doses/week.
Conclusions: This study is the first to model TDF/FTC dosing strategies
utilizing only in vitro and mucosal tissue pharmacokinetic data. This
model predicts available clinical trial data, whereby TDF/FTC is ~70%
effective in women with ≥80% adherence and >90% effective in MSM
with ~30% adherence. We believe a-priori utilization of this novel
paradigm can enhance clinical trial design and outcomes.
Oral Abstract Session 23: Pregnancy Intentions, Safe Conception and PMTCT
OA23.SY
OA23.01
PrEP for Women: Indications and Worldwide
Implementation for Women
Evaluation of HIV-1 Neutralizing Antibodies in
Maternal-infant Transmission in Thailand
Erika Aaron1
Lindsay Wieczorek1,2, Brittani Barrows1,2, Agnès-Laurence
Chenine1,2, Martine Braibant3, Kriengkrai Srithanaviboonchai4,
Panita Pathipvanich5, Shelly Krebs1,2, Nelson L. Michael1,6, Sodsai
Tovanabutra1,2, Jerome H. Kim1,6, Merlin Robb1,2, Victoria Polonis1,6
PrEP has been shown to be efficacious in populations of women and
offers a promising female-controlled method of prevention. Women
continue to be a risk of HIV acquisition due to biological, behavioral,
and cultural factors with unacceptable rates of new infection. Currently
recommended sexual HIV-prevention strategies for women include
abstinence, condom use, and treatment as prevention; all require
the willingness of the male partners and none allows for conception.
HIV-affected couples who want to have children face the challenge of
preventing HIV transmission to the sero-negative partner within a serodiscordant couple. While there are reproductive technologies that can
help HIV-affected couples to safely conceive with minimal risk of HIV
transmission to their partner, for most couples such technologies are
neither geographically nor economically accessible. Periconception PrEP
may be a useful adjunct for serodiscordant couples. This presentation
will describe the benefits and potential role of PrEP for women, the
use of periconception PrEP for HIV-serodiscordant couples, and the
need to improve and scale up the implementation of PrEP in the US
and worldwide.
Military HIV Research Program, Bethesda, MD, United States, 2Henry
M Jackson Foundation for the Advancement of Military Medicine,
Bethesda, MD, United States, 3Université François-Rabelais, Tours,
France, 4Chiang Mai University, Chiang Mai, Thailand, 5Lampang
Regional Hospital, Lampang, Thailand, 6Walter Reed Army Institute of
1
Background: The role of neutralizing antibody (NAb) in mother-to-child
transmission (MTCT) of HIV-1 remains unclear. Previous studies suggest
that maternal NAb might reduce HIV-1 transmission. Higher NAb titers
to MBA, a CRF01_AE strain with an unusually long V2 domain, were
found to correlate with lower rates of intrapartum MTCT. However,
findings from different MTCT studies are inconsistent, and further work
is required to clarify the impact of NAb in MTCT.
Methods: In this study, we evaluated NAb breadth and potency in
plasma from 101 HIV+, ART-naïve mothers (22 transmitters and 79 nontransmitters) collected at delivery, and from 51 of their infants
(16 HIV+ and 35 HIV-) collected two months after birth. Pseudovirus (PV)
assays were employed using a panel of six CRF01_AE isolates, including
MBA and RV144 vaccine strains TH023 and CM244. NAb activity is
reported as ID50 titer or positive area under the curve (+AUC), useful for
evaluating samples with low NAb activity.
Results: Contrary to previously published results, maternal geometric
mean NAb titers and +AUC trended higher for transmitters compared
to non-transmitters for five of the six PV tested (including MBA), with
a significant difference observed for CM244 (p=0.047). Maternal NAb
breadth was also increased in transmitters (p=0.047) and directly
correlated with viral load (p=0.037). As expected, infant NAb +AUC was
increased for HIV+ infants compared to those that did not seroconvert
for two pseudoviruses CM244 (p=0.042) and 644039 (p=0.019). The
relationship between mother and infant NAb activity is currently being
evaluated.
Conclusions: Greater magnitude maternal NAb titers were unexpectedly
associated with MTCT transmission of HIV, but correlated with higher
viral load. Further work is required to understand the development,
specificity, and function of NAb in MTCT of HIV.
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103
Drexler University College of Medicine, United States
1
OA23.02
OA23.03
PMTCT Adherence in Pregnant South African
Women: The Role of Depression, Social
Support, Stigma and Structural Barriers to
Care
Barriers and Promoters to Uptake of
Safer Conception Strategies among HIVserodiscordant Couples with Fertility Intention
in Mbarara, Uganda
Christina Psaros1, Nzwakie Mosery2, Jennifer A. Smit2, Faith
Luthuli3, Janna R. Gordon4, Ross Greener3, Kara Bennett5, David
R. Bangsberg6, Steven A. Safren1
Angela Kaida1, Jasmine Kastner1, Courtney Ng2, Naomi Sanyu3,
Adrine Kusasira3, Jerome Kabakyenga3, David R. Bangsberg3,4,
Lynn T. Matthews4
Massachusetts General Hospital / Harvard Medical School, Psychiatry,
Boston, MA, United States, 2MatCH Research (Maternal, Adolescent
and Child Health Research), Obstetrics and Gynecology, Durban,
South Africa, 3MatCH Research (Maternal, Adolescent and Child Health
Research), Durban, South Africa, 4Massachusetts General Hospital,
Psychiatry, Boston, MA, United States, 5Bennett Statistical Consulting,
Inc., Ballston Lake, NY, United States, 6Massachusetts General Hospital
/ Harvard Medical School, Center for Global Health, Boston, MA, United
States
Simon Fraser University, Faculty of Health Sciences, Vancouver, BC,
Canada, 2Massachusetts General Hospital (MGH), Center for Global
Health, Boston, MA, United States, 3Mbarara University of Science
and Technology, Mbarara, Uganda, 4MGH Center for Global Health &
Division of Infectious Disease, Boston, MA, United States
1
Background: Depression is a robust predictor of non-adherence to
antiretroviral therapy, essential in PMTCT. Women in resource-limited
settings are likely to face additional barriers to PMTCT adherence,
including stigma and structural barriers. While structural barriers may
be circumvented by social support; depression and stigma may make
access difficult. Understanding modifiable factors that contribute to
PMTCT adherence can inform interventions.
Methods: 167 HIV-infected women enrolled in PMTCT (median age 28
years) completed an interview at
> 28 weeks of pregnancy assessing depression, stigma, social support
and structural barriers to PMTCT. An adherence score was created
using principal components analysis on the response to four questions
assessing adherence over the past 30 days. Depression was defined as a
Hopkins score > 1.75 and was examined as a predictor of the adherence
score in a linear regression model. Separate linear regression models
also examined relationships between
(1) social support and structural barriers (income and time spent traveling
to clinic) and
(2) depression and stigma as predictors of social support.
Results: Participants with elevated depressive symptoms had
significantly lower adherence scores
(p< 0.01). Neither income (p=0.10) nor time spent traveling to clinic
(p=0.28) predicted adherence; thus, moderation with social support was
not examined. Depression significantly predicted social support (est=0.46 p< 0.01): those with elevated depressive symptoms had a lower
social support score. Similarly, a higher stigma score was significantly
associated with a lower social support score
(est=-0.09, p< 0.01).
Conclusions: While PMTCT programs are effective, adherence to
these services is suboptimal. Depression may play an important role
in adherence to these behaviors. HIV infected pregnant women with
elevated depressive symptoms may also suffer from low social support
and high stigma; interventions targeting these factors may support
maternal and fetal health.
104
1
Background: We investigated barriers and promoters to uptake of a
safer conception approach to pregnancy among HIV sero-discordant
couples in Mbarara, southwestern Uganda.
Methods: We recruited HIV-infected men and women (index) receiving
antiretroviral therapy (ART) from the Uganda Antiretroviral Rural
Treatment Outcomes cohort who reported an uninfected or unknown
status partner (partner), serostatus disclosure to the partner, and personal
or partner desire for a child within 2 years. We conducted 40 separate
in-depth interviews with 20 couples to explore periconception risks and
awareness of specific safer conception strategies. Data were translated,
transcribed, and analyzed using content analysis.
Results: 12/20 index participants were women, with median age of
36 yrs [IQR 29-41], and median recent CD4 of 433 cells/mm3 [IQR:
277-575]. Median partner age was 34yrs [IQR 30-40]. Awareness of
HIV prevention strategies beyond condoms and abstinence was limited,
however, some participants described timed intercourse and ‘ART
as prevention’ as ways to reduce HIV transmission. Participants were
motivated to learn more about safer conception strategies. Key barriers
included limited couple communication about childbearing plans and
understanding of HIV sero-discordance. Fatalism about eventual HIV
acquisition by the uninfected partner or a sense of protection due to
“strong blood” or “God’s will” were common perceptions that decreased
motivation to practice HIV prevention. Many participants prioritized
pregnancy with minimal perceived options for reducing HIV risk. The
more vulnerable partner (HIV-infected and/or female) was often eager
to pursue pregnancy to secure the relationship, regardless of HIV
acquisition or transmission risks.
Conclusions: Awareness of ART for prevention and high interest in
other safer conception strategies presents opportunity to encourage
mutual status disclosure, contravene normative expectations of eventual
seroconversion, and promote strategies to minimize periconception HIV
risks.
OA23.04
“I Would Say it Does Concern Me and on the
Other Hand it Doesn’t.” Perceptions of South
African Learners’ Experiences with Sex,
Pregnancy, and HIV
Cecilia Milford1, Lizzie Moore1, Mags Beksinska1, Muriel Kubeka1,
Kedibone Sithole1, Sibusiso Sibiya1, Faith Smangele Luhthuli1,
Jennifer Smit1
MatCH Research, Department of Obstetrics and Gynaecology,
University of the Witwatersrand, Durban, South Africa
1
Background: HIV/AIDS, sexually transmitted infections (STIs) and
teenage pregnancy are concerns for South Africa’s youth. Adolescent
pregnancy is a major cause of interrupted schooling and drop-out despite
pregnant learners being protected by law. Incomplete education and
early pregnancy are risk factors for HIV acquisition. This study reports
on perceptions of learners’ experiences with sex, pregnancy, and HIV.
Methods: Focus groups were held with male and female learners (n=41,
4 groups), parents (n=19, 2 groups), educators (n=11, 2 groups) and
community members (n=19, 2 groups) recruited through two schools
in eThekwini District, KwaZulu-Natal, South Africa. Discussions were
transcribed, translated and data coded. Results were organised according
to key themes and NVivo used to facilitate data analysis.
Results: Almost half the learners (n=17), aged 16-21, had initiated
sex, most common age of first sex was 15 (n=5). Four learners had
been pregnant. Substance use, transactional sex and low/inconsistent
condom use were the main risk factors for pregnancy and STIs. Although
learners knew about HIV, some were not concerned about it, “there is
something you can use to reduce it”, however stigma was a barrier to
accessing HIV-related services. While teachers discussed HIV with
learners, across groups, most felt that parents should provide advice
on abstinence, protection during sex and monogamy. However some
parents lacked information and others feared discussing HIV with their
children. Teenage pregnancy was reportedly common in schools, mostly
unplanned but some perceived to access government grants. Pregnancy
led to drop-out and gaps in schooling.
Conclusions: Teenage learners are practicing unprotected sex despite
being educated about HIV and pregnancy. Barriers to accessing services
put them at risk. There is a need for improved access to services, better
access to information for parents, and improved relationships with
parents to address gaps and influence behaviour.
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105
Oral Abstract Session 24: Mucosal Responses
OA24.01
OA24.02
Vaccine Induced Responses in a SIV Model
Can Impact Challenge Outcomes
Local HIV-specific IgA Antibody Production in
the Penile Urethra Mucosal Compartment
Megan Wise1, Michele Kutzler1, Natalie Hutnick1, Zina
Moldoveanu2, Meredith Hunter3, Jian Yan4, Bapi Pahar3, Devin
Myles1, Amir Khan4, David Montefiori5, Michael Betts1, Niranjan
Sardesai4, Jiri Mestecky2, Preston Marx3, David Weiner1
Kadryn Kadasia1, Joseph Politch1, Matt Schoen2, Amy Chung2,
Galit Alter2, Deborah Anderson1
University of Pennsylvania, Philadelphia, PA, United States, 2University
of Alabama at Birmingham, Birmingham, AL, United States, 3Tulane
National Primate Research Center, Covington, LA, United States, 4Inovio
Pharmaceuticals, Inc., Blue Bell, PA, United States, 5Duke University,
1
Background: The use of cytokine gene adjuvants to tailor the immune
response is a strength of EP DNA vaccination. This has been established
through the recent HVTN080 trial which demonstrated the potency of
IL-12 with EP delivered DNA to drive T cell responses. However, for an
HIV vaccine it is important to also induce strong humoral responses. To
address the possible role of serum and vaginal IgA in HIV acquisition
we utilized an adjuvanted DNA vaccine previously shown to drive IgA
induction in a non-human primate vaginal challenge model.
Methods: Groups of 5 Indian rhesus macaques received a pSIVmac239
gag/pol and pSIVe660 gp120 alone, or with plasmids - pCCL25 (TECK),
pCCL27 (CTACK) or pCCL28 (MEC), genetic adjuvants, at weeks 0, 6,
12, 18 and 48. Animals were challenged with 500 TCID SIVsmE660
intravaginaly twice a week for two weeks for 4 challenges.
Results: We observed higher vaginal IgA titers in gene adjuvanted
animals compared with DNA vaccine alone. Following challenge, we
observed an overall protection rate of 68% for all vaccinated animals.
However, this protection rate was different for each vaccination regiment.
Animals vaccinated with CCR10Ls, (CCL27 and CCL28,) exhibited robust
control of set point viremia and chronic viremia (p< 0.05) with 89% of
animals controlling infection compared with only 40% in unadjuvanted
animals and 14% in naïve challenge controls. However, CCR9L (CCL25)
vaccinated animals resisted challenge in 60% of animals. Irrespective of
vaccine group, animals that controlled viremia had the highest vaginal
IgA and IgG levels post-vaccination.
Conclusions: Inclusion of immune plasmid adjuvants encoding
mucosal chemokines in EP DNA vaccine regiments can improve
challenge outcomes. Collectively these adjuvant approaches likely have
importance for the development of next generation DNA vaccines and
the data illuminates the need for continued research into the role of
vaginal antibodies and protection from viral infection in NHP models.
106
Boston University, School of Medicine, Boston, MA, United States,
Ragon Institute of MIT, MGH and Harvard, Cambridge, MA, United
States
1
2
Background: Whereas the genital mucosa serves as the first immune
barrier to sexually transmitted pathogens, little is known about the
production and role of antibodies in the human mucosal environment.
Previous studies have shown an abundance of IgG+ and IgA+ plasma cells
associated with penile urethral glands suggesting that this is a site of
local immunoglobulin (Ig) production in men. We compared HIV-specific
Ig isotypes (A, G) and subclasses (G1, G3) in blood plasma (BP), seminal
plasma (SP) and urethral preejaculatory (PE) secretions from HIV-infected
men and controls to determine whether a distinct population of HIVspecific antibodies is present in the genital mucosa during HIV infection.
Methods: A multiplex bead-based Luminex assay incorporating a panel
of seven HIV-specific antigens was used to compare titres, isotypes, and
subclasses of HIV-specific antibodies in PE, SP and BP from nine HIVinfected men and nine healthy controls. We also conducted antibodydependent cell phagocytosis (ADCP) and cytotoxicity (ADCC) assays to
evaluate antibody function in the different compartments.
Results: There was an abundance of HIV-specific IgG in all three fluids,
with similar titres, subclasses and specificity profiles. Similarly, ADCC
and ADCP activity did not differ between the three sample types. In
contrast, a subset of PE samples from HIV-infected men had significantly
higher (5 - 50X) anti-gp41 IgA titres than found in SP and BP, providing
evidence for local urethral production of HIV-specific IgA during HIV
infection.
Conclusions: IgG HIV antibody profiles in male genital secretions reflect
those in the systemic circulation, whereas IgA HIV antibodies may be
locally produced in the urethral mucosa. More research is needed on
strategies to elicit urethral antibody production with vaccines, and the
function of IgA antibodies in HIV prevention.
OA24.03
OA24.04
Targeting Mucosal Fc-Fc Receptor Interactions
as Vaccine Strategy against Mucosal HIVtransmission
DNA and Protein Co-immunization Improves
the Magnitude, Longevity, and mucosal
Dissemination of Immune Responses
Magdalena Sips1,2, Marina Krykbaeva1, Brittany Bowman1,
Thomas Diefenbach1, Douglas Kwon1, Peter Brouckaert2, Galit
Alter1
George N. Pavlakis1, Jinyao Li1, Antonio Valentin1, Rashmi Jalah1,
Vainav Patel1, Margherita Rosati1, Viraj Kulkarni1, Candido
Alicea1, Diego A. Vargas-Inchaustegui2, Yongjun Guan3, David
Venzon4, Niranjan Sardesai5, Timothy R. Fouts6, Abraham Pinter7,
Marjorie Robert-Guroff2, David C. Montefiori8, Xiaoying Shen8,
Georgia D. Tomaras8, Barbara K. Felber1
States, 2Ghent University, Ghent, Belgium
1
National Cancer Institute at Frederick, Frederick, MD, United States,
National Cancer Institute, NIH, Bethesda, MD, United States, 3Institute
of Human Virology, Baltimore, MD, United States, 4National Cancer
Institute, NIH, Biostatics and Data Management Section, Bethesda, MD,
United States, 5Inovio Pharmaceuticals, Inc., Blue Bell, PA, United States,
6
Profectus Biosciences, Inc., Baltimore, MD, United States, 7Public
Health Research Institute, University of Medicine and Dentistry of New
Jersey, Newark, NJ, United States, 8Duke University Medical Center,
Durham, MD, United States
1
Background: Determining the quality and longevity of vaccine-induced
immune responses is essential for improving the prospects of AIDS
vaccines. DNA and protein (inactivated viral particles) co-immunization
regimen induced systemic and mucosal Ab responses, which correlated
with slower virus acquisition upon challenge, and potent T cell responses
providing protection against chronic viremia. We are evaluating different
regimens of DNA&protein vaccines using purified SIV or HIV-1 Env to
further dissect humoral and cellular responses including magnitude,
breadth and mucosal dissemination.
Methods: Macaques were vaccinated using DNA&protein coimmunization regimen in the presence of IL-12 DNA, coinjected into the
same muscle. Humoral and cellular responses were monitored in blood
and different tissues.
Results: Co-immunization strategy of DNA&protein induced rapid and
high humoral responses while maintaining robust cellular responses
typically obtained with DNA vaccines. The vaccine induced Ab against
both homologous and heterologous Env; high binding titers against
scaffolded V1/V2 env region; efficient dissemination to mucosal sites;
high Env-specific IgG in saliva and Env-specific IgG and IgA in rectal
mucosa. Analysis of cellular responses revealed the presence of cytotoxic
memory T cells against several viral proteins. These cellular responses
disseminated systemically as demonstrated by their presence not only
in blood and lymphoid tissues, but also in bone marrow, liver, lung
(effector site) and, importantly, rectal and vaginal mucosa. The longevity
of the cellular responses induced by this co-immunization regimen was
significantly improved, with SIV-specific T cells detected >5 yrs after the
vaccination.
Conclusions: Intramuscular DNA&protein co-delivery increases the
magnitude and longevity of systemic and mucosal humoral immune
responses in immunized macaques and is proposed as a practical and
efficient method for human vaccination.
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107
Background: To ultimately end the AIDS pandemic, effective means of
preventing transmission by its principal genital and rectal routes must
be developed. Non-neutralizing antibody (nNab) responses, which may
be easier to induce than broadly neutralizing antibodies via vaccination,
have shown some protection, pointing toward a role for nNab functions,
including Fc-Fc receptors (FcRs) interactions. The protective efficacy of
Fc-effector function is critically dependent on innate immune cells;
therefore we sought to define cell repertoires expressing FcRs in the
female genital tract (FGT) and intestinal tract (IT).
Methods: Fixed tissue sections (rectum, vagina, cervix, uterus, lymph
node) were stained for natural killer (NK) cells, macrophages, neutrophils
and Fc receptors, FcγR: I, II, III and FcαR. HIV+ biopsy samples from IT
and lymph nodes were examined for changes in FcR repertoires. Flow
cytometric evaluation of FcR+ cells was performed on freshly isolated
cells from enzymatically digested colon and cervical tissues.
Results: NK cells were very infrequent in IT and FGT. Moreover, none
of the FcR was found on NK cells, suggesting that mucosal NK cells
have limited capacity to mediate antibody-driven-effector functions. In
contrast, macrophages and neutrophils were present at much higher
frequencies in IT and FGT and were detected close to epithelial layers.
They robustly expressed FcRs with macrophages expressing FcγRI,
FcγRII, FcαR but not FcγRIII and neutrophils expressing FcγRIII and FcαR.
Changes in expression pattern in HIV-infected subjects suggest specific
antibody therapeutic opportunities for harnessing Fc-FcR interactions.
Conclusions: The ability of nNabs to provide protection at mucosal
barriers is centrally linked to FcR+ cells available within vulnerable
tissues. Surprisingly, our data suggest that non-ADCC mediated
mechanisms, such as phagocytosis and neutrophil activation, are more
abundant and potentially represent important mechanisms by which
vaccine induced nNabs may offer protection.
2
OA24.05
OA24.06 LB
Antibody Isotypes Differ in their Capacity to
Bind, Capture and Aggregate HIV-1 Virions
Transcriptional Signatures of Viral Control in
HIV-1 Infected South African Women
Sandra G. Okala1, Deborah F. King1, Paul M. Rogers1, Robin J.
Shattock1
Nikki L. Gentle1,2, Sarah Djebali3, Neil A. Martinson4, David
Spencer5, Roderic Guigo3,6, Caroline T. Tiemessen1,2
Imperial College London, Medicine, London, United Kingdom
1
Background: Recently, Env-specific monomeric (m)IgA was correlated
with increased risk of HIV-1 infection in the RV144 vaccine trial and
inhibited IgG effector functions. In contrast, mucosal dimeric (d)IgA has
been associated with resistance to infection in highly exposed uninfected
individuals, and viral aggregation by Env specific antibodies in mucosal
secretions has been often proposed as an alternative mechanism to
block HIV-1 infection at the portal of viral entry. The current study was
designed to determine whether anti-HIV-1 IgG1, mIgA2 and dIgA2 with
the same epitope specificity differ in their ability to bind, capture and
aggregate HIV-1 BaL.
Methods: Bio-Layer interferometry was used to measure kinetics
parameters of the different forms of b12, CH31, 2F5 and 7B2 mAbs to
soluble HIV-1 BaL gp140 Env. Virus capture by the panel of mAbs was
quantified by ELISA and antibody mediated viral aggregation (AMVA)
was determined using Nanoparticle Tracking Analysis.
Results: Interestingly, IgGs captured more virions than both mIgAs
and dIgAs with the same epitope specificity. Although capture did not
correlate with binding affinity (Kd), data indicates that virions capture
correlated with association rate constant (kon). No relationships was
found between binding affinity and AMVA, however a significant
correlation was observed between the number of binding sites and the
proportion of aggregates, highlighting improved aggregation capacity of
dimeric mAbs (P= 0.0108). Further, the data demonstrated that AMVA
was dependent on epitope accessibility with a classical prozone effect.
Conclusions: Overall, our findings indicate that antibody isotype
influences effector functions, with greater capacity of IgGs to capture
HIV-1 particles and Env specific dIgAs to induce viral aggregation. Thus,
the ability of an antibody-based vaccine to prevent HIV-1 infection may
be dependent on the isotype of the antibody, and the effector functions
most relevant to the biological compartment.
108
University of the Witwatersrand, Faculty of Health Sciences,
Johannesburg, South Africa, 2National Institute for Communicable
Diseases, Centre for HIV and STIs, Johannesburg, South Africa,
3
Centre for Genomic Regulation, Bioinformatics and Genomics
Group, Barcelona, Spain, 4University of the Witwatersrand, Perinatal
HIV Research Unit, Johannesburg, South Africa, 5Right to Care,
Johannesburg, South Africa, 6Universitat Pompeu Fabra, Barcelona,
Spain
1
Background: The characterization of host genetic factors that allow
small subsets of individuals to naturally suppress HIV-1 viral replication
in the absence of antiretroviral treatment remains a priority, as an
understanding of the immune mechanisms employed by these
individuals to control viral replication could provide insights into
potential therapeutic targets.
Methods: Therefore, in order to identify genes involved in the control
of HIV-1 viral replication during chronic infection, mRNA was extracted
from peripheral blood mononuclear cells isolated from 14 Black female
South African HIV-1 controllers. Paired-end RNA sequencing of 100 bp
fragments was performed using the HiSeq 2000 and the data obtained
were processed using the GRAPE analysis pipeline. Differences in gene
expression were evaluated using the R/Bioconductor package, DESeq;
with genes exhibiting at least a 2-fold difference in expression and
p-values of p< 0.001 considered to be differentially expressed.
Results: Genes found to be differentially expressed between the eight
individuals with viral loads below 400 copies/ml and six individuals with
viral loads above 400 copies/ml included several previously implicated
in the control of chronic viral infections. These include LAG3, a regulator
of T cell responsiveness, and genes involved in the regulation of the
interferon type I antiviral response (for example IFI6, IFIT3, IFI44 and
OASL). Several of the identified genes also encoded as yet functionally
uncharacterized large intergenic non-coding RNAs, whose impact on
HIV-1 viral control remains to be evaluated.
Conclusions: Collectively, these data describe a transcriptional signature
of immune activation in chronic HIV-1 infection, which suggests the
involvement of innate immune mechanisms previously shown to display
substantial sex-based differences. However, whether these mechanisms
directly regulate HIV-1 viral replication, or rather are activated in
response to increased viral loads, remains to be established.
Oral Abstract Session 25: Adjuvants and Immunogens
OA25.01
Adjuvant Dependent Mucosal V2 Responses
and RAS Activation in Vaccine Induced
Protection from
SIVmac251 Acquisition
RAS, a signal transducer that facilitates cross talk among B-cells, T-cells
and antigen presenting cells, was demonstrated to be a biomarker of
vaccine efficacy in the alum group.
Conclusions: These data highlight the importance of the quality of
the mucosal antibodies to V2 in protection and suggest that activation
of RAS may constitute a novel approach to improve vaccine efficacy
against HIV.
Monica Vaccari1, Shari N. Gordon1, Slim Fourati2, Luca
Schifanella1,3, Mark Cameron2, Brandon F. Keele4, Xiaoying
Shen5, Georgia Tomaras5, Erik Billings6, Mangala Rao6,
Namal P.M. Liyanage1, Diego A. Vargas-Inchaustegui7, Steve
Whitney8, Melvin N. Doster1, Nicolo Binello1, Poonam Pegu1,6,
David C. Montefiori9, Kathryn Foulds10, David S. Quinn10, Mitzi
Donaldson10, Frank Liang10, Karin Lore10, Mario Roederer10,
Richard Koup10, Adrian McDermott10, Zhong-Min Ma11, Miller
Christopher11, Tran B. Phan12, Donald N. Forthal12, Matthew
Blackburn1, Francesca Caccuri1, Guido Ferrari9, Marjorie RobertGuroff7, Silvia Ratto-Kim6, Jerome Kim6, Nelson Michael6, Sanjay
Phogat13, Susan W. Barnett14, James Tartaglia13, David Venzon15,
Donald M. Stablein16, Rafick-Pierre Sekaly2, Genoveffa Franchini1
1
National Cancer Institute, Bethesda, MD, United States, 2Vaccine &
Gene Therapy Institute, Port St. Lucie, FL, United States, 3Universita
degli Studi di Milano, Milan, Italy, 4National Cancer Institute SAIC,
AIDS and Cancer Virus Program, Frederick, MD, United States, 5Duke
Human Vaccine Institute, Durham, NC, United States, 6Walter Reed
Army Institution, U.S. Military HIV Research Program, Silver Spring,
MD, United States, 7National Cancer Institute, Immune Biology of
Retroviral Infection Section, Bethesda, MD, United States, 8ABL,
Rockville, MD, United States, 9Duke University, Durham, NC, United
States, 10Vaccine Research Center, Bethesda, MD, United States,
11
California National Primate Research Center, Davis, CA, United
States, 12University of California, Irvine School of Medicine, Irvine,
CA, United States, 13Sanofi Pasteur, Swiftwater, IL, United States,
14
Novartis Vaccines and Diagnostics Inc., Cambridge, MA, United
States, 15National Cancer Institute, Biostatistics and Data Management
Section, Bethesda, MD, United States, 16The EMMS Corporation,
Rockville, MD, United States
Background: The RV144 HIV vaccine trial resulted in limited, but
significant protection, from HIV acquisition. Serum antibodies directed
to the Env variable regions 1 and 2 (V1/V2) inversely correlated
with the risk of HIV-1 infection and sieve analysis demonstrated
immunologic pressure on two regions of the V2. Prior macaque studies
demonstrated that a similar vaccine for SIV (ALVAC-SIV) induced
protection from SIVmac251 acquisition in a low dose neonatal challenge
model, but not in adult high dose challenge models.
Methods: We challenged ALVAC-SIV/gp120/alum vaccinated adult
macaques intrarectally with repeated low doses of SIVmac251 in a study
powered to allow benchmarking against the results of RV144. An
additional arm of the study evaluated these vaccines together with the
oil-in-water emulsion MF59 adjuvant.
Results: We found that alum protected macaques from SIVmac251
acquisition while MF59 did not despite its ability to elicit higher
systemic T-cell and antibodies responses. MF59 altered homing of
antibody producing cells and increased the frequency of CXCR3+
plasmablasts in blood that positively correlated with anti-envelope
IgA serum levels and phagocytosis. Alum, in contrast, increased the
frequency of plasmablasts expressing the mucosal integrin a4b7 that
positively correlated with IgA responses to cyclic V2 in rectal mucosa.
In the alum group mucosal IgG to cyclic V2 correlated with lower risk
of SIVmac251 acquisition. However, mucosal IgG to linear and cyclic V2
correlated with an increased risk of SIVmac251 acquisition in the MF59
group. The two adjuvants modulated distinct signaling pathways and
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OA25.02
OA25.03
IgG Glycosylation Is Programmed and
Remembered after Immunization with TLR
Stimulating Adjuvants
CD4-induced Epitopes Are Exposed on
Cell-bound HIV-1: The Key to Fc Receptor
Mediated Humoral Immunity?
Alison E. Mahan1, Hamid Mattoo2, Kendall Dionne1, Jacquelynne
Tedesco1, Shiv Pillai2, Galit Alter1
Meron Mengistu1, George K. Lewis1, Anthony L. DeVico1
States, 2Harvard Medical School, Center for Cancer Research,
Charlestown, MA, United States
1
Background: The N-linked glycan in the CH2 domain of IgG potently
affects antibody effector functionality by mediating its affinity for
innate immune cell receptors. In particular, glycosylation is important
for antibody dependent cellular cytotoxicity (ADCC) and complement
deposition, as well as general inflammation. Vaccine correlates analyses
support an important role for effector functions, especially ADCC, in
protection from HIV infection. Therefore, understanding how to program
IgG-glycans through vaccination may prove to be of critical importance.
Methods: Mice were immunized with NP-antigen formulated with a
variety of adjuvants, including TLR4, TLR5 and TLR7/8 agonists mixed
with alum. At 28, 48 and 60 days post immunization, transcriptional
profiles of antigen-specific B cells were analyzed using microarray and
targeted qPCR. In addition, glycosylation of antigen-specific IgGs was
analyzed by capillary electrophoresis.
Results: We observed that animals that received TLR7/8 agonist adjuvants
produced more inflammatory IgG-glycans and this corresponded to
transcriptional levels of important glycosylation enzymes, particularly
the galactosylation enzyme B4GALT1. Additionally, the effector function
modifying sugar fucose and its enzyme transcript, FUT8, were decreased
by the TLR5 agonist adjuvant, suggesting that IgG effector functionality
can be tuned by specific signals during immunization. Importantly, by
boosting some animals at 28 days with antigen alone, we observed
that these glycosylation profiles are remembered since these animals
maintained the same IgG glycan profile with or without repeated
administration of adjuvant.
Conclusions: These data are the first to provide evidence that
glycosylation can be tuned and remembered after immunization with
TLR agonist adjuvants. This is important for understanding how HIV
vaccines can be designed to elicit antibodies with good effector function
glycan profiles in antigen-specific IgGs.
110
Medicine, Baltimore, MD, United States
1
Background: The partially successful RV144 vaccine trial produced
non-neutralizing antibodies that mediate ADCC against HIV-1. These
antibodies recognize the CD4-induced (CD4i) C1 region of gp120.
However, such findings are enigmatic in view of previous arguments that
CD4i epitopes are hidden on viral trimers before, and during interaction
with host cell. It is therefore critical to understand the antigenicity of
these conserved epitopes that become exposed during productive
viral replication, to provide important guidance for designing antiviral
strategies such as vaccines to raise protective antibody responses.
Methods: We studied the antigenicity of conserved epitopes by
visualizing their exposure on single HIVJRFL virus particles as they interact
with target TZM-bl cells using confocal microscopy. Epitope exposure
was probed by visualizing the binding of neutralizing antibodies b12
and 2G12, and CD4i antibodies A32, 17b and C11.
To reconcile the question of CD4i antigenicity, we examined the location
of CD4i antibody attachment with 20nm precision using superresolution
microscopy.
Results: CD4i antibodies can access their cognate epitopes on
TZM-bl cell-bound HIVJRFL virions. Patterns of exposure varied with
antibody; however, in general the level of CD4i exposure was similar
to neutralizing epitopes b12 and 2G12. CD4i epitopes appear distal to
the virus-cell contact site, where they can be accessed by antibodies
involved in ADCC.
Conclusions: The patterns of CD4i epitope exposure are consistent
with the ADCC activities of cognate antibodies against bound virions.
CD4i antibodies were able to gain access to their targets due to the
unexpected epitope exposure on gp120, distal to the site of contact with
cell surface CD4. These findings indicate that HIV-1 exhibits a diversity
of epitope exposure upon attachment that may provide unique insights
for understanding how humoral immunity impacts HIV infection.
OA25.04
OA25.05
Structural Evolution of HIV-1 gp120 Glycan
Recognition by the PGT121 Lineage of Potent
Broadly Neutralizing Antibodies
Refocussing Antibody Responses by Chemical
Modification of Vaccine Antigens
2
1
3
Dept of Integrative Structural & Computational Biol, The Scripps
Research Institute, IAVI Neutralizing Antibody Center, Scripps CHAVIID, La Jolla, CA, United States, 2IAVI Neutralizing Antibody Center,
Scripps CHAVI-ID, Dept Immunology & Microbial Science, The Scripps
Research Institute, La Jolla, CA, United States, 3Dept of Cell and
Molecular Biology, The Scripps Research Institute, La Jolla, CA, United
States, 4Dept of Integrative Structural & Computational Biol, The Scripps
Research Institute, La Jolla, CA, United States, 5Dept of Microbiology &
Immunology, Weill Medical College of Cornell University, New York,
NY, United States, 6Dept of Cell and Molecular Biology, The Scripps
Reaserch Institute, La Jolla, CA, United States, 7IAVI Neutralizing
Antibody Center, Scripps CHAVI-ID, Dept Immunology & Microbial
Science, The Scripps Research Institute, Ragon Institute of MGH, MIT
and Harvard, Cambridge, MA 02139, La Jolla, CA, United States, 8Dept
of Integrative Structural & Computational Biol, The Scripps Research
Institute, IAVI Neutralizing Antibody Center, Scripps CHAVI-ID, Skaggs
Institute for Chemical Biology, La Jolla, CA, United States
1
Background: The HIV-1 envelope glycoprotein trimer is the sole
target of the neutralizing antibody response and, therefore, the
prime platform for vaccine design. The PGT121 lineage of antibodies
(PGT121-124; PGT133-134) exhibits exceptionally potent and broad
neutralizing activity against HIV-1 at serum concentrations achievable
by vaccination. Recently, the crystal structure of PGT122 in complex
with BG505 SOSIP.664 gp140 provided a view of how an affinitymatured antibody from the PGT121 family recognizes gp120. PGT124,
a bnAb from the same family, appears to represent an alternative branch
in the antibody maturation process. Therefore, we have an opportunity
to investigate the different ways in which high affinity recognition of a
complex epitope involving glycans and protein surfaces can evolve from
the same germline precursor.
Methods: Towards this end, we used X-ray crystallography and EM to
unveil the molecular details of the PGT124-gp120 interaction, and deep
sequencing, virus neutralization, calorimetry (ITC), and glycan arrays to
add additional functional and biochemical insights.
Results: The crystal structure of PGT124 in complex with the gp120 at
3.3Å resolution reveals a novel mode of recognition involving primarily
the glycan at position N332 and a “GDIR” protein motif at the V3 loop
base. We also discovered that residues on PGT124 that recognize
this epitope are conserved throughout the PGT121 lineage and may
represent the initial requirement for selection and subsequent evolution
of this antibody family. Interestingly, residues on PGT122 that recognize
additional protein regions and N-linked glycans appear to evolve
separately and are conserved only on one branch of the maturation tree.
Conclusions: The PGT121 family shows divergent glycan recognition
profiles during antibody maturation, presumably in response to virus
escape and sequence evolution. This information should enable the
design of better scaffolds and variants of native-like SOSIP trimers
leading to a successful vaccine.
Torben Schiffner1, Karolis Leonavicius2, Heiko Schuster2, Helen
J. Kim3, Leopold Kong1,3, Regis Saliba2, Florian Brod1, Frank
Wegmann1, Po-Ssu Huang4, Guillaume B. Stewart-Jones5,
William R. Schief3,4,6, Andrew B. Ward3, John P. Moore7, Rogier W.
Sanders7,8, Benjamin G. Davis2, Quentin J. Sattentau1
University of Oxford, Sir William Dunn School of Pathology, Oxford,
United Kingdom, 2University of Oxford, Department of Chemistry,
Oxford, United Kingdom, 3IAVI Neutralizing Antibody Center at The
Scripps Research Institute, San Diego, CA, United States, 4University of
Washington, Washington, WA, United States, 5University of Oxford, The
Weatherall Institute of Molecular Medicine, Oxford, United Kingdom,
6
States, 7Weill Cornell Medical College, New York, NY, United States,
8
University of Amsterdam, Amsterdam, Netherlands
1
Background: The HIV-1 envelope glycoprotein (Env) has developed
several immune-evasion mechanisms to avoid the induction of
neutralizing antibodies, including immuno-dominant non-neutralizing
epitopes, conformational flexibility of conserved epitopes, and
spontaneous subunit dissociation. Here, site-specific immuno-silencing
by glycan masking and chemical fixation of native-like Env trimers are
explored to overcome these obstacles.
Methods: Immunogens, including “next-generation“ BG505 SOSIP.664
trimers, were chemically modified in vitro by either site-directed glycan
addition or chemical cross-linking, and effects on antibody recognition
were characterized in vitro by ELISA and SPR. Immunogenicity was
tested in mice and refocussing of antibody responses was analysed by
cross-competition with monoclonal antibodies.
Results: In vitro, glycan masking led to selective reduction in binding of
antibodies recognizing epitopes containing, or proximal to, modification
sites (lysine residues) whereas binding of antibodies to epitopes devoid
of lysines remained unchanged. Chemical cross-linking of native-like
gp140 trimers led to reduced binding of non-neutralizing antibodies
including V3-loop directed antibodies, which further significantly
increased the existing differential in antibody binding between
neutralizing and non-neutralizing antibodies.
Immunization with modified antigens led to reduced immunogenicity
of “silenced” epitopes compared to unmodified controls. In contrast,
some epitopes that were unaffected by chemical modification showed
significantly increased cross-competition with sera from immunized
animals.
Conclusions: The chemical modifications developed here resulted
in improved antibody recognition profiles in vitro and led to selective
refocussing of antibody responses in vivo. Thus, chemical modification
of vaccine antigens to stabilize antigen and refocus B cell responses
presents an attractive tool for vaccine immunogen design.
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111
Fernando Garces , Devin Sok , Leopold Kong , Ryan McBride ,
Helen J. Kim4, Karen F. Saye-Francisco2, Jean-Philippe Julien1,
Yuanzi Hua4, Albert Cupo5, John P. Moore5, James C. Paulson6,
Andrew B. Ward1, Dennis R. Burton7, Ian A. Wilson8
1
OA25.06 LB
Native-like BG505 SOSIP.664 Trimers Induce
Autologous Tier-2 NAbs against Complex
Epitopes in Rabbits and Macaques
John P Moore1
Weill Medical College, New York, NY, United States
1
Background: Our goal is to induce bNAbs by using soluble, recombinant
gp140 mimics of the native Env trimer. At present, the only way to make
true trimer mimics is to ensure that the gp140 proteins are cleaved
between the gp120 and gp41 subunits, and then stabilized via SOSIP
mutations. Our team has described the production and properties of
native-like BG505 SOSIP.664 trimers, including their high-resolution
X-ray and cryo-EM structures.
Methods: As no native-like, soluble recombinant trimer has previously
been evaluated as an immunogen, we tested the BG505 SOSIP.664
trimers in rabbits and macaques. NAb titers were quantified using the
TZM-bl cell assay, and their specificity assessed by several techniques,
including the use of competitor (“dump-in”) antigens.
Results: The BG505 SOSIP.664 trimers induce strong and consistent
NAb responses to the autologous Tier-2 BG505.T332N virus (20/20
rabbits, 3/4 macaques), as well as heterologous Tier-1 NAb responses.
Cross-reactive Tier-2 NAbs were not induced. Comparator, non-native
BG505 Env proteins performed much worse, exemplified by an
uncleaved gp140 that induced autologous Tier-2 NAbs in 0/4 rabbits.
The Tier-1 and Tier-2 NAb titers were not correlated among the animals,
implying that they are entirely different responses. A variety of “dumpin” competitors were used to characterize the NAb epitopes. Tier-1, but
not Tier-2, NAbs were substantially depleted by simple V3 peptides. The
most prevalent Tier-2 response was influenced by C3 sequences present
in escape variants that emerged in the BG505 HIV-1-infected infant. In
one rabbit the key Tier-2 NAb epitope was a composite of the V1/V2/
V3 regions.
Conclusions: In infected people, bNAbs evolve from a narrow-specificity
autologous Tier-2 response. To induce the Tier-2 breadth necessary for
an effective vaccine, we will explore several approaches including the
use of new, native-like SOSIP.664 trimers based on subtype B and C
sequences. GMP-grade BG505 trimers are now being developed for
human trials.
112
Oral Abstract Session 26: Microbicides and Multipurpose Prevention Technologies
OA26.01
OA26.02
Introduction of the SILCS Diaphragm as a
Multipurpose Technology in South Africa:
Potential Users, Perceived Benefits, and
Barriers to Use
A Simple Intravaginal Ring Pump for
Sustained Vaginal Release of ARV
Microparticles and Macromolecular Agents
Ryan Teller1,2, Rachna Rastogi2, Patrick Kiser1
Cecilia Milford1, Letitia Rambally1, Muriel Kubeka1, Lizzie Moore1,
Mags Beksinska1, Maggie Kilbourne-Brook2, Jennifer Smit1
MatCH Research, Department of Obstetrics and Gynaecology,
University of the Witwatersrand, Durban, South Africa, 2PATH, Seattle,
WA, United States
Northwestern University, Evanston, IL, United States, 2University of
Utah, Salt Lake City, UT, United States
1
Background: SILCS, a single-size diaphragm designed to improve
women’s options for non-hormonal barrier contraception, is being
evaluated as a reusable delivery system for microbicide gel, allowing
SILCS to be a multipurpose prevention technology (MPT). In the context
of high unplanned pregnancies and HIV prevalence in South Africa (SA),
a health systems assessment was conducted to identify opportunities
and challenges for future introduction of SILCS as a MPT in SA. Potential
future users, perceived benefits, and barriers to use of SILCS were
identified and described.
Methods: Key informant interviews were held nationally (with
regulatory authority, policymakers, healthcare providers, trainers,
trialists, advocacy groups; n=31) and 3 focus groups were held with
potential users (1 male, 2 female; n=24) at a primary healthcare clinic
in eThekwini District, KwaZulu-Natal. Discussions were transcribed and
translated. Data were coded, results organised according to key themes,
and NVivo used to facilitate data analysis.
Results: Respondents described the advantage of SILCS as a MPT―“it
protects from so many things.” It was seen as a woman-initiated method
which can be inserted covertly when condom negotiation is difficult.
Participants described multiple potential user types―those who want
to avoid hormonal contraceptives, find it difficult to access health care,
educated/mature women, not afraid to insert vaginal products and
interested in birth spacing. Barriers to use included misunderstandings
of vaginal anatomy, fit (“Will it not disappear here inside?”), concerns
about semen spillage, potentially painful insertion, and partner response
during sex. There were questions about SILCS care, durability, and use
in multiple rounds of sex.
Conclusions: Potential SILCS users include a variety of women, but
uptake may be based on individual preferences and needs. Most barriers
to use can be addressed via healthcare provider training and education
of potential users and their partners.
Background: Intravaginal ring technology is generally limited to
releasing low molecular weight species that can diffuse through the IVR
elastomer. To increase the diversity of the drugs that can be delivered
from IVR, we sought to create a controlled delivery technology that
could uncouple the mechanism of drug release from the interaction of
the antiretroviral with the elastomer and provide near zero order release
of any stable molecule. We designed an IVR pump that contains pellets
of a mixture of micronized drug and hydroxypropyl cellulose (HPC) that
are contained in the hollow core of the IVR. In this system orifices control
the hydration rate of the pellet and flux of the drug-containing HPC gel
that is forced through the orifice by polymer swelling and which then
distributes in the vaginal canal.
Methods: Each IVR-pump contained ARV/HPC pellets within polyether
urethane tubing containing orifices that were sealed by induction
welding. We formulated several of the leading PrEP ARV: TDF,
tenofovir, maraviroc, and IQP-0528 and several macromolecules. We
evaluated IQP-0528 IVR-pumps in sheep for drug release rate, and drug
concentration and distribution in the vaginal canal.
Results: Altering the type of swelling polymer, drug loading, orifice
design and the mass of pellets can control the drug release rate and
duration. Formulations for high molecular weight species like IgG and
linear polymers could be obtained that were nearly zero order for a
month. Less controlled release profiles were achieved with more watersoluble drugs including maraviroc, tenofovir and TDF since the drugs
possess much higher diffusivity in the hydrated polymer matrix. We
observed mg per day release rates in sheep with an average IQP-0528
concentration in vaginal fluid of 270 µg/mL (~105x the IC90).
Conclusions: This device provides an adaptable platform for vaginal
drug delivery. We have been able to deliver impressive quantities of
hydrophobic drugs as microparticles and high molecular weight agents
for a month in animal models.
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113
1
OA26.03
OA26.04
Development of Reservoir Vaginal Rings
Containing Dapivirine or Hormonal
Contraceptive Steroids
Engineering a “Virus Trap and Safety Net”
Microbicide
Stella E. Aniagyei1, Jill M. Steinbach1
Clare McCoy , Peter Boyd , Susan Fetherston , Ian Major ,
Diarmaid Murphy1, Andrew Brimer3, Jonathon Holt3, Brid Devlin3,
Wendy Blanda3, Tiffany Derrick3, Chris Gimour3, Jeremy Nuttall3,
Karl Malcolm1
1
1
2
1
Kingdom, 2University of Ulster, School of Pharmacy and Pharmaceutical
Sciences, Coleraine, United Kingdom, 3International Partnership for
Microbicides, Silver Spring, MD, United States
1
Background: By extending the use duration of vaginal rings (VRs), the
overall production cost per patient per month can be greatly reduced.
Matrix-type ring designs produce an initial burst of drug release, and
concentrations during the burst must be maintained within safe limits.
This constrains both the maximum drug load and, in turn, the use
duration of a matrix ring. By contrast, reservoir-type VRs comprise a
drug-loaded reservoir surrounded by a rate-controlling non-medicated
sheath, such that initial burst release is minimised and constant daily
release rates are achieved. In this way, greater drug loadings and longer
use periods are possible. Here, we report the development of reservoir
VRs formulations containing dapivirine (DPV) and various contraceptive
steroids.
Methods: Reservoir VRs were manufactured with injection molded cores
containing ethinyl estradiol (EE), etonogestrel (ETO), levonorgestrel (LNG)
or DPV. Addition- and condensation-cured silicone cores were tested.
All cores were overmolded with a blank 1.5 mm addition-cured silicone
sheath. In-vitro release was assessed for up to 60 days.
Results: VRs comprising addition cured silicone cores and blank addition
cured silicone sheath layers provided zero order release of DPV (95 µg/
day) and LNG (30 µg/day), but not EE (detectable release on days 2 to
10 only) or ETO (no detectable release at any time). Rings comprising
a condensation-cured silicone core provided zero order release of DPV
(119 µg/day), EE (328 µg/day, days 4 to 18), ETO
(422 µg/day, days 4 to 18) and LNG (69 µg/day).
Conclusions: Reservoir-type VRs offering zero order release kinetics
have potential as a multipurpose prevention technology (MPT) product
for contraception and HIV-prevention with use duration of several
months. However, the contraceptive hormones require careful selection
of the silicone polymer system to ensure adequate release, while DPV is
readily released from both types tested.
114
University of Louisville, Bioengineering, Louisville, KY, United States
1
Background: Multipurpose drug delivery systems (MDDS) have the
potential to impart a variety of attributes to microbicide delivery. We
have previously developed a safe and effective microbicide, using
poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) that encapsulate
short interfering (siRNA) targeting host receptors, to significantly increase
survival in a murine model of HSV-2 infection; and have more recently
developed surface-modified NPs that enhance cell internalization by
70%. Building upon these abilities in our new lab, we seek to create a
proof-of-concept MDDS that integrates some of the protective attributes
of mucus, including structure and pathogen adhesion, to target the
integral stages of HIV-1 and HSV-2 infection. To achieve this, our MDDS
incorporates 3 components:
(1) a biological lectin, Griffithsin (GRFT), to immobilize and debilitate
HSV-2 and HIV upon entry (“trapping”); and “safety nets” of:
(2) polymeric electrospun fibers (EFs) and
(3) NPs to provide prolonged release of encapsulated antiviral drugs and
siRNA, respectively.
Methods: EFs were electrospun with different solvents and compositions,
containing our model small molecule antiviral, Acyclovir (ACV). A
portion of EFs were surface modified with our “trapping” lectin, GRFT.
Virus assays were conducted to test the efficacy of ACV EFs and blank
GRFT-EFs against HSV-2 infection in vitro.
Results: EFs with a range of diameters were produced, depending on
solvent selection and electrospinning conditions. We achieved high
ACV encapsulation, with cumulative release spanning 20-60%. EFs
encapsulating ACV inhibited HSV-2 infection by >90%, while GRFT-EFs
immobilized HSV-2. We are currently expanding this platform to inhibit
both HSV-2 and HIV infection.
Conclusions: We have complemented our siRNA NP and surfacemodified NP microbicide platform, to create novel unmodified and
GRFT-EFs for use in a MDDS. We are excited to further develop and
combine these technologies to eventually capture, prevent, and treat
STIs in one platform.
OA26.05
OA26.06 LB
A Single Dose Novel Formulation of Maraviroc
Using Electrospun Fabrics Designed for Instant
and Multi-day HIV Prevention
Mass Spectrometry Imaging of Hair
Strands Allows for Evaluation of Long Term
Antiretroviral Adherence
Cameron Ball1, Shih-Feng Chou1, Yonghou Jiang1, Kim A.
Woodrow1
Corbin Thompson1, Elias Rosen2, Mark Bokhart2, Heather Prince1,
Craig Sykes1, David C. Muddiman2, Angela D.M. Kashuba1
Background: Current microbicide formulations fail to provide both
instant and multi-day sustained ARV delivery with a single dose. Such
products could be particularly useful for delivering maraviroc (MVC),
which is required at high doses for efficacy but absorbed rapidly in the
vagina. To address this need, we electrospun a composite fabric that
provides both a rapid burst and a multi-day sustained release of MVC.
Fully tunable sustained release is achieved via novel core-shell fiber
architectures with high drug loading, and layering with rapid-release
fibers allows independent control over the ratio of burst to sustained
release.
Methods: MVC was formulated into coaxially electrospun fibers with
core-shell nano-architectures for sustained delivery. Release of MVC was
measured under sink conditions with multiple release media. We probed
the mechanism for sustained release from core-shell fibers by measuring
material physical and chemical properties, hydration, and mass loss.
Finally, core-shell fabrics were combined with rapidly dissolving MVC
fibers using either alcohol welding or physical pressure to create soft,
single dose fabrics for both rapid and sustained MVC release.
Results: Core-shell fibers were loaded with up to 37.5 wt% MVC and
provided near linear release for 5 days. Adjusting the drug loading and
the ratio of shell-to-core content in fibers provided direct control over the
dose and timing of drug release, respectively. The primary mechanism
for sustained drug delivery from core-shell fibers, unlike core-shell IVRs,
was controlled surface wetting. Combining rapid and sustained delivery
fabrics into single doses provided full control over both pericoital (20
min) and sustained (5 days) release.
Conclusions: These electrospun MVC composites are the first
microbicides to realize fully adjustable biphasic release with a single
dose. Translatability to other drugs and ease of manufacturing scale
up make this microbicide platform suitable for simultaneous rapid and
sustained multipurpose prevention.
University of North Carolina at Chapel Hill, Chapel Hill, NC, United
States, 2North Carolina State University, Raleigh, NC, United States
1
Background: Successful pre-exposure prophylaxis requires antiretroviral
(ARV) adherence. Therapeutic drug monitoring provides accurate, but
short term adherence data. Hair strand (HS) analysis with traditional LC/
MS provides long term adherence information, but fails to discriminate
ARV presence at the time of HIV exposure. Mass spectrometry imaging
(MSI) offers the ability to visualize ARV exposure in tissues, allowing for
identification of distinct distribution patterns. Here, as proof of concept,
we use MSI to visualize tenofovir (TFV), emtricitabine (FTC), and efavirenz
(EFV) in HS from HIV positive subjects.
Methods: HS (20-30) were taken from 5 HIV positive, virologically
suppressed subjects receiving Atripla® for > 1 year. HS incubated in
TFV, FTC, and EFV for 24 hours or HS never exposed to ARVs served
as positive and negative controls, respectively. After collection, HS were
analyzed using an infrared matrix-assisted laser desorption electrospray
ionization (IR-MALDESI) source coupled to a Thermo Q-Exactive mass
spectrometer. Signal intensity between HS from a single subject was
compared to assess intra-subject variability. MSI data were analyzed
using MSiReader software.
Results: MSI experiments demonstrate continuous EFV signal along HS
for all 5 subjects. MS/MS analysis and the use of a negative control
confirmed EFV specificity. Variability in signal intensity was similar
within (~ 2.5 fold) and between (~ 4 fold) subjects. TFV and FTC were
not visualized in HS from dosed subjects, though FTC and EFV were both
detected in the positive control sample.
Conclusions: EFV distribution in HS demonstrates consistent ARV
exposure in these subjects, and is in agreement with their suppressed
viral loads. Confirmation of EFV signal with MS/MS and low intra-subject
variability showcase the accuracy and precision of this method. We show
for the first time that MSI of HS is a promising approach for measuring
ARV adherence. Future studies will examine ARV hair distribution
patterns in non-suppressed patients.
www.hivr4p.org
115
University of Washington, Bioengineering, Seattle, WA, United States
1
Oral Abstract Session 27: PrEP: Self-testing, Safety and Modeling
OA27.01
OA27.02
Selection of Rilpivirine Resistant HIV-1 in
a Seroconverter on Long-acting Rilpivirine
(TMC278LA) from the Lowest Dose Arm of the
SSAT 040 Trial
Uptake of HIV Self-testing among People
Receiving PrEP in Kenya
Kerri J. Penrose1, Urvi M. Parikh1, Kristen A. Hamanishi1,
Constantinos Panousis1, Laura Else2, David Back2, Marta Boffito3,
Akil Jackson3, John W. Mellors1
University of Pittsburgh, Pittsburgh, PA, United States, 2University
of Liverpool, Liverpool, United Kingdom, 3SSAT Research, Chelsea &
Westminster Hospital, London, United Kingdom
1
Background: Rilpivirine (RPV) is a potent second-generation NNRTI
whose injectable long-acting formulation (TMC278LA) is a candidate for
PrEP. A participant in the lowest dose arm of a single dose TMC278LA
PK study unexpectedly seroconverted 84 days (d) post-injection. We
evaluated plasma samples for residual RPV and HIV-1 drug resistance
after seroconversion.
Methods: SSAT 040 evaluated plasma PK of TMC278LA following a
single IM dose of 300, 600 or 1200mg in 60 low-risk HIV-negative
women. RPV plasma concentrations were assessed post-injection at
regular intervals. Plasma HIV-1 RNA levels (VL) and drug resistance
by standard genotype and allele-specific PCR were determined
retrospectively in the participant who seroconverted. Recombinant
infectious virus containing full-length RT sequences from plasma was
tested for drug susceptibility in TZM-bl cells.
Results: HIV-1 infection occurred in 1 of 20 participants in the 300mg
arm. Post-seroconversion testing illustrates selection of K101E after
infection: d84 (seroconversion) VL 175,060 copies/ml, < 0.1% K101E,
7.5ng/ml plasma RPV; d115 (TDF/FTC/DRV/r initiation) VL 664,925
copies/ml, 19% K101E, 4.1ng/ml plasma RPV; d151 VL 6,204 copies/
ml, K101EK mixture, 13.8ng/ml plasma RPV; d199 VL 3,558 copies/
ml, < 0.1% K101E, 6.0 ng/ml plasma RPV. Plasma-derived HIV-1 clones
(d115) containing K101E had 4-fold decrease in susceptibility to RPV,
compared to d115 clones without K101E, and exhibited cross-resistance
to ETR (4-fold), NVP (8-fold), and EFV (4-fold).
Conclusions: A 300mg single dose of TMC278LA did not prevent HIV1 infection in one participant in the SSAT 040 trial, but sustained low
levels of RPV (less than the protein adjusted IC90) post-infection selected
RPV resistant HIV-1. This is a unique instance of well-documented
infection with wild-type HIV-1 and subsequent selection of resistant
virus by persistent drug exposure post-infection. Evaluation of different
dose regimens of TMC278LA is ongoing to optimize drug levels for its
use as a PrEP agent.
116
Kenneth Ngure1,2, Renee Heffron2, Nelly Mugo2,3, Elizabeth
Irungu4, Njambi Njuguna4, Lawrence Mwaniki4, Kerry
Thompson5, Connie Celum2,6,7, Jared M. Baeten2,6,7
Jomo Kenyatta University of Agriculture and Technology, Nairobi,
Kenya, 2University of Washington, Global Health, Seattle, WA, United
States, 3Kenya Medical Research Institute, Center for Clinical Research,
Nairobi, Kenya, 4Partners in Health Research and Development,
Thika, Kenya, 5University of Washington, Seattle, WA, United States,
6
University of Washington, Epidemiology, Seattle, WA, United States,
7
University of Washington, Medicine, Seattle, WA, United States
1
Background: Implementation of pre-exposure prophylaxis (PrEP) by
HIV uninfected people will require routine HIV testing to minimize
PrEP use during acute HIV infection and the potential development
of antiretroviral resistance. HIV self-testing could provide an efficient
method to increase testing frequency with minimal increase in cost and
participant burden.
Methods: In the Partners Demonstration Project, an open-label study of
PrEP for HIV prevention among high risk HIV discordant couples, HIV
testing occurs and PrEP prescriptions are filled at quarterly clinic visits. In
this sub-study at the Thika, Kenya site, HIV self-testing training and kits
are offered to HIV uninfected participants using PrEP for use during the
two months between each quarterly clinic visit.
Results: As of April 2014, 120 HIV uninfected partners (93 females and
27 males) have enrolled in the HIV self-testing sub-study. The median
age is 30 years (interquartile range, IQR: 26-35) and number of years
of education is 10 (IQR: 8-12). Among the 58 participants with at least
one follow-up visit to date, 89.7% reported conducting HIV self-testing
at least once and 92.3% of these reported that using the self-test kit
was easy or very easy.17.3% of participants reported that they selftested when they opened a new bottle of PrEP but the majority (78.9%)
reported that self-testing did not coincide with a specific event. The
majority (57.7%) of participants reported testing alone while 38.5% of
participants reported that their study partner was with them. More than
half (59.5%) of participants reported sharing the self-test results with the
study partner while 36.5% did not share the results with anyone. All
self-test results were HIV negative.
Conclusions: Within a PrEP demonstration project conducted in a
peri-urban African setting, HIV uninfected partners successfully
used HIV self-testing with ease. Self-testing may be a feasible, cost
effective, adjunctive method to increase the frequency of HIV testing
in conjunction with PrEP use.
OA27.03
OA27.04
The Better it Is, the More it Will Be Used - The
Synergistic Relationship between Product
Efficacy, Level of Uptake and Overall Impact
Calcutta HIV Model Projections and the
Impact of PrEP
1
2
London School of Hygiene & Tropical Medicine, Global Health and
Development, London, United Kingdom, 2Wits Reproductive Health and
HIV Institute (WRHI), Johannesburg, South Africa, 3University of Bristol,
School of Social and Community Medicine, Bristol, United Kingdom
1
Background: Mathematical models show the important role product
efficacy, user uptake and adherence play in the impact of new HIV
prevention technologies (NPTs). However, these models rely on uptake
assumptions that at best are based on expert opinion, levels of use in
trials or of existing technologies, and none are able to consider how
uptake may be implicitly related to how ‘good’ the product is.
Methods: This study uses predicted product uptake from a choice
experiment (CE) in South Africa to model microbicide effectiveness:
average population protection provided by microbicides with different
efficacies. It incorporates differential uptake of products with different
efficacies and the degree to which condom users say they will switch to
the microbicide. We compare with projections assuming 30% uniform
uptake among non-condom users only.
Results: For a population of women with 20% condom use at baseline,
we predict introduction of a moderately effective microbicide (55% ) will
only increase the population protection provided over an average sex
act from 17% to 23%, with 16% and 11% uptake among women who
do not and do use condoms, respectively. For 60% condom use average
protection is 53%, only 1.5% higher than without microbicides. The
uniform uptake model predicts 30% average protection for a moderately
effective microbicide at low condom use and 58% protection for the
60% condom use scenario. If the product is 95% efficacious, added
microbicide protection is 31% at low condom use versus 17% at high
condom use. Here the uniform uptake model predicts 23% and 11%
added protection, respectively.
Conclusions: Microbicide impact assuming uniform uptake is likely to be
overestimated for moderately effective products while underestimated
for highly effective products. Ignoring this differential effect is likely
to lead to biased models and inefficient allocation of resources. In the
absence of observed uptake, this study proposes the use of hypothetical
CE data to strengthen our impact models.
Zindoga Mukandavire1, Kate Mitchell1, Smarajit Jana2, Peter
Vickerman1,3
London School of Hygiene and Tropical Medicine, Social and
Mathematical Epidemiology Group, London, United Kingdom,
2
Sonagachi Research & Training Institute, Kolkata, India, 3University
of Bristol, School of Social and Community Medicine, Bristol, United
Kingdom
1
Background: Despite low national HIV prevalence in India, HIV
prevalence remains high among high risk groups. We assessed the
potential impact of pre-exposure prophylaxis (PrEP) use for HIV
prevention among female sex workers (FSWs) in Calcutta, India.
Methods: A mathematical model was developed to simulate the
transmission of HIV among FSWs and their commercial clients in
Calcutta. We parameterised the model using data from Calcutta, with
any data gaps being filled with data from Bangalore or Mumbai. The
model was fit to FSW HIV prevalence from Calcutta for 1992 to 2010.
The model was analysed to estimate the efficiency and impact of PrEP
among FSWs assuming PrEP use starts in 2014. We considered different
efficacy and coverage scenarios for PrEP.
Results: Our model projections suggest that with existing interventions,
FSW HIV prevalence is on the decline in Calcutta. The different PrEP
intervention scenarios suggest that PrEP use could result in rapid and
substantial decreases in HIV prevalence and incidence compared to the
baseline scenario of no PrEP, with a 60% reduction in HIV incidence
achievable over 20 years with a PrEP efficacy of 60% and coverage of
60%. However, the efficiency of PrEP is low with about 40 years of PrEP
being required per life year gained in the most optimistic PrEP scenario
(60% coverage and efficacy).
Conclusions: PrEP interventions could be important for controlling HIV
in FSW driven epidemics in Calcutta and India in general, but may not
be cost-effective in many scenarios.
www.hivr4p.org
117
Fern Terris-Prestholt , Matt Quaife , Sinead Delany-Moretlwe ,
Helen Rees2, Charlotte Watts1, Peter Vickerman1,3
1
OA27.05
OA27.06 LB
The Potential Impact of Long-acting PrEP
on HIV Transmission, Mortality and Drug
Resistance in KwaZulu-Natal, South Africa:
Model-based Analyses
A Phase 1 Open Label Safety, Acceptability,
Pharmacokinetic, and Pharmacodynamic
Study of Intramuscular TMC278 LA (the
MWRI-01 Study)
Robert Glaubius1, Greg Hood2, Ume Abbas1
Ian McGowan1, Aaron Siegel2, Kathy Duffill2, Cory Shetler2,
Charlene Dezzutti1, Nicola Richardson-Harman3, Kaleab Abebe1,
David Back4, Laura Else4, Amy Herrick5, Peter Williams6, Khaja K
Rehman2, Ross D. Cranston1
Cleveland Clinic, Infectious Disease and Quantitative Health Sciences,
Cleveland, OH, United States, 2Pittsburgh Supercomputing Center,
Pittsburgh, PA, United States
1
University of Pittsburgh School of Medicine, Pittsburgh, PA, United
States, 2Magee Women Research Institute, Pittsburgh, PA, United States,
3
Alpha StatConsult LLC, Damascus, MD, United States, 4University
of Liverpool, Liverpool, United Kingdom, 5University of Pittsburgh
Graduate School of Public Health, Pittsburgh, PA, United States,
6
Janssen Research and Development, Beerse, Belgium
1
Background: Long-acting injectable rilpivirine (RPV) pre-exposure
prophylaxis (PrEP) could be pivotal for optimizing PrEP effectiveness for
HIV prevention. The impact of RPV PrEP on HIV transmission, mortality
and the spread of drug resistance are unknown.
Methods: We examine the scale-up of RPV PrEP, ART and male medical
circumcision (MMC) using a mathematical model of the HIV epidemic
in KwaZulu-Natal (KZN). We compare a baseline scenario of ART and
MMC scaled up to 80% coverage by 2017 to three scenarios of ART and
MMC plus ten years of PrEP (90% efficacy; 35% cross-resistance) rollout
starting in 2015:
a) 10% overall coverage of susceptible adults (20% of KZN’s HIV budget
at $165 per person-year of PrEP), either unprioritized or
b) prioritized to women aged 20-29; or
c) 80% coverage of high-risk adults (< 2% of the HIV budget).
Outcomes include infections averted and drug resistance prevalence over
ten years, and lifetime life-years (LY) lived, cost and cost-effectiveness.
We classify scenarios as very cost-effective if the incremental cost per
LY saved is less than South Africa’s per capita GDP of $7,500, and costsaving if costs decrease while life-years increase.
Results: Unprioritized PrEP averts 6.8% of infections over ten years
and saves 1.9 million LY in the PrEP-exposed cohort ($605/LY saved).
Age-prioritization improves PrEP’s impact, averting 12.6% of infections
and saving 3.5 million LY ($113/LY saved). Risk-prioritized PrEP reduces
costs by 0.7%; it averts more infections (8.4%) and saves more LY (2.3
million) than unprioritized PrEP while using
< 10% as many person-years of PrEP. Drug resistance decreases with
unprioritized or age-prioritized PrEP (1%; 1.7%), and increases by 2.5%
with risk-prioritized PrEP compared to baseline
(358,000 cases at 2025).
Conclusions: RPV PrEP is potentially very cost-effective, and may be
cost-saving when prioritized to high-risk persons. Drug resistance from
PrEP is limited compared to ART, though risk-prioritized PrEP may
increase overall resistance.
118
Background: Long acting (LA) injectable antiretroviral agents are
being evaluated as a potential strategy for pre-exposure prophylaxis
of HIV infection. This study was undertaken to characterize the safety,
acceptability, pharmacokinetic (PK), and pharmacodynamic (PD) profile
of TMC278 LA.
Methods: Healthy HIV-1 seronegative participants were enrolled into 2
cohorts. 12 women and 6 men received an intramuscular dose of either
1200 mg (Cohort 1; N=18) or 600 mg (Cohort 2: N=18) of TMC278
LA. Plasma and tissue (rectal [RT], cervical [CT], and vaginal [VT]) were
collected before and after exposure to TMC278 LA. Participants were
followed for 4 months after receiving TMC278 LA. Safety, acceptability,
multicompartmental PK, and PD (ex-vivo explant challenge with HIV1BaL) were monitored throughout the study.
Results: Thirty six participants were enrolled. There were 183 adverse
events of which 179 (97.8%) were Grade 1/2. Two Grade 3 and 2 Grade
4 events were unrelated to study product. The mean acceptability score
was 7 out of a maximum of 10. Geometric mean (GM; 90% CI) plasma
TMC278 concentrations at Day 28 post-dose for the 1200 mg and 600
mg cohorts were 53 (38-67) ng/mL (female) / 43 (23-63) ng/mL (male)
and 28 (19-37) ng/mL (female) / 17 (9-24) ng/mL (male) respectively. For
the 1200 mg dose the GM tissue: plasma ratios at Day 28 were 0.68
(VT), 0.77 (CT), 1.21 (female RT), and 1.27 (male RT). Exposure to TMC278
LA was associated with significant suppression of viral replication in RT
(p < 0.0001) that persisted for up to four months following exposure to
TMC278 LA, but viral suppression was not seen in VT or CT.
Conclusions: Single dose administration of TMC278 LA was safe and
well tolerated with dose dependent plasma PK exposure. The TMC278
tissue: plasma ratio for RT was approximately two-fold higher than VT
or CT. This is the first study to demonstrate prolonged suppression of
explant viral replication following systemic antiretroviral exposure.
Multiple dosing studies of TMC278 LA are planned.
Oral Abstract Session 28: Treating and Preventing: the Role of ARVs
OA28.01
OA28.02
High Initiation and Adherence among High
Risk African HIV Discordant Couples in a
Demonstration Project of PrEP and ART for
HIV Prevention
Evaluation of a Risk Score Tool to Identify
Higher-risk HIV-1 Serodiscordant Couples for
Antiretroviral-based HIV-1 Prevention
University of Washington, Global Health, Seattle, WA, United
States, 2Kenya Medical Research Institute, Nairobi, Kenya, 3Makerere
University, Kampala, Uganda, 4Kabwohe Clinical Research Center
(KCRC), Kabwohe, Uganda, 5Jomo Kenyatta University of Agriculture
and Technology, Nairobi, Kenya, 6Fred Hutchinson Cancer Research
Center, Seattle, WA, United States, 7Massachusetts General Hospital,
1
Background: Pre-exposure prophylaxis (PrEP) and antiretroviral therapy
(ART) have high efficacy for HIV prevention among heterosexual
African HIV discordant couples. Assessing initiation and adherence to
antiretroviral-based HIV prevention strategies outside of clinical trial
settings is a priority.
Methods: Enrollment of high-risk HIV discordant couples into an openlabel PrEP and ART demonstration project in Kenya and Uganda began
in November 2012. Without PrEP and ART, anticipated HIV incidence
in this cohort would be ≥5% per year. ART is offered per national
guidelines and PrEP is offered as a ‘bridge’ to ART, provided between
study enrollment and the time when the HIV-infected partner is likely
to have achieved viral suppression. ART adherence is assessed through
biannual HIV RNA measurements and MEMS caps are used to assess
PrEP adherence.
Results: As of April 2014, 714 high risk couples were enrolled, 67% with
an HIV infected woman, and 42% with an HIV infected partner eligible
to initiate ART. At enrollment, 96% of HIV uninfected participants
initiated PrEP and 92% of ART-eligible participants planned to start ART.
To date, 51% of couples have had ≥6 months of expected follow-up
and retention through 6 months is 88% for HIV uninfected and 93%
for infected participants. Among HIV uninfected participants on PrEP,
77% took ≥80% of expected PrEP doses by MEMS between enrollment
and their 6 month clinic visit. Travel and relationship dissolution are
common reasons for missing PrEP doses, reported at 27% and 18% of
visits with missed doses. Among HIV infected participants initiating ART
at enrollment, 85% had a plasma HIV RNA concentration < 400 copies/
ml by the 6 month visit. Six months after enrollment, 85% of couples
were using PrEP and/or ART: 45% PrEP only, 14% ART only, and 26%
both PrEP and ART.
Conclusions: PrEP and ART initiation and early adherence are high
among high-risk African HIV discordant couples participating in an
open-label demonstration project of PrEP and ART for HIV prevention.
1
Kenyatta National Hospital, Nairobi, Kenya, 2University of Washington,
Seattle, WA, United States, 3Kenya Medical Research Institute, Nairobi,
Kenya, 4Jomo Kenyatta University of Agriculture and Technology, Ruiru,
Kenya, 5Makerere University, Kampala, Uganda, 6Kabwohe Clinical
Research Center (KCRC), Kabwohe, Uganda, 7Fred Hutchinson Cancer
Research Center, Seattle, WA, United States
Background: Antiretroviral therapy (ART) and pre-exposure prophylaxis
(PrEP) significantly reduce HIV transmission within heterosexual HIV
serodiscordant couples. To maximize impact and minimize costs, ART
and PrEP interventions should be offered to couples at highest risk of
HIV transmission.
Methods: The Partners Demonstration Project is an open-label,
prospective cohort study of PrEP and ART for HIV prevention among high
risk HIV serodiscordant couples in Kenya and Uganda. We evaluated the
feasibility of using a validated empiric risk score (Kahle et al JAIDS 2013)
that uses a combination of variables (age, number of children, male
circumcision status, plasma HIV levels, condom use, marital status) to
identify couples at highest risk for HIV transmission.
Results: Since November 2012, 1217 couples have screened and 714
enrolled. Of the screened couples, 274 (23%) scored 0-4 (anticipated
HIV incidence of ≤ 2% per year), 493 (41%) scored 5-6 (anticipated
HIV incidence of ~5% per year) and 450 (37%) scored ≥ 7 (anticipated
HIV incidence of ≥7% per year). Couples scoring ≥5 were eligible for
enrollment and 76% entered the study. The median age of the HIV
uninfected partner was 29.5 [IQR 26, 36] years and the HIV seronegative
partner was male in 67% of partnerships. Over half (57%) had no
children, 92% had unprotected sex in the month prior to screening and
34% of HIV susceptible men were uncircumcised. Among HIV infected
members of the couples, the median CD4 count was 436 [IQR 274,634]
cells/mm3 and 41.3% had a plasma viral load >50,000 copies/ml.
Conclusions: An empiric risk score derived from observational analyses
of African HIV serodiscordant couples identified higher risk HIV
discordant couples for a demonstration project of PrEP and ART for HIV
prevention. Three-quarters of the higher risk couples who were eligible
have enrolled. Risk scores could be useful for recruitment of higher risk
persons and couples into future HIV prevention trials and demonstration
projects.
www.hivr4p.org
119
Renee Heffron1, Connie Celum1, Nelly Mugo2, Elly Katabira3,
Elizabeth Bukusi2, Stephen Asiimwe4, Kenneth Ngure5, Nulu
Bulya3, Josephine Odoyo2, Edna Tindimwebwa4, Deborah
Donnell6, Jessica E. Haberer7, Lara Kidoguchi1, Jennifer Morton1,
Jared M. Baeten1, for the Partners Demonstration Project Team
Elizabeth M. Irungu1, Renee Heffron2, Nelly Mugo2,3, Kenneth
Ngure2,4, Elly Katabira5, Nulu Bulya5, Elizabeth Bukusi2,3,
Josephine Odoyo3, Stephen Asiimwe6, Edna Tindimwebwa6,
Deborah Donnell2,7, Connie Celum2, Jared M. Baeten2, Partners
Demonstration Project Team
OA28.03
OA28.04
Facing the Realities of HIV Universal Test and
Treat: Early Lessons Learnt from the Delivery
of the HPTN071/ PopART Study Intervention
in Zambia
HIV Transmission in Discordant Couples in
Non Research Settings: Rwanda Experience
Kwame Shanaube1, Sam Griffith2, Musonda Simwinga1, Ephraim
Sakala1, Chepela Ngulube1, Sandra Simbeza1, Martin Mutonyi1,
Sarah Fidler3, Richard Hayes4, Helen Ayles1,5, HPTN071-PopART
study Team
1
ZAMBART Project, Lusaka, Zambia, 2FHI 360, Science Facilitation
Department, Washington, NC, United States, 3Imperial College,
Department of Infectious Disease Epidemiology, London, United
Kingdom, 4London School of Hygiene & Tropical Medicine, Department
of Infectious Disease Epidemiology, London, United Kingdom, 5London
School of Hygiene & Tropical Medicine, Department of Clinical
Research, London, United Kingdom
Background: The HPTN071 (PopART) is a 5-year community randomized
study of a combination HIV prevention package in 21 communities in
Zambia and South Africa. HPTN071 is one of the first studies to evaluate
a combined HIV prevention package (including universal HIV testing
and ART for all HIV+ individuals irrespective of CD4 count (UTT)) on HIV
incidence at community-level.
Methods: 21 communities were randomly assigned to one of three study
arms. The study intervention consists of door to door voluntary UTT
with the offer of immediate ART to all individuals who test positive, plus
other HIV preventive methods. Community HIV care providers (CHiPs)
are responsible for the delivery of the intervention and linkage to care.
Multi-disciplinary groups have been closely engaged in designing and
developing the study protocol and include community representatives,
community advisory boards, policy makers, government, ministries of
health, care and service providers, funders and research teams.
Results: Trial initiation was preceded by two years of preparatory work.
Early rapid formative research to identify local catalysts and barriers
that could influence intervention proved invaluable, as have regular
door to door and community mobilization activities. Study preparation
required strengthening existing health care systems and improvement
of supply chain management. Hiring and training of approximately 430
community based staff to deliver the intervention was done. Mechanisms
to best deliver some clinic activities-routinely and in optimal time- are
still being identified. Regular feedback on progress and challenges at all
levels through sharing of process data is useful. Synchronization with
other ongoing HIV preventive initiatives in these communities has been
occurring.
Conclusions: Initiating a complex combination-prevention trial at a
population level, that is integrated within current DoH facilities and
therefore sustainable, requires multi-disciplinary approaches, time and
effort.
120
Etienne Karita1, Rachel Parker2, Sabin Nsanzimana3, Placidie
Mugwaneza3, Roger Bayingana1, Robertine Sinabamenye1, Gisele
Umvirigihozo1, Nuri Ahmed1, Kristin Wall4, Amanda Tichacek2,
Eric Hunter2, Susan Allen2
Project San Francisco, Kigali, Rwanda, 2Emory University, School of
Medicine, Atlanta, GA, United States, 3Rwanda Biomedical Center,
Kigali, Rwanda, 4Emory University, Rollins School of Public Health,
Atlanta, GA, United States
1
Background: Most new HIV infections in sub-Saharan Africa occur
through heterosexual transmission in stable couples. The HPTN 052 trial
provided evidence that antiretroviral treatment (ART) significantly reduces
the risk of transmission among discordant couples (DC). However, little
is known of its effectiveness in “the real world”. The objective of our
study was to measure the risk of HIV transmission among DC followed
in public health centers (PHC) in Kigali, Rwanda.
Methods: The Rwandan Ministry of Health has established Couples
Voluntary Counseling and Testing (CVCT) for HIV as a national policy
in antenatal care. Since November 2011, Project San Francisco (PSF) is
providing mentorship to 20 high volume PHC in Kigali for the follow-up
of DC. In this program, HIV-infected partners are offered ART, and HIVnegative partners are offered risk reduction counseling and repeat HIV
testing on a quarterly basis.
Results: From November 2011 to March 2014, 3025 DC were registered
in the 20 PHC. Of these, the index partner in 1367 (45%) were on ARV.
1827 (60%) couples (1070 M+F- and 757 M-F+) had at least one followup visit, of whom 940 were on ARV at enrollment and 467 initiated
ART during follow-up. During follow-up, 39 previously HIV- women and
10 men seroconverted. Viral linkage analysis was performed on 16 out
of the 20 transmission pairs in whom the index partner was on ART,
and 81% were linked. The overall HIV incidence rate was 0.71 per 100
person-years (95% CI: 0.41, 1.13) when the index partner was on ARV,
and 5.33 (95% CI: 3.54, 7.70) when the index partner was off ARV.
Conclusions: Our program data show that in the “real world” if CVCT
and routine follow-up are well established and ART can be offered, ART
is associated with an 87% reduction in the risk of HIV transmission from
discordant partnerships. However, as transmission from treated index
partners is being observed, ongoing risk reduction and ART adherence
counseling should be re-emphasized in “Treatment as Prevention”
programs.
OA28.05
Mathematical Modelling to Estimate the
Impact and Cost-effectiveness of TasP, PrEP
and Condom Promotion for Serodiscordant
Couples in Nigeria
Kate M. Mitchell1, Aurélia Lépine1, Fern Terris-Prestholt1,
Emmanuel Alhassan2, John Idoko2, Peter Vickerman1,3
London School of Hygiene and Tropical Medicine, Global Health and
Development, London, United Kingdom, 2National Agency for the
Control of AIDS, Abuja, Nigeria, 3University of Bristol, Bristol, United
Kingdom
1
Background: In Nigeria, only 35% of those in need receive antiretroviral
therapy (ART). We used mathematical modelling to estimate the impact
and cost-effectiveness of treatment as prevention (TasP), pre-exposure
prophylaxis (PrEP) and condom promotion for discordant couples in
Nigeria.
Methods: A deterministic model of HIV transmission within discordant
partnerships and to/from external partners was parameterised using
data from Nigeria and other African settings. The impact (infections
averted (IA) and disability-adjusted life-years (DALYs) averted) and
incremental cost-effectiveness ratios (ICER) were estimated for offering
condom promotion, PrEP to HIV-negatives and/or TasP to HIV-positives,
compared with a baseline scenario where ART was offered at current
national guidelines (CD4< 350 cells/µl) to all HIV positive partners. Full
costs (US$2012) were estimated from a provider perspective, with future
costs and impacts discounted. Frontier analysis was used to determine
optimal intervention combinations.
Results: Large benefits came from offering ART to all HIV positive partners
at current national guidelines compared with current ART coverage (36
IA (23% of infections), 1600 DALYs averted over the lifespans of 1000
couples and their external partners). Condom promotion was the most
cost-effective strategy to provide after offering ART at national guidelines
(US$650/DALY), with the addition of TasP with condom promotion being
the next best investment (ICER US$1050/DALY). The addition of PrEP was
not cost-effective, but provided substantial additional impact in terms of
IA (an additional 17 IA when added to TasP + condom promotion).
Conclusions: The best first intervention for discordant couples in Nigeria
would be offering ART at current national guidelines to all HIV-positives.
Additional impact could be efficiently achieved from condom promotion
and TasP for HIV-positives. PrEP would avert additional infections, but
is not cost-effective in terms of DALYs averted once other interventions
are in place.
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121
Oral Abstract Session 29: T Cell Immunity
OA29.01
OA29.02
Dendritic Cells from HIV-1 Neutralizers
Efficiently Induce the Generation of CXCR5+
CXCR3+ PD1Lo CD4 T Cells with B Cell Helper
Function
Massive Activation, Expansion and
Subsequent Apoptosis of CD8 T-cells in Acute
HIV Infection
Enrique Martin-Gayo1, Taylor Hickman1, Zhengyu Ouyang1,
Rafael Cubas2, Madelene Lindqvist1, Daniel Kauffman1, Elias
Haddad2, Bruce Walker1,3, Mathias Lichterfeld4, Xu G. Yu1
Ragon Institute of MIT, MGH and Harvard, Boston, MA, United States,
Vaccine and Gene Therapy Institute (VGTI) of Florida, Port Saint Lucie,
FL, United States, 3Howard Hughes Medical Institute (HHMI), Boston,
MA, United States, 4Infectious Disease Division, Massachusetts General
Hospital, Boston, MA, United States
1
2
Background: Cellular mechanisms supporting the evolution of
broadly neutralizing antibody activity (bNAbs) against HIV-1 are poorly
understood. Here, we investigated if conventional dendritic cells (cDC)
from HIV-1 patients who naturally develop bNAbs have superior ability
to prime naïve CD4 T cells into Follicular helper (TFH)-like cells.
Methods: cDCs isolated from healthy persons, HIV-1+ chronic
progressors (CP) and controllers with and without detectable bNAbs in
plasma (Neutralizers and Non Neutralizers, respectively) were co-cultured
with identical allogeneic naïve B cells and T cells. TFH phenotypic and
functional characteristics of primed T cells were compared to peripheral
TFH-like cells isolated from human blood.
Results: cDCs from all patient cohorts similarly induced the generation of
PD1hi CXCR3+ TFH-like cells. However cDCs from Neutralizers promoted
more efficiently the development of PD1lo CXCR3+ TFH-like cells and
subsequent maturation of B cells. Both PD1hi and PD1lo cells expressed
TFH markers such as ICOS, Bcl6 and CXCR5, but PD1lo cells preferentially
expressed central-memory and T memory stem cells markers, and had
improved abilities to survive in vitro, as opposed to PD1hi cells that
were highly susceptible to apoptosis and exhibited an effector-memory
phenotype. Interestingly, upon Ag stimulation, PD-1lo TFH-like cells
isolated from peripheral blood differentiated into PD1hi TFH-like cells.
Importantly, PD1lo and PD1hi TFH-like cells were both detectable in
peripheral blood and lymphoid tissues, but higher proportions of PD1lo
TFH-like cells were found in the blood of HIV controllers who developed
neutralizing breadth.
Conclusions: cDCs from Neutralizers have increased abilities to prime
the generation of PD1lo TFH-like cells, which serve as progenitors of
effector PD1hi TFH-like cells, but also provide help to B cells. Higher
proportions of PD1lo TFH-like cells in the blood of HIV-1 neutralizers
suggest an important role of these cells for supporting the development
of bNAbs
122
Zaza Ndhlovu1,2, Philomena Kamya2, Nikoshia Mewalal1,
Thandeka Nkosi1, Nasreen Ismail1, Amber Moodley1, Krista Dong1,
Thumbi Ndung’u1, Bruce Walker2
University of KwaZulu-Natal, HIV Pathogenesis Programme, Durban,
South Africa, 2Ragon Institute of MGH, MIT and Harvard, Cambridge,
MA, United States
1
Background: The initial CD8 T cell response following acute HIV
infection suppresses virus replication, but viremia persists in the vast
majority of people. We used pre-infection and very early HIV infection
longitudinal samples (ranging from day 1 to day 160 post Fiebig I) to
interrogate the dynamics and fate of the initial T cell response, during
the initial rise and subsequent decline in plasma viremia.
Methods: High risk uninfected women in KwaZulu Natal, South Africa
were screened twice weekly by HIV RNA, as part of a comprehensive
HIV prevention program. Pre-infection and post infection blood samples
were evaluation by intracellular cytokine staining (Cd4, CD8, Ki67, Bcl2, CD38, HLA-DR) as well as IFN-gamma ELIspot, and class I tetramer
staining assays.
Results: Acute HIV infection rapidly induced massive activation and
expansion of HIV-specific CD8 T cells such that more than 90% became
activated by day 14 days post Fiebig I, upregulating CD38, HLA-DR,
and Ki67. In contrast, Influenza and CMV-specific CD8 T cells were
not activated. At peak activation HIV-specific CD8 T cells selectively
expressed high levels of the pro-apoptotic marker CD95, low antiapoptotic molecule BCL-2 and failed to upregulate the IL-7 receptor.
Overnight in vitro incubation of peak activation CD8 T cells resulted in
high levels of spontaneous cell death compared to cells collected after
the resolution of T cell activation. Furthermore, activated CD8 T cells
were also less responsive to ex-vivo stimulation compared to samples
collected after resolution of activation, suggesting maximal in vivo
activation.
Conclusions: These data indicate that primary HIV infection induces
massive CD8 T cell expansion, which declines despite persistence of
detectable viremia. The disappearance of early responses may be
attributed to persistent elevation of antigen-stimulation driving virusspecific cells towards apoptosis. Therefore strategies aimed at blocking
apoptosis might be a viable option for rescuing or strengthening early
responses.
OA29.03
OA29.04
Reduction in Breadth and Not
Polyfunctionality or Proliferative Capacity
of CD8+ T Cells Is Associated with Loss of
Virologic HIV Control
Paradigm-violating HLA Class II-restricted
CD8 T-cells in HIV-infection
University of KwaZulu Natal, HIV Pathogenesis Programme, Durban,
South Africa, 2Ragon Institute, Massachusetts General Hospital Harvard
Medical School, Boston, MA, United States, 3AIDS Research Institute
IrsiCaixa, Hospital Universitari Germans Trias i Pujol, Barcelona, Spain,
4
University of Oxford, Department of Paediatrics, Oxford, United
Kingdom
1
Background: CD8+ T cell properties associated with virologic control
of HIV-1 infection are not fully understood. In cross-sectional studies,
T-cell factors associated with control include breadth of responses to
Gag, T cell polyfunctionality and proliferative capacity. However, events
responsible for the loss of control in some individuals remain unknown.
We characterized longitudinally the immunological features that may
explain divergence in disease outcome in 35 HIV-1 C-clade chronically
infected individuals with protective alleles.
Methods: Participants were enrolled in Durban, South Africa. Viral loads,
CD4 counts and HLA typing performed by standard methods. HLAs
A*74:01, B*57, B*58:01, B*81:01 were considered protective. HIV-specific
CD8+ T-cell immune responses were quantified using the interferon
gamma (IFN-g) enzyme-linked immunosorbent spot (ELISPOT) assay
after stimulation with overlapping peptides (OLP) covering the whole HIV
proteome. T cell polyfunctionality and proliferation upon stimulation
with Gag peptide pool was assessed by flow cytometry and CFSE assay
respectively.
Results: At baseline, 15% were viremic controllers (VCs) (< 2,000 RNA
copies/ml) and 11% were progressors (P) (> 100,000 RNA copies/
ml). Over time, 35% of VCs lost viral control. There was no significant
difference in the overall magnitude or breadth of HIV-1 CD8+ T cell
responses between P and VCs at baseline but VCs had higher breadth
of Gag than P (p=0.0038). Among VCs that subsequently lost viral
control, we noted a drop in the overall breadth of HIV-1 CD8+ T cell
responses (p=0.0059). No significant differences were noted in T cell
polyfunctionality or proliferation.
Conclusions: Our data indicates that sustained HIV-1 control in C-clade
infected patients with protective alleles is related to the breath of HIV-1
CD8+ T-cell responses against Gag. The loss of virologic control is related
to a reduction in the total breath of CD8+ T cell responses in the absence
of differences in polyfunctionality and proliferation.
States, 2La Jolla Institute for Allergy and Immunology, La Jolla, CA,
United States, 3US Military HIV Research Program/WRAIR, Silver Spring,
MD, United States, 4Oregon Health & Science University, Portland, OR,
United States
1
Background: Classically, CD8 T cells recognize short peptides presented
by a single HLA class I molecule, while CD4 T cells recognize longer
peptides bound to HLA class II. However, Hansen et al Science 2013
recently reported the induction of unconventional class II-restricted CD8
T cell responses in RhCMV/SIV strain 68-1 vector vaccinated macaques
(reviewed in Ranasinghe & Walker, Nat Biotech 2013). The detection of
class II-restricted CD8 T cells in vaccinated macaques raises a critical
question: do similar paradigm-violating class II-restricted CD8 T cell
responses exist in natural HIV infection?
Methods: We detected a class II-restricted CD8 T cell population in a
treatment-naive HIV-infected individual. The response was identified
using a novel ĆD8 HLA-DR ELISpot´. In this assay, CD8 T cells
exclusively targeted a single peptide, Gag41, presented by LCL stably
expressing human recombinant HLA class II DRB1*11 (in N=1/22
DRB1*11 subjects). Antibody blocking of HLA class I and II, and flow
cytometric staining with a class II DRB1*11-Gag41 tetramer confirmed
the class II CD8 restriction.
Results: Strikingly, the DRB1*11-Gag41 tetramer specifically bound
12% of total CD8 T cells directly ex vivo (with no T cell expansion). The
class II-restricted CD8αβ T cells exhibited a highly differentiated TEMRA
memory phenotype. They also expressed high levels of Perforin and
Granzyme B cytolytic proteins, and a distinct polyfunctional profile when
compared intra-patient to the conventional class I-restricted B57-KF11
CD8 TEM response. Our data suggests the class II-restricted CD8 response
is qualitatively and quantitatively distinct from conventional class I- CD8
T cells.
Conclusions: In our exploratory study, we have demonstrated the first
proof-of-principle detection of a large, unequivocal class II-restricted Gagspecific CD8 T cell response in an HIV-infected individual. Elucidating
class II-restricted HIV-specific CD8 T cells that violate immunologic
paradigms is likely to be important in future HIV vaccine design.
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123
Catherine K. Koofhethile1, Zaza Ndlhovu1,2, Nasreen Ismail1,
Zanele Mncube1, Lungi Maphumulo1, Mary van de Stok1,
Christina Thobakgale1, Julia G. Prado3, Bruce B.D. Walker1,2,
Phillip J.R Goulder1,4, Thumbi Ndung’u1
Srinika Ranasinghe1, Isaiah Davis1, Damien Soghoian1, Pedro
Lamothe1, John Sidney2, Alessandro Sette2, Mary Carrington1,
Hendrik Streeck3, Louis Picker4, Daniel Kaufmann1, Bruce Walker1
OA29.05 LB
OA29.06
Anti-viral CD8 T-cells with B-cell Follicle
Homing Potential Contribute to Vaccinemediated Enhanced Control of Pathogenic SIV
Infection
A Novel T-cell Vaccine Eliciting T-cell
Specificities Associated with Control of HIV-1
In Humans Is Highly Immunogenic in Mice
and Macaques
Geetha Mylvaganam1, Daniel Rios2, Ifor Williams2, Vijayakumar
Velu1, Rama Amara1
Beatriz Mothe1,2, Xintao Hu3, Anuska Llano1, Margherita Rosati3,
Alex Olvera1, Viraj Kulkarni3, Antonio Valentin3, Candido Alicea3,
Niranjan J. Sardesai4, Muntsa Rocafort1, Manel Crespo5, Jorge
Carrillo1, Andrés Marco6, James I. Mullins7, Lucy Dorrell8, Tomáš
Hanke9, Bonaventura Clotet1,2,10, George N. Pavlakis3, Barbara K.
Felber3, Christian Brander1,2,11
Emory University, Yerkes National Primate Research Center, Atlanta,
GA, United States, 2Emory University, Department of Pathology, Atlanta,
GA, United States
1
IrsiCaixa AIDS Research Institute-HIVACAT, Hospital Germans Trias
i Pujol, Badalona, Spain, 2Universitat de Vic – Universitat Central de
Catalunya, Vic, Spain, 3Human Retrovirus Section-National Cancer
Institute, Frederick, MD, United States, 4Inovio Pharmaceuticals, Inc.,
Blue Bell, PA, United States, 5HIV Unit, Hospital de la Vall d’Hebrón,
Barcelona, Spain, 6Centres Penitenciaris BCN, Barcelona, Spain,
7
University of Washington, Seattle, WA, United States, 8MRC Human
Immunology Unit, Weatherall Institute of Molecular Medicine,
University of Oxford, The John Radcliffe, Oxford, United Kingdom,
9
Jenner Institute, University of Oxford, Oxford, United Kingdom,
10
Universitat Autònoma de Barcelona, Barcelona, Spain, 11Institució
Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain
1
Background: The germinal center (GC) resident T follicular helper cells
(Tfh) represent a significant fraction of total pool of HIV/SIV infected
cells during chronic infection and HAART. The GC are generally thought
to exclude CD8 T cells and anti-viral CD8 T cells with potential to
migrate to GC (follicular CD8) may enhance HIV/SIV control and reduce
viral reservoirs. Here we studied the follicular CD8 in a cohort of DNA/
MVA vaccinated rhesus macaques (RM) that controlled or did not control
a pathogenic SIV infection.
Methods: RM were vaccinated with a DNA/MVA SIV vaccine and
challenged intrarectally with SIVmac251. Animals with viral load below
1,000 copies at set point were defined as controllers. All controller RM
(n=19) were vaccinated and non-controller RM (n=18) consisted of both
vaccinated and unvaccinated.
Results: Post challenge, we observed an aberrant enrichment of SIV+ PD1hi CD4 T cells in the LN and rectum of non-controllers but not controllers.
The enhanced viral control was associated with higher frequency of Gag
CM9 Tet+ CD8 T cells in the LN of controller RM compared to noncontroller RM. This was not evident in blood. Interestingly, a significant
fraction of anti-viral CD8 T cells in the controller RM co-expressed
CXCR5 (required for homing to B cell follicles/GC). The frequency of Tet+
CXCR5+ granzyme B+ cells was also higher in the LN of controller RM
and higher frequencies correlated with lower Tfh and enhanced viral
control. Immunofluorescence staining revealed co-localization of CD8
T cells with PD-1bright cells in IgD- GC, a phenomena not observed in
the non-controller RM. Impressively, the CXCR5+ CD8 T cells from the
controller RM restricted the anti-CD3 driven expansion of CM9 peptide
pulsed Tfh cells in vitro suggesting their killing potential.
Conclusions: Our results reveal a novel subset of anti-viral CD8 T cells
that may contribute to enhanced control of pathogenic SIV infection by
infiltrating to GC of lymphoid sites and limiting SIV replication in Tfh in
a vaccine setting.
124
Background: A top-down strategy was used to search for beneficial viral
targets in large human immunogenicity data and to identify potential
decoy targets that should be avoided in future vaccine designs.
Methods: Through the identification of beneficial T cell responses in
more than 1000 individuals, the HIVACAT T-cell immunogen (HTI) was
designed to contain 16 HIV-1 protein segments of 10-70 amino acids in
length, covering >50 optimal defined CD4+ and CD8+ T-cell epitopes with
>40 different HLA restrictions, without overrepresentation of B27/B57/
B58 restricted epitopes. Heterologous prime-boost regimens combining
DNA (D) and MVA (M) expressing HTI were assessed in C57BL/6 mice
and four Indian rhesus macaques. Cellular responses were characterized
using IFN-γ ELISPOT and intracellular cytokine assays.
Results: In C57BL/6 mice, DNA.HTI induced broad CD4+ and CD8+ T-cell
responses to all segments within Gag, Pol, Vif and Nef. These responses
were strongly increased by using heterologous regimens consisting of
3x DNA.HTI prime followed by MVA.HTI boost compared to 3 or 4x
DNA.HTI only (median magnitude DDDM of 3,051 vs 1,401 and 1,353
SFC/106 splenocytes in DDD and DDDD groups respectively, p=0.0087).
In rhesus macaques, DDD (administered IM in combination with
macaque IL-12 DNA as molecular adjuvant using in vivo electroporation)
induced 0.4-1.5 % of specific T cells that persisted over a period of
4.5 months. MVA.HTI boosted the responses by 3- to 20-fold, reaching
0.4-3.2 % IFN-γ T-cells. DDDM induced central and effector memory
responses with a significant fraction of the vaccine induced IFN-γ+CD8+
T cell being either CD107a+ or GzmB+.
Conclusions: HTI delivered in a DDDM regimen was highly immunogenic
in mice and macaques. The responses were CD8+ and CD4+ effector T
cells with cytotoxic potential as well as CD4+ central memory T cells
indicating long-term immunological persistence. These data justify
further testing of the HTI approach in human clinical trials.
Oral Abstract Session 30: Antibody Functions and Protection
OA30.01
OA30.02
Synthetic Nucleic Acid Antibody Prophylaxis
with Electroporation Drives Biologically
Relevant Anti-HIV-1 Envelope Responses in
vivo
Insights on Synergistic Antibody-dependent
Cellular Cytotoxicity (ADCC) Activity Mediated
by Mutant Human Monoclonal Antibodies
against HIV-1 Env
Kar Muthumani1, Seleeke Flingai1, Megan Wise1, Colleen Tingey1,
Kenneth E. Ugen2, Niranjan Y. Sardesai3, Joseph J. Kim3, David
B. Weiner1
Chiara Orlandi1, Anthony L. DeVico1, Yongjun Guan1, George
K. Lewis1
Background: Monoclonal Ab’s have demonstrated therapeutic utility
against several malignancies and infectious diseases. A drawback of
this strategy is the time-consuming and expensive process requiring
purification and scale up production of the Ab’s for clinical use. A method
to produce antibodies in vivo would be significant improvement for this
platform. It would be important if these Ab’s could be administered with
out induction of vector serology allowing repeated administrations.
Furthermore, delivery in a non-permanent fashion would also have
advantages.
Methods: Here we report development of new synthetic optimized
plasmid vector/improved EP encoding Abs genes for delivery in vivo.
This strategy allows for in vivo synthesis and serum expression of such
ex vivo developed antibodies.
Results: An “enhanced and optimized” DNA plasmid generates
immunoglobulin heavy and light chains (Fab) of an established
neutralizing anti-HIV monoclonal antibody (VRC01). We demonstrate
that the serum of transfected animals exhibited the ability to bind to
HIV envelopes in ELISA and FACS analysis against diverse isolates and
this serum possessed HIV neutralizing activity equivalent to the “native”
VRC01 antibody in vivo. In vivo delivery seroconverted the animals with
in a few hours and neutralizing activity lasted for weeks. Antibodies
exhibited a broad neutralization profile. This technology has important
advantages for in vivo antibody production which could compliment or
circumvent the need for standard antigen based vaccination, particularly
in situations where there is difficulty in generation of protective antibody
responses by immunization.
Conclusions: This is the first study we are aware of using synthetic DNA
plus EP delivery to produce circulating bioactive antibody responses
in a living animal. The study has implications for prophylactic and
therapeutic strategies for HIV and other important diseases especially
in resource limited settings where antibody therapy is cost prohibitive.
University of Maryland, Institute of Human Virology, Baltimore, MD,
United States
1
Background: Several reports showed the key role of ADCC in protection
against HIV-1 infection as it inversely correlates with progression in
natural infection and with protection in non-human primate and human
vaccines. Of note, optimal ADCC activity is the result of efficient antibody
bridging between infected cells and FcγR-bearing effectors. We report
that mAb pairs attenuated for FcγR binding by mutagenesis mediate
potent synergistic ADCC when admixed.
Methods: We engineered the human mAbs C11, N5-i5 and N12-i2,
specific for HIV-1 Env highly conserved epitopes, to exhibit three levels
of anti-HIV ADCC strength: wild type, LALA variant and Fab fragment.
ADCC and binding potency were assessed with Gp120 sensitized
CEM NKr CCR5 target cells and the affinity to FcγRs by Elisa. We also
developed a high throughput screening of ADCC-mediating antibodies
combinations, quantifying the synergy with Chou-Talalay method.
Results: Our ADCC assay revealed stronger activity for gp120 region C1
specific C11 and N5-i5 mAbs then the Co-Rbs N12-i2 mAb. Interestingly,
LALA mutations entirely abrogated only N12-i2 cytotoxicity, while
C11 and N5-i5 LALA retained weak ADCC activity at the highest
concentrations tested. As expected, Fab fragments did not induce any
target cells killing. Binding assays showed no difference in the affinity of
all mAbs variants for their viral antigens on target cells, meaning that the
diverse cytotoxicity potency did not reside in the Fab regions. Moreover,
the higher ADCC potency of C11 and N5-i5 wt and LALA more likely
was due to different levels of binding to FcγR. Finally, we report for the
first time that pairs of LALA mAbs synergize strongly for ADCC with
patterns unique to each mAb pair.
Conclusions: These data show that the antigen binding nature can
overcome the loss of energy for IgG-FcγR interaction in LALA mutants.
The findings provide a new experimental tool to clarify antigen-antibody
aggregation contribution to Fc-mediated effector function and for
passive immunization studies design.
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125
University of Pennsylvania School of Medicine, Pathology and Lab.
Medicine, Philadelphia, PA, United States, 2University of South Florida
Morsani College of Medicine, Department of Molecular Medicine,
Tampa, FL, United States, 3Inovio Pharmaceuticals, Inc., Blue Bell, PA,
United States
1
OA30.03
OA30.04
The Impact of Antiretroviral Treatment on
HIV-1-Specific Broadly Neutralizing Antibody
Responses
Topical Application of Broadly Neutralizing
Monoclonal Antibodies Reduces HIV Infection
of Mucosal Tissue
Nonhlanhla N. Mkhize1,2, Maphuti Madiga1, Raveshni Durgiah1,
Elin S. Gray1, Penny L. Moore1,2,3, Sengeziwe Sibeko3, Salim
Abdool Karim3, Lynn Morris1,2,3, CAPRISA Acute Infection Study
Team
Yanille Scott1, Kevin Whaley2, Charlene S. Dezzutti3,4
National Institute for Communicable Diseases of the NHLS, Virology,
Johannesburg, South Africa, 2University of the Witwatersrand,
Johannesburg, South Africa, 3CAPRISA/University of Kwa Zulu Natal,
Durban, South Africa
1
Background: Initiation of antiretroviral therapy (ART) in HIV-infected
individuals is associated with control of plasma viremia and improved
CD4 counts which impacts on immune responses. We identified
individuals from the CAPRISA 002 cohort with broadly cross-neutralizing
(BCN) antibodies, who all initiated treatment within 4-6 years of infection.
In this study we compared the neutralization titers and antibody
specificties present before and after initiating ART.
Methods: Total IgG was isolated (to remove traces of antiretroviral
drug) using Protein G columns from plasma collected just prior to and
within 2 years of ART from 6 women with BCN antibodies. These BCN
antibodies were shown to target an N160-dependent epitope, a glycan
at position 332 and in the MPER region on Env. Purified IgG was tested
for neutralization against a panel of pseudoviruses (7 clade C, 4 clade B
and 3 clade A) in TZM-bl cells. Neutralizing antibody specificities were
mapped using pseudoviruses with mutations in key residues within each
epitope. VSV-G was used as a control.
Results: All plasma samples had IgG-associated neutralizing activity,
with pre-ART and post-ART IgG fractions neutralizing the same viruses
in the panel. However, there was a significant decline in neutralization
titers in the post-ART longitudinal samples. None of the IgG fractions
neutralized VSV-G, indicating no residual ARV drug in these samples.
Antibody epitope mapping data suggested that IgG antibodies isolated
pre-ART and after ART targeted the same epitopes. There was a
significant decrease in viral loads and an increase in CD4 cell count as
early as 6 months post-ART initiation.
Conclusions: Despite the low antigenic stimulation following ART,
neutralizing antibodies were still present in most individuals with BCNs,
even after 12 months of therapy. However, the decline in neutralization
titers with preservation of antibody specificities during ART treatment
suggests that the maintenance of robust BCN antibody responses is
largely dependent on viral load.
126
University of Pittsburgh Graduate School of Public Health, Pittsburgh,
PA, United States, 2Mapp Biopharmaceutical, Inc., San Diego, CA,
United States, 3University of Pittsburgh School of Medicine, Pittsburgh,
PA, United States, 4Magee-Womens Research Institute, Pittsburgh, PA,
United States
1
Background: Broadly neutralizing monoclonal antibodies (nAbs) reduce
HIV transmission in vitro and in animal models of HIV transmission.
However their efficacy in preventing human mucosal transmission of
HIV remains unknown. During sexual transmission of HIV, semen is
the inoculum and previous studies show that semen contains factors
that modulate HIV infection in vitro. We theorize that topically applied
nAbs can reduce HIV infection of ectocervical and colonic mucosa in the
context of semen.
Methods: nAbs, 4E10, VRC01, PG16 and PG9, were evaluated against
HIV-1JR-CSF ex vivo in polarized human ectocervical and colonic tissues.
Tissues were treated with nAbs and then inoculated apically with
virus in the presence or absence of 50% whole human semen. Viral
replication in tissues was monitored by HIVp24 ELISA for up to 21 days
post inoculation. Data are presented as the median HIVp24 (pg/mL) and
95% confidence interval of 3-7 tissues from individual donors. ANOVA
and post hoc multiple comparison procedures were used to compare
nAb and semen treatment groups.
Results: 1.5µM nAbs were required to protect ectocervical tissue
compared to 0.03µM nAbs in colonic tissue ex vivo. Treatment of
ectocervical tissue with 1.5µM nAbs resulted in protection of 90%, 100%,
80% or 30% of tissues by PG16, PG9, VRC01 and 4E10, respectively.
Treatment with 0.03µM PG16, PG9 or VRC01, resulted in protection of
100% of colonic tissues, but only 50% with 4E10. Our combined results
show that nAb potency follows the order PG16>PG9>VRC01>>4E10,
consistent between cell-based assays and ex vivo tissue studies. Similar
levels of viral inhibition were observed in tissues treated with nAbs in
the presence of semen.
Conclusions: Collectively, these data suggest nAbs are effective in the
presence of semen and should be considered as a non-chemotherapeutic
option for the prevention of HIV. Importantly nAbs could be considered
for use in persons already infected with HIV and seeking ways to
mitigate transmission to their partners.
OA30.05
OA30.06 LB
Isolation of Monoclonal Antibodies from a
SHIV-AD8 Infected Rhesus Macaque with
Broad Neutralizing Activity
Heterogeneous Conformation of HIV-1
Envelopes on Individual Virions
1
1
Diseases, National Institutes of Health, Bethesda, MD, United States,
2
Institutes of Health, Bethesda, MD, United States, 3Columbia University,
Department of Biochemistry and Molecular Biophysics, New York City,
NY, United States
1
Background: To assess the ability of immunogens to elicit broadly
neutralizing antibodies (bNAbs) against HIV-1, an appropriate animal
model is necessary. It has been previously established that the CCR5tropic SHIV-AD8-EO molecular clone is a pathogenic and mucosally
transmissible virus in Rhesus macaques. Furthermore, some macaques
infected with this SHIV develop bNAbs; however the mechanism of
eliciting bNabs in this model remains unclear.
Methods: One macaque, JG7, slowly developed breadth on a multiclade 21-virus panel from weeks 47-131 post-infection. Frequency
of antigen-specific germinal center B cells and T-follicular helper (TFH)
cells were evaluated at the early wk47 time-point as well as somatic
hypermutation in these B cell receptors (BCRs). Monoclonal antibodies
were isolated and characterized by ELISA binding and neutralization
profile.
Results: By wk47, JG7 had high viral load, and high levels of antigenspecific TFH and IgG+ B cells in the germinal center. A fingerprint analysis
of sera neutralization data suggested a CD4-induced (CD4i) specificity
at the early wk47 time-point. BCR sequencing revealed a large clonal
family distinguished by high V-gene mutation frequency and long heavy
chain CDR3s of 31 amino acids. Four BCRs were cloned and found to
recapitulate most of the serum breadth at this time. Furthermore, these
monoclonals competed with 17b for binding to gp120 and had enhanced
neutralization activity in the presence of soluble CD4, confirming the
CD4i prediction.
Conclusions: Our findings support the hypothesis that in the presence
of high antigen levels and T cell help, Rhesus macaques can develop
cross-neutralizing anti-HIV antibodies with high levels of mutation and
long CDRH3s akin to bNAbs isolated from HIV infected humans. These
data highlight the utility of the SHIV model for studying development of
bNAbs in Rhesus macaques.
Anush Arakelyan1, Debbie King2, Jean-Charled Grivel1, Robin J.
Shattock2, Leonid Margolis1
NIH/NICHD, Section of Intercellular Interactions, Bethesda, MD, United
States, 2Imperial College, London, United Kingdom
1
Background: HIV-1 has 7-15 envelope spike glycoproteins (Env) per
virion consisting of a trimer of non-covalently linked gp120-gp41
heterodimers. These “functional” spikes are essential to virion binding
and fusion, however it is unclear as to number required to mediate
efficient fusion. Indeed other non-functional Env conformations,
including uncleaved precursors (gp160), aberrant oligmers, monomers
and gp41 stumps devoid of gp120 are also thought to be displayed on
virions. The extent to which functional and non-function forms of Env
are co-displayed on individual virions is currently a matter of debate.
Recently, we developed a new technique, Flow Virometry that allows the
study of proteins on the surface of individual virions. We have applied
this technique to probe the conformation of Envs on the surface of
individual particles using a panel of anti-env antibodies that discriminate
between different conformations of these molecules.
Methods: HIV-1 virions of BaL strain grown in PM1 CD4 T cells were
captured with 15 nm magnetic nanoparticles (MNPs) coupled to one
of several monoclonal antibodies recognizing particular conformations
of Env. Captured virions were then stained with fluorescent anti-Env
antibodies different from the capture antibody and separated from free
antibodies on a magnetic column. A range of neutralizing and nonneutralizing antibodies targeting functional and non-function Env forms
were used to stain virions and their representation on individual viral
particles assessed by flow virometry.
Results: Our data indicate a non-uniform distribution of functional and
non-functional Env within a viral population, where differential patterns
of antibody staining revealed virion populations that were homogenous
or mosaic with respect to functional and non-functional forms of Env.
Conclusions: These finding have important implications for
understanding antibody binding and neutralization, as well as other
antibody effector functions.
www.hivr4p.org
127
Rebecca M. Lynch , Takuya Yamamoto , Patrick Wong , Ivelin
Georgiev1, Rodrigo Matus-Nicodemos1, Stephen Schmidt1,
Rajeev Gautam2, Zizhang Sheng3, Lawrence Shapiro3, Yoshiaki
Nishimura2, Malcolm A. Martin2, John R. Mascola1, Richard A.
Koup1
1
Poster Discussions
Poster Discussion 01: Community Engagement and Advocacy
PD01.01
PD01.02
Supporting Participant Adherence through
Structured Engagement Activities in the MTN020 (ASPIRE) Trial
Pre-screening Strategies to Enhance Accrual
and Retention in the ASPIRE Trial at the
Wits Reproductive Health and HIV Institute,
Johannesburg
Katie Schwartz1, Patrick Ndase2, Kristine Torjesen1, Ashley Mayo1,
Rachel Scheckter1, Ariane van der Straten3, Lydia Soto-Torres4,
Thesla Palanee5, Jared Baeten2
1
FHI 360, Durham, NC, United States, 2University of Washington,
Seattle, WA, United States, 3Women’s Global Health Imperative;
RTI International, San Francisco, CA, United States, 4NIAID/DAIDS,
Bethesda, MD, United States, 5Wits Reproductive Health & HIV Institute,
Background: Sub-optimal product use in previous HIV prevention
trials highlights the need for strategies to foster participant adherence
to protocol requirements. Early in ASPIRE, participant engagement
activities were implemented across all 15 study sites to enhance
participant commitment within the prevention trial setting.
Methods: Monthly tracking tools, site presentations and assessment
visit summaries were reviewed to describe engagement activities,
lessons learned and staff feedback. Proportion of ASPIRE participants
engaged is reported from event attendance logs. Participant feedback is
taken from staff debriefing reports of qualitative interviews.
Results: Across ASPIRE sites, 76% of enrolled participants attended
at least one of 199 staff-facilitated events held between 3/13-4/14.
Events fell into 6 broad categories: milestone/retention events, social
events, group education, male involvement events, couples workshops
and holiday events. Support was fostered through peer educators,
motivational speakers and shared experiences. Discussion topics included
ring experiences, contraceptive options, adherence, impact of HIV, trial
disclosure and community rumors. Social activities included movies,
games and employment/life skills training. Lessons learned include
designing events to be flexible and participant-centered, ensuring a mix
of attendees, while maintaining participant confidentiality and comfort
in group settings. Staff reported increased rapport with participants and
improved team morale. Preliminary data from 28 participant interviews
suggest that participants perceive engagement events as having a
positive impact on personal commitment to trial and product use, as
well as improved staff relations.
Conclusions: Meaningful participant engagement is essential to
HIV prevention trials with long follow-up and ongoing product use.
Monitoring of participant engagement activities in ASPIRE will help
inform design of future participant engagement and product support
strategies in prevention trials.
POSTER DISCUSSIONS
128
Pranitha Ramchuran1, Krishnaveni Reddy1, Patience Ramalo1,
Nombulelo Maseko1, Amukelani Vuma1, Lizzy Gama1, Sylvia
Sibeko1, Nosipho Duba1, Helen Rees1, Thesla Palanee1
1
Background: The ASPIRE trial aims to assess the safety and effectiveness
of Dapivirine Vaginal Ring for the prevention of HIV infection in women.
In clinical trials, participant experiences at screening, visit waiting times
and management of clinic flow impact accrual and retention significantly.
To optimize these factors and improve operational and cost efficiencies
in ASPIRE, novel pre-screening (PS) strategies were implemented.
Methods: These strategies include recruitment of known HIV negative
participants from Primary Health Care clinics within the catchment area,
provision of voluntary counselling and testing on-site prior to the conduct
of ASPIRE screening procedures and the implementation of a sitespecific PS checklist. The PS checklist lists basic ASPIRE eligibility criteria
as well as documents/information required for screening (identification
document, proof of contraception, details of 3 contacts). This tool is
administered by community health workers (CHWs) during recruitment
and allows determination of potentially eligible participants prior to their
arrival at the clinic for screening procedures. It is also reviewed by the
study administrators at the clinic when potential participants present for
study screening in the event that information has changed.
Results: The implementation of the above PS strategies results in the
screening of only potential participants who meet basic ASPIRE eligibility
criteria and reduces the number of participants who are screened out
at a later stage. In addition, the PS Checklist reduces the frequency of
split visits due to inadequate documentation/information by alerting
participants to bring in all required information at their scheduled
screening visit.
Conclusions: These reductions in the number of unsuccessful/split
screening visits reduce the clinic workload and participant waiting
times. As a result, participant study experience is improved and study
operational and cost efficiencies are optimized. This in turn impacts
positively on accrual and retention.
PD01.03
PD01.04
Why Should I Take Drugs for your Infection:
Outcomes of Formative Research on Use of
PrEP in Nigeria
Consultation beyond the CAB: Engaging the
Greater Community in Rectal Microbicide
Clinical Trial Design and Planning
John Idoko1, Morenike O. Ukpong2, Nancin Yusufu Dadem3,
Grace O. Kolawole3, James Anenih4, Emmanuel Alhassan5
Clare Collins1, Cindra Feuer2, Jerome T. Galea3, Pedro Gonzales4,
Brian Kanyemba5, Udom Likhitwonnawut6, Jonathan Lucas7,
Steve Miralles3, Benjamin Perkins8, Jim Pickett9, Pongpun
Saokhieo10, Wipas Wimonsate11
Background: The study aimed to explore public opinion about PrEP
including community interests and perceptions about the use of PrEP.
Methods: The study used a mixed method approach. This included
conduct of telephone interviews, in-depth interviews, focus group
discussions, consultative meetings with critical stakeholders engaged
in HIV prevention, treatment, care and support programs in Nigeria
and an online survey to explore opinions about the design of the
PrEP demonstration project in Nigeria. Information required include
appropriate target group for the use of PrEP, community interest and
perception about the use of PrEP as HIV prevention tool, how best to
communicate about PrEP; concerns about PrEP use by sero-discordant
couples; and suggestion for the design and implementation of the PrEP
demonstration project
Results: HIV sero-discordant couples were identified as the appropriate
target group for the use of PrEP for a demonstration project in Nigeria.
Most respondents felt PrEP would bring about added gains to the
country by reducing HIV incidence and scaling-up of HIV prevention
for key affected populations. Electronic and print media were identified
as important means for massive public education about PrEP to prevent
stigma and create awareness about PrEP. Stigma was identified as a
major concern and a potential barrier for the uptake and use of PrEP
by HIV sero-discordant couples. Most times, identified index partner
is the sero-discordant relationship is a HIV positive female diagnosed
during pregnancy. In a chauvinistic society like Nigeria, HIV negative
male spouses may resist enrolling on a PrEP program because of the HIV
positive status of the female partners.
Conclusions: PrEP uptake and use by HIV sero-discordant couples in
Nigeria may face challenges which are surmountable. A lot depends on
appropriate actions taken by multiple players including motivation of
HIV negative male partners to use PrEP, and effective public education
programs to address stigma.
Microbicide Trials Network, Pittsburgh, PA, United States, 2AVAC:
Global Advocacy for HIV Prevention, New York, NY, United States,
3
Epicentro, Lima, Peru, 4IMPACTA, Lima, Peru, 5Desmond Tutu HIV
Foundation, Cape Town, South Africa, 6Consultant, Chiang Mai,
Thailand, 7Microbicide Trials Network, Durham, NC, United States,
8
Fenway Health, Boston, MA, United States, 9International Rectal
Microbicide Advocates, Chicago, IL, United States, 10Research Institute
for Health Sciences, Chiang Mai, Thailand, 11Thailand MOPH - U.S. CDC
Collaboration, Bangkok, Thailand
1
Background: Poor communication with key stakeholders and lack of
community support and engagement can jeopardize HIV prevention
research. This was demonstrated by earlier PrEP trials in Cambodia and
Cameroon which were halted in response to pressure from advocates
and activists. Indeed, these events might not have escalated to crisis
levels had community and civil society representatives been invited
to take part in the protocol development process or if they had been
engaged before and during the course of these trials.
Methods: Prior to the launch of the first ever Phase II rectal microbicide
clinical trial (MTN-017), the Microbicide Trials Network (MTN) partnered
with trial sites, some that were new to the MTN, and advocacy
organizations to hold a series of civil society consultations. Six face-toface consultations were held during the study’s protocol development
stage in South Africa, Thailand, Peru and the U.S.
Results: Consultation attendees (n= 181) gave feedback on an early
draft of the MTN-017 study design and through in-depth discussions
provided unique insight about regional and local social norms and
practices, which resulted in significant changes to the study’s design
and implementation plan. For example, the protocol team heard from
community stakeholders that the initial study design was confusing and
that the eligibility criteria should include transgender women.
Conclusions: The MTN-017 consultations moved beyond the traditional
model focused on Community Advisory Boards (CABs) by securing
feedback from individuals who were more closely representative of
potential study participants and the community at large, benefitting the
communities, trial sites and the MTN. This approach exemplifies the
tenets of Good Participatory Practice Guidelines (GPP) for Biomedical
HIV Prevention Trials jointly developed by AVAC and UNAIDS, which
maintain effective communication and meaningful community
engagement are essential for the successful and ethical conduct of HIV
prevention trials.
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129
POSTER DISCUSSIONS
National Agency for the Control of AIDS, Abuja, Nigeria, 2Institute of
Public Health, New HIV Vaccine and Microbicide Advocacy Society,
Ile-Ife, Nigeria, 3Jos University Teaching Hospital, Department of AIDS
Prevention Initiative Nigeria (APIN), Jos, Nigeria, 4National Agency for
the Control of AIDS, Strategic Knowledge Management, Jos, Nigeria,
5
National Agency for the Control of AIDS, Resource Mobilisation, Abuja,
Nigeria
1
Poster Discussions
PD01.05
The Impact of a Decade of Investment in HIV
Prevention Research in Uganda
Emmanuel Mugisha1, Daniel Murokora2, Bahati Prince Ngongo3,
Leslie Nielsen4, Bonnie Bender3
Independent Consultant, Country Director PATH, Kampala, Uganda,
Independent Consultant, Uganda Women’s Health Initiative, Kampala,
Uganda, 3International AIDS Vaccine Initiative, Nairobi, Kenya,
4
International AIDS Vaccine Initiative, Kampala, Uganda
1
2
Background: We reviewed the outcome of 10 years of investment
in HIV new prevention technology (NPT) research including vaccine,
microbicides and Pre-exposure prophylaxis (PrEP) in Uganda to identify
areas of impact.
Methods: In-depth interviews and focus group discussions were held
with 19 key informants using the Research Impact Framework (London
School of Hygiene and Tropical Medicine) to identify where HIV NPT
investment has had the greatest impact. Secondary data was collected
from four key research institutions in Uganda.
Results: Investment in NPT research from 2001-2012 has had a positive
impact in Uganda. Respondents highlighted areas of high impact such
as scientific contributions, including data demonstrating efficacy of
Voluntary Medical Male Circumcision and PrEP; improved understanding
of key populations epidemiology and increased scientific productivity as
evidenced by increased publications. There have been significant gains
in skills development and expansion of expertise; more than 1,000
clinicians, nurses, and counselors trained in Good Clinical Practice,
specialized training of more than 800 lab scientists and technicians
and sponsorship of more than 50 higher degrees. Improved clinical
trial infrastructure and standards of community engagement were also
highlighted.
The investment impact was less than anticipated in the area of health
system strengthening. Health benefits were limited to volunteer
populations and to facilities in the vicinity of trial sites. In a few cases,
improvements were not sustainable, expiring once projects were
completed. Some respondents perceived international collaborations as
being driven by external priorities resulting in upstream research and
early product development being deprioritized.
Conclusions: There have been clear benefits to HIV NPT research
investment in Uganda. Future investments should consider how to
incorporate objectives of local investigators, including upstream
research, and ensure the sustainability of health system strengthening.
POSTER DISCUSSIONS
130
Poster Discussion 02: Correlates of Protection in Highly Exposed Seronegative People
PD02.01
PD02.02
Genital Proteome Correlates of Highly HIV-1
Exposed Uninfected African Women
Mx2 Expression Is Associated with Reduced
Susceptibility to HIV Infection in Highly
Exposed HIV Seronegative Sex Workers from
Nairobi, Kenya
University of Washington, Global Health, Seattle, WA, United States,
University of Manitoba, Winnipeg, MB, Canada, 3University of
Washington, Seattle, WA, United States, 4University of Manitoba,
Winnipeg, WA, United States, 5University of California at San Francisco,
San Francisco, CA, United States, 6Kenya Medical Research Institute,
Nairobi, Kenya
1
2
Background: Identification of biologic factors robustly associated with
resistance to HIV-1 infection among highly HIV exposed seronegative
(HESN) individuals is a priority. We quantified HIV exposure among
HIV-1 uninfected women in HIV serodiscordant couples and evaluated
protein abundances as potential biomarkers of HIV resistance by
comparing HESN versus low exposure (non-HESN) women.
Methods: 33 vaginal swabs from 10 HESN (16 swabs) and 10 nonHESN (17 swabs) women in the placebo arm of the Partners PrEP Study
had tandem mass spectrometry (MS) to assess frequency of 435 human
and 247 bacterial proteins. We assessed differences by generalized
estimating equations (GEE) adjusting for bacterial vaginosis, and
calculated false discovery rates (FDRs).
Results: Of HESN women, 100% reported unprotected sex versus 0%
of non-HESN and had HIV infected partners with higher median plasma
HIV RNA log10 copies/mL (5.0 [interquartile range (IQR):4.9,5.1]) than
non-HESN women (1.6 [IQR:1.6,1.9]). Based on these factors, HESN had
>100-fold higher risk of infection than non-HESN. Hierarchical clustering
of differentially abundant proteins distinguished HESN from non-HESN
with 88% and 76% sensitivity and specificity, respectively. Nine proteins
had FDR< 5%, including elafin, azurocidin and β2 integrin. Elafin was 8
times more abundant among HESN (95% CI=[4.0-16.0];FDR=8.9x10-7),
while β2 integrin was 4 times less abundant among HESN (95% CI=[1.99.2];FDR=0.046)..
Conclusions: The vaginal proteome differs in HESN and non-HESN
women. Consistent with an earlier study, elafin, a mucosal anti-protease
that may inhibit HIV, is more abundant among purportedly HIV resistant
women; β2 integrin, which may facilitate cell-to-cell transfer of HIV, was
decreased in HESN despite high exposure. Further research is needed
to clarify if these factors mark HIV exposure or host resistance to HIV.
Derek R. Stein1, Souradet Y. Shaw2, Max Abou3, Stuart J.
McCorrister4, Garrett R. Westmacott4, Francis A. Plummer5, T.
Blake Ball3
University of Manitoba, Medical Microbiology, Winnipeg, MB, Canada,
University of Manitoba, Community Health Sciences, Winnipeg,
MB, Canada, 3Public Health Agency of Canada, National HIV and
Retrovirology Laboratories, Winnipeg, MB, Canada, 4Public Health
Agency of Canada, Mass Spectrometry and Proteomics Core Facility,
Winnipeg, MB, Canada, 5Public Health Agency of Canada, Winnipeg,
MB, Canada
1
2
Background: Recent studies have identified Mx2 as a novel HIV-1
innate restriction factor that inhibits proviral integration. In these same
studies Mx1 was found not to have an effect on HIV-1 in vitro. An initial
proteomic study of immune cells from highly exposed HIV seronegative
individuals (HESN) enrolled in the Pumwani sex worker cohort identified
Mx1 and Mx2 as potential correlates of HIV protection. A detailed
population level analysis of the role Mx1 and Mx2 contribute to reduced
susceptibility to HIV infection was performed.
Methods: Peripheral blood mononuclear cells (PBMC) were isolated
from 102 HESN women and 100 high-risk negative controls enrolled
in a Nairobi-based sex worker cohort. Whole cell lysates were prepared
and assayed for Mx1 and Mx2 expression by commercial ELISA.
Bivariate and multiple linear regression analyses were conducted to
control for age, pregnancy, menopause, contraception, recent infections,
and medication use.
Results: Mx2 was significantly overexpressed in HESN women
compared to high-risk negative controls (p=0.027). No associations with
Mx1 expression were observed. After multiple linear regression analyses
accounting for age, menopause, pregnancy, Depo-Provera use, recent
infections and medication usage, Mx2 expression was still significantly
over-expressed in the PBMC of HESN women (p=0.04). Multivariate
analysis also indicated that HESN women who use Depo-Provera have
significantly higher levels of Mx2 than controls.
Conclusions: This is the first epidemiological report of Mx2 and its
association with altered susceptibility to HIV infection. These data
corroborate findings from the in vitro HIV infection studies identifying
Mx2 as a potent innate HIV restriction factor and suggests this molecule
might have utility in HIV prevention strategies.
POSTER DISCUSSIONS
Romel D. Mackelprang1, Kenzie Birsie2, Mary J. Emond3, Adam
Burgener4, Blake Ball2, Jared Baeten3, Connie Celum3, Craig R.
Cohen5, Nelly R. Mugo6, Jairam R. Lingappa3, Partners PrEP Study
Team
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131
Poster Discussions
PD02.03
PD02.04
Low Immune Activation in the Ectocervix of
HIV-exposed Seronegative Commercial Sex
Workers
Plasma and PBMC miRNA Profile in Sexually
HIV-1 Exposed Seronegative Individuals
Kristina Broliden , Anna Gibbs , Taha Hirbod , Terry B. Ball ,
Rupert Kaul3, Joshua Kimani4, Annelie Tjernlund1
1
1
1
2
Karolinska Institutet, Stockholm, Sweden, 2University of Manitoba,
Winnipeg, MB, Canada, 3University of Toronto, Toronto, ON, Canada,
4
University of Nairobi, Nairobi, Kenya
1
Background: Heterogeneity in susceptibility to HIV infection has been
demonstrated in HIV-exposed seronegative (HESN) individuals and
resistance against infection may partly be attributed to a low mucosal
immune activation state.
Methods: Cervical tissue samples were collected from HESN female
sex workers (FSW) and HIV seropositive FSW (HIV+FSW) from the
Pumwani cohort of FSW, Nairobi, Kenya. HIV seronegative lower risk
women (HIV- LR) from the same geographical region were also included
as controls. Protein and mRNA expression of selected immune markers
were assessed by immunohistochemistry and qPCR, respectively, and
compared between the study groups. The thickness of the ectocervical
epithelium was analysed on sequential tissue sections.
Results: The mRNA levels of the cell makers CD3, CCR5, HLA-DR,
DC-SIGN, Langerin, CD69 and of the pro-inflammatory cytokine
markers IFN-alpha, IFN-gamma, TNF-alpha, IL-6, IL-17 and IL-22 were
comparable between the HESN FSW and the HIV- LR study groups. Most
of these markers were however significantly lower in the HESN FSW
study group as compared to the HIV+FSW study group. Furthermore,
the HESN FSW group had significantly lower mRNA expression as well
as protein expression of CD4+ cells as compared to the other two groups
and they had significantly more mRNA and protein expression of CD8
as compared to the HIV- LR group. The ectocervical epithelium of the
HESN FSW samples was generally thicker than that of the other two
study groups.
Conclusions: Despite active commercial sex work these relatively
resistant HESN FSW had a low immune activation state as assessed in
ectocervical tissue samples. They also had relatively low numbers of
HIV target cells (CD4+ cells) and a thicker epithelium than the control
groups. This mucosal immune phenotype may be beneficial in the
resistance against HIV infection.
POSTER DISCUSSIONS
132
Mara Biasin1, Sara Yahyaei1, Mariacristina De Luca1, Irma Saulle1,
Federica Gnudi1, Salomè Ibba1, Micaela Garziano1, Angela
Berzi1, Veronica Rainone1, Daria Trabattoni1, Sergio Lo Caputo2,
Francesco Mazzotta2, Mario Clerici3
University of Milan, Biomedical and Clinical Sciences, Milan, Italy, 2S.
Maria Annunziata Hospital, Florence, Italy, 3Don Gnocchi Foundation,
Milan, Italy
1
Background: MicroRNAs (miRNAs) are small 20- to 24-nt non-coding
RNAs involved in the post-transcriptional regulation of gene expression
which play important defensive roles in several viral infections. Global
expression profiles of cellular miRNAs have identified alterations of
specific miRNAs post-HIV-1 infection both in vitro and in different patient
cohorts suggesting potential roles for miRNA in pathogenesis and
disease progression. We therefore decided to verify if natural resistance
to HIV-1 infection observed in seronegative individuals repeatedly
exposed to HIV-1 (HESN) through unprotected sexual intercourse could
be secondary to a different expression of their miRNA profile.
Methods: expression levels of 25 miRNAs selected according to their
proven anti-HIV-1 properties were analyzed in plasma, basal PBMC and
in in vitro HIV-1 infected macrophages isolated from 30 HESN, 30 HIV
seropositive subjects (HIV+) and 30 healthy controls (HC).
Results: In plasma the expression of mir-155, mir-382, mir-28 and mir198 was significantly augmented in both HIV+ and HESN compared to HC
probably as a consequence of viral exposure. Conversely the expression
of mir-223 and mir-150 in plasma was significantly increased only in
HESN and this result was also confirmed in basal PBMC suggesting
a protective effect for these miRNAs in resistance to HIV-1 infection.
Furthermore, the expression of mir-150 was significantly increased in
HESN macrophages following HIV-1 infection.
Conclusions: mir-223 and mir-150 can target the 3’UTR of HIV-1
transcripts, and they have already been identified as anti-HIV-1 miRNAs.
The higher expression of these miRNA in HESN samples could therefore
represents a key protection mechanism against HIV infection.
PD02.05
The Role of Hormones in Natural Protection
against HIV-1 in the Kenyan HIV-exposed
Seronegative Cohort
Aida Sivro1, Ruey-Chyi Su1, Max Abou2, Joshua Kimani1,3, Walter
Jaoko3, Adam Burgener1, Frank Plummer1,2, T. Blake Ball1,2,3
University of Manitoba, Medical Microbiology, Winnipeg, MB, Canada,
National Microbiology Laboratory, Public Health Agency of Canada,
Winnipeg, MB, Canada, 3University of Nairobi, Medical Microbiology,
Nairobi, Kenya
1
2
POSTER DISCUSSIONS
Background: Studies of populations at high risk of HIV-1 infection
have identified subsets of individuals who are HIV-exposed but remain
seronegative (HESN). Work is in progress to understand the mechanisms
contributing to the observed natural protection against HIV-infection. Sex
hormones, which play crucial roles in regulating immune function, are
also shown to contribute to increased prevalence of sexually transmitted
infections in women. Hormonal contraceptive use and pregnancy, a
state of high estrogen and progesterone levels, have been associated
with increased risk of HIV-1 acquisition. Our earlier study showed
association of three polymorphisms in interferon regulatory factor 1
(IRF1) gene with HIV-resistant phenotype and changes in IRF1 regulation.
This study examined the plasma estrogen, progesterone, cortisol and
prolactin hormone levels with respect to IRF1 polymorphisms and HESN
phenotype.
Methods: Samples used for this study were obtained from the Majengo
Sex Worker Cohort in Nairobi, Kenya. Plasma hormone levels were
measured using Millipore Milliplex Hormone Magnetic Bead kits (Human
Pituitary and Steroid/Thyroid Panels) and analyzed on the BioPlex-200.
Results: Significantly lower prolactin levels were observed in the plasma
samples from women with protective IRF1 genotypes when corrected for
age, use of hormonal contraceptives and menstrual cycle. Interestingly,
levels of all four hormones examined were significantly lower in the
plasma samples from HESN women (with or without the protective IRF1 genotypes) compared to HIV-Susceptible women from the same sex
worker cohort.
Conclusions: These results indicate that hormonal regulation of the
immune environment could be one of the key contributing factors to
the natural resistance against HIV-infection observed in the Kenyan
Majengo cohort. Understanding how hormone levels manipulate the
microenvironment of the female genital tract could greatly contribute
to the development of improved immunization strategies against HIV-1.
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133
Poster Discussions
Poster Discussion 03: Preclinical and Clinical Vaccine Trials
PD03.01 LB
PD03.02
CD8+ T-cell Mediated HIV Inhibition after
Vaccination with a DNA/Recombinant Ad5
(rAd5) HIV Vaccine Is Similar to that Seen in
Treated HIV Infection
DNA/MVA and Protein-based SIV Vaccine
Regimens Delivered to Rhesus Macaques
with Novel Adjuvants Fail to Elicit Neutralizing
Antibodies with Breadth
Nicole Frahm1,2, Bryce A. Manso1, Stephen C. De Rosa1,3, Christina
Ochsenbauer4, Shelly Karuna1, Magdalena Sobieszczyk5, Scott
Hammer5, Edith Swann6, Barney Graham7, Peter Gilbert1,8, Margaret
Juliana McElrath1,2,3, the NIAID HIV Vaccine Trials Network
Megan K. Murphy1, Katherine S. Wetzel2, Katie M. Kilgore1, Stacey
A. Smith1, Samantha L. Burton1, Sharmila Reddy1, Nicholas
Francella2, Donald L. Sodora3, Guido Silvestri1, Kelly S. Cole4,
Francois Villinger1, James E. Robinson5, Bali Pulendran1, Ronald
G. Collman2, Rama R. Amara1, Cynthia A. Derdeyn1
Disease Division, Seattle, WA, United States, 2University of Washington,
Department of Global Health, Seattle, WA, United States, 3University of
Washington, Department of Laboratory Medicine, Seattle, WA, United
States, 4University of Alabama at Birmingham, Birmingham, AL, United
States, 5Columbia University, New York, NY, United States, 6Division
of AIDS, National Institute of Allergy and Infectious Diseases, National
Institutes of Health, Bethesda, MD, United States, 7Vaccine Research
Center, National Institute of Allergy and Infectious Diseases, National
Institutes of Health, Bethesda, MD, United States, 8University of
Washington, Department of Biostatistics, Seattle, WA, United States
1
POSTER DISCUSSIONS
Background: HVTN 505, a phase 2b trial assessing if a DNA/rAd5 HIV
vaccine prime/boost regimen reduces HIV acquisition or viremia post
infection, halted vaccinations in 2013 due to efficacy futility, though
it induced substantial CD8+ T-cell responses. We examined whether
vaccine-induced T cells in HVTN 505 were unable to inhibit virus
replication, as a potential reason for their inability to control viremia
post infection.
Methods: CD8+ T cells from 48 participants (38 vaccine, 10 placebo)
were tested pre-immunization and at peak immunogenicity (4 weeks
post rAd5 boost) for their ability to inhibit HIV-1 replication in autologous
CD4+ T cells infected with a NanoLuc®-secreting JR-CSF infectious
molecular clone. Relative light units (RLU) were measured at day 7, Δlog
inhibition of virus (difference of log RLU in the presence vs. absence of
CD8+ T cells) is reported for an effector:target ratio of 5:1. Responses
were considered positive if above the mean + 3SD of baseline and
placebo samples (1.05 log). Seven subjects with chronic HIV infection on
antiretroviral treatment (ART) served as controls.
Results: Viral inhibition by CD8+ T cells was significantly greater in
vaccinees at peak immunogenicity than pre-immunization (p< 0.0001);
no changes over time were observed in placebo recipients (p=0.7). The
response rate was 61%, with median Δlog inhibition of 1.9 logs (range
1.2-2.6) in responders. This level of inhibition is similar to that in HIVinfected subjects requiring ART to control viremia (median 2.1 logs,
range 1.7-2.8; p=0.06).
Conclusions: The DNA/rAd5 vaccine in HVTN 505 led to significant
induction of CD8+ T-cell responses able to inhibit HIV-1 replication in
vitro, but the magnitude in responders was comparable to that observed
in treated HIV infected subjects requiring ART to control viremia. It is
thus likely that vaccination must elicit responses of higher magnitude
(such as those observed in elite HIV controllers in similar assays) to drive
an overall reduction in viral load post infection.
134
Emory University, Atlanta, GA, United States, 2University of
Pennsylvania, Philadelphia, PA, United States, 3Seattle Biomedical
Research Institute, Seattle, WA, United States, 4University of Pittsburgh,
Pittsburgh, PA, United States, 5Tulane University, New Orleans, LA,
United States
1
Background: Antibodies that neutralize diverse HIV-1 strains are likely
to be important for a protective vaccine. To this end, pre-clinical SIVbased nonhuman primate immunization regimens have been designed
to enhance antibody-mediated protection. Here we investigated whether
neutralizing antibody breadth was elicited by four different DNA/MVA
and protein-based vaccination regimens that included novel adjuvants
such as GM-CSF and TLR ligands, but expressed the same SIVmac239
envelope (Env) glycoprotein.
Methods: A novel panel of 14 diverse SIV Envs described in
previous studies was generated. Neutralization and inhibition assays
were performed using Env pseudoviruses in the Tzm-bl assay. CD4
independent entry was evaluated using 293T cells expressing rhesus
CCR5 with or without CD4.
Results: SIV-infected plasma and monoclonal antibodies targeting V3,
CD4-induced structures, and the CD4 binding site neutralized four of the
Envs, designated as tier 1. Three tier 1 Envs were also CD4 independent,
and only one carried a recently described signature of neutralization
sensitivity (45T/47R) in the gp120 C1 region. The remaining 10 SIV
Envs had varying levels of neutralization resistance. We tested sera
from 91 immunized rhesus macaques for neutralization breadth against
the SIV Env panel. The pooled vaccinated monkey sera from each of
the four trials neutralized only the sensitive tier 1 Envs, regardless of
immunization regimen, and there was no difference between serum
from animals that became infected or remained protected after 12
mucosal challenges within each trial.
Conclusions: These results indicate the SIVmac239 Env immunogen
was the primary determinant of the neutralizing antibody profile,
suggesting that elicitation of antibodies with greater neutralization
breadth will likely require modification or careful selection of the Env
immunogen itself. In addition, this panel of SIV Envs provides a useful
tool for evaluating the neutralization capacity and breadth of antibodies
elicited by SIV-based vaccines.
PD03.03
PD03.04 LB
Antigenicity and Immunogenicity of Disulfidestabilized HIV-1 Envelope Glycoprotein
Oligomers
Clinical Safety and Immunogenicity of Two
HIV Vaccines SeV-G (NP) and Ad35-GRIN in
HIV-uninfected, Healthy Adult Volunteers
Yu Feng1, Hao Jiang1, Jidnyasa Ingale2, Krisa McKee3, John
Mascola3, Richard Wyatt1,2
Etienne Karita1, Omu Anzala2, Brian Gazzard3, Philip Bergin4, Julien
Nyombayire1, Gloria Omosa5, Akil Jackson6, Rosine Ingabire1, Gina
Ouattara5, Harriet Park7, Anne Gumbe5, Kundai Chinyenze5, Sabrina
Welsh7, Carl Verlinde7, Lorna Clark4, Paramesh Chetty8, Mumtaz
Booley8, Jean Bizimana1, Bashir Farah5, Peter Hayes4, Devika
Zachariah7, Kristen Syvertsen7, Michele Fong Lim7, Len Dally9,
Burc Barin9, Makoto Inoue10, Hiroto Hara10, Takashi Hironaka10,
Tsugumine Shu10, Mamoru Hasegawa10, Tetsuro Matano11, Eddy
Sayeed7, Christopher Parks7, Jim Ackland12, Patricia M. Fast7, Jill
Gilmour4, Josephine H. Cox7, Angela Lombardo7, Dagna Laufer7
Background: Our previous study suggests that YU2 gp120 trimers,
deleted of their major variable regions and partially stabilized in the
CD4-bound state display better elicitation of CD4 binding site (CD4bs)directed neutralizing antibodies compared to core and full-length
trimers. The elicited neutralizing antibodies target elements exposed
only on Env of sensitive viruses and cannot neutralize more resistant
isolates. Structural studies reveal a “layered” gp120 architecture in which
three topologically separate and structurally plastic layers act as a shapechanging spacer, facilitating movement between the outer domain and
gp41 to allow movement among alternative conformations required
for virus entry and immune evasion. A recent report demonstrates that
disulfide bond (C65-C115) inserted between layers can lock gp120 in a
CD4 bound state.
Methods: Here, we integrated the disulfide bonds C109-C428 (inner
and outer domain), C65-C115 (layers 1 and 2) and C95-C484 (layers 2
and 3), into the YU2 gp120 trimers.
Results: These stabilized Envs displayed improved stability and
antigenic profiles. The substituted cysteines eliminated binding of most
non-neutralizing CD4bs antibodies while retaining recognition by most
broadly neutralizing CD4bs antibodies. Armed with this promising
antigenic profile, we immunized guinea pigs and non-human primates
with these disulfide-stabilized Envs with or without conjugation onto
liposome particles. The parental trimers once again elicited Tier 1
neutralization, however, introduction of the C109-C428 rendered
antibodies unable to neutralize Tier 1 viruses. However, the stabilized
Envs elicited improved binding antibodies toward the CD4bs and, in
addition, conjugation onto liposomes shifted the immune responses to
the CD4bs.
Conclusions: These results suggest that structure-guided stabilization
can be used to better expose conserved epitopes of gp120 and the
immune responses can be shifted onto the conserved epitopes by
particulate display or by other stabilization strategies.
Project San Francisco, Rwanda Zambia HIV Research Group, Kigali,
Rwanda, 2Kenya AIDS Vaccine Initiative-Kangemi, University of
Nairobi, Nairobi, Kenya, 3Chelsea and Westminster Healthcare
National Health Service Foundation Trust, London, United Kingdom,
4
International AIDS Vaccine Initiative-Human Immunology Lab,
Imperial College, London, United Kingdom, 5Kenya AIDS Vaccine
Initiative University of Nairobi, Nairobi, Kenya, 6Chelsea and
Westminster Healthcare National Health Service Foundation Trus,
London, United Kingdom, 7International AIDS Vaccine Initiative,
New York, NY, United States, 8International AIDS Vaccine Initiative,
Johannesburg, South Africa, 9EMMES Corporation, Rockville, MD,
United States, 10DNAVEC Corporation, Tsukuba, Japan, 11University of
Tokyo, Tokyo, Japan, 12Global BioSolutions, Melbourne, Australia
1
Background: Development of vaccines that stimulate sustained
humoral and/or cellular immunity at mucosal HIV entry points is critical
in the quest for an HIV vaccine. To achieve this goal, we are investigating
replication-competent viral vectors for mucosal delivery that might
mimic the efficacy of live-attenuated viral vaccines.
Methods: A prototype HIV vaccine based on replication-competent
Sendai virus expressing subtype A Gag [SeV-G(NP)] was manufactured.
SeV-G(NP) was administered intranasally (IN) in heterologous prime
boost (P/B) combinations with an Adenovirus-35 encoding subtype
A Gag, RT, Integrase and Nef (Ad35-GRIN at 1x10^10 vp) given
intramuscularly (IM) or in a homologous regimen, all at 0 and 4 months:
Groups A and B were dose escalations (2x10^7, 2x10^8 CIU SeV-G(NP),
respectively) followed by Ad35-GRIN, Group C: Ad35-GRIN followed by
2x10^8 CIU SeV-G(NP); Group D: two doses of 2x10^8 CIU SeV-G(NP).
Sixty-five HIV uninfected adults were enrolled in Kenya, Rwanda and the
United Kingdom. Systemic and mucosal cellular and humoral immune
responses were assessed. We present here preliminary Interferongamma (IFN-y) ELISPOT and serum Gag-p24 binding antibody data.
Results: All vaccine regimens were well-tolerated with no vaccine-related
SAEs. Systemic HIV-specific IFN-y ELISPOT responses were seen in all
recipients but 1 of the heterologous P/B regimen of SeV-G(NP) followed
by Ad35-GRIN. In Groups A and B the magnitude and response rate of
Gag specific T-cell responses indicated that the SeV-G(NP) provided a
strong priming effect (‘hidden prime’), superior to that seen with other
primes such as DNA. Systemic IgG and IgA Gag-p24 antibody responses
were detected in recipients of the heterologous P/B regimen of Ad35GRIN followed by SeV-G(NP).
Conclusions: The combination of IN SeV-G(NP) and IM Ad35-GRIN was
well tolerated. The SeV-G(NP) vaccine appears to enhance both cellular
and antibody responses to Ad35-GRIN. The order of vaccination appears
to determine which immunogenic response is stimulated.
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135
POSTER DISCUSSIONS
International AIDS Vaccine Initiative, IAVI Neutralizing Antibody Center
at TSRI, La Jolla, CA, United States, 2The Scripps Research Institute, La
Jolla, CA, United States, 3National Institute of Allergy and Infectious
Diseases, Vaccine Research Center, Bethesda, MD, United States
1
Poster Discussions
PD03.05 LB
Learning from the Success of HPV Vaccines to
Develop HIV Vaccines that Break B Cell Selftolerance
John T. Schiller1, Bryce Chackerian2
National Cancer Institute, Laboratory of Cellular Oncology, Bethesda,
MD, United States, 2University of New Mexico School of Medicine,
Department of Molecular Genetics and Microbiology, Albuquerque,
NM, United States
1
Background: Even a single dose of an HPV vaccine composed of L1
virus-like particles (VLPs) consistently induces high titer and durable
neutralizing antibody (nAb) responses that afford high vaccine efficacy
against incident HPV infection.
Methods: We believe that the exceptionally potent response induced by
this non-infectious subunit vaccine is largely due to the dense repetitive
display of epitopes on the surface of the VLPs.
Results: Correspondingly, we found that ordered repetitive display of
central self-antigens on VLPs, at a spacing of less than 150Å, efficiently
breaks self-tolerance, resulting in high titer auto-antibodies. Self-antigen
display with this spacing can even reactivate anergic B cells induced by
self-tolerance mechanisms. It follows that HPV VLPs, which have typical
virus epitope spacing of 50-100Å, should be able to induce nAb through
immunoglobulin gene developmental pathways that involve self-reactive
intermediates. Since the majority of naïve B cells are self-reactive, this
ability likely makes a critical contribution to the consistently potent nAb
responses after even a single VLP dose.
Conclusions: HIV has an exceptionally low number of receptor binding
determinants on its surface. The paucity of envelope spikes likely
contributes to its exceptionally low transmission rate. The spacing of HIV
envelope spikes has been estimated to be 230Å, and so we postulate
that neutralizing antibodies to HIV, unlike typical viruses, cannot
develop through self-reactive intermediates. This might explain why
nAb, especially broadly cross-neutralizing ones, are produced through
highly convoluted developmental pathways. In sacrificing transmission
efficiency for delayed induction of nAb, HIV may have evolved an
Achilles heal that could be exploited by vaccines that can generate nAbs
through self-reactive intermediates. We believe that development of
vaccines displaying HIV envelope epitopes at high density should be a
major focus HIV vaccine development.
POSTER DISCUSSIONS
136
PD04.01
PD04.02
VOICE-C Participant Narratives of Rape:
What they Mean for Female-initiated HIV
Prevention Products
“He Said, She Said.” Exploring Couples’
Sensory Perceptions and Experiences
with Vaginal Gels & Film: Implications for
Microbicide Development
Miriam A. Hartmann1, Elizabeth T. Montgomery1, Jonathan
Stadler2, Nicole Laborde1, Ariane van der Straten1,3, on behalf of
the VOICE-C team
RTI International, Women’s Global Health Imperative, San Francisco,
CA, United States, 2University of the Witwatersrand, Wits Reproductive
Health and HIV Institute, Faculty of Health Sciences, Johannesburg,
South Africa, 3UCSF, Center for AIDS Prevention Studies, Department of
Medicine, San Francisco, CA, United States
1
Background: Numerous recent trials have sought to develop femaleinitiated methods of HIV prevention; in part to address barriers to
women’s prevention rooted in unequal gender norms. Understanding the
gender-related context in which these products may be used, including
levels of violence against women, is critical. MTN-003C (VOICE-C), a
qualitative sub-study of the MTN-003 (VOICE) HIV prevention trial,
examined socio-cultural barriers and facilitators to product use within
the Johannesburg site.
Methods: We conducted and analyzed focus group discussion (FGD),
in-depth interview (IDI), and serial ethnographic interview data from 102
female VOICE participants, 22 male partners, 17 community advisory
board (CAB) members, and 23 community stakeholders. Violence was
not a designated interview topic; however it arose through discussions
of community context, interpersonal relationships, and product use. For
this analysis, all textual data coded as “violence” was systematically
reviewed.
Results: The data revealed the prominence of sexual violence in
women’s lives. Rape was discussed in 2/4 FGDs with CAB members, 2/3
FGDs with stakeholders and among a quarter of interviews/FGDs with
VOICE female participants; two of whom described personal experiences
of rape. These narratives demonstrated a pervasive perception that
women are vulnerable to rape and that this threat contributes to
their susceptibility to HIV. The possibility of rape was used to reframe
their HIV risk as external to their own or their partner’s behavior and
was ultimately used to rationalize the importance of female-initiated
products in HIV prevention.
Conclusions: Fear or experience of rape is pervasive in this community,
reflecting underlying gender inequalities, which in turn are likely to
influence how HIV prevention products are perceived and used. While
the actual impact on product use in VOICE is uncertain, results illustrate
how women, in contexts of high sexual violence, may utilize existing
unequal gender norms to negotiate their use.
Kathleen M. Morrow1, Rochelle K. Rosen2, Joseph L. Fava3, Lisa
Rohan4, Erna M. Kojic5, David Friend6, David Katz7, Robert
Buckheit8
Miriam Hospital & Alpert Medicial School of Brown University, Centers
for Behavioral and Preventive Medicine, Providence, RI, United States,
2
Miriam Hospital & Brown University School of Public Health, Center
3
Miriam Hospital, Center for Behavioral and Preventive Medicine,
Providence, RI, United States, 4Magee Womens Research Institute,
Pittsburg, PA, United States, 5Miriam Hospital & Alpert Medical School
of Brown University, Division of Immunology, Providence, RI, United
States, 6CONRAD, Arlington, VA, United States, 7Duke University,
Biomedical Engineering and Ob/Gyn, Durham, NC, United States,
8
ImQuest BioSciences, Inc., Frederick, MD, United States
1
Background: Perceptibility science, the objective measurement of user
sensory perceptions (sensations) and experiences (USPE) of formulation
performance during use, is a new concept. For vaginal gels, sets of
rheological and other biophysical properties, including measures of
spreading and retention, may critically impact user experiences. For
vaginal films, other properties, e.g., disintegration time or puncture
strength, may play additional roles.
Methods: In a mixed methods study, 24 monogamous HIV-/STIheterosexual couples completed 3 formulation evaluation visits (100%
retention). Following vaginal sex, participants (ppts) independently rated
USPEs for 2 volumes of HEC gel (2 mL; 4 mL) and 1”x2” vaginal film.
8 USPE scales were scored at each visit for each participant. Pairwise
comparisons between females (F) and males (M) were conducted on
each form.
Results: There were no significant differences between F and M USPE
evaluations for low volume gel. For high volume gel, there was a
significant difference in Initial Penetration scores, with F>M (4.3 v 3.8,
p=.012; large effect size (ES) d=.78). Perceived Leakage scores were also
higher in F than M (2.4 v 2.1; medium ES d=.53), and near significant
(p=.08). For film, a significant difference was seen in the Stimulating
scale score with M>F (2.0 v 1.4, p=.014; large ES d=.77). All other USPE
scale score comparisons (21 of 24) across F & M were strikingly similar.
Both F & M ppts reported greater sensory experiences overall with gels
than films. High volume gel was slightly favored in both genders (F=11,
M=10). Contrasts occurred in choices between low volume gel and film:
more F chose low volume gel (F=9) and more M chose film (M=9).
Conclusions: A non-optimized user experience could compromise an
optimized drug and its delivery. Perceptibility, for both members of a
sexual dyad, should be considered when designing and advancing
potential vaginal (and rectal) formulations for HIV/STI prevention and
multipurpose prevention technologies.
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137
POSTER DISCUSSIONS
Poster Discussion 04: Behavioral and Social Sciences
Poster Discussions
POSTER DISCUSSIONS
PD04.03
PD04.04
Successfully Addressing Challenges to
Implementing a Multinational SMS-based
Reminder and Data Collection System in a
Biomedical HIV Prevention Trial
Reimagining HIV Testing in an Era of ART
Martina Brostrom1, Ruben Granich1, Somya Gupta1, Badara
Samb1
UNAIDS, the Joint United Nations Programme on HIV/AIDS, Geneva,
Switzerland
1
William Brown III1,2, Rebecca Giguere1, Mobolaji Ibitoye1, Alex
Carballo-Diéguez1, Ross D. Cranston3
Columbia University, HIV Center for Clinical and Behavioral Studies,
New York, NY, United States, 2Columbia University, Biomedical
Informatics, New York, NY, United States, 3University of Pittsburgh,
Department of Medicine, Pittsburgh, PA, United States
1
Background: Adherence to product use in a clinical trial is critical
to assessing safety and efficacy. Real-time data collection can help
monitor adherence. The ubiquity of Short Message Service (SMS) allows
researchers to collect adherence data and provide behavioral reminders
remotely in real-time. MTN-017 is a phase 2 safety and acceptability
study of oral Truvada tablets and rectally-applied tenofovir gel. To
monitor adherence in MTN-017, we sought to
1) identify an SMS system that allowed for daily reminders and collection
of data on product use,
2) implement it in four countries and five languages, and
3) develop a centralized data management system with an automated
backup system.
Methods: We assessed feature availability of several SMS systems based
on ideal criteria, including keyword response, email capability, participant
identification and grouping, text message scheduling, multiple language
operating system, and international SMS capability. After identifying the
optimal SMS system, we systematically implemented it in each country,
working with IT staff at clinical research sites.
Results: We successfully addressed 24 critical challenges pre- and
post-implementation. Solutions included: developing a federated
SMS-system architecture to mitigate SMS message costs and manage
data access; using secure email protocols to centralize data backup,
developing several programming syntaxes to facilitate daily data
analysis, developing a calendar template for reporting SMS behavior
to sites, ambiguation of text message language to increase privacy, and
standardizing operating systems and hardware to minimize variability
in system performance. Other solutions and metrics for estimating cost
effectiveness will be discussed.
Conclusions: Our tests and continued use have allowed us to identify
factors that should be consistent across countries to ensure smooth
implementation and operation of SMS as an adherence reminder system
and real-time data collection modality.
138
Background: UNAIDS launched Treatment 2015 to accelerate treatment
access through the concepts of innovation, speed and focus. In
2013,12,9 million of 35 million people living with HIV globally were
receiving treatment. UNAIDS has proposed a 90-90-90 target for 2020
- 90% of people living with HIV tested, 90% of those tested on ART
and 90% of those on ART virally suppressed. Ensuring treatment and
viral suppression for people living with HIV will require special efforts
to ensure that people living with HIV but who do not know their statushave access to HIV testing.
Methods: We reviewed the national strategies of 10 high impact
countries in Africa and identified gaps and strategic opportunities for
innovating HIV testing.
Results: Harnessing the therapeutic and preventive benefits of ART, will
entail a massive scale up and reorientation of testing efforts in countries
where HIV testing deficits limit access to ART. An estimated 55% of
people living with HIV in SSA are unaware of their HIV status. Data
from select countries of people ever tested in SSA, where treatment
coverage is 37%, ranges from very low to moderate or high and with
some countries having a significant gender gap (where the proportion
of men ever tested is low but the range among women is moderate
or high). Often testing coverage is mismatched with the epidemic
realities of countries and testing policy is not in aligned with treatment
expansion targets. This calls for revised programmatic targets for testing
and treatment, high yield strategies and new benchmarks for quality HIV
care, with an attention to location, gender and age.
Conclusions: HIV testing programmes in many countries are inadequatethey were conceptualized when AIDS was an emergent and frightening
phenomenon- now they need to address mature epidemics, where there
is approaching universal access to treatment which allows HIV infection
to be non-life threatening, and where the concentrations of HIV in the
most affected populations in different contexts is much more known.
Key areas for innovation include self-testing, multi-disease community
campaigns and PoC testing for CD4 and VL for first line service delivery.
POSTER DISCUSSIONS
PD04.05
The Need for Demonstration Projects to
Ensure Key Populations Gain Access to New
HIV Prevention Biomedical Tools in South
Africa
Brian Kanyemba1, Ben Brown1, Maaza Seyoum2
Desmond Tutu HIV Foundation, IIDMM University of Cape Town,
Cape Town, South Africa, 2International AIDS Vaccine Initiative (IAVI),
1
Background: Sero-discordant couples and Key Populations(KP) including, but not limited to, men who have sex with men (MSM), sex
workers (SWs) and people who inject drugs (PWID) - urgently need
new, effective HIV prevention methods. Numerous studies have shown
that pre-exposure prophylaxis (PrEP) and treatment as prevention (TasP)
offer effective HIV prevention to these vulnerable groups. Prior to the
2013 South African AIDS Conference, a pre-conference satellite was
conducted to gain key stakeholders’ perspectives on the implementation
of demonstrations projects for KP.
Methods: After a plenary session that reviewed current HIV prevention
research and advocacy, 50 delegates were divided into four groups to
debate opportunities and challenges for demonstration projects. The two
key questions were: 1) Which HIV prevention research methods should be
prioritized for demonstration projects among KP in SA; and 2) What will
the main challenges be? The session was divided into four 15-minute
focus groups, where the delegates circulated through topics. Facilitators
captured key findings.
Results: There was strong agreement that PrEP and TasP should
be available to MSM, SW, and PWID in SA. In order to achieve this
goal, demonstration and pilot projects would be a valuable source
of data on the implementation of ARV based prevention outside the
clinical trial setting. The key recommendations focused on the need to
implement demonstration projects that are tailored to priorities, needs
and circumstances of diverse KP groups. Such projects should answer
questions on adherence, the impact of stigma in health care settings,
and access to care despite structural barriers such as criminalization of
some KP.
Conclusions: Stakeholders who participated in the session highlighted
need for PrEP access among KP in SA (and other African countries) in
order to begin addressing effectiveness, acceptability, adherence and
behavioral issues. Research, policy, and advocacy agendas, should
consider these in the future.
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139
Poster Discussion 04:
Behavioral and Social
Sciences
Poster Discussions
Poster Discussion 05: Glycans and Antibody Effector Functions
POSTER DISCUSSIONS
PD05.01
PD05.02
Exploring the Influence of N-linked Glycans
on the Molecular Dynamics of HIV-1 gp120 in
the Context of Viral Coreceptor Tropism
Variable Dependence on Glycan Recognition
within a Lineage of V1V2-directed HIV
Neutralizing Antibodies
Natasha T. Wood1, Elisa Fadda2, Oliver C. Grant3, Robert J.
Woods4, Simon A. Travers1
Nicole A. Doria-Rose1, Ryan S. Roark1, Penny Moore2, Michael
J. Ernandes1, Jinal N. Bhiman2,3, Chaim A. Schramm4, Krisha
McKee1, Sijy O’Dell1, Mark Louder1, Salim S. Abdool Karim3,
Lawrence Shapiro4, Lynn Morris2,3, John R. Mascola1
University of the Western Cape, South African National Bioinformatics
Institute, Cape Town, South Africa, 2National University of Ireland
Maynooth, Chemistry, Maynooth, Ireland, 3National University of
Ireland Galway, Chemistry, Galway, Ireland, 4University of Georgia,
Complex Carbohydrate Research Centre, Athens, GA, United States
1
Background: The dense glycan shield of the HIV-1 gp120 surface
protein protects the virion from recognition and neutralisation by the
host immune system, plays a role in binding to target cell chemokine
receptors (coreceptors), and forms part of viral epitopes for broadly
cross-neutralising antibodies. The composition and distribution of these
N-linked glycans also vary depending on the cell-type, stage of infection,
and HIV subtype.
We have previously applied molecular simulation techniques to show
that the spatial dynamics of the gp120 glycoprotein are significantly
influenced by the composition of the underlying protein sequence as
well as the presence of glycans at specific N-linked glycosylation sites.
Methods: To continue this work and further explore the effect of the
glycan composition and distribution on HIV coreceptor tropism, we have
modelled the structures of three pairs of viral clones from three patients.
Each sequence pair consisted of a CCR5- and CXCR4-tropic virus, which
had been clonally phenotyped, and represented subtype A, C, and D
infections. Oligomannose and complex glycans were linked to N-linked
glycosylation sites of each structure based on the reported predominant
glycan-type at specific positions along the gp120 glycoprotein. We
subsequently applied a molecular dynamics approach, using GROMACS,
to identify HIV-1 coreceptor tropism-associated structural features
introduced by glycans on the surface of gp120.
Results: Our preliminary results align with our previous work where
the presence of glycans had a substantial impact on the dynamics of
the V3 loop. Further analysis reveals the degree to which the glycan
composition and density around key regions of HIV-1 gp120 impact the
tropism-associated dynamic characteristics of the protein.
Conclusions: These results present a unique view on how the glycanprotein, as well as the glycan-glycan, interactions of HIV-1 gp120 may
modulate the infectivity and immunogenicity of the virion.
140
National Institutes of Health, Vaccine Research Center, NIAID,
Bethesda, MD, United States, 2University of the Witwatersrand and
the NICD, Johannesburg, South Africa, 3Centre for AIDS Programme of
Research In South Africa (CAPRISA), Congella, South Africa, 4Columbia
University, New York, NY, United States
1
Background: The study of HIV donors with broad and potent neutralizing
antibodies can inform rational vaccine design and immunization
strategies. We previously identified 12 antibodies from a single lineage
in donor CAP256 that target V1V2 with breadth up to 60% in clades
A and C, and very high potency on many strains. The antibodies varied
in dependence on glycan. We sought to isolate additional lineage
members and to understand variation in epitope recognition.
Methods: Antibodies were isolated via 14-day B cell culture with IL-2, IL21, and CD40L. Initial RTPCR was performed using IgG-specific primers.
For weeks 119 and 206, wells were re-interrogated using IgA-specific
primers. 5 additional antibodies were isolated from week 119 and 4
from week 206, and reconstituted as IgG1 and IgA1. The IgGs were
assayed for neutralization on TZM-bl cells on a panel of 49 autologous
and heterologous Env-pseudoviruses, as well as viruses with mutations
in known epitopes.
Results: Several B cell culture wells yielded both IgG and IgA amplicons
with the same VDJ sequence, suggesting that class-switching occurred
during culture. Nine new antibodies of the CAP256-VRC26 lineage
were identified, several of which had breadth similar to the previous
best members of the lineage. Two from week 206 lack the characteristic
tyrosine sulfation signal, and are also less broadly neutralizing. All
depend on residues in V2, with variable and strain-specific effects of
glycans. We examined the impact of removing the glycan at N160
which is critical for the PG9 class: some lost neutralization; some were
unaffected; and CAP256-VRC26.13, and two nearly identical antibodies
cloned independently, showed gain of neutralization, the opposite of
PG9.
Conclusions: We have expanded the CAP256-VRC26 clonal family. Its
members show diversity in recognition of specific determinants within
the V1V2 epitope, particularly the glycans, which vary in the virus as a
means of escape from antibody pressure.
l
PD05.03
PD05.04
Polyfunctional Non-neutralizing Fc-antibody
Responses in Acute HIV Infection Predict
Spontaneous Viral Control
HIV-1 Env-specific IgA/IgG Ratio Is Related
to Antibody Dependent Cellular Cytotoxicity
(ADCC) Responses Observed during Acute/
Early HIV Infection
Amy W. Chung1, Matthew K. Schoen1, Hannah Robinson1, Anna
Licht1, Elizabeth Tkachenko1, Davey M. Smith2, Susan J. Little2,
Douglas D. Richman2, Galit Alter1
States, 2University of California San Diego, San Diego, CA, United States
1
Maria Julia Ruiz1, Yanina Ghiglione1, Juliana Falivene1, María
Eugenia Socías2, Natalia Laufer1,3, Pedro Cahn2,3, Omar Sued2,3,
María Magdalena Gherardi1, Horacio Salomon1, Ana Maria
Rodriguez4, Gabriela Turk1
Universidad de Buenos Aires/CONICET, Buenos Aires, Argentina,
2
Fundación Huesped, Buenos Aires, Argentina, 3Hospital Juan A.
Fernández, Unidad Enfermedades Infecciosas, Buenos Aires, Argentina,
4
Instituto de Inmunología, Genética y Metabolismo (INIGEM),
Universidad de Buenos Aires-CONICET, Buenos Aires, Argentina
1
Background: Analysis of the moderately protective RV144 vaccine trial
revealed that vaccination induced non-neutralizing antibodies (Abs)
capable of mediating polyfunctional, highly co-ordinated Fc-effector
responses. Additionally, non-neutralizing Fc-effector functions have been
associated with delayed disease progression and strength of Fc-effector
activity in acute infection correlates with decreased acute viremia. Thus
we hypothesized that non-neutralizing Fc-effector functions in acute
infection may contribute to spontaneous HIV viral control.
Methods: IgG was purified from plasma of 11 acutely infected chronic
(CRO) subjects and 9 spontaneous controllers (CTR) at 4, 12, 24 and 48
weeks post infection. Non-neutralizing Fc-activity: Ab dependent cellular
cytotoxicity (ADCC), Ab dependent cellular phagocytosis (ADCP), Ab
dependent NK cell activation and Ab dependent cellular virus inhibition
(ADCVI), along with HIV-specific IgG subclass profiles were characterized
across all time points.
Results: Despite higher observed ENV-specific bulk IgG titers in CRO
subjects, similar levels of ADCC, ADCP and Ab NK activation were
detected from the two cohorts. However CTRs developed significantly
higher ADCVI activity than CROs at later timepoints. Subclass analysis
revealed that ADCVI activity was correlated with ENV-specific IgG3,
which was maintained at high levels within CTRs, in comparison to
CROs, which progressively lost their IgG3. In addition, CTRs were able
to induce highly co-ordinated polyfunctional Fc-effector functions, while
in contrast, inversely correlated Fc-responses were detected from CRO
subjects at early timepoints.
Conclusions: This data suggest that spontaneous controllers of HIV
infection are able to induce highly co-ordinated Fc-effector responses,
along with maintained high levels of IgG3, that may contribute to HIV
control. Thus vaccine strategies able to induce high and sustained levels
of polyfunctional Fc-effector responses along with IgG3 may provide
protection from and after infection.
Background: Immune correlates analysis of the RV144 vaccine trial
revealed that high levels of Env specific IgA antibodies (ab) may have
mitigated the capacity of protective vaccine-induced IgG ab to mediate
ADCC.
Aim: To evaluate the relation between Env-specific IgG and IgA ab and
ADCC responses in the context of acute/early primary HIV infection (PHI)
Methods: Plasma from 20 HIV+ subjects enrolled during PHI, obtained
at enrollment (baseline sample) and 1 year post-infection sample (ypi),
10 HAART-naïve chronically infected patients (C) and 7 Elite Controllers
(EC) were used. Percentage of ADCC-killing was determined using the
rapid and fluorometric ADCC assay. Env-specific IgG and IgA ab were
assessed by ELISA. Data was analyzed using non-parametric statistics
Results: Levels of Env-specific IgG ab were significantly higher in PHI ypi
sample compared to baseline (median 2802 vs 24902 ng/ml p=0.006)
and as expected directly correlated with %ADCC-killing both at baseline
(r=0,476 p= 0,039) and ypi samples (r=0,627 p=0,016). Similar direct
correlations were observed in C and EC (r=0,898 p=0,033 and r=0,766
p=0,021 respectively). Conversely, IgA titers declined over the course of
acute infection (baseline vs ypi: 200 vs 35 respectively, p=0,001) and
did not correlate with ADCC responses observed at the ypi sample. Envspecific IgG/IgA ratios were also significantly higher at the ypi sample
compared to baseline (median 556,5 vs 10,39, respectevly, p= 0,0001).
More interestingly, Env-specific IgG/IgA ratios directly correlated with
%ADCC-killing at the ypi sample (r=0,538 p=0,047)
Conclusions: The direct correlation between IgG/IgA ratio and %ADCCkilling observed at the ypi sample indicate that Env-specific IgA ab might
be interfering with the magnitude of IgG-mediated ADCC responses in
subjects undergoing PHI. This result indicates that the elicitation of
Env-specific IgA ab hampers protective ADCC responses not only in
vaccinees but also in natural infection and should be taken into account
in vaccine settings
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141
POSTER DISCUSSIONS
Poster Discussions
PD05.05
POSTER DISCUSSIONS
Fcγ Receptor Functional Variability Impacts on
Mother-to-Child Transmission of HIV-1
Ria Lassauniere1,2, Glenda E. Gray3, Louise Kuhn4, Caroline T.
Tiemessen1,2
National Institute for Communicable Diseases, Centre for HIV and
STIs, Johannesburg, South Africa, 2University of the Witwatersrand,
Faculty of Health Sciences, Johannesburg, South Africa, 3Perinatal HIV
Research Unit, Chris Hani Baragwanath Hospital, Soweto, South Africa,
4
Gertrude H. Sergievsky Centre, College of Physicians and Surgeons
and Department of Epidemiology, Mailman School of Public Health,
Columbia University, New York, NY, United States
1
Background: The study model of maternal-infant HIV-1 transmission
provides insights into what may constitute protective immunity to
HIV-1. Using this model, we investigated the potential role of gene
variants known to influence Fc gamma receptor (FcγR) mediated effector
mechanisms that include antibody-dependent cell mediated cytotoxicity
and phagocytosis.
Methods: We genotyped FcγR functional variants and gene copy
number in 207 HIV-1 infected mothers and their infants: 138 were
non-transmitting mothers and their exposed-uninfected infants; 69
transmitting mothers and their intrapartum or in utero infected infants.
The following functional variants were investigated: FcγRIIa-p.H131R,
FcγRIIb-p.I232T, FcγRIIc-p.T118I, FcγRIIIa-p.F158V, FcγRIIIb-HNA1a/
HNA1b/HNA1c, FCGR2B/C promoter variants, and genetic markers
predicting FcγRIIc expression.
Results: Mothers carrying at least one FcγRIIIa-158V allele had a
68% reduction in the odds of transmitting HIV-1 in utero (OR 0.318,
95%CI: 0.15-0.66, P = 0.002) compared to mothers who did not
carry a 158V allele. When mothers and infants possessed discordant
genotypes, in utero transmission of HIV-1 was associated with the
FcγRIIIa-158FF genotype in the mother (OR: 2.959, 95%CI: 1.197.356, P = 0.021), while in utero acquisition was associated with the
FcγRIIIa-158FV genotype in the infant (OR 2.271, 95%CI: 1.05-4.91, P
= 0.039). In concordant mother-infant pairs, an increased odds of HIV1 transmission/acquisition in utero was independently associated with
homozygosity for the FcγRIIb-232T allele (OR 3.208, 95%CI: 1.0110.16, P = 0.048) and the presence of at least one FcγRIIIb-HNA1c
allotype (OR 2.37, 95%CI: 1.11-5.08, P = 0.035).
Conclusions: Antibody affinity variability conferred by the FcγRIIIaF158V allele, and lower inhibitory activity of FcγRIIb in both mother
and infant was associated transmission and acquisition of HIV-1.
These findings suggest a role for FcγR-mediated effector functions in
transmission and acquisition of HIV-1.
142
PD06.01
PD06.02
To Use or Not to Use Clade Specific Vaccines: A
Central Question
The Tipping Point: Moving from Rhetoric to
Real Milestones for Ending AIDS
Dobromir Dimitrov1, James Kublin1, Scott Ramsey1, Larry Corey1
Emily Bass1, Emily D. Donaldson1, Mitchell Warren1, Kevin Fisher1
Fred Hutchinson Cancer Research Center, Seattle, WA, United States
AVAC, New York, NY, United States
1
1
Background: Recent studies support the hypothesis that immune
responses to HIV-1 are markedly influenced by clade. Vaccines currently
entering efficacy trials are clade C. We discuss whether these vaccines
should be used in other regions before clade-specific prototypes are
constructed and evaluated.
Methods: Mathematical models were used to simulate the impact of
vaccination strategies on the HIV epidemics in South Africa (SA) and
San Francisco (SF). We compared three interventions based upon a 50%
vaccine efficacy (VE) of a clade-specific vaccine, reduced to 20-40% VE
in non-clade matching regions:
i) immediate use of non-matching vaccine;
ii) delayed intervention by developing a clade-specific vaccine and
iii) immediate use of non-matching vaccine replaced by a clade-specific
vaccine when developed.
Results: While implementation of a maximally effective vaccine
provides the greatest population benefits, immediate vaccination with a
20-40% VE may prevent thousands of new infections in SF and millions
in SA over 30 years reducing the HIV prevalence by up to 7 %.
Vaccination with 50% VE delayed for 5 years needs 6 and 12 years
to break-even with immediate vaccination with 20 and 30% VE,
respectively, but is not able to match the 30-year impact of immediate
vaccination with 40% VE. Replacing a vaccine with 30% VE with 50%
VE vaccine after 5 years prevents 5% more infections than delayed
vaccination with 50% VE but these benefits may be lost if vaccinees
reduce condom use.
Conclusions: The immediate use of a non clade matching HIV vaccine
may be desirable if it does not lead to changes in sexual behavior that
result in increased HIV infection risk. The pursuit of higher VE through
clade matching prototypes will hasten the reduction in HIV prevalence
over time if the initial vaccination program does not impact adherence to
subsequent vaccination programs. This study illustrates the importance
of developing biomarkers of HIV-1 acquisition to support more rapid and
economical assessment of clade specific vaccines.
Background: The epidemiologic “tipping point” in the AIDS epidemic
is the point at which the rate of ART treatment scale-up outpaces HIV
incidence. AVAC and amfAR analyzed modeling research and consultated
with top HIV prevention experts to lay out essential steps that must be
taken by national governments, international organizations, civil society,
researchers and technical agencies to steadily reduce annual new HIV
infections and continue to expand HIV treatment access in order to meet
the tipping point.
Methods: The analysis used modelling to build a prevention advocacy
agenda around ending AIDS. The targets set reflect best-case scenario
calculations based on published modeling and epidemiologic data, as
well as analysis provided by experts in the field. Data projections were
cross-checked with modelers and epidemiologists. Modelling data is
tracked and updated to ensure the most recent metrics are used, and
real-time data is included and analyzed as available.
Results: By engaging with modelling data, advocates are able to set
real, measurable targets and hold accountable national and global
agencies to ensure key data points are met, including decreasing new
HIV infections and deaths, as well as specific epidemiologic and policybased milestones tied to global scale-up of critical interventions. Rate of
treatment coverage and the rate at which incidence decreases are key
to the analysis—thus, real-time assessment of results allows advocates
to use informed decision-making to ensure these time-related targets
kept on track.
Conclusions: By using data-driven targets, such as the tipping point,
as an advocacy tool, advocates can translate complex models for
policy makers and donors to ensure continued and timely scale-up of
core interventions and sustained resources. It is essential to continue
scale-up and funding of key interventions and research towards ending
AIDS, establishing a set of targets that advocates can track globally and
nationally ensures movement towards, and beyond, the tipping point.
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143
POSTER DISCUSSIONS
Poster Discussion 06: Policy, Advocacy and Modeling
Poster Discussions
POSTER DISCUSSIONS
PD06.03
PD06.04 LB
Bioethical and Biostatistical Considerations of
Innovative HIV Prevention Trial Designs
Differences in PrEP Knowledge and Use
in U.S. MSM Users of a Popular Sexual
Networking Site Surveyed in August 2013
and January 2014
Darpun D. Sachdev1,2, Ryan Whitacre3, Susan P. Buchbinder2,4
Center for AIDS Prevention Studies (CAPS), UCSF, San Francisco, CA,
United States, 2San Francisco Department of Public Health, Bridge
HIV, San Francisco, CA, United States, 3University of California at San
Francisco, San Francisco, CA, United States, 4University of California
at San Francisco, Department of Epidemiology and Biostatistics, San
Francisco, CA, United States
1
Background: Given the efficacy of pre-exposure prophylaxis (PrEP),
there is ongoing debate regarding whether future placebo-controlled
biomedical HIV prevention trials are ethically justifiable. Several
innovative trial designs under consideration to test new biomedical HIV
prevention modalities provide PrEP as part of the standard of prevention
or use PrEP as an active control, however, little is known regarding the
ethical considerations of these designs.
Methods: We purposively sampled and conducted in-depth interviews
with 2 biostatisticians and 3 bioethicists, each with >10 years
of experience in HIV prevention trials. We sought to understand
potential HIV vaccine or long-acting PrEP trial designs that were under
consideration and preferences toward the designs. We analyzed
interview transcripts using constant comparative methods to inductively
develop and refine themes.
Results: Several innovative HIV prevention trial designs were elicited
including factorial designs, pre-randomization PrEP run-in periods, noninferiority studies and adaptive trials. Respondents agreed that PrEP
should be offered in future trials unless there is ethical justification
precluding inclusion. Adaptive trial designs, which allow for modification
of a trial in response to study data, were highly acceptable to both
biostatisticians and bioethicists due to their potential to: 1) reassess
clinical equipoise during the course of the study, 2) utilize informed
consent to apprise participants of prespecified trial design changes, and
3) save time and resources while also assessing the number of clinical
endpoints necessary for regulatory approval.
Conclusions: Adaptive trial designs may allow for scientifically rigorous
studies to evaluate the efficacy of new biomedical HIV prevention
modalities while also maximizing benefits to trial participants. Ongoing
interviews with investigators and regulators will evaluate for the “real
world” limitations of these designs.
144
Kenneth H. Mayer1,2, Catie Oldenburg3,4, David S. Novak5,
Douglas Krakower1,6, Matthew J. Mimiaga1,4,7
Fenway Health & Harvard University, The Fenway Institute, Boston,
MA, United States, 2Beth Israel Deaconess Medical Center, Infectious
Disease Division, Boston, MA, United States, 3Fenway Health, The
Fenway Institute, Boston, MA, United States, 4Harvard School of Public
Health, Epidemiology, Boston, MA, United States, 5On-Line Buddies,
On-Line Buddies Research Institute, Cambridge, MA, United States,
6
Beth Israel Deaconess Medical Center, Division of Infectious Diseases,
Boston, MA, United States, 7Massachusetts General Hospital / Harvard
Medical School, Psychiatry, Boston, MA, United States
1
Background: Pre-exposure prophylaxis (PrEP) has been recommended
for HIV prevention for at risk men who have sex with men (MSM). Prior
reports have suggested slow uptake.
Methods: U.S. MSM members of a sexual networking site were invited to
complete a survey in August, 2013 (Wave 1: W1) and January, 2014 (W2)
that included a variety of questions related to HIV prevention practices.
Comparisons of those who responded at each time point were conducted
using Fisher’s exact tests in order to assess differences in PrEP knowledge
and uptake between W1 and W2. Study design did not permit assessment
of whether MSM responded once or at both time points.
Results: Although more MSM responded in W1 than W2 (6,683 vs.
4,759), they were similar in median age (45 y.o.), race/ethnicity
(about 86% White), educational attainment (about 2/3 had at least a
college degree) and regional geographic distribution (almost ¼ from
the Southeast, with the next largest numbers being in the Northeast,
Great Lakes region or Midwest-about 15% each). The majority reported
engaging in condomless anal sex (CAS) in the prior 3 months (58.7% in
Wave 1 and 74.1% in Wave 2). A higher proportion of MSM in W2 had
heard of PrEP than in W1 (43.8% vs. 27.3%, p< 0.001). Among MSM
reporting CAS in the prior 3 months, PrEP interest increased between W1
and W2 (54.1% vs. 60.4%, p=0.002) and use also increased (2.0% vs.
3.1%, p=0.004). PEP knowledge increased (38.9% and 49.7% in W1
and W2, p< 0.001), but PEP utilization among MSM reporting CAS in
the prior 3 months did not significantly increase (4.3% in W1 and 4.7%
in W2, p=0.44).
Conclusions: Although recent increases in PrEP interest and use among
U.S. MSM accessing a sexual networking site were detected, the
majority of those who could benefit have not availed themselves of this
prevention modality, with the majority of at risk respondents not being
knowledgeable about PrEP. Further educational efforts are needed for
U.S. MSM about PrEP.
POSTER DISCUSSIONS
PD06.05 LB
Guidelines on Post Exposure Prophylaxis for
HIV: Recommendations for a Public Health
Approach
Nathan Ford1
HIV/AIDS, World Health Organization, Geneva, Switzerland
1
WHO has recently updated recommendations on the use of antiretroviral
drugs as post exposure prophylaxis (PEP) to prevent HIV infection.
The guidelines are based on a public health approach that considers
feasibility and effectiveness across a variety of settings. In producing
these guidelines the key principles of availability, acceptability,
accessibility and quality have been considered. In contrast to previous
published WHO guidelines, these guidelines consider all exposures
and provide recommendations for all populations. In doing so no
distinction between occupational and non-occupational settings has
been made with the aim of simplifying guidance for PEP prescribing.
These guidelines promote the approach that individuals should be
offered PEP if the exposure constitutes a significant risk of transmission
and the same drug regimen should be prescribed irrespective of
exposure source. Recommendations for preferred regimens, simplifying
prescribing approaches and supporting adherence are also provided.
www.hivr4p.org
145
Overview of Poster Sessions
Posters Presented on Wednesday, 29 October
P01: Acute Seroconversion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 148
P02: Community Engagement and Advocacy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150
P03: Correlates of Protection and Exposure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
P04 : Correlates of Protection in Highly Exposed Seronegative People. . . . . . . . . . . . . . . 158
P05: Family Planning. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
P06: Financial Incentives. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
P07: Good Participatory Practices and Community Involvement. . . . . . . . . . . . . . . . . . . . 165
P08: HIV Care. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169
P09: HIV Testing and Counseling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171
P10: Immunogens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180
P11: Informed Consent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
P12: Innate Immunity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190
POSTERS
P13: Key Populations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 199
P14: MPT Development. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208
P15: Novel Formulations, Agents and Microbicides. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213
P16: Passive Antibody Functions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
P17: Post-exposure Prophylaxis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232
P18: Preclinical Evaluation of Vaginal Films and Gels. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233
P19: PrEP Implementation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 237
P20: PrEP: Resistance, Modeling and Acceptability. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241
P21: Product Acceptability and Adherence. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 243
P22: Resistance and Treatment Failure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 246
P23: Retention and Adherence in Trials. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 249
P24: T-Cell (CD4 & CD8) Responses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 258
P25: Transmission and Viral Diversity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 268
P26: Vaccine Clinical Trials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 274
146
Overview of Poster Sessions
Posters Presented on Thursday, 30 October
P27: Adjuvants. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 284
P28: Behavioral and Social Sciences . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 287
P29: Circumcision and Acceptability. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 288
P30: Condoms: Attitudes, Use and How to Increase. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 291
P31: Drug Transporters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 294
P32: Ethics and the Law. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 296
P33: Evaluation of Novel Compounds in Cell-Based Systems. . . . . . . . . . . . . . . . . . . . . . . 298
P34: Glycans and Antibody Effector Functions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 304
P35: HIV Drug Resistance in vitro. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 311
P36: HIV Incidence and Prevalence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 313
P37: Immunogenetics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 319
P39: Molecular Epidemiology. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 327
P40: Mucosal Immune Activation and Inflammation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 333
P41: Novel Vaccine and Prevention Concepts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 349
P42: Participation in Trials: Willingness, Benefits and Challenges . . . . . . . . . . . . . . . . . . . 364
P43: Policy, Advocacy and Modeling. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 370
P44: Preclinical Evaluation of Biomedical Agents in Animal Models Tissues Explants . . . 378
P45: Pregnancy and Prevention of Mother to Child Transmission . . . . . . . . . . . . . . . . . . . 384
P46: PrEP Trials: Preparing for Demos, Participant Experiences . . . . . . . . . . . . . . . . . . . . . 388
P47: Resource Tracking and Economics. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 391
P48: Risk Perception. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 395
P49: Sexually Transmitted Infections. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 399
P50: Tenofovir Gel: Acceptability and Adherence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 406
P51: Therapeutic Vaccine and Viral Latency. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 410
P52: Treatment as Prevention: Initiation, Adherence, and Monitoring on ARV. . . . . . . . . . 412
P53: Vaginal Rings: Baseline Characteristics, Impact on Condoms and Flora . . . . . . . . . . . 416
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147
POSTERS
P38: Innovations in Vaccine and Microbicides Studies in Lab and Monitoring. . . . . . . . . 323
Posters
Posters 01: Acute Seroconversion
P01.01
P01.02
Recall of Acute Retroviral Symptoms in a
Multicentre Cohort Study in Africa
Long-term Follow up of HIV Seroconverters
from the VOICE Trial - An Analysis of Data
from MTN-015
Eduard J. Sanders1,2, Kimberley A. Powers3, Etienne Karita4,
Anatoli Kamali5, William Kilemba6, Susan Allen7, Eric Hunter7,
Omu Anzala8, Pat Fast9, Matthew A. Price9,10
of Oxford, Headington, United Kingdom, 3University of North Carolina,
Chapel Hill, NC, United States, 4Project San Francisco, Kigali, Rwanda,
5
Medical Research Council/Uganda Virus Research Institute, Entebbe,
Uganda, 6Zambia Emory Research Project, Lusaka, Zambia, 7Emory
University, Atlanta, GA, United States, 8Kenya AIDS Vaccine Initiative
Institute of Clinical Research, Nairobi, Kenya, 9International AIDS
Vaccine Initiative, New York, NY, United States, 10University of California
at San Francisco, San Francisco, CA, United States
1
POSTERS
Background: Remarkable differences exist for reported acute retroviral
symptoms (ARS) in African adults when acquiring HIV-1 infection,
notably for fever (range: 19-78%), or diarrhea (range: 2-42%). This
may lead to under-appreciation of the clinical presentation and delayed
diagnosis of patients with acute HIV-1 infection (AHI). We sought to
determine whether reporting of ARS was associated with timing of
ascertainment of signs and symptoms and clinical research centre (CRC)
at nine collaborating CRC´s in Africa.
Methods: Adults with acute or early HIV-1 infection in a multicenter HIV1 incidence study were enrolled in a sub-study assessing ARS. Estimated
date of infection (EDI) was based on a positive plasma viral load or
p24 antigen test prior to seroconversion, or the mid-point between a
negative and positive HIV-1 serologic test. 11 ARS signs and symptoms
were assessed at sub-study enrollment. We used log-binomial regression
to estimate the prevalence of ARS signs and symptoms by recall period
(i.e. period 1= enrollment ≤ 42 days after EDI and period 2 = enrollment
> 42 and ≤ 90 days after EDI) and CRC.
Results: Among a total of 124 and 118 participants evaluated in period
1 and 2, respectively, 66.0 (95% confidence interval (CI): 59.2-72.7%),
and 49.4 (95% CI: 43-55.7%, p=0.0006) reported having had at least
one sign or symptom. The most common symptoms of ARS were fever,
headache, myalgia/arthralgia, and fatigue. With the exception of fever,
diarrhea, and oral ulcers, the prevalence of all signs or symptoms
was statistically significantly greater in those who were evaluated in
period 1 as compared to those evaluated in period 2 (Figure). There
was considerable variability in the prevalence of signs and symptoms
by CRC.
Conclusions: In this multicenter African cohort, the prevalence of ARS
symptoms of patients evaluated within 6 weeks following EDI was
higher for all symptoms except for fever, diarrhea and oral ulcers as
compared to patients who were evaluated between 6 weeks and 3
months of the EDI.
148
Sharon Riddler1, Marla Husnik2, Arendevi Pather3, Thesla
Palanee4, Gonasagrie Nair5, Felix Mhlanga6, Marthinette
Taljaard7, Clemensia Nakabiito8, Anamika Premrajh3, Lisa Levy9,
Vanessa Elharrar10, Gita Ramjee3, Jennifer Balkus11,12, MTN-015
Protocol Team
University of Pittsburgh, Pittsburgh, PA, United States, 2Statistical
Center for HIV/AIDS Research & Prevention (SCHARP), Seattle, WA,
United States, 3Medical Research Council, Durban, South Africa, 4Wits
Reproductive Health and HIV Institute, University of the Witwatersrand,
Johannesburg, South Africa, 5CAPRISA/University of Kwa Zulu Natal,
Durban, South Africa, 6University of Zimbabwe - University of California
San Francisco Collaborative Research Programme, Harare, Zimbabwe,
7
The Aurum Institute, Klerksdorp, South Africa, 8Makerere University Johns Hopkins University Research Collaboration, Kampala, Uganda,
9
FHI 360, Research Triangle Park, NC, United States, 10Division of AIDS,
NIH, Bethesda, MD, United States, 11Fred Hutchinson Cancer Research
Center, Seattle, WA, United States, 12University of Washington, Seattle,
WA, United States
1
Background: In 2008, the Microbicide Trials Network (MTN) initiated
a multi-site, long-term observational study of women who become HIV
infected during microbicide trials participation to assess the impact of
investigational products on HIV disease progression, treatment response,
and clinical and behavioral outcomes (MTN-015). Here we report on
women enrolled from the VOICE trial.
Methods: MTN-015 recruited eligible VOICE participants from 15 sites
in South Africa, Zimbabwe and Uganda. Visits occurred at enrollment
and at 1, 3, 6 months after detection of HIV infection, then every 6
months thereafter. CD4+ T cell count, HIV-1 RNA, clinical history, and
risk reduction counseling are completed at each visit. We evaluated the
effect of the interventions (oral TDF, oral FTC/TDF, and vaginal tenofovir
gel) on HIV disease progression using Cox proportional hazard models
to assess time to CD4< 350, ART initiation, and first AIDS-defining event
as separate outcomes.
Results: MTN-015 enrolled 255/356 (72%) of VOICE seroconverters;
93% were from South Africa. Median age at MTN-015 enrollment was
23 years; median time from detection of HIV infection to enrollment
was 2.1 months. To date, median follow-up is 22.9 months (cumulative
457 person-years). At enrollment, median CD4+ and HIV-1 RNA were
539 cells/mm3 and 4.4 log10 copies/ml. We observed no significant
difference by VOICE study arm with regard to time to CD4< 350, ART
initiation, or AIDS-defining events. ART was initiated by 49 women (3
prior to MTN-015 enrollment); 82% met local CD4 guidelines for ART
initiation and 16% started ART due to pregnancy. Initial ART regimens
were NNRTI based, most commonly EFV/TDF/3TC (61%); only 2 (4%)
included d4T. AIDS-defining events occurred in 11 (4%) women.
Conclusions: Long term follow-up of participants who seroconvert in
prevention trials is feasible, acceptable and is an important component
of the assessment of ART-based prevention modalities.
Posters 01: Acute Seroconversion
P01.03 LB
VOICES: A Qualitative Study with MSMs in
South Florida May 2014
Gregory Edwards1, William Green2, Marsha A. Martin3,4, William
DiStefano5, Leonard Jones6
Flowers Heritage Foundation, Oakland, CA, United States, 2Broward,
Health Care Services Section, Broward County, FL, United States, 3Get
Screened Oakland, Office of the Mayor, Oakland, CA, United States,
4
Urban Coalition for HIV/AIDS Prevention Services, Washington, DC,
United States, 5Get Screened Oakland, Oakland, CA, United States,
6
Human Services Section-Ryan White Part A, Broward County, FL,
United States
1
POSTERS
Background: In Broward County, FL 21,100 persons lived with HIV
disease in 2011, and 3,334 (15.8%) were unaware, with the largest
number in Ft. Lauderdale. More than 1000 new cases of HIV were
reported, an increase of 25% from 2010. New cases affected persons
between the ages of 40 and 49. MSMs make up approximately 70%
of all infections.
Methods: A multifaceted approach assessed the epidemic among gay,
bisexual, and other MSMs: community advisory workgroup, literature
review, 25 key informant interviews, 17 focus groups with men of
known and unknown HIV serostatus, 4 focus groups with transgender
women and consumer surveys. The study sought to answer the following
question: What are the socio-sexual-cultural circumstances that lead to
increased risk for and transmission of HIV among gay, bisexual men and
transgender women?
Results: Young and older MSMs report and experience social
isolation, disaffiliation and limited choices for socialization and healthy
relationships. HIV infection is felt to be ‘just a matter of time.’ For
some gay men who have been living with HIV for more than 15 years,
HIV messages are not relevant to their experiences. The availability
and accessibility of venues and multi-day circuit parties that promote
sexual play and risk-taking behaviors also contribute to the spread of
HIV among gay and bisexual men, and transgender women. Online
‘hookups’ that advertise ‘bareback sex play’ create the perception that
it is no longer a socio-sexual cultural norm to use protective barriers
during sexual intercourse. Gay, bisexual MSMs living with HIV, many
of whom experience shame, low self-esteem and isolation have very
limited venues for positive self expression and health promotion.
Conclusions: The Broward County HIV epidemic is influenced by sociosexual-cultural economic dynamics: anonymous sex, alcohol, and
crystal meth used to ease hardships associated with a lack of social and
communications skills, loneliness, isolation, and exhaustive vigilance
create “the perfect storm” of HIV infection.
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Posters
Posters 02: Community Engagement and Advocacy
P02.01
P02.02
Managing Communication and Facilitating
the Exchange of Ideas in a Large, Multi-site
Clinical Trial: Experiences from the ASPIRE
Study
A Rectal Revolution Takes a Village:
Developing an Educational Video about Rectal
Microbicides
Clare Collins1, Barbara Friedland2, Jim Pickett3
Rachel Scheckter1, Katie Schwartz1, Ashley Mayo1, Jared Baeten2,
Thesla Palanee3, Lydia Soto-Torres4, Kristine Torjesen1
1
FHI 360, Durham, NC, United States, 2University of Washington,
Seattle, WA, United States, 3Wits Reproductive Health and HIV Institute
(WRHI), Johannesburg, South Africa, 4National Institute of Allergy and
Infectious Diseases, Bethesda, MD, United States
POSTERS
Background: Streamlined communication and coordinated strategies
are essential for large multi-site clinical trials to ensure high-quality
implementation and to minimize burden from large volumes of
disaggregated input across multiple protocol team partners.
Methods: A review of approaches to improve coordination and enhance
communication in the MTN-020 study (ASPIRE) was conducted to
develop qualitative and quantitative descriptions of these activities.
Results: Multiple strategies have been used in ASPIRE to streamline
communication and coordination across sites and the protocol team. Six
in-person trainings, designed to incorporate adult learning techniques
and establish common approaches, were conducted prior to study
start. Additional workshops and trainings have continued throughout
implementation and 38 assessment visits have been conducted to
date. Assessment visits focus on strengthening site-specific systems and
communication strategies. A written study communication plan was
developed at study outset that included use of weekly priority e-mails
with sites to minimize communication burden of important information
and action items. Communication was streamlined through monthly
or twice-monthly implementation calls with individual site leaders and
weekly management team calls to share key updates. Cross-site learning
was fostered through cadre-specific listservs, semiannual in-person
meetings, monthly site-led protocol team calls, and quarterly calls to
strengthen adherence counseling and qualitative data collection. The
ASPIRE communication strategy has led to sharing of innovative sitedriven approaches, such as participant engagement strategies, as well as
early identification and quick resolution of implementation issues, such
as participant and community myths and misconceptions.
Conclusions: Strategies to streamline communication and coordination
in ASPIRE to improve efficiency and quality of study implementation
could inform implementation of other large multi-site clinical trials.
150
Microbicide Trials Network, Pittsburgh, PA, United States, 2Population
Council, New York, NY, United States, 3International Rectal Microbicide
Advocates, Chicago, IL, United States
1
Background: Rectal microbicides (RMs) are new products being
developed to prevent HIV transmission through receptive anal
intercourse. Given that most people are unfamiliar with RMs, educational
materials that are accessible, understandable and appealing to multiple
communities are essential. To address this need, the Population Council,
International Rectal Microbicide Advocates (IRMA) and the Microbicide
Trials Network (MTN) developed a 14-minute video on RMs, “The Rectal
Revolution is Here: an Introduction to Rectal Microbicide Clinical Trials,”
in conjunction with the launch of an international Phase II RM clinical
trial.
Methods: The video, which combines animation and documentarystyle footage, was developed with the guidance of a Video Advisory
Committee (VAC), comprised of 12 individuals from international trial
sites, non-affiliated researchers, and IRMA advocates. The VAC provided
input and expertise on all aspects of the video, from script content to
translations to final edit. An early cut of the video was viewed by over
80 participants at Microbicides 2012 who gave feedback via a survey,
and was tested in 13 focus group discussions at five sites in English,
Spanish, and Thai among more than 100 gay men, other men who have
sex with men, and transgender women.
Results: Feedback from the surveys and focus groups resulted in several
significant changes to clarify key messages, including the addition of
information about HIV testing, the cutting of a main character as well as
some interview footage, and a change in music. The final video has been
made available to trial sites and advocates, and online via YouTube.
Conclusions: The extensive year-long process with multiple inputs used
to develop the video resulted in concrete suggestions to improve it, and to
make it relevant and accessible for intended audiences. By incorporating
this vital feedback, video producers were able to develop a culturally
competent educational video that was inclusive and responsive, and is
being used extensively.
P02.03
P02.04
Seven Steps to Strengthen Community
Engagement in HIV Preventions Trials in
Durban
How Community Education Tools Facilitated
Understanding of the ASPIRE Vaginal Ring
Study: Kampala Experience
Neetha Shagan Morar1
Patrick Ndawula1, Teopista Nakyanzi1, Juliane Etima1, Samuel
Kabwigu1, Flavia K. Matovu1, Sophie C. Nanziri1, Doreen
Kemigisha1, Stella Nanyonga1, Rhonda White2, Cheryl Cokley2,
Clemesia Nakabiito1
1
Collaboration, Kampala, Uganda, 2FHI 360, Duham, NC, United States
Background: Effective community engagement is the cornerstone
of cultivating ownership, trust and acceptance of research within
communities. Researchers at the six sites of the HIV Prevention Research
Unit (HPRU) in Durban interact with a diverse range of urban and semirural communities where trials are conducted. We describe the outcomes
of the seven steps in community engagement while conducting
microbicide and other HIV prevention trials at HPRU.
Methods: Using a 7 step cyclical process, community engagement
began a decade ago at the research sites. The process involved:
(1) conducting a situational analysis at entry;
(2) education and outreach;
(3) networking and partnerships with stakeholders;
(4) selection of community advisory boards (CABs);
(5) input on protocol and study procedures;
(6) participant engagement and
(7) dissemination of study outcomes.
Results: Community partnerships with stakeholders and CABs have
been sustained for over a decade. Ongoing engagement fostered
trusting relationships and facilitated acceptance of trials by community
and participants. Education from stakeholders on community culture and
social dynamics helped researchers address community perceptions of
HIV prevention and microbicides. CAB members observed the destruction
of blood samples and this transparency enhanced their understanding
of ethics in research and dispelled the myth that researchers sold blood.
Trial participants volunteered to publicly share their experiences during
the dissemination of trial results to stakeholders. They joined CABs
and are currently advocates of HIV prevention research by promoting
adherence and retention.
Conclusions: Stakeholders, CABs and trial participants assisted in
developing trust between communities and researchers, who were
transparent and engaged them throughout the research lifecycle.
Education and information enabled stakeholders to make informed
decisions and facilitated their understanding of trial results and
acceptance of the conduct of trials in their communities.
Background: UNAIDS Good participatory practice guidelines for
biomedical HIV prevention trials recommends that sufficient trial
information, such as study objectives, procedures, risks, benefits, and
what is expected of participants, is provided for potential participants
to make informed decisions. We describe how ASPIRE community
education tools increased study awareness and enabled literacy
regarding research, reproductive health, family planning, and HIV
prevention.
Methods: Kampala site began community education about ASPIRE in
July 2012. Community education tools are used to raise awareness and
knowledge about the study. Tools like the ASPIRE Community Education
flip chart and fact sheets; contain graphic images to support learning in
low literacy populations. Visual aids, such as IUCD and pelvic models
and a demonstration vaginal ring, are also used. These tools have
helped to dispel anxieties and rumors, provide accurate information,
and reduce potential barriers to participation. Following sensitization
sessions in which these educational tools are used, a comprehension
assessment is administered to randomly selected session attendees. The
assessment is invaluable in identifying information that study educators
need to clarify to enable informed decision-making. For example, when
assessment results identified limited understanding of the placebo
concept, educators adjusted their messaging to improve learning.
Results: Through March 2014, 7,611 women were educated about
ASPIRE; 6,013 (79%) completed a comprehension assessment. 5,893
(98%) responded that one should be HIV negative to participate in
the study; 5,592 (93%) identified 3 HIV prevention methods; (81%)
indicated the study product to be tested; and 4,690 (73%) named 2
reliable family planning methods. However only 1,924 (32%) explained
the placebo concept.
Conclusions: Community educational tools are effective in facilitating
potential participants’ understanding of research, reproductive health,
and family planning and HIV prevention.
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151
POSTERS
1
Posters
P02.05
P02.06
Health Science Journalists Are Bridge Partners
as HIV Prevention Research Moves Ahead
Bridging the Gap: Uniting Participants and
Staff for Improved ASPIRE Study Metrics at
Wits Reproductive Health and HIV Institute in
Hillbrow
Bobby Ramakant1, Badri N. Saxena2,3
Citizen News Service (CNS), Lucknow, India, 2Microbicides Society of
India, Delhi, India, 3ICMR Microbicides Expert Group, Government of
India, Department of Health Research, Delhi, India
1
POSTERS
Background: Health science journalists routinely focus on évent
reporting´ news and seldom get opportunity to dwell deep into health
science research, understand science as it moves ahead. Media owners
also are reluctant to invest in opportunities that further understanding of
their health science journalists and editors.
Methods: CNS supported by its partner Microbicides Society of India (MSI)
worked with CNS Correspondents team and identified those interested
in HIV prevention research and/or were based in countries where HIV
prevention research is moving forward. CNS Annual Health Fellowship
programme organized audio/ video calls with CNS Fellows every month
on specific health science research issues and Fellows interviewed incountry experts and regional/ global experts to develop in-depth news
feature articles to keep readers abreast of progress on HIV prevention
research. CNS Correspondents and Fellows also joined AVAC or IRMA
webinars whenever appropriate to further their own understanding on
key developments in HIV prevention science. CNS also supported its
correspondents to go to HIV or other related conferences in countries,
regions or globally to do key informant interviews with scientists who
were doing HIV prevention research, other advocates, trial participants,
among others and develop these news feature articles published and
syndicated through CNS.
Results: Over 100 HIV prevention research scientists, advocates, trial
participants were interviewed by CNS Correspondents during 20122014 and articles got published and syndicated by CNS to online and
print media in Asian and African nations. These news feature articles
were also disseminated through social media platforms, conference
websites, official conference newspapers, among others.
Conclusions: - important for media house owners to invest in
opportunities for health science journalists to understand scientific
research and follow it consistently
- important for media to be involved as key partners of HIV prevention
research
152
Krishnaveni Reddy1, Helen Rees1, Thesla Palanee1
1
Background: ASPIRE is a Phase 3 study assessing safety and
effectiveness of an intravaginal Dapivirine ring in preventing HIV
infection. At Wits RHI, we recognize that to improve product adherence
and visit retention metrics, understanding the importance of meeting
study objectives and being committed to the HIV prevention agenda
must be optimized amongst both participants and staff. To achieve this,
novel strategies were implemented to secure participant and staff buyin.
Methods: Standard practice for encouraging adherence and retention
entails staff counseling participants regarding these metrics at study
visits followed by telephonic retention/adherence reminders. Although
effective, we note that achieving study metrics is not a concern for most
participants and routine visit contact is insufficient to develop thorough
study objective understanding and build commitment and trust
needed to impact these metrics positively. We have found that study
understanding, commitment and shared study metric accountability can
be encouraged among participants and staff by frequent professional
but social interaction. Informal participant-staff engagement activities
(e.g. tea/retention parties, movie days) are hosted on visit-free days with
the aim of bridging the communication gap and establishing a shared
commitment to the ASPIRE “1 team-1 goal” motto. Discussions include
prior and current trial information, study objectives, adherence and
retention progress and participants’ contributions to these metrics.
Results: These activities remind participants of their impact on the
HIV prevention field and their health. The non-judgmental atmosphere
results in engaged discussion, disclosure of concerns and addressing
critical issues. Participants develop support networks and peer-educate
each other during visit waiting times.
Conclusions: These activities have facilitated building of participant-staff
rapport and potentially increased study commitment. Consequently,
communication, product adherence and retention are enhanced.
P02.07
P02.08
Engaging the Community and Stakeholders
to Develop Microbicide Communication
Materials in Kenya
Factors that Influence Trainees’ Acquisition of
Knowledge about Biomedical HIV Prevention
Research: Experience from the Field
Elizabeth Ryan1, Emily Bockh1, George Githuka2, Caroline
Mackenzie3, Alice Olawo3, A. Cornelius Baker1, Elizabeth Tolley4
Oluwatosin Bamidele Alaka1, Morenike Oluwatoyin Ukpong2,
Florita Durueke1, Augustina Obiajulu Amuamuziam1
FHI 360, Washington, DC, United States, 2Ministry of Health, Nairobi,
Kenya, 3FHI 360, Nairobi, Kenya, 4FHI 360, Durham, NC, United States
1
Background: In 2010 CAPRISA 004 provided proof of concept that
peri-coital, vaginal use of tenofovir 1% microbicide gel can reduce
HIV acquisition among women. Communication campaigns will play
a key role in generating demand and educating women on correct
use. Communicating about Microbicides with Women in Mind used a
participatory process to create and pretest a suite of communication
materials for potential users, partners and health care providers in
Kenya. The project is part of a larger USAID-funded effort to support
microbicide introduction if/when tenofovir gel is proven effective.
Methods: The project conducted numerous engagement activities. A
national policy consultation was held to identify priority audiences for
microbicide communication in Kenya. A Project Advisory Committee
(PAC), led by the National AIDS Control Programme and consisting of local
stakeholders, was formed. Audience consultations were conducted with
young women, female sex workers (FSWs), women in relationships, and
men, and were followed by a national message development workshop
(MDW) to draft messages and materials. Participants from the policy
consultation and MDW were reconvened to review research results,
discuss final material changes, and provide input into a communication
strategy and materials adaptation guide.
Results: Engagement activities resulted in a well-received and effective
suite of materials, including logos, posters, radio spots, animated TV
storyboards, a brochure, counseling algorithms, and educational
flipcharts. These materials were refined through two round of pretesting
and a study to assess their impact on microbicide interest, attitudes
and condom migration. The PAC provided substantive contributions
throughout.
Conclusions: Stakeholder engagement at all levels, and from various
sectors, is critical for ensuring communication strategies and materials
are relevant, effective, and well-received. The systematic process
followed by this project could benefit future HIV prevention projects.
New HIV Vaccine and Microbicide Advocacy Society, Programmes,
Lagos, Nigeria, 2New HIV Vaccine and Microbicide Advocacy Society,
Coordinator / Programmes, Lagos, Nigeria
Background: The New HIV Vaccine and Microbicide Advocacy Society
teachers training showed that capacity building of teachers in Lagos
was an important mechanism to enable students understands HIV
prevention. In view of this, NHVMAS structured an annual training
to support sustained capacity building programmes for teachers in
secondary schools.
Methods: NHVMAS undertook a two days training of 23 secondary
school teachers in Lagos State using a curriculum ratified by the State
Ministry of Education. A 12 item questionnaire pre and post test was
administered. Analysis of the pre and post test was conducted so see
how participants faired on acquiring new knowledge.
Results: There was a significant difference in the pre and post test
scores (57.6% vs66.5%; P=0.001). The cohort of trainees had a good
understanding of HIV/AIDS &STIs. However very few persons understood
the concept of effectiveness: only 19.2% understood about the concept
of effectiveness of NPT, and 14.1%HIV vaccine respectively. About 76%
of participants were aware of the potential of microbicides to prevent HIV
infection but only 21.5% of participants understood the mechanisms of
action of microbicide in preventing HIV infection. Only 14% understood
its potential role for STI and pregnancy prevention.
Post training, participants significantly gained knowledge and
understanding of these concepts: 38.2% understood about the concept
of effectiveness of NPT, and 66.6%HIV vaccine respectively.
An average deficient gap of -18.4% and -23.4% was seen in the pre and
post test evaluation in answering questions on; the mechanism of action
of microbicide and test for safety of microbicide, however, in the second
training only -10% and 0% respectively.
Conclusions: The study participants seem to have challenges with the
understanding of the concept of HIV vaccine research. There may be a
need to design a training curriculum that focuses attention on building
skills and knowledge about HIV vaccine.
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POSTERS
1
Posters
P02.09
Media Engagement and Reporting on
Nigerian - Canadian Collaboration on AIDS
Vaccine Research Study in Jos Plateau State
Wika Gofwen1, Godwin Odemijie2, Florita Durueke3, Chuks
Okonkwo3, Morenike Ukpong3
Plateau Radio Television Corporation, News and Current Affairs, Jos,
Nigeria, 2FRCN, Federal Radio Corporation of Nigeria, News and Current
Affairs, Abuja, Nigeria, 3New HIV Vaccine and Microbicide Advocacy
Society, Microbiology, Ibadan, Nigeria
1
POSTERS
Background: The Media is a critical stakeholder in ensuring public
education about biomedical HIV prevention technology and can
also play roles in setting the research agenda.The Nigerian Canadian
Collaboration on AIDS Vaccine (NICCAV) media engagement programme
developed model of engaging journalists to ensure it achieves its set
objective of building the capacity of the media to effectively educate the
general public in Jos Metropolis and other stakeholders on HIV vaccine
research and the NICCAV project specifically.
Methods: A community based organisation with expertise on
community and media engagement in research was engaged on the
project to facilitate a systematic engagement of journalists. Specifically,
the project through consultation with the Nigeria Union of Journalists
(NUJ), identified 30 health reporters from the print and electronic media
that were enrolled in the media programme, at the state and national
level who equally have their capacity enhanced on (understanding and
communicating HIV prevention research) to enable them disseminate
culturally and linguistically appropriate information on HIV vaccine
research. Media roundtables discussions were held to provide update
on the clinical study.
Results: The training did show significant increase in knowledge among
the journalists, as they identified new knowledge and skills gained.
The pre-test and post-test analysis did show significant change in the
mean values (p value=0.04). Media roundtables provided update on
the clinical study. There was significant increase in the volume of media
publications on vaccine research and NPT. Comparative analysis of
vaccine and NPT reporting from journalists engaged on the vaccine
study was significantly higher than other health reporters that were not
enrolled in the project.
Conclusions: Investment in a structured media engagement programme
on NPT research will facilitate media reporting and advocacy on NPT
as well as mobilize community and relevant stakeholders support for
vaccine and NPT research.
154
Posters 03: Correlates of Protection and Exposure
P03.01
P03.02
HIV-specific T and NK-cell Responses Do
Not Correlate with Protection from Sexual
Acquisition of HIV
Type II Interferon Stimulated Genes were
Tightly Regulated in Kenyan Commercial
Sex Workers who Exhibit Natural Resistance
towards HIV Infection.
University of Washington, Seattle, WA, United States, 3Kenya Medical
Research Institute, Nairobi, Kenya, 4University of Manitoba, Winnipeg,
MB, Canada
1
2
Background: The existence of HIV-specific immune responses has
been observed in several cohorts of HIV-exposed seronegative (HESN)
subjects, yet the ability of these responses to protect against incident
HIV infection has not been directly evaluated.
Methods: Within a prospective cohort of African male and female HESN
who were at high risk of HIV infection as a result of having an HIV
infected heterosexual partner, we conducted a nested case-control study
to assess HIV-specific immune responses among 176 subjects, including
29 who acquired HIV (cases) and 147 who remained HIV uninfected
(controls). Peripheral blood mononuclear cells were collected at the visit
prior to HIV acquisition. HIV-specific immune responses were detected
by intracellular cytokine staining using global potential T-cell epitope
11mer peptide pools for Gag and Env, with positive responses defined
as dual production of IFN-γ and CD107a for CD8+ T-cell responses, IFN-γ
and TNF-α for CD4+ T-cell responses, and IFN-γ and CD107a for NK
responses. Logistic regression was used to assess if the frequency and
magnitude of immune responses were associated with protection from
HIV acquisition.
Results: Response rates for CD8+ T-cells were 17.2% and 6.8% in cases
and controls, respectively, to Gag, 11.1% and 10.6% to Env (p>0.05).
Similarly, no statistically significant differences between cases and
controls were observed for CD4+ T-cell responses (Gag: 3.4% vs. 3.4%,
Env: 0.0% vs. 2.8% p>0.05) and NK cell responses (Gag: 0.0% vs.
3.4%, Env: 0.0% vs. 6.3%, p>0.05). Furthermore, the magnitude and
breadth of responses among responders did not differ between cases
and controls.
Conclusions: Among HESN with a known HIV positive partner, no
relationship was found between T-cell and NK cell responses prior to
HIV infection and protection from sexual acquisition of HIV. Our results
do not support the hypothesis that natural HIV-specific responses reduce
susceptibility to HIV infection in HESN.
Marcel Gluchowski1, Xiaoqiong Yu2, Renee Douville1, Joshua
Kimani3, Frank A. Plummer4,5, T. Blake Ball5,6, Ruey-Chyi Su5
1
University of Winnipeg, Biology, Winnipeg, MB, Canada, 2Yangquan
Municipal Center for Disease Prevention and Control, Yangquan, China,
3
University of Nairobi, Department of Medical Microbiology, Nairobi,
Kenya, 4Public Health Agency of Canada, National Microbiology
Laboratories, Winnipeg, MB, Canada, 5University of Manitoba, Medical
Microbiology & Infectious Diseases, Winnipeg, MB, Canada, 6Public
Health Agency of Canada, National HIV & Retrovirology Laboratories,
Winnipeg, MB, Canada
Background: Interferon stimulated genes (ISGs) are shown to mediate
critical anti-viral responses. Cell lines overexpressing ISGs showed
reduced viral replication, including HIV. HIV-1 infection was shown
to induce a distinct subset of ISGs that are regulated by interferon
regulatory factor-1 (IRF-1). However, low endogenous IRF-1 level was
found to associate with reduced susceptibility to HIV-infection among
Kenyan commercial sex worker (CSW). Here, we assessed whether
reduced IRF-1 level, observed in highly HIV-exposed, yet seronegative
(HESN) CSW, also correlate with reduced expression and responsive
potential of ISG genes.
Methods: Endogenous ISG mRNA level in peripheral blood mononuclear
cells (PBMC) of HESN CSW and control CSW were assessed using
quantitative RT-PCR. The responsive potential of these ISGs to type II
interferon (IFN) was assessed by stimulating PBMC with IFN-g for 3h;
optimized to observe transient IRF-1 response in HESN and persisting
in controls.
Results: Not all ISGs are constitutively expressed. Ex vivo HESN PBMC
expressed 22 of 67 ISGs examined, while CSW controls expressed 31.
Endogenous level of 11 ISGs differed between the 2 groups. ISGs of
anti-viral function(IRF1, CXCL10, TRAILR1, ISG56, IFNb, COX2, MXA,
TNFa) were significantly lower in the HESN, while the ISGs signaltransducing function(IFIT3, MDA5, IFI6) were notably higher in most
HESN. Exogenous IFN-g stimulation resulted in significant increases
(e.g., CXCL-10, IFN-b, IRF-1, IFIT3, STAT-1) and decreases (e.g., TRAILR1,
p16p14, BclXL, BCL2) of some ISGs. Overall, robust increases of CXCL10, IRF-7 & IFIT3 were found in HESN with less change in other ISGs.
Conclusions: Together, baseline level of ISGs with anti-viral,
proinflammatory function was reduced in HESN, with IFN-g responsive
ISGs genes being tightly regulated to mount robust, yet transient
responses, suggesting an important role for these ISGs in maintaining
the HESN phenotype against HIV-infection.
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155
POSTERS
Laura Pattacini1, Jared M. Baeten2, Katherine K. Thomas2, Tayler
R. Fluharty1, Pamela M. Murnane2, Deborah Donnell1,2, Elizabeth
Bukusi2,3, Allan Ronald4, Nelly Mugo2,3, Jairam R. Lingappa2,
Connie Celum2, Juliana M. McElrath1,2, Jennifer M. Lund1,2,
Partners PrEP Study Team
Posters
P03.03 LB
P03.04 LB
HIV-1 of Children with Slow Disease
Progression Escapes Autologous
Neutralization and Triggers Development of
Cross-neutralizing Responses
Modulation of RAS Pathways as a Biomarker
of Protection against HIV and as a Means to
Improve Vaccine Efficacy
Stefania Dispinseri1, Mariangela Cavarelli1, Anna Maria Plebani2,
Marianne Jansson3,4, Gabriella Scarlatti1
San Raffaele Scientific Institute, Milan, Italy, 2Foundation IRCCS cà
Granda Ospedale Maggiore Policlinico Milan, Milan, Italy, 3Lund
University, Lund, Sweden, 4Karolinska Institutet, Stockholm, Sweden
1
POSTERS
Background: One-quarter of HIV-1 infected children has a disease
progressing towards AIDS within the first year of age, whereas the
majority of them progress slowly over several years. With the purpose
to assess the role of the humoral immune response underlying different
patterns of disease we studied the kinetics of antibody responses to
HIV-1 and childhood vaccine antigens in slow and rapid progressors
followed from birth to 5 years of age.
Methods: Autologous and heterologous neutralizing capacity of plasma
obtained during follow-up of 25 children was tested with PBMC- and/
or TZMbl-assays, against up to 5 primary viruses for each child or 3
pseudotyped viruses (Tier 1 subtype B and Tier 2 subtype A), respectively.
The seroreactivity to HIV-1 gp41, autologous gp120 V3-loop peptides,
and tetanus and diphtheria toxoid was assessed by ELISA.
Results: Newborns displayed antibodies towards an immunodominant
HIV-1 gp41 epitope, which were of maternal origin, but did not
neutralize the transmitted virus. The child’s neutralizing antibodies
(Nabs) developed usually within 1 year of age to its own early virus
concomitantly with autologous V3-loop directed antibodies in slow,
but rarely in rapid progressors. Autologous Nabs persisted throughout
disease progression and induced continuous emergence of escape
variants. Heterologous Nabs developed within two years independently
of disease progression, but their subsequent increase in potency and
breadth was a preferential trait of slow progressors. Kinetics of antibody
responses to the immunodominant gp41 antigens and childhood
vaccine antigens was preserved independently of disease progression.
Conclusions: Persistent autologous Nabs triggering viral escape and
increase in breadth and potency of heterologous Nabs are exclusive trait
of slow progressors. Thus, immune-competence seems to impact the
ability to develop potent Nabs, suggesting that early transmitted viruses
of slow progressors represent an attractive target for vaccine design,
which aims to induce cross-Nabs.
Slim Fourati1, Monica Vaccari2, Shari N. Gordon2, Luca
Schifanella2, Mark Cameron1, Brandon F. Keele3, Xiaoying
Shen4, Georgia D. Tomoras4, Erik Billings5, Mangala Rao5, Amy
W. Chung6, Karen Dowell7, Chris Bailey-Kellogg7, Eric Brown8,
Margaret E. Ackerman8, Namal P.M. Liyanage2, Diego A. VargasInchaistegui9, Stephen Whitney10, Melvin N. Doster2, Nicolo
Binello2, Poonam Pegu2, David C. Montefiori11, Kathryn Foulds12,
David S. Quinn13, Mitzi Donaldson13, Frank Liang12, Karin
Loré12, Mario Roederer12, Richard A Koup12, Adrian McDermott12,
Zhong-Min Ma14, Christopher J Miller14, Tran B Phan15, Donald
N. Forthal15, Matthew Blackburn2, Francesca Caccuri2, Guido
Ferrari11, Devon Thompson16, Marjorie Robert-Guroff9, Silvia
Ratto-Kim5, Jerome H. Kim5, Nelson L. Michael5, Sanjay Phogat17,
Susan W. Barnett18, James Tartaglia17, David Venzon19, Donald M.
Stablein20, Galit Alter6, Rafick-Pierre Sekaly21, Genoveffa Franchini2
Vaccine & Gene Therapy Institute of Florida, Port Saint Lucie, FL,
United States, 2National Cancer Institute, Animal Models and Vaccine
Section, Bethesda, MD, United States, 3National Cancer Institute,
AIDS and Cancer Virus Program, Frederick, MD, United States, 4Duke
Human Vaccine Institute, Durham, NC, United States, 5U.S. Military HIV
Research Program (MHRP), Walter Reed Army Institute of Research,
Silver Spring, MD, United States, 6Ragon Institute of MGH, MIT and
Harvard, Boston, MA, United States, 7Dartmouth College, Computer
Science, Hanover, NH, United States, 8Dartmouth College, Thayer
School of Engineering, Hanover, NH, United States, 9National Cancer
Institute, Immune Biology of Retroviral Infection Section, Bethesda,
MD, United States, 10Advanced BioScience Laboratories, Inc., Rockville,
MD, United States, 11Duke University Medical Center, Durham, NC,
United States, 12National Institutes of Health, Vaccine Research Center,
Bethesda, MD, United States, 13Karolinska Institutet, Stockholm,
Sweden, 14University of California, California National Primate Research
Center, Davis, CA, United States, 15University of California, Irvine
School of Medicine, Irvine, CA, United States, 16Advanced BioScience
Laboratories, Inc, Rockville, MD, United States, 17Sanofi Pasteur,
Swiftwater, PA, United States, 18Novartis Vaccines and Diagnostics Inc.,
Cambridge, MA, United States, 19National Cancer Institute, Biostatistics
and Data Management Section, Bethesda, MD, United States, 20The
EMMES Corporation, Rockville, MD, United States, 21Case Western
Reserve University, Pathology, Cleveland, OH, United States
1
Background: Adjuvants modulate the immune response and can
improve the immunogenicity of vaccines. Immunogenicity studies in
humans suggest that MF59 is a more efficient adjuvant than alum as it
triggers enhanced B and T cell immune responses.
Methods: We performed a study (P162MRV144) in macaques using
repeated challenges with SIVmac251 powered to benchmark the results
of RV144 vaccine that showed limited efficacy in a phase III clinical
trial in Thailand. We compared the efficacy of ALVAC-SIV/gp120 vaccine
administered with alum or MF59 adjuvants to placebo in the SIV mac251
model. Transcriptional profiling of blood from 54 immunized macaques
before vaccination (pre-vax), after immunization with ALVAC-SIV alone
(post-1st) and after immunization with ALVAC-SIV combined with the
protein/adjuvant MF59 or Alum immunization (post-3rd) was performed.
Differential expression analysis and pathway enrichment analysis was
used to identify genes and pathways associated with protection. A naïve
Bayes classifier was build to predict vaccine protection.
Results: We found that alum protected macaques from SIVmac251
acquisition (log-rank test: p=0.0205), confirming the main results
156
P03.05 LB
Association of HIV-1 Env Antibody Responses
with Postnatal Mother-to-Child Transmission
of HIV-1 via Breastfeeding
Genevieve Fouda1, Justin Pollara1, Erin McGuire1, Wesley
Rountree1, Glenn Overman1, Gerald Tegha2, Deborah
Kamwendo2, Jacob Kumwenda2, Julie Nelson3, Larry Liao1,
Sascha Ellington4, Caroline King4, Denise Jamieson4, Charlie Van
der Horst3, Athena Kourtis4, Georgia Tomaras1, Guido Ferrari1,
Sallie Permar1
Duke Human Vaccine Institute, Durham, NC, United States, 2University
of North Carolina Malawi Project, Lilongwe, Malawi, 3University of North
Carolina, Chapel Hill, NC, United States, 4Centers for Disease Control and
Prevention, Atlanta, GA, United States
1
Background: Immune factors in milk may contribute to protect HIVexposed breastfed infants who do not acquire HIV-1 despite high levels
of mucosal virus exposure. To identify humoral immune responses
associated with protection against postnatal transmission, we evaluated
milk and plasma humoral responses in Malawian HIV-infected women
from the control arm of the Breastfeeding, Antiretrovirals and Nutrition
study, who did or did not transmit HIV to their infants via breastfeeding.
Methods: Transmitting (n=22) and non-transmitting women (n=65)
were matched on peripheral CD4 count and postpartum visit. Envspecific IgG and IgA responses were assessed in available milk (n=87)
and plasma samples (n=42) using a binding antibody multiplex assay.
Antibody dependent cell-mediated cytotoxicity (ADCC) was measured
against a postnatally-transmitted HIV-1 Env variant, and neutralization
was measured against a clade C tier 1 virus. Multivariable logistic
regression models adjusting for plasma viral load and CD4 count were
used to compare responses between groups.
Results: The magnitude of IgG responses against the Env antigens,
including gp70 B CaseA2 V1V2, was similar between transmitters and
non-transmitters. However, there was a trend towards an association
between lower transmission risk and plasma IgG against gp70 B CaseA2
V1V2 with a valine to lysine mutation at position 169 before but not after
adjusting for multiple comparisons (OR=0.39, p=0.09, false discover
rate (FDR) =0.46). Lower risk of transmission was also associated with
high magnitude milk, but not plasma IgA against certain Env antigens,
that was not significant after adjusting for multiple comparisons (B con
gp140 IgA: OR=0.57, p=0.007, FDR=0.1). There was no association
between ADCC or tier 1 neutralization responses and transmission.
Conclusions: These hypothesis generating results indicate that further
studies to examine the role of mucosal IgA and plasma V1V2-specific
IgG responses in protection against postnatal HIV-1 transmission may
be warranted.
www.hivr4p.org
157
POSTERS
of the RV144 study. However MF59 did not protect macaques from
SIVmac251 acquisition (log-rank test: p=0.562), despite MF59’s ability
to elicit higher systemic T-cell and antibodies responses. Association
between the changes in transcriptional profiles and risk of SIVmac251
acquisition resulted in the identification of 12-gene expression signature
able to predict prior to vaccination protection by ALVAC+alum (ROC:
Accuracy=65.2%, p≤0.05). Seven of the twelve genes of the signature
were related to Ras signaling.
Conclusions: System biology revealed that RAS, a signal transducer
that facilitates cross talk among B-cells, T-cells and antigen presenting
cells, as a biomarker of vaccine efficacy in the ALVAC+alum treated
animals. These data suggest that activation of RAS may constitute a
novel approach to improve vaccine efficacy against HIV.
Posters
Posters 04: Correlates of Protection in Highly Exposed Seronegative People
P04.01
P04.02
ABSTRACT WITHDRAWN
Molecules Involved in the Vitamin-D Pathway
Correlate with Higher mRNA Expression
of Anti-HIV Molecules in HIV Exposed
Seronegative Individuals
Wbeimar Aguilar-Jiménez1, Manuel G. Feria1, Eliuth D. Arcia1,
Wildeman Zapata1,2, Maria T. Rugeles1
Grupo Inmunovirología. Universidad de Antioquia UdeA, Medellin,
Colombia, 2Grupo Infettare. Universidad Cooperativa de Colombia,
Medellin, Colombia
1
POSTERS
Background: Vitamin D (VitD) is an endogenous immunomodulator that
could protect from HIV-1 infection. We recently found that high levels
of VitD and its receptor are associated with natural resistance to HIV1 infection; up-regulation of the anti-inflammatory cytokine IL-10, and
the induction of defensins in mucosa might be part of the mechanisms
involved in this association. However, several other molecules might be
involved in this protection.
Methods: To further explore this issue, a case-control study using
samples of 58 HIV-1-exposed but seronegative (HESN) individuals, and
59 non-exposed healthy controls (HCs) was carried out. mRNA relative
units (RU) of APOBEC3G, ELAFIN, TRIM5alfa, SAMHD1 and Catelicidin
in peripheral blood mononuclear cells (PBMCs), oral and genital mucosa
were quantified by qRT-PCR. Circulating VitD and mRNA levels of VDR
were previously reported and used for correlations.
Results: HESNs had significantly higher mRNA RU of the antiviral
molecules APOBEC3G and ELAFIN in PBMCs and SAMHD1 and
Catelicidin in oral mucosa compared to HCs. Positive correlation between
VDR and ELAFIN mRNA in PBMCs of HESNs was found.
Conclusions: These results suggest that high levels of APOBEC3G,
ELAFIN, SAMHD1, and Catelicidin are associated with natural resistance
to HIV-1 infection. VitD may be up-regulating the expression of ELAFIN
as observed for other anti-HIV-1 molecules. However, further studies are
required to define causal associations.
158
Posters 05: Family Planning
P05.01
P05.02
Contraceptive Use and Pregnancy Incidence
among VOICE Participants in Uganda
Stop and Save Children from HIV! Rural-urban
Differentials, Barriers and Motivators of Longterm Contraception in Zimbabwe, Mixed
Methods Results
Collaboration, Kampala, Uganda, 2Magee Women’s Research Institute,
Pittsburgh, PA, United States, 3Johns Hopkins University School of
Medicine, Baltimore, MD, United States, 4University of Washington,
Seattle, WA, United States, 5University of Zimbabwe, Harare, Zimbabwe,
6
Fred Hutchinson Cancer Research Center, Seattle, WA, United States
1
Background: Recent HIV prevention trials have required use of effective
contraceptive methods to fulfill eligibility for enrolment. We compared
pregnancy rates in participants enrolled in the VOICE (MTN-003) trial
who initiated depot medroxyprogesterone acetate (DMPA) or combined
oral contraceptives (COCs) at enrollment (new users) relative to those
already on DMPA or COCs.
Methods: Data were analyzed from HIV-1 seronegative participants
enrolled in Uganda. Prior to enrolment, information on contraceptive
type and initiation date was obtained; contraception was provided
to new users. Participants received urine pregnancy tests at monthly
follow-up visits. Cox proportional hazards models stratified by baseline
contraceptive method were used to compare pregnancy incidence among
new users (initiated ≤ 60 days prior to enrollment) and established users
(initiated >60 days prior to enrolment). Participants were censored after
the first observed pregnancy on study.
Results: Of 322 women enrolled, 296 were COC or DMPA users; 82
(28%) were new users (50 [61%] initiated DMPA and 32[39%] initiated
COCs) and 214 (72%) were established users. Overall pregnancy
incidence was 13.35 per 100 person-years [p-yrs] (49/367 p-yrs),
with an incidence in DMPA users of 5.39 per 100 p-yrs (13/241 p-yrs)
compared to 28.62 per 100 p-yrs (36/126 p-yrs) in COC users. In new
DMPA users, pregnancy incidence was 10.20 per 100 p-yrs versus 3.48
per 100 p-yrs in established DMPA users (adjusted hazard ratio [aHR] =
2.84; 95% confidence interval [CI] 0.92 - 8.74). Similarly, in new COC
users, pregnancy incidence was 42.67 per 100 p-yrs versus 23.67 per
100 p-yrs in established COC users (aHR 2.08; 95% CI 1.02 - 4.23).
Conclusions: New contraceptive users in VOICE had an increased
pregnancy risk. Pregnancy incidence was high among both new and
established COC users. New DMPA and all COC users participating in HIV
prevention trials may benefit from intensive contraceptive counseling
and provision of less user-dependent methods.
Munyaradzi Kenneth Dodzo1,2, Munyaradzi Paul Mapingure1,
Noah Taruberekera3
Population Services International Zimbabwe, Research and Metrics,
Harare, Zimbabwe, 2University of Zimbabwe, Population Studies,
Harare, Zimbabwe, 3Population Services International, Research and
Metrics, Johannesburg, South Africa
1
Background: Socio-economic and biologic vulnerabilities predispose
women to HIV infection, leading to higher maternal risk and infection of
babies. Contraception lowers HIV incidence, viral load, pregnancy and
vertical transmission. Long-acting reversible contraceptives (LARCs) implants and IUDs - help overcome access barriers, common in poverty,
emergency or abuse cases. In Zimbabwe, their prevalence is 2% but
CPR is 60%. PSI seeks their full potential by integrating them in HIV
prevention, sexual and reproductive health. We aim to identify barriers,
facilitators and correlates of LARCs by place of residence.
Methods: We used two studies. One involved 1185 women (15 - 49
years) in a national, multi-stage household survey, using UNIANOVA in
SPSS to analyse data. Correlates were built from multiple items scored
1 to 5. Qualitative data were analysed thematically from 44 in-depth
interviews with adult men and women. 24 women were using LARCs
and 12 were not. 4 men had partners using LARCs and 4 were not.
Results: Urban women, 6%, were more likely to use LARCs than rural,
3%, p< 0.05. Dual use of LARCs and condoms was higher among urban,
5%, than rural women, 3%, p< 0.05. Partner support was higher among
urban women, 24%, than rural, 18%, p< 0.001. Scaled availability was
better among urban (3.92) than rural (3.69), p< 0.001 as was social
support - urban (2.81) vs. rural (2.62), p< 0.001 and self-efficacy - urban
(3.07) vs. rural (2.56), p< 0.001. Qualitative data showed that LARCs
spell peace of mind, low maintenance costs, flexibility and freedom from
parenting duties. But they interfere with menses, link with infertility and
threaten womanhood. Fear of pain, infection and side effects are other
barriers. Insertion or removal skills portray them as elitist, costly and
inaccessible.
Conclusions: Prevalence of LARCs is low and myths abound. PSI must
promote awareness and knowledge about LARCs. Program must create
opportunity, improve ability and increase motivation, especially among
rural women.
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159
POSTERS
Carolyne A. Akello1, Katherine E. Bunge2, Clemensia Nakabiito1,
Brenda G. Mirembe1, Mary G. Fowler1,3, Anupam Mishra4,
Zvavahera Mike Chirenje5, Jeanne Marrazzo4, Connie Celum4,
Jennifer E. Balkus4,6
Posters
P05.03
P05.04
Capacity Strengthening and Training
of Government Nurses on Long-acting
Reversible Contraceptive (LARC) Methods in
Kigali, Rwanda
Strategies for Successful Promotion and
Implementation of Contraceptive Method Mix
in MTN-020/ASPIRE: The Why and How at
Kampala Site
Rosine Ingabire1, Etienne Karita1, Nurilign Ahmed1, Roger
Bayingana1, Julien M. Nyombayire1, Robertine Sinabamenye1,
Jean N. Nduwamungu1, Honoree Uwamahoro1, Sarah Strunk1,
Amanda Tichacek1,2, Susan Allen1,2
Betty Kamira1, Clemensia Nakabiito1, Justine Kamya Nsangi1,
Joselyne Nabisere1, Christine Nagawa1, Samuel Kabwigu1, Fred
Kapaata1, Flavia K. Matovu1
Projet San Francisco, Rwanda Zambia HIV Research Group, Kigali,
Rwanda, 2Emory University, School of Medicine, Department of
Pathology and Laboratory Medicine, Atlanta, GA, United States
MU-JHU Research Collaboration, Kampala, Uganda
1
1
POSTERS
Background: In Rwanda, adult women 15-49 have higher HIV
prevalence rate (4%) compared to men (2%). Each year, nearly half (47%)
of all pregnancies in the country are unintended. Preventing unintended
pregnancies among HIV positive women through family planning (FP) is
the lowest cost perinatal HIV prevention strategy and contraceptive use
is associated with longer survival in HIV positive women. HIV negative
women enrolled in clinical trials should also avoid pregnancy.
Methods: Project San Francisco (PSF) provided training in Long Acting
Reversible Contraceptives (LARC), the copper IUD and Jadelle implant in
6 Kigali government clinics to enhance access to and uptake of the most
effective family planning methods. From June 2009- May 2014, PSF
conducted monitoring & evaluation of LARC services in Kigali Rwanda.
The overall numbers of nurses trained, provision of LARC methods at the
government clinics, and LARC provision were measured.
Results: PSF conducted 19 LARC training sessions with 59 nurses
trained in IUD/Implant insertions. From 2009-2012, these nurses
provided 5,281 LARC methods (996 IUDs and 4,285 Implants). During
this period, the percentage of family planning clients who requested
LARC methods increased annually in the 6 target clinics: IUD provision
among FP clients increased from 1% in 2009 to 1.2% in 2010, 5%
in 2011 and 9% in 2012. Implant insertions increased from 8.6% in
2009 to 7.1% in 2010 to 21% in 2011 and 37% in 2012. PSF continues
to provide technical assistance for the provision of LARC methods and
between June 2012- March 2014, 12,937 (3,009 IUDs and 9,928
Implants) have been provided.
Conclusions: When nurses are comfortable and competent with
inserting LARC methods, they are more likely to successfully promote
and provide these methods to both HIV positive and negative clients. In
turn, these clients are very likely to opt for LARC when they are available.
Established LARC services in government clinics enhance prong 2 of
PMTCT and readiness for prevention trials.
160
Background: To enroll in ASPIRE; participants must be on an effective
method of contraception ranging from COC, DMP, Implants, IUCD and
sterilization. In Uganda, there is a low contraceptive prevalence and a
high unmet need (43%) of use. This was reflected in the VOICE study
where uptake for Long Acting Reversible Contraceptive (LARC) was only
6%. In an attempt to enhance Contraceptive Method Mix (CMM) in
ASPIRE, the Kampala team describes how this was made successful.
Methods: Preparatory activities started prior to site activation with
formation of the Contraceptive Action Team (CAT) under the leadership
of the MTN Contraceptive Steering Committee with the aim of exploring
misconceptions, identifying barriers, and increasing LARC uptake.
Following this, 2 members were declared contraceptive experts, and
charged with spearheading CMM. These liaised with existing f/planning
clinics to train study counselors on counseling messages and clinicians
in inserting and removal of LARCs. We started linkages with the
community and satellite clinics to demystify myths and misconceptions
using flip charts and models and the community found this exciting. All
methods were made available onsite, assigned a nurse for logistics and
created a room for providing the LARC.
Results: There is an increase in the use of IUCDs and implants and a
decrease in injectables and pill use after the inception of CAT. Previously,
contraception was limited to COCs and DMPA. LARCs (Implants, IUCD)
and tubal ligation were being offered at the nearby Hospital. Now all
methods are available on-site except tubal ligation.
Conclusions: Method Mix and expansion of LARC methods is possible
in a low resource limited setting. For success, information is key, ongoing
refresher training, regular evaluations, peer support, good political will
ongoing participant care and counseling are all important.
P05.05
P05.06
Contraceptive Preference among HIV High
Risk Women Pre-screened for a Phase III
Vaginal Microbicide Trial in South-Western
Uganda
Knowledge and Use of Modern Family
Planning Methods in Fishing Communities
along Lake Victoria, Uganda
Medical Research Council/ Uganda Virus Research Institute, Uganda
Research Unit on AIDS, Entebbe, Uganda, 2MRC/UVRI, Uganda
Research Unit on AIDS, Entebbe, Uganda, 3International Partnership for
Microbicides, Paarl, South Africa
1
Background: Women’s contraceptive choices vary according to the type
of relationships they have and other aspects of their life. We describe
the preference of modern contraceptive methods among HIV high risk
women screened to participate in a Phase III vaginal microbicide trial.
Methods: Medical Research Council/Uganda Virus Research Institute,
Uganda Research Unit on AIDS, in collaboration with International
Partnership for Microbicides (IPM), is evaluating safety and efficacy
of a dapivirine vaginal ring among healthy HIV negative women in a
multicentre Phase III trial (The Ring Study). One of the study eligibility
criteria is being on a stable form of contraception in order to reduce
the pregnancy rate. As part of the screening process, women are
identified from townships and fishing communities within 50 km
from the centre. Data on use of modern contraception methods (Pills,
Injectables, Implants, IUCDs, Surgical) are collected, and confirmed by
a contraceptive card or presence of an implant or IUCD strings. Women
who are willing are provided by the research centre with contraception
of choice (pills, injectables) or referred to a provider (Implants, IUCDs,
Surgical). The proportion of women per method used was determined.
Results: A total of 489 women, with a mean age of 28 years, were
pre-screened (age range 18 to 44 years). Most (306, 63%) were already
using contraception: 55% were on Injectables, followed by Implants
(28%), IUCDs (8%), Pills (7%) and only 2% had undergone a Surgical
method. Of the women who were not on any method, 106 (58%)
accepted Injectables, 40 (22%) preferred Implants, 33 (18%) oral Pills
and 4 (2%) chose IUCDs.
Conclusions: There is high contraceptive use among HIV high risk
women and the majority prefer Injectables. This is encouraging and
will hopefully reduce discontinuation of volunteers from the trial due
to pregnancy.
1
UVRI-IAVI HIV Vaccine Program, Entebbe, Uganda, 2Kabwohe Clinical
Research Center (KCRC), Kabwohe, Uganda, 3International AIDS Vaccine
Initiative, New York, NY, United States, 4Makerere University College of
Health Sciences, School of Public Health, Kampala, Uganda
Background: Interventions geared towards reduction of unwanted
pregnancies and elimination of mother to child transmission of HIV have
led to the integration of family planning (FP) and HIV-care services into
the general health care package. Fishing communities are another key
population for HIV infection in Uganda with incidence and prevalence
~3-6 times higher than estimated national rates. These communities are
being targeted for HIV prevention trials where FP is a requirement. There
is paucity of data on the knowledge and use of modern FP including
hormonal, barrier and permanent methods in fishing communities.
Methods: Data are accrued from a baseline survey of 2191 HIVuninfected individuals aged 18-49 years enrolled between September
2011 and March 2013 from 8 fishing communities. At the 12-months
post baseline visit, data on knowledge and use of family planning were
collected through a semi structured questionnaire.
Results: About three quarters (1688/2191, 77%) of the volunteers
provided data on FP. Knowledge about family planning was high, 87%
(1477/1688). Just above a third, 35% (595/1688) reported current use
of at least one modern FP method. Use of modern FP by a respondent
and/or their sexual partner was more frequently reported by females
than males (41.6% vs. 29.2%, p< 0.001). Factors associated with higher
use of modern FP were: increasing level of education (p=0.02), being
currently married (p< 0.001) and currently having other sexual partner/s
(p< 0.001). Reasons for non-use of modern FP methods were: desire
for more children (30.6 %), currently not being in a sexual relationship
(27%), fear of side effects (12.2%), currently pregnant/breast feeding
(7.8%) being young (7.1%) and partner refusal (5.2%).
Conclusions: Knowledge of modern FP methods among fishing
communities along Lake Victoria, Uganda is high but use is still low.
High fertility desires, not being in a sexual relationship and fear of
perceived side effects are key barriers to use of modern FP.
www.hivr4p.org
161
POSTERS
Sylvia Kusemererwa1, Andrew Abaasa2, Emanuel Aling2, Margaret
Kalibbala2, Sarah Nakato2, Juliet Kyomugisha2, Neliette Van
Niekerk3, Annalene Nel3, Anatoli Kamali2
Annet Nanvubya1, Juliet Mpendo1, Ali Ssetaala1, Julius Ssempiira1,
Annet Nalutaaya1, Mathias Wambuzi1, Steven Asiimwe2, Leslie
Nielsen3, Fredrick Makumbi4, Noah Kiwanuka1,4
Posters
P05.07
P05.08
Introduction of Long Acting Reversible
Contraception (LARC) in a Clinical Trial Setting
in Kwazulu-Natal
Combination Prevention for PMTCT Prongs 1
& 2: HIV Testing for Couples and Long Acting
Reversible Family Planning/dual Method Use
Promotion in Zambia
Arendevi Pather1, Jayajothi Moodley1, Naureen Cele1, Vermala
Juggernath1, Gita Ramjee1, MTN 020
Medical Research Council of South Africa, HIV Prevention Research
Unit, Durban, South Africa
1
POSTERS
Background: Although debatable, recent studies have shown a possible
increased risk of HIV acquisition with injectable hormonal contraception.
In KwaZulu-Natal (KZN), the epicenter of the HIV epidemic, injectable
contraception is widely used. Based on the recommendation by the
World Health Organization (WHO) to increase contraceptive options, the
Microbicide Trials Network (MTN) formed a Contraceptive Action Team
(CAT), tasked to improve the contraceptive options at MTN research sites.
As part of this initiative, the Medical Research Council (MRC) Clinical
Research Sites (CRSs) embarked on a drive to provide Intrauterine
Devices (IUDs) and implants at the CRSs.
Methods: Challenges included lack of LARC knowledge; myths and
misconceptions about LARC, participant fears about partner objection
and LARC side effects; preference to remain on current method, provision
of training to clinical staff and accessing LARC. Strategies used included:
community education and awareness conducted during community
meetings and outreach campaigns; ongoing training provided to clinical
staff on LARC insertions in collaboration with the KZN Department of
Health (DoH); accessing of LARC from private suppliers and current
negotiation with DoH; counselling and education on LARC during waiting
room discussions at the CRSs and individual contraception counselling
sessions. At these discussions, participant fears about partner objection
and fear of side effects of LARC were also addressed.
Results: We have received positive feedback from the wider community
regarding LARC. Community programmes ensure that myths and
misconceptions about LARC are dispelled. Clinical staff have been
trained on insertion of LARC at three CRSs.
Conclusions: The CAT initiative has been successful in introducing and
implementing LARC within the MRC CRSs in KZN. Despite this, ongoing
efforts are still required to increase the number of clinical staff trained
on IUD and implant insertions at the CRSs and to increase the utilisation
of LARC.
162
Bellington Vwalika1,2, Inambao Mubiana3, William Kilembe1, N
Munir1, Kathleen Wu1, N Ahmed1, Ashey Appiagey1, B Getachew4,
R Parker4, J Seed5, Danielle Dang6, Amanda Tichacek5, Kristin
Wall5, Susan Allen5
1
Zambia Emory HIV Research Project, Lusaka, Zambia, 2University of
Zambia, Lusaka, Zambia, 3Zambia Emory HIV Research Project, Ndola,
Zambia, 4Zambia Rwanda HIV Research Group, Atlanta, GA, United
States, 5Rwanda Zambia HIV Research Group, Atlanta, GA, United
States, 6Rwanda Zambia HIV Research Group, Lusaka, Zambia
Background: Prongs 1 and 2 of PMTCT focus on prevention of
heterosexual HIV and prevention of unplanned pregnancy in HIV+
women but receive little attention in the HIV prevention world. Couples’
HIV counseling and testing (CVCT) reduces new heterosexual infections.
Long acting reversible contraceptives (LARC), in particular the IUD
and the implant, are effective modern methods that are not available
to most African women with or at risk of HIV. The UK Department for
International Development has funded a program to integrate CVCT,
family planning promotion, and LARC for clinic visitors and couples in
the community with the goal of averting13,900 new HIV infections.
Methods: Formative research with providers; clinic clients in family
planning, infant vaccination, and ART clinics; and couples seeking
CVCT informed development of training and promotional materials for
nurses, counselors, and community promoters. A program dovetailing
simultaneous ramp-up of supply and demand was implemented in 50
clinics in 6 Zambian cities in Lusaka, Southern and Copperbelt Provinces.
Results: 199 nurses from government clinics were LARC-trained and
306 CVCT counselors, community health workers and LARC clients/peer
educators were trained in couples’ family planning counseling (CFPC)
and promotion. From March 2013 to March 2014, 37,281 couples
received CVCT and 14,564 of those also received CFPC. In this same
time frame, 1269 IUD and 6977 Jadelle were inserted, and 357 IUD
and 1906 Jadelle were removed after a mean of 2.6 years of use. Dual
method use (barrier + LARC) was emphasized with discordant couples.
Conclusions: Combining HIV testing with family planning and targeting
couples has the potential to prevent many new heterosexual and perinatal
infections. Both LARC training and LARC methods are sustained once
administered. With a concerted effort and coordination of supply and
demand, the IUD and implant can be integrated in government clinics and
HIV programs. The combination is feasible, cost effective and sustainable.
Posters 06: Financial Incentives
P06.01
P06.02
Economic Incentives to Reduce HIV Risks
among Male Sex Workers: A Randomized
Pilot in Mexico
Unanticipated Impact of Financial Incentives
on HIV Patients and Providers: Findings from
a Qualitative Substudy (HPTN 065)
Omar Galarraga1, Sandra G. Sosa-Rubi2, Carlos J. Conde-Glez3,
Luis Juarez-Figueroa4, Andrea Gonzalez5, Florentino BadialHernandez5, Sergio Bautista-Arredondo2, Caroline Kuo6, Don
Operario6, Kenneth H. Mayer7
Theresa Gamble1, Jill Stanton1, Elizabeth Greene1, Jamilah Taylor1,
Allison P. Pack1, Victoria Shelus1, Elizabeth E. Tolley1, Sheldon T.
Brown2, Wafaa El-Sadr3,4, for the HPTN 065 Study Team
Background: We test economic incentives for HIV risk reduction among
male sex workers (MSW). N=265 participants were randomized into 4
groups.
Methods: In Treatment 1, they received a medium conditional incentive
($50 USD/each time) only if they were free of new, curable STI at months
6 and 12. In Treatment 2, they received a high incentive ($75 USD/
each time) if they were free of new, curable STI at months 6 and 12.
Controls did not receive incentive even if free of STI. In Unconditional
Incentive, they received medium incentive ($50 USD at months 6 and
12) regardless of STI status. All groups received inconvenience fees
($10 USD/each time at baseline, month 6 and month 12). All incentives
were in the form of food and grocery vouchers. We recruited from
Alameda Central and Clínica Condesa (largest HIV clinic in Mexico City>
9,000 patients). All eligible men took part in a standard HIV education
session after completing baseline measures and before randomization.
All received HIV/STI testing; those infected were offered treatment (for
curable STI).
Results: At baseline, mean age was 25, 76% were single, and 35%
completed high school. On average they had 6 sexual partners per
week, and 83% had used condoms with last three clients. Most were
street-based (70%), but worked at bars/clubs also (30%). Most had at
least one STI (63%): 19% had active syphilis, 11% had chlamydia; and
HIV prevalence was 38%. The average median transaction price per
sexual act was USD$25 with 40% higher price for unprotected sex.
Conclusions: Results provide strong rationale for structural intervention
for MSW. High HIV rates and higher payments for unprotected sex justify
economic intervention which has been demonstrated to be feasible and
acceptable. Preliminary efficacy on STI outcomes, condom use, number
of partners, and transaction prices will be presented at proposed session.
1
FHI 360, Durham, NC, United States, 2James J. Peters VA Medical
Center, Bronx, NC, United States, 3Columbia University Mailman School
of Public Health, ICAP, New York, NY, United States, 4Harlem Hospital,
Background: The HPTN 065 (TLC-Plus) study evaluated the effect of
providing quarterly $70 financial incentives (FI) to HIV‑infected patients
on antiretroviral therapy (ART) with HIV RNA< 400 copies/mL to
encourage adherence. Nineteen HIV care sites randomized to the FI
intervention dispensed 39,359 FI gift cards over two years in the Bronx,
NY and Washington, DC.
Methods: We conducted semi-structured interviews with 75 patients
from 14 sites and 12 site investigators (SIs) (mostly clinicians); three
focus groups were conducted with 12 staff members from 10 sites. Data
were analyzed to determine the effect of the FI on aspects other than
adherence.
Results: While the purpose of the FI was to influence ART adherence,
patients and staff reported more engagement in care (improved visit
adherence, patient/provider relationships, and general preventive care)
and increased opportunities for HIV and general health education.
Patients reported that the FI increased their ability to manage their HIV,
improved their attitudes towards the disease and its treatment, and
caused other behavior change (e.g. motivation not to drink so as to
remember to take their ART). Many patients also reported that the FI
met a real financial need. During the program, patients felt cared for
by staff, and staff felt good about doing something positive for their
patients. Conversely, for some patients, the gift cards invoked a sense
of entitlement (i.e. patients felt they deserved, rather than earned, the
gift card), reminded them of their HIV status, and may have devalued
their efforts to remain adherent. Especially at the beginning of the
intervention, some clinics experienced changes in clinic flow and had
difficulty managing the influx of patients.
Conclusions: The use of FI intended to influence ART adherence can
affect both patients and providers in unanticipated ways. While some
effects were, at least transiently, negative, most of the additional findings
were positive, with the potential to improve HIV care and overall patient
health.
www.hivr4p.org
163
POSTERS
Brown University, Health Services Policy & Practice, Providence, RI,
United States, 2National Institute of Public Health, Health Economics,
Cuernavaca, Mexico, 3National Institute of Public Health, Infectious
Disease Research Center, Cuernavaca, Mexico, 4National Institute of
Public Health, Mexico City, Mexico, 5Clinica Condesa, Mexico City,
Mexico, 6Brown University, Behavioral & Social Science, Providence,
RI, United States, 7Fenway Health & Harvard University, Global Health,
1
Posters
Posters 06: Financial Incentives
P06.03
P06.04
The Impact of Implementing a Financial
Incentive Program for Viral Suppression
on the Clinic Environment: A Qualitative
Substudy of HPTN 065
Understanding of Viral Load among
Participants Receiving Financial Incentives for
ART Adherence: Findings from a Qualitative
Substudy of HPTN 065
Elizabeth Greene1, Theresa Gamble1, Elizabeth E. Tolley1, Allison
P. Pack1, Jill Stanton1, Jamilah Taylor1, Victoria Shelus1, Jason
Leider2, Wafaa El-Sadr3,4, Bernard Branson5, for the HPTN 065
Study Team
Elizabeth Greene1, Jamilah Taylor1, Allison P. Pack1, Jill Stanton1,
Victoria Shelus1, Elizabeth E. Tolley1, Lawrence J. D’Angelo2,
Wafaa El-Sadr3,4, Theresa Gamble1, for the HPTN 065 Study Team
FHI 360, Durham, NC, United States, Jacobi Medical Center, New
York, NY, United States, 3Columbia University Mailman School of Public
Health, ICAP, New York, NY, United States, 4Harlem Hospital, New York,
NY, United States, 5Centers for Disease Control and Prevention, Atlanta,
GA, United States
1
2
POSTERS
Background: HPTN 065 (TLC-Plus) studied the effect of providing
quarterly $70 financial incentives (FI) to HIV-infected patients on antiretroviral therapy (ART) with HIV RNA< 400 copies/mL to encourage ART
adherence. Nineteen HIV care sites randomized to the FI intervention
dispensed a total of 39,359 FI gift cards over two years in the Bronx, NY
and Washington, DC.
Methods: Semi-structured interviews were conducted with 75 patients
from 14 sites and 12 site investigators (SIs) (mostly providers); three
focus groups were conducted with 12 staff members from 10 sites. Data
were analyzed qualitatively to determine the impact of the FI program
on clinic environment.
Results: The majority of patients interviewed noticed no effect on their
clinic experience due to the FI program. SIs and staff reported that the
study-imposed timing of quarterly viral load tests necessary to qualify
for FIs often conflicted with typical clinical care schedules and caused
clinics to become busier as patients visited frequently or adhered to
appointments. SIs saw a benefit to this (better engagement in care), but
it also increased demands on staff time and space and interfered with
clinic flow. Some SIs and staff found determining eligibility, tracking and
disbursing gift cards to be logistically challenging. Some staff also had
difficulty dealing with patients who felt “entitled” to FIs and discussing
eligibility for FIs in semi-private spaces. SIs and staff described a
learning curve as processes were developed to improve logistics, clinic
flow, and FI and patient tracking, and as patients and providers became
educated about program requirements. Most SIs attributed smooth
implementation to well-trained and dedicated staff and provider buy-in.
Many indicated that positive patient interactions with the FIs outweighed
implementation challenges.
Conclusions: A large scale FI program posed challenges for providers
and staff that were mitigated by effective systems and education without
negative effects on patients’ clinic experiences.
164
1
FHI 360, Durham, NC, United States, 2Children’s National Medical
Center, Washington, DC, United States, 3Columbia University Mailman
School of Public Health, ICAP, New York, NY, United States, 4Harlem
Hospital, New York, NY, United States
Background: HPTN 065 (TLC-Plus) evaluated the effect of providing
quarterly $70 financial incentives (FI) to HIV-infected patients on
antiretroviral therapy (ART) to encourage ART adherence. Viral load
(VL) suppression, defined as HIV RNA< 400 copies/mL, was used as a
marker for ART adherence. Nineteen HIV care sites were randomized to
the 2-year FI intervention in the Bronx, NY and Washington, DC.
Methods: We conducted semi-structured interviews with 75 patients
ages 14-72 from 14 sites and 12 site investigators (SIs) (mostly clinicians).
Qualitative data were analyzed to determine perceptions about VL.
Clinical data were collected to determine patients’ VL suppression status.
Results: Based on their descriptions, patients’ understanding of the
meaning of VL or their specific VL results was variable; almost half
misunderstood VL. Many confused it with CD4 count, some thought VL
should ideally be high, and some admitted lack of understanding despite
efforts from providers. Several used the term ‘undetectable’ incorrectly,
suggesting they had heard this term but did not grasp its meaning.
More patients recognized that ART adherence would result in a good
VL, even though some were unaware of the definition of a “good” value.
While some patients said their understanding of VL improved during the
study, over half said that it did not. In contrast, most SIs thought that
many of their patients understood their VL results, and several felt the
study increased understanding of VL. There was no association between
observed viral suppression and accurate understanding of VL or its
relationship to ART adherence.
Conclusions: Many patients did not accurately understand VL even
though it was the outcome that qualified them to receive the FI.
Clinicians thought patients understood VL when often they did not.
However, accurate understanding of VL was not always necessary to
comprehend that VL improved with ART adherence, nor was it related to
viral suppression among patients interviewed.
Posters 07: Good Participatory Practices and Community Involvement
P07.01
P07.02
Ethics of HIV Prevention in Research Literacy
Implementing Good Participatory Practice
(GPP) in HVTN505 - The HIV Vaccine Trials
Network (HVTN) Experience
New HIV Vaccine and Microbicide Advocacy Society (NHVMAS),
Programme, Lagos, Nigeria, 2New HIV Vaccine and Microbicide
Advocacy Society (NHVMAS), Administrative, Lagos, Nigeria,
3
Journalists Against AIDS (JAAIDS) Nigeria, Programme/Administrative,
Lagos, Nigeria
1
Background: When NHVMAS studied the methodology section of
the 2007 and 2010 IBBSS survey reports, it was clear that though the
methodology was sound, there was a gap in the ethics of the conduct of
trials as there was no evidence of community engagement in the design
of the survey. Members of the communities engaged with these surveys
are becoming concerned with their continued participation in surveys
that do not translate to interventions for them. While NHVMAS makes
effort at increasing awareness of the public, researchers and policy
makers to this clause of the HIV research policy, it also wants to focus
on building the capacity of the community.
Methods: NHVMAS undertook three days training for gatekeepers
(IDU, FSW and MSM) communities in seven sites where the IBBSS 2014
survey was to take place. The training focused on empowering the
community gatekeepers as trainers on what they need to know about
research, its importance, informed consent, confidentiality and so on
and how they can also transfer same to their peers. Participants learnt
about the new and existing HIV prevention tools such as microbicides,
HIV vaccines, medical male circumcision and female condoms. An
interactive approach was engaged. Analysis of the pre and post test
was conducted to compare knowledge of participants on topics treated
before and after training.
Results: There was difference in the pre and post test (45.4vs 56.2;
p=1.07626E-05). One of the impact of the training was the demand
made by community members during the planning of the IBBSS 2014
survey for a pre-planning consultation with the community of MSM
before conclusion on study design. There have been other reports of
community members demanding for informed consent and ethics
clearance prior to conduct of research with community members.
Conclusions: When community gatekeepers become more research
literate, they can become actively engaged with research design. Also
empowered to make demands for changes in research processes that
could address their community needs.
Gail B. Broder1,2, James P. Maynard1,3, Shelly T. Karuna1,4, Chuka
Anude5, Magda E. Sobieszczyk6, Scott M. Hammer6, HVTN 505
Protocol Team of the NIAID-funded HIV Vaccine Trials Network
Disease Division, Seattle, WA, United States, 2HIV Vaccine Trials
Network, Community Engagement Unit, Seattle, WA, United States,
3
HIV Vaccine Trials Network, Communications and Community
Engagement Units, Seattle, WA, United States, 4HIV Vaccine Trials
Network, Clinical Development Unit, Seattle, WA, United States,
5
DAIDS/NIAID/NIH, Vaccine Clinical Research Program, Bethesda,
MD, United States, 6Columbia University College of Physicians and
Surgeons, Division of Infectious Diseases, Department of Medicine,
1
Background: HVTN has valued community involvement in its operations
since it began, and continued this practice with HVTN505, a Phase 2B
trial in MSM and transwomen. During HVTN505 implementation, results
from the iPrEx trial and subsequent FDA approval of Truvada® for preexposure prophylaxis (PrEP) led the HVTN to consider changes to HIV
prevention services offered to participants (ppts). Several community
consultations informed our actions that addressed the GPP principles of
transparency and the standard of HIV prevention.
Methods: Community members were part of the protocol team from
the outset, contributing to study design, informed consent materials,
engagement and recruitment efforts, and communications planning.
Post-iPrEx, HVTN505 ppts were surveyed about their intent to use
PrEP. Consultations were held with Global Community Advisory Board
members representing all network sites and with a wider group of
community stakeholders to discuss the iPrEx results and standards of
HIV prevention. Community input was sought on the advisability of
providing PrEP for trial ppts interested in using it, and on the mechanisms
of that provision. Post-FDA approval, a second consultation was held
regarding implementation of such a plan.
Results: Stakeholders felt strongly that ppts should be educated about
PrEP and counseled about how to access it. The study was amended
and incorporated the potential for 20% uptake in PrEP use among ppts.
As discussions in the field continue about the changing standard of HIV
prevention, HVTN is working to make Truvada® available free of charge
to interested HVTN505 ppts. This evolving discussion will continue to
inform future trial designs.
Conclusions: The HVTN embraces the GPP guidelines and continues its
efforts to fully implement them. Involving the community at all levels
is key in the HVTN’s structure and all of its trials. HVTN505 provided
a unique opportunity to engage the community at a time when the
conversation about HIV prevention was evolving particularly rapidly.
www.hivr4p.org
165
POSTERS
Obiajulu A. Amuamuziam1, Morenike O. Ukpong1, Florita C.
Durueke1, Oluwatosin B. Alaka1, Olayide Akanni2,3
Posters
P07.03
P07.04
Increasing the National Impact and Uptake
of Good Participatory Practice Guidelines
for Biomedical HIV Prevention (GPP): Three
Country Case Studies
Innovation in Stakeholder Engagement:
Piloting a Monitoring and Evaluation Toolkit
Stacey Hannah1, Julius Ecuru2, Udom Likhitwonnawut3, Catherine
Slack4, Ntando Yola5
1
AVAC, New York, NY, United States, 2Uganda National Council of
Science and Technology, Kampala, Uganda, 3Thai NGO Coalition
on AIDS, Chiang Mai, Thailand, 4HIV AIDS Vaccines Ethics Group,
Pietermaritzburg, South Africa, 5Desmond Tutu HIV Foundation, Cape
Town, South Africa
POSTERS
Background: The Good Participatory Practice Guidelines (GPP) provide
a normative framework for stakeholder engagement in HIV prevention
trials. GPP has been applied at site, network and sponsor levels to
enhance trial-specific and general site operations. GPP contains
recommendations, not requirements, and no entity was established to
ensure implementers’ fulfillment of practices. There is increasing interest
in incorporating GPP into national trial oversight processes as one
method of strengthening positive impact. We review case studies from
three key countries for HIV prevention research.
Methods: The Uganda National Council of Science and Technology
(UNCST) oversees regulatory approval of research nationally. In 2011,
UNCST initiated incorporation of GPP into revision of national ethics
guidelines for clinical research. In South Africa, recommendations
for national GPP implementation were developed through a series
of consultations with research staff and stakeholders. Efforts are also
ongoing to consider select high-impact activities for Research Ethics
Committees reviewing engagement plans in protocols. In Thailand,
implementation of GPP at research centers led to formation of a national
CAB, development of a national GPP plan within the Ministry of Public
Health and incorporation of GPP into ethics review.
Results: Adoption of minimum requirements for GPP in clinical research
has helped establish the guidelines as standard practice in several
countries and work toward greater impact of these efforts. Cases
described provide models for adaptation and use in additional countries
engaged in prevention research.
Conclusions: GPP was developed to enhance the ethical and effective
implementation of HIV prevention research. National adoption and other
strategies to increase impact are critical to realizing this aspirational
goal. The field may benefit by replication of these models in additional
countries.
166
Stacey Hannah1, Stephanie Seidel2, Sinazo Pato3, Monique Oliff4
1
AVAC, New York, NY, United States, 2Global Alliance for Tuberculosis
Research, New York, NY, United States, 3Wits Reproductive Health &
HIV Institute, Johannesburg, South Africa, 4Independent Consultant,
Mombasa, Kenya
Background: The Good Participatory Practice Guidelines (GPP) for HIV
prevention trials were adapted for TB drug trials and are being used
generically in additional settings. As practices expand, increasing
focus rests on GPP impact. A cross-field, cross-disease monitoring and
evaluation (M&E) toolkit was developed; it has been piloted widely,
using a variety of methodologies, research settings, and respondents.
Methods: Pilot sites were identified both from centers conducting
novel combination TB drug trials and those conducting biomedical HIV
prevention trials, all with significant history of engagement programs.
Example pilot sites included the Aurum Institute and FACTS Consortium,
both in South Africa, and the Kenya Medical Research Institute in Kilifi,
Kenya.
Program staff piloted a series of mapping and data collection tools using
quantitative and qualitative methods to survey targeted clinical and
engagement activities through the trial lifecycle. A web-based interface
for data analysis was also tested. Tools were assessed for effectiveness
in documenting and analyzing engagement activities aimed to improve
the research process and stakeholder outcomes.
Results: Indicators of success, relevant data points and data collection
methodology were refined. Respondents helped revise key parameters
of engagement work, including community advisory board activities,
small group/individual consultation, communications and outreach
to stakeholders beyond trial community. The pilot helped refine
links between engagement and clinical outcomes such as participant
adherence.
Conclusions: Incorporation of feedback from sites with leading GPP
experience led to development of an effective, relevant, user-friendly
toolkit for monitoring and evaluation of community and stakeholder
engagement activities. Wide use of the toolkit is needed to build an
evidence base and to better understand the impact of this work on
research and stakeholder-related outcomes.
P07.05
P07.06
Implementing Community Involvement in
National Institutes of Health (NIH) HIV/AIDS
Clinical Trials Networks
Research Participants Skills Development as
Peer Educators in their Communities
Neetha Shagan Morar1
Rona Siskind , Neetha S. Morar , Russell D. Campbell , Jeffrey
Schouten3
1
2
3
Division of AIDS, Workforce Operations, Communications and
Reporting Branch, Bethesda, MD, United States, 2South African Medical
Research Council, HIV Prevention Research Unit, Westville, South Africa,
3
Fred Hutchinson Cancer Research Center, Office of HIV/AIDS Network
Coordination, Seattle, WA, United States
1
Background: Researchers and funders in HIV clinical prevention and
therapeutic research have long recognized the value of including
community stakeholders in research planning and implementation to
ensure its integrity, relevance and acceptance. Early on, NIH-funded HIV/
AIDS clinical research has required community engagement. In 2009
the “Recommendations for Community Involvement in HIV/AIDS Clinical
Trials Research” were developed to help guide these efforts. With similar
principles, the Recommendations complement the Good Participatory
Practice (GPP) guidelines (developed by UNAIDS/AVAC 2007, updated
2011); both are used across the NIH HIV/AIDS Clinical Trials Networks.
Methods: NIH, the Office of HIV/AIDS Network Coordination (HANC)
and communities from across the NIH HIV/AIDS Clinical Trials Networks
worked to develop these Recommendations. Focusing on the community
advisory board (CAB) model, the Recommendations delineate roles/
responsibilities of staff and community throughout each stage of the
research process, from initiation through results dissemination and site
closure, taking the social and cultural context into consideration.
Results: The NIH HIV/AIDS Clinical Trials Networks and sites use the
Recommendations, GPP and other local guidelines to develop CABs or
equivalent groups to engage global and local communities. This allows
for community input into the scientific agenda, study design, eligibility,
informed consent and recruitment materials. Dissemination of research
results continue to be improved to reach the broader community. Better
training on the Recommendations and GPP is needed and is currently
underway.
Conclusions: The NIH Clinical Trials Networks and sites have successfully
engaged community using the Recommendations and GPP guidelines,
adapting them as needed, to be responsive to community suggestions
and concerns. Both encourage the use of multiple engagement
strategies to develop community partnerships and more comprehensive
approaches to community engagement.
Background: Peer education is a global community-based HIV
prevention intervention to improve health. In 2005, researchers initiated
a peer education programme, nested within HIV prevention clinical
trials. This paper describes the influence of this programme on skills
development of trial participants, as they exercised agency in their role
as peer educators.
Methods: Qualitative methods using round table discussions and semi
structured interviews were used to collect data from 22 women on their
experiences as peer educators. From June to November 2009, data
was collected, audio-recorded and transcribed by trained interviewers.
Content thematic analysis was used to identify themes which included
peer educators experiences, opinions of training and skills development.
Results: Experience and training within the peer education programme
advanced skills development of the peer educators. Using the newly
acquired information on HIV prevention, sex and health promotion,
they provided health-related advice and knowledge to members of their
family and community. The experience improved their self-esteem and
confidence as they developed communication skills to discuss HIV, sex
and research procedures with their partners and family members. The
peer programme was empowering and improved their knowledge of
HIV and research.
Conclusions: Peer experiences contributed to skills development where
peer educators were able to conduct outreach and education sessions.
It enhanced their agency as they developed confidence to engage their
family and community on HIV prevention and research related issues.
Training and partnering with clinical trial participants as peer educators
is an effective and sustainable community based approach that develops
their skills and enhances agency among women.
www.hivr4p.org
167
POSTERS
1
Posters
P07.07
P07.08
Challenges for Community Engagement in
HIV Research in Thailand
Skills Training of Kenyan Health Care
Providers Attending to Men who Have Sex
with Men Improved Services Two Years Post
Training
Niwat Suwanphatthana1
Thai NGO Coalition on AIDS, Muang, Thailand
1
POSTERS
Background: Thailand has been involved in several breakthroughs in
HIV research for many years including RV144, iPrEx and HPTN 052.
These are multisite trials involving general population or specific
populations.
Community Advisory Board (CAB) is the main mechanism for community
engagement of these trials. Most CABs have limited autonomy in
providing recommendations due to power relationship between
researchers and CABs. Research institutions handled CAB formation with
limited inputs from communities. CAB inputs are reduced to reviewing
inform consent documents. Most institutes don’t have structured plan
to strengthen CAB capacity. CABs are not educated about their role and
responsibility resulting in confusion and conflict of interest.
Methods: Promotion of meaningful community engagement in
HIV prevention
The Thai NGO Coalition on AIDS (TNCA) with supports from AVAC is
promoting community engagement including:
1. Good Participatory Practices (GPP) promotion for research institutions
and stakeholder
2. GPP integration in national policy and mechanism
Results:
1. Stakeholder consultations on the 2nd edition of GPP
2. GPP trainings of trainers for research institutions and a lessons
learned workshop
3. The Thai National AIDS Management Center (NAMc) and the
Department ofDisease Control with inputs from TNCA organized
series of workshops on HIV research and strategic programing
involving key stakeholders and targeted populations.
4. The Thai National AIDS Committee established the Sub-committee
of Biomedical HIV Prevention recommended by NAMc.
5. TNCA formed a National CAB consisted of members of existing CABs
in the country to enhance CAB capacity in HIV and clinical research.
Conclusions:
1. GPP is integrated in the national AIDS plan for biomedical HIV
research.
2. Ethical committees and research institutes become familiar with
GPP. Some research institutes develop stakeholder engagement
plans based on GPP.
168
Elise Maria van der Elst1, Bernadette Kombo1, Evans Gichuru1,
Anisa Omar2, Helgar Musyoki3, Susan Marie Graham4, Adrian D.
Smith5, Don Operario6, Eduard Joachim Sanders7
Kenya Medical Research Institute, HIV/AIDS Social Science, Kilifi,
Kenya, 2Kilifi County, Ministry of Health, Kilifi, Kenya, 3National AIDS
and STI Control Programme, National MARPs Programme, Nairobi,
Kenya, 4Washington University, Infectious Disease Medicine, Seattle,
WA, United States, 5Oxford University, Department of Public Health,
Oxford, United Kingdom, 6Brown University, Public Health, Providence,
RI, United States, 7Kenya Medical Research Institute, Epidemiology,
Kilifi, Kenya
1
Background: Men who have sex with men (MSM) are often at high risk
for HIV acquisition, and access to and quality of health services within
this population are negatively affected by hetero-normative attitudes.
A recently developed online MSM-healthcare education program www.
marps-africa.org helped reduce homoprejudice among healthcare
providers (HCP) in Kenya. In this study we used qualitative methods
to explore the provision of MSM healthcare services two years posttraining.
Methods: We held 8 focus group discussions (FGD) with 54 participants
overall; 3 included HCP, 2 included district AIDS-coordinators, and
3 included local LGBT-members. Participants discussed availability,
acceptability and accessibility of HIV-healthcare for MSM. HCP also
discussed changes in health service practices after completing the
training. FGD were recorded, transcribed verbatim, and analyzed using
the grounded theory approach.
Results: HCPs described improvements in their service provision to
MSM-patients post-training. They felt more empowered and comfortable
asking male patients about same-sex behavior, which contributed to
nonstigmatizing HIV-service environments for MSM. Four unique
dimensions for improving MSM utilization of healthcare were identified:
1) HCP expressed a clear need for pre-service MSM sensitivity training,
including the need for national guidelines to manage sexually
transmitted anal infections (STI);
2) AIDS-coordinators advocated for reporting tools specifically developed
for MSM in terms of program implementation and capacity building;
3) MSM described a need for programs to reduce stigma, especially
among male HCPs;
4) All felt that the government should endorse improved health-skillstraining for MSM patients.
Conclusions: Positive impacts of this skills training were reflected in
HCP attitudes two years post-intervention. Scaling-up of efforts will
rely on continued policies to include MSM in healthcare, programs to
reduce stigma in health settings, and directives guiding MSM-STI-service
delivery.
Posters 08: HIV Care
P08.01
P08.02
Choosing Appropriate Interventions and
Defining Locally Achievable Standard of
Care for HIV Prevention Trials in Africa: A
Multidisciplinary Approach
The Continuum of HIV Care in Peru - Where
Are we Now? Key Lessons from an Estimation
in the Context of Very Limited Data
Rwanda Zambia HIV Research Group, Emory University, Pathology &
Laboratory Medicine, School of Medicine, Atlanta, GA, United States,
2
Rwanda Zambia HIV Research Group, Emory University, Zambia
Emory HIV Research Project, Lusaka, Zambia, 3Rwanda Zambia HIV
Research Group, Zambia Emory HIV Research Project, Ndola, Zambia,
4
Rwanda Zambia HIV Research Group, Zambia Emory HIV Research
Project, Lusaka, Zambia, 5Emory University School of Public Health,
Epidemiology, Atlanta, GA, United States, 6Emory University, Economics
and Political Science, Atlanta, GA, United States
1
Background: Per capita incomes (PPP) in most African countries are <
$2000/year and per capita expenditures on health < $100. The cost per
HIV infection averted (HIA) should be less than per capita income. These
metrics should inform what interventions are appropriate to test and
what local standards of care (SOC) should include. To reduce AEs and
unplanned pregnancy, trials should also provide other health services.
Methods: Phase I-III trials conducted at the Rwanda Zambia HIV
Research Group (RZHRG) were assessed to identify common preventable
infectious diseases reported as AEs, and pregnancies. Separately, a
non-RZHRG study protocol for a large phase III trail of ART treatmentas-prevention (TasP) was reviewed by a team with 25+ years of HIV,
development, and ethics research in Africa. SOC services were evaluated
and ethical concerns were identified.
Results: In 5 RZHRG trials, malaria, helminth infections, and diarrhea
were common: in one trial, 6/24 HIV- participants were diagnosed with
urinary schistosomiasis. Bed nets, chlorine, hand soap, and helminth
treatment could prevent most of these AEs. Contraception (FP) was
provided and there were no pregancies.
In the TasP trial, the majority of participants will have a spouse. Voluntary
HIV Testing and Counseling (VTC) is a trial procedure and Couples’ VCT
(CVCT) should also be, as recommended by WHO. The protocol refers to
´family testing´ but this is not a prevention strategy. CVCT is associated
with a reduction in new HIV infections and should be explicitly included
in protocols and procedures. HIA for TasP should be compared with HIA
CVCT, MC, and FP.
Conclusions: Both trials and participants would benefit from low-cost
screening and treatment services for endemic diseases such as bed
nets, routine deworming, soap and chlorine as well as provision of
contraceptives. TasP trials should analyze costs and have a rationale for
testing interventions with HIA > PPP. Excluding locally affordable HIV
prevention services including CVCT from trial SOC is unethical.
Cayetano Heredia University, Unit of Health, Sexuality and Human
Development, Lima, Peru, 2University of California at Los Angeles,
Program in Global Health, Lima, Peru, 3University of California at Los
Angeles, School of Medicine, Los Angeles, CA, United States
1
Background: The role of universal access to treatment in combination
HIV prevention has been established. Estimation of coverage across
steps in a continuum of care cascade (CCC), a key approach to monitor
progress and identify barriers, is novel in Peru and most of Latin America.
We aimed to build an estimation of the CCC for Peru, identify potential
barriers defining losses across the process, and establish data gaps.
Methods: We drew from data from both peer-reviewed publications and
official estimates (i.e. Global Fund Evaluation Reports, Government and
UNAIDS bulletins) to estimate coverage for the CCC steps in Peru. We
assessed whether a single CCC was appropriate, as opposed to specific
CCC by sub-populations. Then we used the most plausible parameters to
generate estimates of the cascade steps. We provide recommendations
for initial policy changes and further research.
Results: Of 75,000 adults estimated to be living with HIV in Peru
in 2013, 22,000 were women. Tested primarily in connection with
reproductive health services, they generally showed a better CCC
profile than men, with earlier access to and higher retention in care,
and more frequent viral suppression after 6 months. Among men, data
are available primarily for MSM and male-to-female transwomen (TW).
Key barriers across the CCC include: low HIV testing rates (only 27% of
HIV+ know their status) and late registration in care (as reflected in CD4
counts around 100 at ARVT initiation) . Only 18% were virally supressed.
Conclusions: The CCC for women in Peru shows progress towards
universal access, reflecting success of programs to prevent mother to
child transmission, and possibly better health seeking. For MSM/TW,
active promotion and facilitation of biannual HIV testing is needed to
improve serostatus awareness, linked with easier access to care, with
strategies to avoid losses to follow up prior to ARVT initiation. Data for
other men are extremely limited. An integrated, reliable information
system to monitor progress is urgently needed.
www.hivr4p.org
169
POSTERS
Susan Allen1, William Kilembe2, Mubiana Inambao3, Bellington
Vwalika4, Shabir Lakhi4, Kristin Wall5, Amanda Tichacek1, Gordon
Streeb6
Carlos F. Caceres1, Kelika Konda2, Alfonso Silva-Santisteban1,
Ximena Salazar1, Lottie Romero1, Segundo Leon1, Jeffrey D.
Klausner3
Posters
Posters 08: HIV Care
P08.03
HIV Care Continuum among MSM in Latin
America Using Online Sexual Networking: Is
Engagement in Care Related to Sexual Risktaking?
Jessica F. Magidson1, Katie B. Biello2,3, Steven A. Safren1,2, Joshua
G. Rosenberger4, David S. Novak5, Kenneth H. Mayer2,6, Matthew
J. Mimiaga1,2,3
Massachusetts General Hospital / Harvard Medical School, Psychiatry,
Boston, MA, United States, 2The Fenway Institute, Boston, MA, United
States, 3Harvard School of Public Health, Boston, MA, United States,
4
George Mason University, Department of Global and Community
Health, Fairfax, VA, United States, 5OLB Research Institute, Online
Buddies, Inc., Cambridge, MA, United States, 6Beth Israel Deaconess
Medical Center, Harvard Medical School, Boston, MA, United States
1
POSTERS
Background: The HIV/AIDS epidemic in Latin America is concentrated
among men who have sex with men (MSM). Yet, it has been difficult to
characterize the HIV care continuum in this population, as many may
not self-identify as MSM. Further, given recent evidence of reduced
likelihood of sexual HIV transmission in the context of viral suppression,
examining whether HIV-infected individuals not in care are engaging in
HIV transmission risk behavior is important for secondary prevention.
Methods: The current study recruited via an online sexual networking
website to reach men who may not self-identify as MSM but are seeking
male sex partners online--a population at high risk for HIV acquisition and
transmission. Primary aims were to examine 1) the HIV care continuum
in this sample; and 2) whether HIV-positive individuals not in care had
higher rates of unprotected anal intercourse (UAI) than those in care
at each step of the care continuum. Surveyed 29,787 active members
of an MSM sexual networking site in Latin America on HIV testing
and HIV diagnosis, receipt of medical care and ART for HIV treatment,
ART adherence, and UAI in the past three months. Percentages were
calculated and logistic regressions were performed.
Results: Overall, 74.1% reported ever being tested for HIV and 9.0%
reported HIV diagnosis. Of the HIV-positive individuals, 20.0% reported
not being in medical care, 29.1% not receiving ART, and 55.3% reported
suboptimal adherence to ART. HIV-positive individuals not in care
reported greater UAI compared to those in care (OR=1.40; 95%CI=1.101.78). Those not ART adherent reported greater UAI than those who
were adherent (OR=2.02; 95%CI=1.60-2.56).
Conclusions: Findings estimate the HIV care continuum among a large
sample of MSM in Latin America using the internet to meet sex partners. A
key limitation of this study was not having a measure of viral load. Despite
this, findings still suggest that secondary prevention for HIV-positive MSM
in Latin America not in care or ART adherent are warranted.
170
Posters 09: HIV Testing and Counseling
P09.01
P09.02
Impact of South-to-South Technical
Assistance from Rwanda & Zambia in Couples
Voluntary HIV Counseling and Testing (CVCT)
Achievements in 19 Countries
VCT Acceptance and High Level of HIV Stigma
and Discriminatory Attitudes towards HIV
Infected People among Male Prisoners in
Northern Thailand
Nurilign Ahmed1, Ashley Appiagyei2, Annie Mwaanga2, Cortney
Robinson2, Robertine Sinabamenye1, Elias Gudo2, Helman
Banda2, Muyunda Mulenga3, Bella Mutumwinka1, Bella
Siangonya3, William Kilembe4, Mubiana Inambao3, Etienne
Karita1, Susan Allen5
Suwat Chariyalertsak1,2, Chonlisa Chariyalertsak3, Kriengkrai
Yotruean3, Chutima Charuwat3, Kriengkrai Srithanaviboonchai1
Background: CVCT is recommended by WHO for HIV prevention and is
associated with reduction in HIV, STI and unplanned pregnancy. Fewer
than 5% of African couples have been jointly tested and it is thus critical
that CVCT be expanded. The RZHRG Center of Excellence provides
training and technical assistance (TTA) to countries and partners wishing
to implement CVCT as standard of care in ongoing service delivery or
research activities.
Methods: Primary data from RZHRG was analyzed to describe TTA
events from 2009-2014. TA events were classified as high-level
advocacy (including policy, workshops and study tours of RZHRG
sites), technical assistance for needs assessments, data management or
other program implementation needs and training (of trainers, service
providers, community-level promoters and others). Data from recipient
countries was analyzed to investigate resulting country-adapted CVCT
models, as well as changes in uptake of CVCT.
Results: RZHRG conducted 61 TA events in 19 countries ranging from
development of national strategic plans and international workshops to
CVCT curriculum adaptations and trainings. Countries such as Botswana
and Uganda have demonstrated higher success in CVCT implementation
through the roll out of a national CVCT strategy. Increased uptake of
CVCT led to increased identification of discordant couples. Countries such
Ghana and Ivory Coast have made strong commitments to expand CVCT
services and are making good progress to that end. Through RZHRG TA,
several countries including Namibia and South Africa have an extensive
number of trainers across levels and sectors available to scale-up CVCT
training efforts. Kenya, Mozambique, Tanzania and Swaziland have sent
high level observers to Rwanda and Zambia.
Conclusions: Through TTA, recipient countries were able to acquire tools
and skills and multiple levels, which enabled them to more effectively
plan for and implement CVCT. This program increased capacity of
countries to prioritize and expand locally tailored CVCT activities.
Background: Although prisoners should be a venue for providing HIV
testing, few studies have explored the factors that may influence HIV
VCT acceptance especially on HIV stigma and discriminatory attitudes
related to PLHIV among prisoners.
Methods: This study aims to measure HIV-related stigma and
discrimination among male prisoners in one prison in Chiang Mai,
Northern Thailand. Only male ethnic minority prisoners were invited
to participate by using self-administered questionnaires for those who
can read and write Thai and by face-to-face interview for prisoners
who have no education. The questionnaire developed by the Global
Stigma and Discrimination Indicator Working Group aims to measure
three domains of this phenomenon included fear of HIV transmission,
manifestations of HIV-related stigma, and discriminatory attitudes
towards PLHIV. Participants who responded affirmatively to any question
within any one domain were considered to possess HIV stigma believe
and discriminatory attitudes. The data were also disaggregated by age,
education, incarcerated period, type of prosecution and HIV VCT uptake.
Results: There were 282 prisoners with average age of 31 years old. The
domain receiving the highest affirmative responses was manifestations
of HIV-related stigma (87.6%), followed by discriminatory attitudes
towards PLHIV (52.8%), and fear of HIV transmission (48.2%). The
subjects who never received an HIV testing compared to ever had HIV
VCT (RR 1.34, CI=1.03-1.76) and had no more than primary education
compared to had higher education (RR 1.34, CI=1.05-1.71) revealed
more fear of HIV contraction. The less educated prisoners also responded
poorly discriminatory attitudes (RR 1.56, CI=1.18-2.06) compared to the
higher educated group. Other factors and incarcerated period were not
associated with affirmative responses in either domain.
Conclusions: Stigma-reduction program in prison are urgently needed
to gain HIV VCT and reduce their risk behavior of HIV infection during
and after discharged from the facility.
www.hivr4p.org
171
POSTERS
Project San Francisco, Kigali, Rwanda, 2Zambia Emory HIV Research
Project, Lusaka, Zambia, 3Zambia Emory HIV Research Project,
Ndola, Zambia, 4Zambia Emory Research Project, Lusaka, Zambia,
5
Emory University, School of Medicine, Department of Pathology and
Laboratory Medicine, Atlanta, GA, United States
1
Research Institute for Health Sciences Chiang Mai University, Chiang
Mai, Thailand, 2Faculty of Medicine Chiang Mai University, Community
Medicine, Chiang Mai, Thailand, 3Chiang Mai Public Health Office,
Chiang Mai, Thailand
1
Posters
P09.03
P09.04
HIV Test: Which Is your Best? An Italian
Survey on Testing Preferences among MSM
Test for Triage: A New Approach to
Community-based HIV Testing and
Counselling
Giulio Maria Corbelli1,2, Sandro Mattioli1, Stefano Pieralli1, Michele
Degli Esposti1, Roberto Cascioli1, Valerie Taccarelli1
Plus Onlus, Bologna, Italy, 2European AIDS Treatment Group, Bruxelles,
Belgium
1
POSTERS
Background: HIV testing opportunities in Italy are frequently limited to
the hospital setting. Other strategies could improve HIV testing uptake,
especially in targeted populations.
Methods: We aimed at better understanding about testing preferences
in the Italian MSM population. An internet-based survey was conducted
between March 10 and April 3, 2014. The survey was promoted on Plus
onlus social networks and gay websites.
Results: 348 questionnaires were collected. Responders were 88%
male, aged 25-34 (35%) or 35-44 (25%), homosexual (81%) or bisexual
(9%). Half or them had an HIV test within 2 years (56%) 18% never
tested for HIV. 61% had more than 2 sexual partners in the past year.
Reported condom use in the past year was: always 39%, always but
once 11%, sometimes 27%, never 14% (10% had no penetrative
sexual intercourse). Most known places to have an HIV test are hospital
(95%), STI clinic (58%) and chemical analysis laboratory (54%); most
used: hospital (73%), STI clinic (30%), laboratory (22%); 5 responders
reported having had a self-test at home. Preferred places is self-testing
at home (53%), hospital (36%), pharmacy (32%) and headquarter of an
organization (31%). Most known testing method is draw blood from vein
(97%), which is also most used (80%) but the least preferred (31%) while
saliva (65%) and finger prick (56%) are the preferred choices. Physicians
are the preferred operator (54%) followed by self-testing (46%), nurses
(46%) and peer-volunteers (39%). The ideal HIV test should be: reliable
(86%), with no medical prescription (75%), free (63%), rapid (55%), with
no personal information collected (45%), with the opportunity to speak
with a peer-counselor (36%).
Conclusions: Awaiting for the results and bureaucratic obligations
represent the major barriers to HIV test in Italy. Home-testing and
community-based testing seem to be among the best ways to offer new
opportunities for HIV test though home testing will not allow any kind of
support for newly diagnosed people.
172
Rachel Baggaley1, Kathryn Curran1, Cheryl Johnson1, Anita
Sands2, Martina Brostrom3, Vladanka Andreeva4
WHO, World Health Organization, Department of HIV/AIDS, Geneva,
Switzerland, 2WHO, World Health Organization, Essential Medicines
and Health Products, Geneva, Switzerland, 3UNAIDS, the Joint United
Nations Programme on HIV/AIDS, Geneva, Switzerland, 4UNAIDS, the
Joint United Nations Programme on HIV/AIDS, Bangkok, Thailand
1
Background: Despite recent achievements in scaling-up HIV testing and
counselling (HTC), two major challenges remain:
1) increasing the number of people who know their HIV status,
particularly populations at highest risk for HIV, and
2) ensuring correct HIV test results are delivered. WHO recommends
community-based HTC (CBHTC) in generalized epidemics and for key
populations in all epidemics.
However, in some settings, there are regulatory barriers and lack of
support for use of HIV rapid diagnostic test (RDTs) at point-of-care by
community providers. There are also concerns that quality of testing and
accuracy of results may be sub-optimal in many settings.
Methods: A “test for triage” approach represents a new, simple way to
support CBHTC. Community providers conduct a single RDT. Individuals
with a reactive test result are immediately linked to a facility for further
HIV testing (starting at the beginning of the national testing algorithm)
and assessment for treatment. Individuals with a non-reactive test result
are given their results, referred for appropriate HIV prevention services
and recommended for re-testing according to national guidelines.
Results: This new approach encompasses both (1) expansion of CBHTC
performed using a single RDT and (2) emphasis of immediate linkage
for HIV diagnosis and treatment assessment at health facilities. The
simplification of CBHTC to a single RDT followed by linkage of people
who have a reactive test result to further testing with a new specimen
and a “second reader” at a facility could reduce operator errors, thus,
leading to more reliable HIV test results, especially for low prevalence
populations (Figure 1). Supportive HTC policies, clear messages for
clients and providers to understand test results and strong linkage-tocare are critical to the success of this approach.
Conclusions: This simplified approach to CBHTC may address current
challenges in HTC. It could be implemented along with efforts to
strengthen quality assurance.
P09.05
P09.06
Opportunities for HIV Prevention among
Couples in Durban, South Africa
Effectiveness and Impact of Promotion of
Health Rights by Sex Workers for Reducing
HIV Prevalence in the Eastern of Democratic
Republic of Congo
Rwanda Zambia HIV Research Group, Lusaka, Zambia, 2HIV
Pathogenesis Program, Durban, South Africa, 3Medical Research
Council of South Africa, Durban, South Africa, 4Rwanda Zambia HIV
Research Group, Ndola, Zambia, 5Emory University, Atlanta, GA, United
States, 6Simon Fraser University, Burnaby, BC, Canada, 7Rwanda
Zambia HIV Research Group, Atlanta, GA, United States
1
Background: Couples’ Voluntary Counseling and Testing (CVCT) reduces
HIV transmission by up to 60% among discordant couples but uptake is
low in a priority setting. Rwanda Zambia HIV Research Group (RZHRG)
in collaboration with the HIV Pathogenesis Programme (HPP) at the
University of KwaZulu-Natal (UKZN) conducted a pilot project to expand
of CVCT and to assess opportunities to implement HIV prevention
strategies that target couples at local clinics in Durban, South Africa.
Methods: In February 2013, trainers from RZHRG conducted CVCT
training and supervision for counselors and health promoters at five
clinics around Durban. Client level indicators including age, gender,
pregnancy status, ART status, and sero‐status of both partners, were
collected for all couples as a component of routine CVCT service
operation. Descriptive analyses are presented here.
Results: A total of 20 counselors and 28 promoters completed training.
Out of 908 couples (1816 individuals) that underwent CVCT, 50% of
men and 62% of women had been previously tested alone; only 4%
had been tested as a couple. Prevalence of HIV was 42% and prevalence
of discordant couples was 29% (19% F+M-, 10%F-M+). Almost half of
the couples were within the age group 20 to 29 years. This group had
the highest prevalence of HIV and contributed the highest proportion
of discordant couples. Only 21% of discordant couples had the positive
partner on ARVs (13% M-F+, 8 % M+F-) and 30% of the concordant
positive couples with one or both partners on ARVs. In 95 couples (10%),
the woman was pregnant. Of these, 14 (15%) were discordant couples
where the woman was the negative partner and had never been tested
as couples before; only in one of these couples was the man on ARVs.
Conclusions: The burden of HIV in Durban is high. CVCT, an intervention
that targets the largest risk group in sub-Saharan Africa, would greatly
benefit the couples in Durban as an HIV prevention strategy and as
an entry point to care and treatment services aimed at reducing HIV
incidence.
Mambo Amisi Modeste1,2
Humanitarian Action for Health and Community Development, Public
Health, Bukavu, Democratic Republic of the Congo, 2Institut National
de Sante Public, Epidemiologie, Kinshasa, Democratic Republic of the
Congo
1
Background: Since 1994, the Eastern Democratic Republic of Congo
has been a theater of armed conflict, violence, and serious human rights
violations. Two out of three women have experienced the violence
personally. While the whole community suffers, sex workers and MSM
transgender people are particular victims of discrimination, rape and
sexual violence. The 2011 annual report of the National Program for
fighting against HIV/ AIDS, indicated that a 4.7% HIV prevalence
among sex workers in Eastern Democratic Republic of Congo. This
information drove the sex workers gathered at the AHUSADEC NGO to
initiate a program in 2012, “Club mutual pleasure of sex workers,” with
the goal of promoting prevention and sharing information about STIs
and HIV by sex workers to sex workers, the community and customers,
and to promote human rights and advocacy in the towns of Bukavu and
Goma in the Eastern Democratic Republic of Congo.
Methods: We’re organized into six solidarity communities. 276 received
training in peer education on HIV/AIDS prevention. Through our
voluntary counseling and testing center, we promote education, training
and sensitization for HIV testing. In the conflict zone we educated and
trained an army group and women about HIV prevention.
Results: In 2013
- 276 sex workers were sensitized about human rights and trained as
peer educators in STI-HIV/AIDS prevention;
- 308,600 male and 1,803 female condoms used by 27 army group
member in the conflict zone;
- 104 sessions conducted by sex workers peer educators on IST-VIH/
SIDA and human right with 27 army groups in Eastern of D.R. Congo;
- 2769 soldiers from army groups know their HIV status through
UNHCR and VCT of AHUSADEC;
- 503 lubricants distributed;
- 74 soldiers from army groups who are HIV + received ARV treatment
through the UN mission HIV section;
- 439 were identified as having sex with condoms in February, 793 in
July and 3705 in November 2013, an increase of 75%.
Conclusions: The effective involvement of sex workers in the promotion
of human health could reduce the prevalence of HIV / AIDS to 12%
in sentinel areas (Walikale, Masisi, Rutshuru and Minova) in territory
occupied. The very low economic power of sex workers is our challenge.
www.hivr4p.org
173
POSTERS
William Kilembe1, Mammekwa Mokgoro2, Annie Mwaanga1,
Miriam Kamusoko2, Tarylee Reddy3, Elisabeth Dissen1, Jonathan
Davitte4,5, Shumba Phiri Hilda1, Mark Brockman6, Thumbi
Ndung’u2, Susan Allen5,7
Posters
P09.07
P09.08
Engaging Young Adult Clients of Retail
Pharmacies for HIV-1 Testing in Coastal Kenya
Correlates of Voluntary HIV Testing and
Collection of Test Results among Male Clients
of FSWs in Three States of India
Peter M. Mugo1, Henrieke Prins1, Elizabeth Wahome1, Grace
Mwashigadi1, Alexander Thiong’o1, Evanson Gichuru1, Anisa
Omar2, Susan M. Graham1,3, Eduard J. Sanders1,4
1
Kenya Medical Research Institute, Kilifi, Kenya, 2Ministry of Health,
Kilifi, Kenya, 3University of Washington, Seattle, WA, United States,
4
Oxford University, Headington, United Kingdom
POSTERS
Background: Adults in resource-limited countries frequently use retail
pharmacies as the first or only source of treatment for various ailments.
The aim of this study was to assess whether young adult clients of retail
pharmacies can be referred for HIV-1 testing and engaged for HIV-1
prevention research.
Methods: We requested five pharmacies to refer clients meeting
predefined criteria (18-29 years of age and requesting treatment for
fever, diarrhoea, sexually transmitted infection (STI) symptoms or
body pains) to selected study health facilities, where HIV-1 testing and
screening for an ongoing HIV-1 prevention study was offered. Using
multivariable logistic regression, we determined client characteristics
associated with uptake of HIV-1 testing.
Results: From February through July 2013, 1,490 pharmacy clients met
criteria for referral (range of weekly average by pharmacy: 4-35); 377
(25%) reported for screening at a health facility, 353 (24%) were HIV1 tested and 127 (9%) met criteria for the prevention study. Of those
tested 14 (3.9%) were HIV-1 infected. Test uptake varied significantly
by referring pharmacy, and was higher for clients who presented at the
pharmacy without a prescription vs. those with a prescription, and for
clients who sought care for fever or STI symptoms vs. those who sought
care for body pains.
Conclusions: About a quarter of retail pharmacy clients engaged for
HIV-1 prevention research were tested for HIV-1. Clients seeking care
directly at the pharmacy (i.e., without a prescription) and those with fever
or STI symptoms were more likely to take up HIV-1 testing. Engagement
of adult pharmacy clients for HIV-1 testing may identify undiagnosed
individuals and offers opportunities for HIV-1 prevention research.
174
Karikalan Nagarajan1, Sheela Godbole1, Sucheta Deshpande2,
Ramesh Paranjape1
National AIDS Research Institute, Pune, India, 2Public Health
Foundation of India, New Delhi, India
1
Background: Male clients of female sex workers (FSWs) are a high risk
(HRG) and bridging population driving the heterosexual HIV epidemic
in India. National AIDS control programme has prioritized scaling up
of voluntary HIV testing (VT) among HRG´s. Impact of VT, dependent
upon ‘collection of test results’ (CR), is of significance in this bridging
population. We assessed the correlates of VT&CR among male clients in
India to understand its determinants.
Methods: Data were drawn from the cross-sectional IBBA survey of
male clients of FSWs between 2009-2010 in three high prevalence
states of Tamil Nadu, Andhra Pradesh and Maharashtra, India.Informed
consent was obtained. Multivariate logistic regression models were used
to assess the correlates of VT&CR and to identify factors preventing
clients from collecting test results
Results: Of 4803 ‘clients’, 781(16.2%) reported VT, of whom 710
(90.9%) of clients had collected reports (CR). Clients exposed to STI
advertisements [AOR 1.9(CI-1.5-2.4), p< 0.05] and ‘Key’ clinics (branded
STI clinics) [AOR 1.2(CI-0.9-1.5), p< 0.05 respectively] were more
likely to take-up VT&CR. Higher education status(>12 grade) increased
the likelihood of VT&CR uptake [AOR 1.8(CI-1.2-2.7), p< 0.05], while
unorganized laborers and truck/transport workers were less likely to
undergo VT&CR [AOR 0.6(CI-0.5-0.7), p< 0.05 AOR 0.7(CI-0.5-0.9) p<
0.0 respectively]. Higher education was linked with increased likelihood
of collecting results: grades 6-12 and grades > 12 were less likely to
have “not-collected” test reports [AOR 0.4(CI-.2-0.9), p< 0.05 AOR 0.1(CI0.5-0.7), p< 0.05 respectively].
Conclusions: We highlight the overall low uptake of VT among clients of
FSW and that not all test- seekers are collecting reports. Program focus
to improve test-seeking as well as report collection is warranted through
IEC campaigns as they have positively influenced uptake of VT&CR.
Specific interventions among less educated , unorganized laborers and
truckers would help in improving VT and CR among clients.
P09.09
P09.10
Socio-demographic Factors Associated with
Uptake of HIV Counseling and Testing (HCT)
among Nigerian Youth
HCT as a Point of Entry for HIV Prevention
and Treatment - Profile of Men and Women
Presenting at HCT Centers in Durban, South
Africa
Ayodeji Oginni1, Otibho Obianwu2, Sylvia Adebajo2
Population Council, Abuja, Nigeria, 2Population Council, HIV, Abuja,
Nigeria
1
Jessica Lyn Phillip1, Renee Street1, Susie Hoffman2, Kelly
Blanchard3, Theresa Exner4, Elizabeth Kelvin5, Gita Ramjee1,6,
Joanne Mantell2
HIV Prevention Research Unit, Medical Research Council, Durban,
South Africa, 2HIV Center for Clinical and Behavioral Studies and
Columbia University, New York State Psychiatric Institute, New York,
NY, United States, 3Ibis Reproductive Health, Cambridge MA, USA and
Johannesburg, South Africa, Johannesburg, United Kingdom, 4HIV
Center for Clinical and Behavioral Studies, New York State Psychiatric
Institute, New York, NY, United States, 5CUNY School of Public Health
at Hunter College, New York, NY, United States, 6London School
of Hygiene & Tropical Medicine, Department of Epidemiology and
Population Health, London, United Kingdom
Background: HCT is an important gateway for HIV prevention
interventions as it educates sero-negative individuals on HIV preventive
behaviours and enables seropositive individuals to gain access
to treatment, care and support services. We evaluated the sociodemographic factors associated with HCT-uptake among Nigerian youth
aged 15-24.
Methods: Secondary data analysis was conducted on Nigeria´s 2012
National HIV/AIDS & Reproductive Health Survey data. Multivariable logbinomial regression analysis was used to estimate adjusted prevalence
ratio (APR) with 95% Confidence intervals.
Results: Of the 10,091 youth, half were aged 15-19, 66.9% were single,
65.7% were rural dwellers, 20.7% had no education, 46% were students
& 31.1% were employed. About 10.5% ever had HCT & 3.5% tested
positive in the survey. Multivariable analysis revealed that the aged
20-24 [APR=1.67(1.41-1.96)] were more likely to have had HCT than
the aged 15-19. HCT-uptake increased with educational level [primaryAPR=2.29(1.59-3.32); secondary—APR=3.48(2.54-4.77) & higherAPR=6.68(4.66-9.58)]. The non-Catholic [APR=1.60(1.36-1.89)] and
the Catholics [APR=1.85(1.51-2.26)] Christians were more likely to have
had HCT than the Muslims. Those having comprehensive knowledge
of HIV [APR=2.09(1.83-2.39)] were twice more likely to have had HCT.
Students [APR=0.80(0.67-0.94)] were less likely to have had HCT than
the employed. Those from poor-households [APR=0.63(0.51-0.77)]
were less likely to have had HCT than those from average-households.
Conclusions: HCT-uptake among young Nigerians is very low despite
the increased availability of free HCT services in the country. The
fact that being employed and having higher educational level and
household wealth are associated with HCT-uptake suggests that socioeconomic barriers to HCT-uptake persist among young people. The
association with age may be due to age of consent barriers faced by
adolescents. More youth-friendly interventions aimed at increasing HCTuptake among young Nigerians are urgently needed.
Background: HIV Counselling and Testing is an important component
of HIV prevention and treatment. Early identification of HIV-seropositive
status will enable individuals to be linked to HIV care and support.
Pathways to Engagement in HIV Care for Newly-Diagnosed South
Africans was a prospective cohort study that enrolled newly-diagnosed
HIV positive individuals and followed them up for 8 months post
diagnosis to explore the barriers and facilitators of enrolling and staying
in care. To describe the socio-demographic profile of men and women
presenting at PHC clinics for HCT.
Methods: Between November 2010 and May 2012, we screened
2935 individuals from 3 public-sector primary health care clinics in the
Durban, South Africa. Women and men who presented for HCT
(as opposed to antenatal care) were sequentially approached prior to
testing and invited to participate in a screening interview. Eligibility
criteria were: 18 years or older, not cognitively impaired, not pregnant,
not having previously been diagnosed HIV+, willing to provide written
informed consent, to have a witness if illiterate, and to share test results
with study staff.
Results: Of screened participants, 2746 met the eligibility criteria.
Level of education varied, with 38.4% participants having completed
Grade 12. Over 91% were unmarried, of which 79% reported having
a steady partner. Half of the participants (53.4%) indicated that they
were unemployed but were looking for work. Only 16.5% reported
being employed full time. Self-motivation for HIV testing was reported
by 70.4% of participants, with no difference by gender; 17.4% were
motivated by family/friend. However, few reported testing because they
were encouraged by the clinic staff. Prior testing was reported by 46%,
including 57% of women and 33% of men.
Conclusions: Public health benefit would be for HCT programs to tailor
interventions for effective gender-based counselling. HIV prevention
education should take into account motivations for HIV testing.
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175
POSTERS
1
Posters
P09.11
P09.12
The “Worried Well” Among Clients Attending
HIV/AIDS Counselling and Testing Services at
a Clinical Research Centre in SW Uganda
Does HIV Counseling and Testing Change
Sexual Risk Behaviors: Findings from a
Community Health Center (CHC) in North
Central Nigeria?
Richard Rwanyonga1, Andrew Abaasa1, Gershim Asiki1, Benjamin
Twefeho1, Ubaldo Bahemuka1, Emanuel Aling1, Eugene Ruzagira1,
Matthew A. Price2, Anatoli Kamali1
MRC/UVRI, Uganda Research Unit on AIDS, Entebbe, Uganda,
International AIDS Vaccine Initiative, New York, NY, United States
1
2
POSTERS
Background: HIV/AIDS counselling and testing (HCT) provides a
gateway to participation in epidemiological studies. We characterised
participants accessing HCT through the two models (facility based [FHCT]
vs. community based [CHCT]) offered at the MRC/UVRI clinical research
centre in SW Uganda.
Methods: From May 2010-March 2014, we offered HCT to communities
(farmers, fisher folk and small townships) in SW Uganda, as part of
screening for epidemiological studies. FHCT is offered at the MRC/UVRI
clinical research centre in Masaka town where clients voluntarily come
for testing, while CHCT participants are approached in their homes or
work place and invited to a central point (community hub) within their
locality. Both approaches applied the Uganda national HCT guidelines.
Results: A total of 23,536 (53% women) individuals (mean age 29 years
SD 9.3) received HCT; 9,405 (55% women) mean age 28 years (SD 8.5)
FHCT and 14,131 (52% women) mean age 30 years (SD 9.7)CHCT.The
main reasons for seeking FHCT were; perceived risk exposure (87%),
intending to marry (8%) and poor health (4%). CHCT participants were
from fishing communities (78%) and small townships (22%). Over all,
HIV prevalence was 13%; [CHCT (16%) Vs. FHCT (9%), p< 0.01], higher
among women compared to men (14% vs. 12%; p< 0.01), and higher
among those aged >25 years compared to younger age groups (15%
vs. 10%, p< 0.01; CHCT 17% vs. 13 %, p< 0.01; FHCT 11% vs. 7%,
p< 0.01). Fishing communities had higher HIV prevalence compared to
small townships (18% vs. 8%, p< 0.01). HIV prevalence also varied by
the reasons for testing; risk exposure (8.4%), intending to marry (4.6%)
and poor health (28.8%).
Conclusions: Although majority of those in the FHCT tested because
they were worried of their risk, only 8.4% were infected. There is higher
burden of HIV in CHCT especially in fishing communities and therefore
may be more suitable for recruiting into clinical trials.
176
Ibrahim Suleiman1,2, Obianwu Otibho1, Jean Njab1, Ayodeji
Oginni1, George Eluwa1, Sylvia Adebajo1, Babatunde Ahonsi2,
Terfa Kene3, Jide Keshinro4
Population Council, HIV and AIDS, Abuja, Nigeria, 2Population Council,
Abuja, Nigeria, 3Millitary HIV Research Program, Department of
Defense, Abuja, Nigeria, 4Millitary HIV Research Program, Department
of Defence, Abuja, Nigeria
1
Background: HIV counseling & testing (HCT) is essential for prevention,
care & support. It serves as a means of providing HIV prevention
information to people who opt for HCT. The aim of this study was to
determine if HCT uptake influences comprehensive HIV knowledge
(CHK) & reduces risky sexual behaviors among attendees of a key
population focused CHC in Kaduna Metropolis, Nigeria.
Methods: From May 2013 - January 2014, clients who accessed sexual
health services at a CHC in Kaduna were offered HCT. Data on sexual risk
behaviours were collected with the use of a structured HCT client intake
form. HIV sero-status was determined using rapid test according to the
national algorithm. Descriptive & inferential analyses were conducted to
determine the effect of HCT on risk behaviors.
Results: A total of 2092 clients accessed the clinic. Majority of the
clients were male (98.6%); about 70% were MSM. The median age of
the clients was 25 years; above 80% were single; 51% were employed.
About 69% of the clients had CHK; 60% had engaged in transactional
sex (TS) & 49% had ever been tested for HIV. Comparative analysis
between clients with prior HCT uptake & those with no prior HCT uptake,
shows that CHK was higher (86% vs. 53%; p< 0.001) among those
with prior HCT uptake than those with no prior HCT uptake. TS was
lower (43% vs. 75% p< 0.001) while multiple sexual partnerships was
higher (87% vs. 57% p< 0.001) among those with prior HCT uptake
compared to those with no prior uptake. When controlled for age, sex,
occupation, education & marital status, clients with prior HCT uptake
were more likely [Adjusted OR= 4.54 (95%CI=3.64, 5.67)] to have CHK
& less likely [Adjusted OR= 0.29 (95%CI=0.23—0.35)] to engage in TS.
Conclusions: HCT uptake among MSM was low; however access to HCT
has the potential of increasing CHK as well as reducing risky sexual
behaviours among MSM. High quality pre and post-test counselling
should be delivered as a means of providing vital behavior change
communication messages to MSM.
P09.13
P09.14 LB
Young Thai Men who Have Sex with Men and
a Voluntary HIV Testing Service, Bangkok,
2005-2013
“Flash Test” Week in Lyon and Rhône Valley:
An Expanding Access to Rapid Testing, First
Step of the Cascade and Prevention
Wipas Wimonsate1, Supaporn Chaikummao1, Anuwat Sriporn1,
Pikunchai Luechai1, Kesinee Satumay1, Santi Winaitham1, Somsak
Yafant1, Sarika Pattanasin1, Anupong Chitwarakorn2, Timothy H.
Holtz1,3
Jean-Michel Livrozet1,2, Anne-Cécile Delinotte2, Sébastien
Cambau2,3, Mireille Joliot-Vilain2, Colette Coudeyras2,4, Geneviève
Retornaz2,5, Albertine Pabingui2,6, Christine Fernandez2,7, Patrick
Caillon2,8, Aurélie Neveu9, Véronique Ronzière10, Yasmine
Erraïs2,11, Christine Haydont2,12, Christian Chidiac2,13, Pascale
Prin2,14, Jean Paul Godeau15, Julie Biron16, Patrice Redon17, Gérard
Millier2,18, Stéphane Castello19, Sylvaine Boige-Faure20, Christelle
Bibollet21, Pascal Pourtau2,22, Christophe Julien2,23
Thailand-MoPH U.S. CDC Collaboration, Nonthaburi, Thailand,
Ministry of Public Health, Department of Diseases Control, Nonthaburi,
Thailand, 3Division of HIV/AIDS Prevention, U.S. Centers for Disease
Control and Prevention, Atlanta, GA, United States
1
2
Hôpital Edouard Herriot, SMIT-Unité du Pavillon P, Lyon, France,
COREVIH Lyon Vallée du Rhône, Lyon, France, 3ENIPSE, Paris, France,
4
Réseau Virages Santé, Lyon, France, 5Association de Lutte Contre
le Sida, Lyon, France, 6Datiseni, Lyon, France, 7Hôpital de la CroixRousse, CDAG, Lyon, France, 8Hôpital Edouard Herriot, CDAG, Lyon,
France, 9Médecins du Monde, Mission France, Lyon, France, 10Conseil
Général du Rhône, Lyon, France, 11AIDES, Lyon, France, 12Le Mas, Pause
Diabolo, Lyon, France, 13Hôpital de la Croix-Rousse, SMIT, Lyon, France,
14
Centre Hospitalier de Bourg-en-Bresse, Centre de Santé Publique,
Bourg-en-Bresse, France, 15AIDES, Bourg-en-Bresse, France, 16CroixRouge Française, Bourg-en-Bresse, France, 17Centre Saliba, Bourg-enBresse, France, 18Hôpital de Privas, CDAG, Privas, France, 19Agayri,
Bourg-les-Valence, France, 20Mairie de Valence, Direction Santé Famille
Environnement, Valence, France, 21Conseil Général de la Drôme, CDAG
le Forum, Valence, France, 22CRIPS, Lyon, France, 23Agence Régionale
de Santé, Prévention et Promotion de la Santé, Lyon, France
1
Background: Previous studies have shown high HIV incidence among
young men who have sex with men (YMSM: age 14-21 years) in
Bangkok. We investigated the proportion of MSM clients who were
YMSM who utilized the anonymous HIV voluntary counseling and
testing (VCT) services at the Silom Community Clinic (SCC) during 20052013, and the rate of subsequent testing.
Methods: We collected data during counselling sessions held between
September 30, 2005 and November 30, 2013. We used age at first HIV
testing as baseline, and the chi-square test for trend for this analysis. HIV
testing was voluntary but recommended every 6 to12 months.
Results: Between 2005-2013, 6,101 MSM presented for HIV testing. The
median age at first HIV testing was 27 years (range: 14-80 years); 12.5%17.0% were YMSM during any one year. The proportion of clients who
were YMSM who utilized the service during 2005-2013 remained stable
(p-value for trend 0.47). Among 4,352 who tested HIV-negative at first
testing, only 264/692 (38.2%) YMSM had subsequent HIV testing while
1,452/3,660 (39.7%) older MSM had subsequent testing (p-value 0.65).
Conclusions: For the past nine years, the number, but not the proportion,
of YMSM who have utilized the free HIV testing service at SCC has
increased. The proportion of MSM who returned for subsequent testing
did not differ by age group. Specific community efforts must be made to
increase the use of VCT services among YMSM.
Background: The French Minister of Health organized an experimental
week of rapid testing in the 4 French regions with high HIV prevalence.
In Rhône-Alpes, the action took place in September 2013.
Methods: Expanding access to rapid testing for the most exposed
populations to HIV , Men who have Sex with Men ( MSM), migrants
and drugs users ( DU) with:
• innovating actions in new places next to the targeted populations;
• new partnerships between hospitals, Voluntary Testing Centers
(VTC), community based offers and associations;
• communication performed by the National Institute for Prevention and
Health Education: 400 posters, 1000 flyers in gay venues and African
shops and commercials in gay and African press and on radios;
• local coordination by the Regional Committee for HIV ( COREVIH);
• A questionnaire investigating HIV testing history and sexual
behaviors including inconsistent condom use. This questionnaire
was analyzed by the French Institute for Public Health Surveillance
in June 2014.
Results: 49 structures were involved: hospitals, VTC, associations and 71
partnerships were signed.38 new people were trained for rapid testing.
1 049 rapid tests were performed: 60.3% were male, 39.5% female
and 0.2% trans. 60.5% were between 18 and 30 year-old. 74.5% were
born in France and 20.3% in a foreign country:11.8% in sub-Saharan
Africa,4.5% in North Africa.19.6% were MSM, 3.5% DU. 55.8% reported
non-condom use, 6.5% sex for money, 14% drug use in the past 5 years.
64.3% have had an HIV test before and 35.5% in the past 2 years. 27.2%
of people performed a HIV test for the first time. In total, 4 tests were
positive (0.4%):3 migrants and 1 MSM, and the patients were referred to
HIV units due to the links developed before this week.
Conclusions: This week is an original initiative to enhance HIV prevention
and screening in high-risk populations. An important proportion of persons
were HIV tested for the first time. Finally, it allows new partnerships
between community-based associations and referral centers.
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POSTERS
2
Posters
P09.15 LB
P09.16 LB
Assessment of Namibian Pharmacy Practices
of the Sale of Over-the-Counter HIV Rapid
Test Kits in the Private-sector
ARV-based Prevention for Women: New Data
on HIV Testing Behaviors from Potential
Users of Vaginal Microbicides in Mpumalanga,
South Africa
Matthew Rosenthal1, Ismelda Pieterson2, Cheryl Johnson3,
Nersesian Paula4, Mechtild Hulsman5
USAID Namibia, HIV Office, Windhoek, Namibia, Government of
Namibia, Ministry of Health and Social Services, Windhoek, Namibia,
3
World Health Organization, HIV Testing and Counseling, Geneva,
Switzerland, 4AIDSTAR, John Snow International, HIV, Arlington, VA,
United States, 5Consultant, Windhoek, Namibia
1
2
POSTERS
Background: In 2011, the Namibia private sector began the sale of Overthe-Counter (OTC) HIV rapid test kits (RTK) for personal-use. By beginning
2012, volume of RTK sales increased within the general public. In Dec
2012, a team conducted a rapid survey of RTK for personal-use. The
GRN has identified increased HIV Testing and Counseling (HTC) as an
entry point into HIV care and treatment. Despite high HIV prevalence
(13.4%), only 54% of Namibia’s population has received HTC. While,
RTK may offer a new strategy to HTC programs for implementing mixed
HTC models, concern within GRN on unregulated sale and potential
social harms of RTK remain.
Methods: The assessment was conducted over a month. Key informants
(n=68) included private-sector and GRN staff. A survey at 53/112
pharmacies (48%) throughout 6 regions in Namibia was administered
to pharmacists. All of the pharmacies in 5 high prevalence regions
were surveyed and a sample in Windhoek. Survey assessed attitudes,
practices and approaches. 52/ 53 pharmacies participated and 36/52
reported current RTK sales.
Results: Analysis of RTK varied by pharmacy and geography. Some
coastal city pharmacies reported 30 tests sold in the last month prior to
the survey and southern areas reported lower sales (1-3 in same period).
Price and location of tests differed (i.e. behind counter, controlled sales,
on pharmacy floor) and influenced sales volume. Attitudes, practices
and training of pharmacists varied and reflected poor regulation.
Pharmacists (38%) reported having been trained in performing HIV RTK
and 62% reported HTC training, ranging from pre-service to self-training.
Stocking and supplying RTK varied with good lead time (i.e. 1 day in
some locations), but little quality assurance, maintenance and proper
storage. 4 different test kits were sold to the public.
Conclusions: Additional assessments of RTK in Namibia need to be
conducted; including development of regulatory frameworks for RTK in
Namibia and user acceptability, feasibility and quality of RTK.
178
Saiqa Mullick1, Martha M. Brady2, Barbara A. Friedland2, Thabiso
Mango3, Ravikanthi Rapiti4
Wits Reproductive Health and HIV Institute (WRHI), Johannesburg,
South Africa, 2Population Council, New York, NY, United States,
3
Solutions for Innovative Policies, Programs, and Technologies
(Solutions-IPPT), Johannesburg, South Africa, 4Population Council,
1
Background: Use of ARVs, such as Tenofovir (TFV) gel, for preexposure prophylaxis (PrEP) will require regular HIV testing to ensure
users are uninfected before initiating and continuing product use.
Understanding HIV testing practices among potential users is critical
in planning for introduction.
Methods: A cross-sectional descriptive study of women’s healthseeking behaviors and HIV-testing practices was conducted among a
convenience sample of women attending 5 high-volume primary health
clinics (PHC) in Mpumalanga, South Africa in 2014. A quantitative faceto-face interview captured information on frequency and circumstances
around potential HIV exposure, HIV testing, risk perception, awareness
of and willingness to try TFV gel.
Results: A total of 1,227 women completed the survey. 93% of women
reported having been tested at least once and 90% who tested more
than once did so in the past year. The vast majority (over 90%) knew their
HIV status, and the majority of those (79%) disclosed this information
to someone. Although most women were not familiar with microbicides,
after being provided with information 94% said they would be willing
to try TFV gel. Most women (92%) said they would prefer to get the
product from a clinic and 81% preferred receiving it from a nurse. The
majority (82%) believed that a vaginal gel used only around the time
of sex would be easier to use than a daily oral pill. Half said they would
be willing to get tested for HIV every 3 months and an additional 22%
said they would be willing to get tested once a month. However, 68%
indicated that they would only use such a product if it were free.
Conclusions: The data show that many women are already being
tested regularly for HIV, a pre-requisite for provision of ARV-based
prevention. Women’s stated willingness to try TFV gel suggests an
overall positive attitude. Preferences for accessing microbicides from a
nurse at a clinic suggest that delivery of TFV gel from existing primary
health services is acceptable.
P09.17 LB
P09.18 LB
The Cost of Community Based HIV Counseling
and Testing and Linkage to Care in Rural
South Africa: Estimates from the Linkages
Randomized Control Trial
Does Culture Affect Counselling in HIV
Prevention Research?
Colleen Herman1, Annalene Nel1
International Partnership for Microbicides, Clinical Operations, Cape
Town, South Africa
1
Monisha Sharma1, Heidi Van Rooyen2, Connie Celum3, Jared
Baeten3, Carol Levin3, Ruanne Barnabas3
University of Washington, Epidemiology, Seattle, WA, United
States, 2Human Sciences Research Council, HIV/AIDS, STIs and TB,
Pietermaritzburg, South Africa, 3University of Washington, Global
Health, Seattle, WA, United States
Background: Community based HIV testing and counseling (HTC) and
linkage to HIV care has demonstrated high effectiveness. However, HTC
program costs are needed to inform policy decisions.
Methods: We estimated costs of mobile clinic and home-based HTC
with point-of-care (POC) CD4 testing and counselor follow up at home
to encourage linkage. Costs were collected in KwaZulu-Natal in 2013
from the Linkages Study, a randomized trial of community HTC and
linkage to care. Time and motion studies were conducted to separate
research from program activities. Costs were obtained from budgets,
invoices and staff interviews. We assumed task shifting from nurses to
community workers. Program effectiveness was estimated from our pilot
study in a nearby area (N=1272, 30% HIV prev). Total program costs for
HIV+ persons were divided by number virally suppressed at 12 mos to
estimate incremental cost per person virally suppressed.
Results: Program cost of mobile HTC was $5.45 per HIV- person tested
and $8.28 per HIV+ tested. POC CD4 increased cost per HIV+ tested to
$14.78 and POC with follow-up cost $21.78. Home HTC cost $8.22 and
$12.13 per HIV- and HIV+ person tested. Using effectiveness from our
prior study (32% of HIV+ persons were ART eligible at CD4≤350mL,
of whom 69% initiated ART and 70% virally suppressed at 12 mos),
incremental cost for all HIV+ persons for home HTC with POC CD4
and counselor follow up was $126.10 per person linked to care and
$179.19 per person virally suppressed. Mobile HTC with POC CD4 and
follow-up cost $96.62 and $137.30 per person linked to care and virally
suppressed. Assuming similar program effectiveness under the new
ART guidelines of eligibility at CD4≤500mL, costs would decrease to
$100.95 and $77.35 per person virally suppressed through home and
mobile HTC respectively.
Conclusions: Community HTC with POC CD4 and follow-up achieves
high linkage and viral suppression at costs of $137-179 per HIV+ person
virally suppressed. Incremental costs are expected to decrease with new
ART guidelines.
Background: The Ring Study is a Phase III microbicide study, currently
enrolling in communities with a culturally-diverse background in
Southern and Eastern Africa. The ‘person-centred counselling’ (PCC)
approach, which stimulates probing, is used by Research Centre (RC)
counsellors. Participants and counsellors alike are diverse in ethnicity,
cultural background and education. Culture can be described as the blue
print for who we are. It includes patterns, beliefs, values, expectations
and symbols for how people behave, think and feel in a certain social
group. PCC aims at improving retention to study visits and adherence to
study product.
Methods:
• The following counselling is conducted during The Ring Study: ring
use adherence; HIV pre- and post-test counselling; risk reduction;
contraception and visit adherence counselling.
• RC staff attended informal training sessions on understanding and
implementing the PCC approach.
• Regular assessments are conducted to get feedback on the
implementation of the approach; e.g. visiting RCs; engaging with
counsellors and nurses who conduct counselling on site.
• Additional feedback on adherence is obtained subjectively via
questionnaire feedback.
Results:
1. Participants differ and decisions are influenced by cultural norms (e.g.
partner, family, friends and community).
2. Cultural self-awareness is an important counselling aspect as it allows
identification of differences and more empathy and sensitivity in the
counselling relationship.
3. Reticence in probing for in-depth PCC answers.
Conclusions: Cultural norms are powerful and can sway a participant
from her good intentions, even if the principles of the PCC approach are
used effectively. These cultural norms can result in different outcomes
with regard to protocol compliance and ring adherence. Further
qualitative research is recommended to determine if culture affects
counselling in HIV prevention research as this could lead to improved
retention and adherence.
www.hivr4p.org
179
POSTERS
1
Posters
Posters 10: Immunogens
P10.01
P10.02
Improving the Vaccine that Provided
Protection in the RV144 Trial
“Canyon Shielding” of the CD4-Binding Site on
HIV-1 Trimer
Javier F. Morales1, Trevor J. Morin1, Bin Yu1, Gwen P. Tatsuno1,
David L. Alexander1, Sara M. O’Rourke1, Richard Theolis1, Kathryn
A. Mesa1, Phillip W. Berman1,2
Lei Chen1, Peter D. Kwong1
University of California, Santa Cruz, Biomolecular Engineering,
Santa Cruz, CA, United States, 2Gladstone Institute of Virology and
Immunology, University of California, San Francisco, CA, United States
VRC/NIAID/NIH, Bethesda, MD, United States
1
1
POSTERS
Background: Two approaches have highlighted the importance of
antibodies to the gp120 V1/V2 domain in providing protection from
HIV-1 infection. First, the RV144 HIV-1 trial documented a correlation
between non-neutralizing antibodies to the V2 domain and protection.
Secondly, multiple broadly neutralizing monoclonal antibodies (bNmAbs) to glycan-dependent epitopes (GDEs) in the V1/V2 domain have
been isolated from rare infected individuals (elite neutralizers).
Methods: Analysis of the glycan content of the A244 and MN-rgp120
antigens used in the RV144 trial showed that they lacked the glycans
required for the binding of bN-mAbs to the V1/V2 and the V3 domains.
We wondered if we could improve the ability of these immunogens to
elicit both types of protective antibodies by in vitro mutagenesis and by
production in GnTI- cells that limit carbohydrates to mannose-5 glycans.
Results: We found that we could re-engineer monomeric gp120s used
in the AIDSVAX B/E vaccine used in the RV144 trial to bind multiple bNmAbs , e.g. PG9, PGT128, and VRC01 with high affinity. We also found
that we could produce stable disulfide bonded fragments of the V1/V2
domain (scaffolds) that preserved the 4-stranded beta sheet structure
and could bind PG9 and PG9-like antibodies. Immunization studies
showed that these gp120s and scaffolds could significantly enhance
the magnitude of antibody responses that correlated with protection in
the RV144 trial. These immunogens also enhanced the magnitude of
antibodies directed to the PG9 binding site.
Conclusions: Our studies suggest that we can substantially improve the
immunogenicity of the vaccine that provided protection in the RV144
trial. Our goal is to improve the level of protection (31.2%) observed in
RV144 to a level of 60% or more required for regulatory approval. By
improving an existing vaccine with a record of safety in more than 9000
subjects and demonstrated efficacy, years of time and millions of dollars
can be saved compared to developing a new vaccine from scratch.
180
Background: Many viruses use single-headed immunoglobulin domains
as receptors, but restrict access of double-headed immunoglobulins that
comprise the recognition domains of human antibodies. The membranedistal single-headed immunoglobulin domain of CD4, for example, is
recognized by the HIV-1 gp120 envelope glycoprotein, but access to the
site of CD4 binding is restricted for most human antibodies.
Methods: To understand the immunological consequences of doubleheaded versus single-headed recognition, we determined crystal
structures of gp120 in complex with four single-headed antibodies, A12,
C8, D7 and J3, derived from immunized llamas, which target the CD4binding site.
Results: Antibody J3 neutralization breadth approached 98% despite
a targeting precision similar to that of the double-headed antibody
VRC16, which has only ~56% breadth. A12 breadth was 41% despite a
targeting precision below that of b13 and F105, which have only ~10%
breadth. Double-headed A12 lose neutralize activity against twenty
selected viruses.
Conclusions: Overall, HIV-1 neutralization by double-headed antibodies at
the CD4-binding site required substantially higher targeting precision, and
more V-gene affinity maturation than single-headed antibodies, an effect
our results attribute to “canyon shielding” of this important vaccine target.
P10.03
P10.04
Optimization of a Clade A Env Outer Domain
for Enhanced Binding to Germlines of Diverse
VRC01 Class Antibodies
Negative Selection Using CD4-binding Site
Non-broadly Neutralizing Antibodies Yields
Conformationally Homogeneous Clade B and
C SOSIP Trimers
Diseases, National Institutes of Health, Bethesda, MD, United States
1
Background: Effective HIV vaccines need to elicit broadly neutralizing
antibodies (bNab) as an essential component of protective immunity.
Among the known bNAb, those that target the conserved CD4 binding
site (CD4bs) are among the most broad and potent. The VRC01 class
of CD4bs antibodies derive from a VH1-2 germline gene and have a
characteristic short (5AA) CDRL3. These antibodies are highly affinity
matured and when reverted to germline sequence fail to recognize most
HIV Env sequences. Thus, a major challenge is to design immunogens
that would activate the appropriate naïve B cells and generate VRC01
class antibodies. Previous studies have identified few clade C gp140 and
clade B gp120 outer domain constructs (OD) that can engage germline
of a subset of CD4bs antibodies. Here, we aim to identify HIV clade A
OD mutants that can bind to germlines of diverse CD4bs antibodies to
be used as HIV imunogens.
Methods: Clade A OD4.2.2 (PDB code 4I3R) was displayed on the
cell surface using a yeast display system. Mutations in its D loop (274283), which constitutes part of the CD4bs, were introduced by PCR to
construct two libraries: one with randomized sequences and the other
with sequences designed by structure based bioinformatics. The libraries
were screened for binding to germline revertants of VRC01-class
antibodies. Germline binders were sorted with FACS and characterized.
Results: Consistent with previous studies, the original yeast displayed
OD failed to bind to any of the germline revertants of CD4bs antibodies.
Two clones from the randomized D loop library were identified to bind
VRC03 germline (VH1-2). When displayed on lumazine synthase 60mer
nanoparticles, they bound to several VRC01 class germline revertants
including VRC01, VRC07, VRC20, 12A12, CH31 and VRC03.
Conclusions: Optimization of amino acids in loop D of the OD helped to
identify mutants that can bind to diverse VRC01 class germline reverted
antibodies. These OD constructs can thus serve as priming immunogens
to elicit CD4bs bNab.
Javier Guenaga1, Natalia de Val2, Karen Tran1, Karen Satchwell1,
Yu Feng1, Andrew Ward2, Richard Wyatt1
International AIDS Vaccine Initiative (IAVI), Neutralizing Antibody
Center, Scripps Research Institute, La Jolla, CA, United States, 2Scripps
Research Institute, Immunology and Microbial Science, La Jolla, CA,
United States
1
Background: The recently published structures of the clade A-derived
BG505 SOSIP trimers, mimetics of the native HIV envelope glycoprotein
(Env) spike, mark the beginning of new era in HIV structural biology.
Displaying a well-ordered quaternary array, the BG505 SOSIP trimers
display an excellent antigenic profile, discriminating recognition by broadly
neutralizing antibodies (bNAbs) from non-broadly neutralizing antibodies
(non-bNAbs) and represent interesting Env-based immunogens. Even with
this significant advance, obtaining soluble SOSIP trimers derived from
other clades has been challenging. Here, we report the isolation of two
SOSIP trimers derived from clades B and C.
Methods: We modified the 2G12 affinity purification by replacement with
lectin affinity purification, affording a scalable process with mild elution
conditions. Using CD4 binding site-directed (CD4bs) non-bNAbs in a
negative selection purification process, we obtained homogeneous wellordered clade B JRFL and clade C 16055 SOSIP trimers from a mixture of
conformations. We then used EM, bio-layer interferometry and differential
scanning calorimetry to characterize these new Env mimetics.
Results: Following negative selection, by EM we demonstrated that we
achieved nearly complete removal of disordered trimers, recovering
predominantly well-ordered trimers. We obtained EM 3D reconstructions
of the trimers, unliganded, in complex with sCD4 and the bnAbs, VRC01,
VRC03 and the newly identified trimer-specific PGT151. We employed
bio-layer light interferometry to get a full antigenic profile and showed
that the negatively selected homogeneous trimers behave as faithful
mimetics of the native spike, possessing avid recognition by bnAbs and
poor recognition by non-bnAbs.
Conclusions: This study affords a new means to obtain conformationally
homogeneous soluble mimetics of Env derived from two different HIV
clades from a mixture of Env conformations that can potentially expand
to other strains.
www.hivr4p.org
181
POSTERS
Cheng Cheng1, Ivelin Georgiev1, M Gordon Joyce1, Xuejun
Chen1, Adam Bossert1, Wing-Pui Kong1, Peter D. Kwong1, John R.
Mascola1
Posters
P10.05
P10.06
Double Safety Measure for Vaccines Based on
Replication Competent Poxvirus
CD8 T-cell Based HIV Vaccines - Is Targeting
Conserved Epitopes the Answer?
Ying Liu1, Qicheng Zhang1, Shuhui Wang1, Chang Liu1, Zheng
Liu1, Hong Peng1, Yiming Shao1
Shelby L. O’Connor1, Dane Gellerup1, Max Harris1, Ericka Becker1
National Center for AIDS/STD Control and Prevention, Chinese Center
for Disease Control and Prevention, Division of Virology & Immunology,
Beijing, China
University of Wisconsin-Madison, Madison, WI, United States
1
1
POSTERS
Background: Poxvirus is a commonly used viral vector in vaccine
research. There is no pre-existing vector immunity in young population
due to eradication of the smallpox in 1970s. Replication competent pox
vector is more immunogenic than non-replicating one, but may not be
safe in immunecompromised people. In HIV vaccine research, double
safety measures may be needed to deal with rare case of spill over from
vaccinee to AIDS patients. The Vaccinia Virus Tiantan (TV), the Chinese
small pox vaccine, was attenuated and used as vector for recombinant
HIV-1 vaccine (rTV). The rTV has proved to be safe and immunogenic in
phase I /II trails in China.
Methods: To provide a double safety measure, the Herpes Simplex Virus
thymidine kinase gene was inserted into the TV to obtain the replication
competent TV/tk vector. The inhibition of the Ganciclovir (GCV) to the
viral vector was tested in vitro and in vivo. The immunogenicity of TV/tk
vector was also tested and compared with the rTV.
Results: The replication of TV/tk on CEF and Vero cells were significantly
inhibited with EC50 of 15.4 µM and 2.5 µM respectively.GCV could inhibit
the replication of TV/tk in brains of mice and provide 100% protection
to the mice at a dose 80mg/kg/day. If the GCV treatment began before
3 day p.i., mice infected with TV/tk could be protected. The TV/tk/Env
can stimulate strong humoral and cellular responses, similar to that by
TV/Env vector.
Conclusions: The study showed that insertion of HSV-tk gene into TV
vector is a reliable rescue measure to the replication-competent vaccinia
virus vaccination. This strategy can provide a double safety guarantee in
large scale application of such vaccine in situation, where the vaccine target
population is mixed with ineligible immune compromised individuals.
182
Background: Immunodominant CD8 T cell responses emerge in the
first few weeks after HIV/SIV infection, suppress virus replication, and
select for escape variants. Although T cell based HIV vaccines have not
successfully provided sterilizing immunity, vaccine-elicited CD8 T cells
may contribute to the control of replication of breakthrough viruses.
It is critical that a T cell based vaccine generates an effective pool of
memory CD8 T cells that are available to respond during acute HIV
infection and effectively control both acute and chronic virus replication.
Designing a vaccine to elicit these potent CD8 T cells in all vaccinated
individuals, however, is a daunting challenge. Conceptually, a vaccine
that elicits CD8 T cell responses targeting highly conserved virus peptide
sequences would have the most widespread impact, but the efficacy of
T cells targeting these conserved epitopes is unknown.
Methods: We employ a model of SIVmac239Δnef-infected MHCidentical Mauritian cynomolgus macaques to test the hypothesis
that CD8 T cells targeting conserved epitopes are unable to detect
and destroy virally infected cells. Accordingly, we expect that CD8 T
cells targeting epitopes that accumulate variants are more effective at
controlling virus replication. To test this hypothesis, we are creating
variants of live attenuated SIVmac239Δnef designed to elicit CD8 T cells
targeting epitopes that do and do not accumulate variants.
Results: We will determine whether the mutant viruses are ´fit´ and
whether the included variant epitope sequences are no longer detected
by CD8 T cells. In the future, we will determine if acute CD8 T cells
targeting these different categories of epitopes are able to control virus
replication, in vivo.
Conclusions: The conclusions from this study will help identify whether
CD8 T cells that develop during acute HIV infection can be specific for
highly conserved epitopes and whether they can control virus replication.
P10.07
P10.08
Bivalent NYVAC-based Vaccine Candidates
against HIV/AIDS Expressing Clade C Trimeric
Soluble gp140(ZM96) and Gag(ZM96)-PolNef(CN54) as VLPs
Characterization of the Binding Affinity of
Siglec-1 to gp120, gp145, and V2 Loop via
Sialic Acid Binding Motif
Centro Nacional de Biotecnología, CNB-CSIC, Madrid, Spain, 2Centre
Hospitalier Universitaire Vaudois and University of Lausanne, Lausanne,
Switzerland, 3University of Regensburg, Regensburg, Germany, 4The
Biodesign Institute at Arizona State University, Tempe, AZ, United States
1
Background: The generation of vaccine candidates against HIV/AIDS
able to induce long-lasting protective immunity remains a major goal
in the HIV field. The reduced efficacy (31.2%) against HIV infection
observed in the RV144 Thai clinical trial highlighted the need to develop
novel improved poxvirus-based recombinants.
Methods: In the present study, we have generated two novel NYVAC
vectors expressing HIV-1 clade C gp140(ZM96) (NYVAC-gp140) or
Gag(ZM96)-Pol-Nef(CN54) (NYVAC-Gag-Pol-Nef) and defined their
biological characteristics in cultured cells and in mice.
Results: Insertion of the HIV-1 genes in the viral genome does not
affect the replication capacity of the NYVAC recombinants in primary
chick cells and the HIV-1 antigens are correctly expressed and released
from the cells, with Env protein as a trimer (NYVAC-gp140), while in
cells infected with NYVAC-Gag-Pol-Nef, Gag-induced VLPs are abundant.
Electron microscopy revealed that VLPs are accumulated with time at
the cell surface, with no interference with NYVAC morphogenesis. GagPol-Nef expression in human primary monocytes markedly increases
the levels of immunomodulatory molecules, such as cytokines and
chemokines. Both recombinant viruses show an attenuation profile
in immunocompromised BALB/c mice after intracranial inoculation.
Analysis of the immune responses elicited in mice after homologous
NYVAC prime/NYVAC boost immunization shows that the two
recombinant viruses induced polyfunctional Env-specific CD4 or Gagspecific CD8 T cell immune responses. Antibody responses against
gp140 and p17/p24 were also elicited by both NYVAC vectors.
Conclusions: Our findings showed that bivalent NYVAC vectors can be
considered as candidate vaccines against HIV/AIDS.
U.S. Military HIV Research Program (MHRP)/HJF, Laboratory of
Adjuvant and Antigen Research, Silver Spring, MD, United States,
2
Catholic University of America, Department of Biology, Washington,
DC, United States, 3U.S. Military HIV Research Program (MHRP), Walter
Reed Army Institute of Research, Laboratory of Adjuvant and Antigen
1
Background: Although HIV-1 primarily infects T-cells through the
interaction of viral envelope with CD4 and a co-receptor CCR5/CXCR4,
CD4 expresses in low level in macrophages. Recently, sialic acid-binding
Ig-like lectin 1 (Siglec-1, CD169), which is highly expressed on the cell
surface of macrophages, has been identified as a major receptor for HIV1 infection. It is known that Siglec-1 binds to glycans containing sialic
acid. To further understand the mechanism; we determined the binding
affinity/kinetics of monomeric gp120 and trimeric gp145 to assess if
there were differences in their ability to bind to Siglec-1. We further
narrowed down the env-binding motif to the V2 loop of gp120. Finally,
we conducted an inhibition assay in the presence/absence of lactose
(LA), 2, 6’-Sialyllactose (6’-SL) and sialic acid (SA) to identify the nature
of the interactions.
Methods: Siglec-1 was immobilized on a CM5 chip on a Biacore
T200. JRFL and SF162 (clade B) gp120, gp145, and V2 loop proteins
were captured. In additional experiments, these env proteins were
immobilized, while Siglec-1 was injected over that surface. For the
inhibition assays, LA, SL and SA were mixed with the env proteins or
with Siglec-1 and then injected onto the immobilized Siglec-1 or env
proteins, respectively.
Results: The gp120 and gp145 binds to Siglec-1 with high affinity, 4.13
nM (JRFL gp120), 0.28-2.9 nM (JRFL gp145) and 0.44-8.26 nM (SF162
gp145). Importantly, we found that the interaction of env protein to
Siglec-1 took place through the V2 loop, (4.34 nM JRFL V2; 1.96 nM
SF162 V2). The presence of SA completely blocked the interaction of the
env and the V2 loop protein with Siglec-1, suggesting thereby that the
interaction occurred via a SA motif.
Conclusions: The binding of gp120, gp145, and V2 loop to Siglec-1 is a
high affinity interaction. Binding occurs through the V2 loop and requires
sialic acid. This suggests that the vaccine design should aim to induce
antibodies against V2-sialic acid complexes to prevent HIV-1 infections.
www.hivr4p.org
183
POSTERS
Beatriz Perdiguero1, Carmen Elena Gómez1, María Victoria
Cepeda1, Lucas Sánchez-Sampedro1, Juan García-Arriaza1,
Ernesto Mejías-Pérez1, Victoria Jiménez1, Cristina Sánchez1,
Carlos Oscar S. Sorzano1, Julie Delaloye2, Thierry Roger2, Thierry
Calandra2, Ralf Wagner3, Benedikt Asbach3, Karen V. Kibler4,
Bertram L Jacobs4, Giuseppe Pantaleo2, Mariano Esteban1
Hung V. Trinh1, Ousman Jobe1, Guofen Gao2, Carl R. Alving3,
Venigalla Rao2, Mangala Rao3
Posters
P10.09
P10.10
Scalable and Robust Purification of Intact HIV1 gp120 Monomer Subunit Antigens
Transmitter Founder Multi-envelope DNA
Vaccine Induces Potent Cross-clade Cellular
and Humoral Responses in Rabbits and Nonhuman Primates
Yingxia Wen1, Sai Tian1, Christine Linton1, Susan Barnett1,
Andrea Carfi1
1
Megan Wise1, Muthumani Karuppiah1, Janess Mendoza2, Natalie
Hutnick1, Jian Yan2, David Montefiori3, Celia Labranche3, Kate
Broderick2, Matthew Marrow2, Niranjan Sardesai2, David Weiner1
Background: Development of an effective vaccine against HIV-1 is
challenging due to various viral evolutionary mechanisms to evade
human immune system. The partial efficacy of the recent RV144
vaccine efficacy trial in Thailand provides hope for improvements of
vaccine regimens for higher efficacy. Clinical trials in Thailand is planned
to confirm and extend the results of the RV144 trial with the vaccine
strategy of poxvirus vector prime plus envelope protein boost.
Methods: To produce gp120 monomers, we generated CHO stable
cell lines, consistently expressing intact gp120 subunits with high
yield. Simple, scalable and robust antigen purification processes were
developed to generate gp120 proteins with high purity, integrity and
homogeneity.
Results: The ion-exchange based purification strategy enabled the
isolation of gp120 from host cell contaminates and separation of
gp120 monomer from dimer. This purification strategy also significantly
removed contaminated proteinase and inhibited proteinase activity,
leading to intact gp120 monomer with high purity. Purified gp120
monomers were stable, either alone or in combination, and when
formulated with adjuvant Alum and MF59. The antigenicity and
immunogenicity studies are on-going.
Conclusions: Scalable and robust purification process of intact HIV-1
gp120 monomer subunit antigens is developed to produce HIV gp120
antigen for clinic trials.
1
Novartis Vaccines, Cambridge, MA, United States
POSTERS
184
University of Pennsylvania, Philadelphia, PA, United States, 2Inovio
Pharmaceuticals, Inc., Blue Bell, PA, United States, 3Duke University,
Background: It has been reported that guinea pigs vaccinated with
transmitted founder gp140 envelope proteins are able to induce
low neutralizing antibodies with some improved breadth (Liao et al
2013). This general induction of coverage may be ideal for a priming
immunization, establishing a response which is able to be boosted with
the addition of either chronic or consensus envelopes. Since the DNA
vaccine platform is seen as a priming regiment, we aimed to investigate
if broad responses could be induced using primary transmitter founder
(TF) and acutely isolated gp160 immunogens developed from clades A,
B, and C as DNA plasmids.
Methods: Initial studies were performed in rabbits, which were
immunized by EP route with different combinations of synthetically
optimized gp160 immunogens from clades A, B, and C. Animals
received the same amount of total DNA and were immunized at 3 week
intervals. Humoral responses were determined after each immunization.
As a follow on, non-human primates were immunized with clusters of
gp160 DNA at weeks 0, 4, 8, 12 and boosted at week 48.
Results: Rabbits immunized with clusters of clade A gp160 envelope DNA
were able to induce cross-clade binding titers with limited neutralization.
Including TF envelopes from different clades increased binding titers
and neutralization breadth and potency. NHP immunized with clusters
of clade A and B gp160s showed cross-clade cellular responses after
two immunizations. These responses increased after each immunization
and were maintaining into memory. After two immunizations, NHP were
able to induce cross clade Ab binding titers against primary gp120 from
clades A, B and C and tier 1 neutralization.
Conclusions: DNA plasmids encoding TF and acute gp160 immunogens
are expressed and induce a potent immune response in vivo. We observed
for the first time that exposure of the immune system to multiple DNA
env vaccines at one time dramatically alters the immune phenotype
induced resulting in increased magnitude and breath of responses.
P10.11
P10.12
CD4-binding-Site Recognition by VH1-46
Germline-derived HIV-1 Neutralizers
Longitudinal Antibody Development in
SHIVAD8 Infected Non-Human Primate
Priyamvada Acharya1, Tongqing Zhou1, Cinque Soto1, Lei Chen1,
Timothy S. Luongo1, Stephanie Moquin1, Ivelin S. Georgiev1,
Stephen D. Schmidt1, Mark K. Louder1, M. Gordon Joyce1,
Yongping Yang1, Baoshan Zhang1, Johannes Scheid2, Michel C.
Nussenzweig2, John R. Mascola1, Peter D. Kwong1
Zizhang Sheng1, Joseph R. Francica2, Yoshiaki Nishimura3, Stephen
D. Schmidt2, Rebecca Lynch2, Sam Darko2, Zhenhai Zhang1,4,
Frederick Jaeger5, Munir Alam5, Daniel Douek2, John R. Mascola2,
Malcolm A. Martin3, Robert A. Seder2, Lawrence Shapiro1
2
Background: The human immune system generates diverse antibodies
against the binding site for the CD4 receptor in response to infection or
vaccination. Two classes of antibodies that effectively neutralize HIV1 through CD4 mimicry have been described, including those derived
from VH1-2 germline, which include the well-characterized VRC01 class
of antibodies, and those derived from VH1-46.
Methods: We defined structural modes of recognition of three VH146 germline-derived antibodies from 2 donors by crystallizing antigenbinding fragments in complex with extended core version of the HIV-1
gp120. We used surface plasmon resonance and biolayer interferometry
to determine antigen-binding properties, tested neutralization in
a representative panel of 192 viruses, and performed cross-donor
phylogenetic analyses to study antibody evolution.
Results: VH1-46 derived antibodies show a structural mode of CD4
mimicry distinct from that of the VH1-2 derived antibodies. The altered
mode of heavy chain recognition allows the VH1-46 derived antibodies
to accommodate light chain CDR3s of different lengths. Cross-donor
phylogenetic analysis showed that the VH1-46 derived antibodies from
2 donors evolved similarly and indicated that the VH1-46 antibodies
form a class.
Conclusions: Our studies show how antibodies of the VH1-46 class
achieve CD4 mimicry, expand the range of known structural solutions
that permit such heavy-chain mimicry, and reveal how small differences
in germline (e.g. between VH1-2 and VH1-46) can impact mature
antibody recognition.
Columbia University, Department of Biochemistry and Molecular
Biophysics, New York, NY, United States, 2National Institutes of Health,
Vaccine Research Center, Bethesda, MD, United States, 3National
Institute of Allergy and Infectious Diseases, National Institutes of
Health, Laboratory of Molecular Microbiology, Bethesda, MD, United
States, 4Southern Medical University, Division of Nephrology, Nanfang
Hospital, Guangzhou, China, 5Duke University Medical Center, Duke
Human Vaccine Institute, Durham, NC, United States
1
Background: SHIV infected Non-human primate (NHP) is an important
system to study antibody development relevant to HIV-1 infection in
humans. CCR5-tropic SHIV AD8-EO is a clade B virus that has a tier
2 neutralization phenotype and can induce broadly neutralizing
antibodies. However, the developmental characteristics of SHIVAD8
elicited NHP antibodies are unclear.
Methods: Four of eight SHIVAD8 infected Rhesus macaques developed
cross-reactive antibodies (good neutralizers) and others showed weak
Tier1 virus neutralizing activity (poor neutralizers). Peripheral memory
B cells from all animals at four time frames (6-8, 26-32, 52-54, and
91-110 weeks) were sorted into gp120 reactive (gp120+) and nonreactive B cells (gp120-). Antibody heavy chains were then sequenced
by 454-pyrosequencing. Germline V gene was assigned for each
antibody sequence using a newly characterized NHP heavy chain V
gene database. Somatic hypermutation level and CDRH3 length were
then calculated using an in-house bioinformatics pipeline.
Results: Compared with gp120- B cells, VH gene composition analysis
of gp120+ B cells showed enriched VH3-J (human ortholog VH3-23),
VH4-D (human VH4-B) and VH4-A (human VH4/OR15-8) antibodies
from good neutralizers. The somatic hypermutation level of gp120+
B cell antibodies increased gradually from ~4% at week6 to ~10%
at week110. In gp120+ B cells of six animals, more frequent usage of
antibodies with CDRH3 ~20aa in length is observed. In animal DCF1
(good neutralizer), more than ten long CDRH3 (>=28aa) antibody
lineages were elicited at early time points and some continued
expanding through week108 post infection.
Conclusions: SHIVAD8 infection preferably elicited certain VDJrecombined antibodies. The SHIVAD8 infected NHP system captures
characteristics of antibody development in HIV-1 infected humans,
such as longitudinal antibody maturation and elicitation of long CDRH3
antibodies. This supports the idea that SHIVAD8 infected NHP provides a
good model for the study of HIV antibody development.
www.hivr4p.org
185
POSTERS
NIAID, NIH, VRC, Bethesda, MD, United States, Rockefeller University,
1
Posters
P10.13 LB
P10.14 LB
A Novel Trimeric V1V2-Scaffold Immunogen
Induces V2q-Specific Antibody Responses
Chimeric Bovine-V-region and Human-Cregion mAbs with Long and Extensively
Mutated CDHR3 Domains Bind HIV-1 Env
gp140 Trimers, but Not gp120 Monomer
Xunqing Jiang1, Max Totrov2, Constance Williams3, Wei Li4, Shan
Lu4, Shixia Wang4, Susan Zolla-Pazner3, Xiang-Peng Kong1
NYU School of Medicine, Department of Biochemistry and Molecular
Pharmacology, New York, NY, United States, 2Molsoft LLC, San Diego,
CA, United States, 3NYU School of Medicine, Department of Pathology,
New York, NY, United States, 4University of Massachusetts Medical
School, Department of Medicine, Worcester, MA, United States
1
POSTERS
Background: Data from the RV144 vaccine clinical trial revealed that
high levels of antibodies to the first and second variable regions (V1V2) of
gp120 were correlated with the modest protection from HIV-1 infection;
thus V1V2 is a potential target for HIV/AIDS vaccine development. V1V2
is known to harbor three distinct epitope types including V2q defined by
quaternary broadly neutralizing mAbs such as PG9/PG16, V2p defined
by the RV144 mAbs CH58/CH59, and V2i defined by a panel of mAbs,
such as 697-D, targeting a region overlapping the V1V2 integrin-binding
site. It is highly desirable to develop immunogens that can induce
antibody responses specific for these epitope types.
Methods: We engineered a trimeric V1V2 (ZM53 sequence) immunogen
that was designed to mimic Env conformation by inserting it into a
trimeric scaffold (PDB ID 2J9C). We produced this immunogen in 293
cells and tested its antigenicity by ELISA. Rabbits were immunized by the
DNA prime-protein boost regimen with a gp120 DNA (ZM109 sequence)
and the V1V2ZM53-2J9C immunogen. The immunogenicity was then
assessed by antibody competition assays.
Results: We found that the V1V2-2J9C immunogen could bind mAb
PG9, CH58 and 697-D, thus it harbors the V2q, V2p and V2i epitopes.
We also found that this immunogen is highly immunogenic in rabbits.
Direct binding competition ELISA assays showed strong competition
between immune rabbit sera and CH58 or PG9, but not with 697-D,
thus this trimeric V1V2 immunogen induced strong antibody responses
targeted not only the linear V2p epitope type but also the quaternary
V2q epitope type.
Conclusions: Our results demonstrate that structurally constrained
V1V2 scaffolds with Env quaternary conformation can be designed
and synthesized, resulting in elicitation of antibody responses with
specificities overlapping the PG9 epitope region - a first step in
immunogen design to target a major vulnerable site of HIV-1 Env.
186
Behnaz Heydarchi1, Robert Center1, Sri Ramarathinam2,
Christopher Gonelli1, Brian Muller1,3, Charlene Mackenzie1, Jack
Cuthbertson1, Marit Kramski1, Damian F.J. Purcell1
University of Melbourne, Microbiology and Immunology, Melbourne,
Australia, 2Monash University, Biochemistry and Molecular Biology,
Clayton, Australia, 3Reef Pharmaceutical, Melbourne, Australia
1
Background: Bovine immunoglobulins (Ig) typically have variable
third heavy complementarity determining regions (CDRH3) for antigen
engagement that are significantly longer than in humans and other
mammals. We aimed to isolate HIV-1 bovine memory B cells binding HIV1AD8 Env gp140 trimer antigen from a vaccinated cow producing broadly
neutralising antibodies to examine the structure of their HIV Env gp140
antigen binding sites and construct chimeric bovine-human antibodies.
Methods: A cow was vaccinated with HIV-1AD8 Env gp140 trimers over
4 years and antigen-specific memory B-cells. HIV specific memory B
cells were detected in ELISPOT assay and isolated by FACS single-cell
sorting from PBMC. The bovine Ig heavy (H) and light (L) chain variable
(V) regions were amplified from cDNA from anti-CD21+ anti-IgG+
gp140-PE-binding+ cells by nested PCR. Paired H and L chain expression
vectors using human Ig constant regions were made and co-transfected
into 293T cells. Chimeric bovine-human (BH) Ig in supernatant was
screened for HIV-1 gp140 binding in ELISA.
Results: The frequency of HIV specific memory B cells was 1.96%
± 0.31% of total memory B cells on a background of 0.24% in nonimmune PBMC. Ig from 6 of 30 matched chimeric BH H and L chain
plasmid transfections displayed strong binding to HIV-1 AD8 gp140 Env
trimers using direct ELISA, but not gp120 monomers or cleaved gp41.
Two mAbs bound gp140 by mass-spectrometry and western blotting.
Analysis of CDRH3 of the anti-HIV antibodies had a CDRH3 containing
Cys and aromatic residues and were 14-22 amino acids (average size of
18.8 ± 3.1). In addition, the somatic mutation rate in CDRH3 compared
to the DH3 germline gene was 82.35- 90.90% (Average: 87.34±
2.51%). The V-region sequence aligned most strongly with 2F5 and
4E10 patient derived mAbs.
Conclusions: The bovine V-gene CDRH3 size and frequency of somatic
mutations of the isolated bovine Ig-genes raised through vaccination were
comparable with those for human patient’s elite neutralizing antibodies.
Posters 11: Informed Consent
P11.01
P11.02
Developing a Novel Approach to Assess
Stakeholder Views on the Length of Consent
forms for HIV Prevention Research
Regulated Flexibility in Adolescent Informed
Consent Procedures: Ensuring Inclusion of the
Most Vulnerable Populations in HIV Research
Amy Corneli1, Emily Namey1, Monique Mueller1, Ansley Lemons1,
Jeremy Sugarman2
Rachael C. Dellar1, Kshama Haribhai1, Fanelesibonge Ntombela1,
Silvia Maarschalk1, Ayesha Kharsany1, Quarraisha Abdool
Karim1,2
Background: Potential research participants often do not understand
information in consent forms (CFs). Long CFs can be a contributing
factor, yet empirical evidence is limited on the information that should
be included or that could be removed from CFs in HIV-related research
and on the barriers to reducing CF length.
Methods: We are exploring these issues from the perspectives of
stakeholders affiliated with the HIV Prevention Trials Network (HPTN):
participants, investigators and site staff, community and regulatory
representatives, institutional officials, and members of institutional
review boards. We reviewed data collection approaches that could 1)
allow stakeholders to identify CF text as essential or extraneous and
2) identify areas of agreement and divergence across the stakeholder
groups. We also explored approaches that could allow stakeholders to
learn the perspectives of other stakeholders and to confirm or refute
comments attributed to them.
Results: A novel, modified Delphi approach was developed for building
stakeholder consensus on how to shorten CFs. The three-step process
begins with stakeholders highlighting essential or extraneous sentences
of an HPTN CF on a tablet device and describing barriers to reducing CF
length during a face-to-face interview. Next, up to three follow-up, online surveys will be conducted among a sample of participants from each
stakeholder group. During each survey, stakeholders view results from
the previous stage; refine their selections of essential information after
considering all stakeholder responses, particularly for areas of continued
disagreement; and reflect upon the barriers previously identified. The
Delphi process concludes with an on-line group interview in which
stakeholders will give input on a shorter CF built from the results of the
prior steps.
Conclusions: Consensus among key stakeholders about information to
keep and remove from HIV-related CFs is critical for reducing CF length.
Innovative methods should facilitate this process.
CAPRISA, Durban, South Africa, 2Columbia University, Epidemiology,
New York, South Africa
1
Background: Adolescent-focused HIV prevention trials are key to
altering predicted epidemiologic trajectories for HIV in hyper-endemic
settings such South Africa. However, obtaining informed consent for
adolescent participation in clinical and behavioral trials in such settings
presents a number of challenges which can result in exclusion of the
most HIV-vulnerable adolescents from HIV research.
Methods: We assessed the major challenges to obtaining informed
consent from assenting high school students under 18 years, and the
uptake of more flexible informed consent procedures designed to
overcome these challenges, in the behavioural school-based adolescent
trial CAPRISA 007 based in rural KwaZulu-Natal, South Africa.
Results: A total of 2675 students were enrolled into CAPRISA 007
in 2010, of whom 1918 students (72.0%) were under 18 years of
age. Major challenges in obtaining informed consent for assenting
students under 18 years were the significant proportions of students
who were from child-headed households (8.8%), had absent parents
or guardians (11.3%), or had parents or guardians who failed literacy
and comprehension assessments (3.9%). An important pathway
developed and approved in collaboration with University of KwaZuluNatal’s Biomedical Research Ethics Committee to overcome such
challenges and to introduce regulated flexibility in consent procedures
were community groups which were permitted to give proxy consent
for student participation in the trial. These community groups assisted
directly in the consent procedures for 24.0% (461/1918) of students who
might otherwise been excluded from participation.
Conclusions: Community engagement and the development of flexible
and adaptive informed consent procedures are critical for facilitating the
inclusion of the most vulnerable populations in HIV prevention trials
targeted to adolescents.
www.hivr4p.org
187
POSTERS
1
FHI 360, Durham, NC, United States, 2Johns Hopkins University,
Baltimore, MD, United States
Posters
P11.03
P11.04
Assessment of Understanding for Informed
Consent in HIV Vaccine Trials
Determinants of Informed Consent
Comprehension among Fisher Folk Cohort
in HIV Vaccine Preparatory Studies in SW
Uganda
Graham C. Lindegger1, Michael Quayle2, Catherine M. Slack1,
Sagri Singh3, Sabrina Welsh3, Pat Fast3
University of KwaZulu Natal, HIV/AIDS Vaccine Ethics Group
(HAVEG), Pietermaritzburg, South Africa, 2University of KwaZulu Natal,
Psychology, Pietermaritzburg, South Africa, 3International AIDS Vaccine
Initiative (IAVI), New York, NY, United States
1
POSTERS
Background: Informed consent (IC) is accepted as an essential ethical
requirement for clinical trials, including HIV vaccine trials (HVT).
Authentic IC requires that potential participants adequately understand
various trial concepts and the implications of trial participation before
being accepted into trials. While assessment of understanding is usually
done using forced choice questionnaires, there are many limitations to
this method of assessment. This paper reports on a series of studies
done to compare various methods of assessment of understanding,
including true/false questionnaires, vignettes and narrative approaches.
Methods: Three studies have been conducted to date. The methodology
involved quantitatively comparing the levels of measured understanding
on three assessment tools, viz. forced choice questionnaires, trial related
vignettes and narratives of trial participation. The comparisons were
conducted first on a group of potential HVT trial participants in South
Africa. A second study was conducted using a hypothetical HVT trial
on samples of potential trial participants in South Africa and Zambia. A
subsequent study compared measured understanding on these tools on
samples of potential participants in HVT in Uganda and South Africa.
Results: Results of all studies show a significant difference between
measured levels of understanding, with the forced choice questionnaire
having the highest scores, arguably over-estimating levels of
understanding, and open-ended measures the lowest scores.
Conclusions: It is suggested that open-ended methods of assessment
provide more realistic and appropriate measures of understanding,
which should be incorporated in IC procedures for HVT. Reservations
have been expressed about the cost and skill implications of using such
methods, however, suggestions are made for how these measures might
be cost-effectively incorporated into HVT.
188
Elizabeth Mbabazi1, Andrew Abaasa1, Gershim Asiki1, Ubaldo
Bahemuka1, Eugene Ruzagira1, Margaret Nambooze1, Cissy Lilian
Nalubega1, Mathew A. Price2, Anatoli Kamali1
Medical Research Council/Uganda Virus Research Institute, Entebbe,
Uganda, 2International AIDS Vaccine Initiative, New York, NY, United
States
1
Background: Informed consent comprehension is mandatory for
individuals’ decision to enrol and participate in clinical trials. Low
literacy may be a barrier to consent. We report the relationship between
volunteer literacy and comprehension of study information at entry into
a fisher folk HIV vaccine preparedness cohort.
Methods: Community meetings were conducted at fishing sites
along Lake Victoria to sensitize individuals to participate in HIV
vaccine preparedness study. Interested individuals were provided HIV
counselling and testing through which HIV uninfected volunteers were
identified and invited to a study clinic located about 40km away. At
the study clinic, written information on the study objectives, risks,
benefits and procedures was read aloud and explained by a nurse to
each individual. Those unable to read were requested to identify an
independent witness to confirm provision of accurate information.
Assessment of understanding (AoU) of study information was then done
by a different study staff using 8 questions requiring “true” or “false”
responses. Literacy was defined as the individuals’ ability to read the
study information in the local language and write their names. For the
analysis scoring all 8 points correctly was considered as a pass.
Results: Of 584 (60% men) with mean age 28 years enrolled between
Jan 2012 and Mar 2014, 122 (21%) were not literate. A total of 508
(87%) passed the AoU at first attempt. This did not differ by: literacy
status (87% for literate and not literate), gender (86% men vs. 89%
women, p=0.28] or other socio-demographic characteristics (age,
occupation religion, marital status, number of dependants, tribe and
parity) of the individuals.
Conclusions: Literacy status and other demographic factors may not
affect individuals’ comprehension of the informed consent process at
study onset in these communities provided the investigator reads and
explains the study information in the language understood by the
individuals.
P11.05
Participants’ Models of HIV Vaccine Trialrelated Concepts: The Dilemma of Trust for
the Informed Consent Process
Clinton L. Rautenbach1, Graham C. Lindegger1, Catherine M.
Slack1, Peter A. Newman2, Melissa Wallace3
University of KwaZulu Natal, HIV/AIDS Vaccine Ethics Group (HAVEG),
Pietermaritzburg, South Africa, 2University of Toronto, Factor-Inwentash:
Department of Social Work, Toronto, ON, Canada, 3Desmond Tutu HIV
Foundation, Socio-Behavioural and Adolescent Division Leader, Cape
Town, South Africa
1
POSTERS
Background: Informed consent (IC) is a dialogical process between
researchers and potential participants, which should be founded upon
a reciprocal research rapport. A bilateral, dialogical process should
be rooted in an ethos of credibility and trust. This paper will explore
enablers and inhibitors of trust within the IC process in HIV vaccine trials
(HVTs) in South Africa.
Methods: The study employed an open qualitative, process-oriented
exploratory design, using Focus Group Discussions (FGDs) to elicit mental
models or key representations of critical research-related concepts from
key constituencies, including CAB representatives, educators, and site
staff involved in consent processes at an HIV prevention trial research
site. Four FGDs were conducted - each constituency was interviewed
separately - with approximately eight participants per group. Transcribed
FGDs were subjected to thematic analysis by three qualitative researchers.
Results: Findings showed several barriers to credibility and trust around
research, researchers, and key research-related concepts indicated
by community members, which may complicate how concepts are
understood, interpreted or believed. Study respondents reported and
engaged in several strategies to build trust and develop research rapport
- likely to be foundational to research participation. We found socioeconomic, political, gender, race and culture-based mental models
that may compete with or, alternatively, complement explanations of
concepts offered by site-staff (e.g. vaccine-induced seropositivity [VISP]).
Conclusions: Authentic IC should involve a systematic exploration of
these barriers and enablers to trust as part of the on-going IC process,
to complement efforts to promote and assess understanding. We make
a series of recommendations for stakeholders who wish to incorporate
trust-building into their engagement and consent work to enhance
partnering and decision-making for HIV prevention trials.
www.hivr4p.org
189
Posters
Posters 12: Innate Immunity
P12.01
P12.02
Protective Role OF DC-SIGNR in HIV-1
Pathogenesis
Skin Antigen-presenting Cells and
Inflammation for Tailored Immunity to HIV
Vaccines
Omkar Chaudhary1, Sanjeev Kumar1, Muzamil A. Makhdoomi1,
Manju Bala2, Jasbir Singh3, Anjali Hazarika4, Rajesh Kumar5,
Kalpana Luthra1
All India Institute of Medical Sciences, Biochemistry, New Delhi, India,
Apex Regional STD Teaching, Training & Research Centre, Vardhman
Mahavir Medical College & Safdarjung Hospital, Microbiology, New
Delhi, India, 3Kurukshetra University, Biochemistry, Kurukshetra, India,
4
All India Institute of Medical Sciences, Blood Transfusion Services,
Cardiothoracic and Neurosciences Center, New Delhi, India, 5Society for
Promotion of Youth and Masses Center, New Delhi, India
1
2
POSTERS
Background: Dendritic cells (DCs) capture HIV-1 from periphery via DCSIGN/R receptors and transfer to CD4+T cells. The DC-SIGNR is highly
polymorphic and changes in its repeat regions have been suggested to
influence HIV-1 disease progression.
Methods: Blood from 230 seronegative healthy individuals, 200
injecting drug users and 230 patients infected with HIV-1 was collected.
DC-SIGNR polymorphism was performed by Polymerase chain reaction.
The distribution of peripheral blood DCs and their subset frequency were
determined by flow cytometry. DC-SIGNR expression in Peripheral blood
mononuclear cells was determined in HIV-1 antiretroviral naïve patients
and healthy individuals. DCs were cultured from monocytes and infected
with HIV-1 Indian clade C virus. The expression of DC-SIGNR on DCs was
silenced using DC-SIGNR siRNA and the effect of down regulation of
this receptor on HIV-1 infectivity of DCs, Co-stimulatory and signaling
molecules.
Results: The frequency of heterozygous DC-SIGNR 7/5 genotype and
allele 5 was significantly higher in injecting drug users compared to HIV1 infected patients and was associated with high dendritic cells count,
CD4+ T cells and low DC-SIGNR expression, viral load. The expression of
DC-SIGNR was higher in HIV-1 infected patients and inversely correlated
with CD4+ T cell count. DC-SIGNR transfected DCs, the expression of
co-stimulatory molecules and P38 MAPK was significantly reduced.
Silencing of DC-SIGNR expression in DCs followed by infection with HIV1 clade C viruses demonstrated lower levels of p24.
Conclusions: DC-SIGNR 7/5 genotype and allele 5 may have a
protective role in HIV-1 infection. The silencing of DC-SIGNR expression
decreases the gene expression of the co-stimulatory molecules that
in turn may inhibit the DC-T cell interactions needed for progression
of HIV-1 infections. A long-term goal of our study is to develop novel
therapeutic approaches to prevent HIV-1 infection.
190
Behazine Combadiere1, Clement Levin1, Olivia Bonduelle1,
Charles Nuttens1, Mireille Centlivre1, Helene Perrin1, CUT’HIVAC
Consortium
INSERM U 1135 CIMI-Paris and UPMC, Immunity and Vaccination,
Paris, France
1
Background: The EU-funded Ćutaneous and mucosal HIV vaccination´
(CUT´HIVAC) project are proposing alternative routes for vaccinating
against HIV. A considerable part of CUT´HIVAC work has been dedicated
to understanding the mechanisms triggered after HIV vaccination
via the skin for the induction of mucosal immunity by activation of
T follicular helper cells. Skin outer surface protects from pathogens
invasion thanks to skin antigen-presenting cells, which scan their
changing microenvironment based on molecular changes that need to
be identified.
Methods: Using human skin explants model for vaccination as well as
mouse models, we will present our understanding of skin cellular and
molecular networks processes and aims at designing novel therapeutic
approaches.
Results: Langerhans and dermal dendritic cells pay a major role in
dictating the quality of the immune responses in the draining lymph
nodes and in programming mucosal immunity. Skin APC (Langerhans
cells, dermal dendritic cells) surrounding the hair duct use their dendrite
extension to scan their constantly changing microenvironment and
interact to other skin cells (keratinocytes, fibroblasts, skin immune cells
and other antigen presenting cells). In addition, inflammatory cells
collaborate with professional antigen-presenting cells in the induction of
CD8 in the draining lymph node and CD8 memory in the bone marrow
as well as mucosal immunity in the vagina. Our work in mice model also
highlights the importance of skin epidermal DCs in eliciting a strong TFH
and B cell responses. It provides insight in skin vaccination mechanisms
to improve the expansion of the TFH for vaccine efficacy.
Conclusions: Thus, skin routes of immunization allow to target specific
population of antigen-presenting cells and solicit inflammatory cells
that could tailored cellular and humoral immunity for a preventive HIV
vaccine development.
P12.03
P12.04
Protective Role of TLR3 Induced Human Beta
Defensins during Acute HIV-1 Infection
Chronic Untreated HIV-1 Infection Is
Associated with Increased Erythrocyte
Apoptosis and Inflammatory Monocyte
Erythrophagocytosis
Ragon Institute of MGH, MIT, and Harvard, Cambridge, MA, United
States, 2Massachusetts General Hospital, Division of Infectious
Diseases, Boston, MA, United States, 3Heinrich-Pette-Institut, Leibniz
Institute for Experimental Virology, Hamburg, Germany
1
Background: Human beta defensins 1 (HBD1) and 2 (HBD2) are
antimicrobial peptides expressed by epithelial cells and immune cells
in tissue and peripheral blood. They are part of the first line of innate
immune defense against invading pathogens at mucosal sites. Although
beta defensins have been shown to have antiviral activity in vitro, their
expression during the course of HIV-1 infection in vivo has not been
fully characterized.
Methods: We investigated production and activity of HBD1 and HBD2
in intestinal mucosal biopsies and peripheral blood mononuclear cells
(PBMCs) from HIV-1-uninfected individuals (n=10) and subjects with
chronic untreated HIV (n=10), immunologically controlled HIV (n=13)
and acute HIV (n=23) using quantitative PCR and viral inhibition
assays. In vitro studies of the effect of Toll like receptor (TLR) signaling
on defensin expression were conducted using TLR specific ligands,
inhibitors and siRNA technology.
Results: HBD1 expression is highly upregulated (p< 0.0008) in PBMCs
but not in intestinal biopsy sample from acute HIV patients. Monocytes
are the main producers of HBD1 in PBMCs whereas epithelial cells are
the main source in the intestinal tract. HBD1 is upregulated in monocytes
during acute infection (Fiebig stages 2/3) but returns to basal levels in
chronic infection. In vitro studies reveal that HIV induces HBD1 but not
HBD2 transcription in monocytes. HIV-induced HBD1 expression is
dependent on TLR3 but not TLR 4, 5, 7 or 9 activation and independent
of interferon or tumor necrosis factor alpha signaling. Importantly,
recombinant HBD1 blocks HIV replication in activated T cells and
suggests a protective role for defensins during acute HIV infection.
Conclusions: Our study shows that the antiviral peptide HBD1 is
upregulated in monocytes during acute HIV infection in a TLR3
dependent manner. Activation of the TLR3 pathway and upregulation
of beta defensins at sites of infection might be an important defense
mechanism during HIV acquisition and is currently under investigation.
Richard H. Glashoff1, Stanley Loots1, Hayley Ipp1
Stellenbosch University, Medical Virology, Cape Town, South Africa
1
Background: Chronic HIV-1 infection is characterized by inflammation
and also by anaemia. Erythrocyte apoptosis (erythroptosis) may
contribute to anaemia and may also drive production of phagocytic
monocytes/macrophages. In this study we investigated erythroptosis in
asymptomatic, untreated HIV-1 infection and the relationship between
erythrocyte death and phagocytic monocyte activity.
Methods: A total of 44 chronically HIV-1 infected individuals (CD4
count > 200) and 33 matched uninfected individuals were included.
Erythrocytes were stained with annexin V for determining ex vivo
levels of erythroptosis. Erythrocytes were also subjected to oxidative
stress in vitro (5mM hydrogen peroxide), in the presence or absence
of the antioxidant N-acetyl cysteine (NAC). Finally, erythrocytes were
also stained with CFSE and then incubated with purified autologous
monocytes to monitor phagocytosis.
Results: HIV-1 infected individuals had reduced haemoglobin levels
(12.9g/dL vs. 14.1g/dL, p=0.017). This was mirrored in decreased RCC.
Significantly higher erythrocyte annexin V expression was observed
(13.4% vs. 10.4%, p=0.0189). Annexin V expression increased in both
groups when exposed to hydrogen peroxide. NAC significantly reduced
the percentage of apoptotic RBCs in the control group (20±5.1%
to 15±4.4%, p=0.006), but not the HIV-1 group (18.3±5.0% to
16±5.2%, p=0.065). There was a significant increase in the percentage
inflammatory monocytes (CD14+CD16+) in the HIV-1 group (8.3±3.5%
vs. 5.3±3.8% p=0.0054). The expanded inflammatory monocytes were
more erythrophagocytic than classical monocytes, and preferentially
phagocytosed apoptotic erythrocytes (phagocytic index 2.9% vs. 5.2%,
p=0.008).
Conclusions: Significant reduction in haemoglobin levels in untreated
chronic HIV-1 infection was associated with increased erythroptosis.
Inflammatory monocytes displayed enhanced uptake of erythrocytes.
Limiting inflammatory toxicities may counteract the development of
anaemia and associated monocyte changes in HIV-1 infection.
www.hivr4p.org
191
POSTERS
Bjorn Corleis1, Antonella Lisanti1, Christian Korner1, Molly A.
Amero1, Eric S. Rosenberg2, Todd M. Allen1, Marcus Altfeld1,3,
Douglas S. Kwon1
Posters
P12.05
P12.06
Dysfunctional Neutrophil Responses to SIV
Infection
Preliminary Evaluation of Serpins as Potential
Candidate Microbicides
Tiffany Hensley-McBain1, Laura E. Richert-Spuhler1, Michael
Koday1, Jillian Gile1, Brandon F. Keele2, Jacob D. Estes2, Nichole R.
Klatt1
Carolina Herrera1, Natalia Olejniczak1, Frank Plummer2, Robin
Shattock1, Adam Burgener2
Washington National Primate Research Center, University of
Washington, Department of Pharmaceutics, Seattle, WA, United States,
2
Frederick National Laboratory for Cancer Research, AIDS and Cancer
Virus Program, SAIC-Frederick, Inc., Frederick, MD, United States
2
1
POSTERS
Background: Recent studies indicate that individuals with low
neutrophils (PMN) are at increased risk of HIV infection. Also, the RV144
vaccine trial implicated non-neutralizing antibodies and associated Fcmediated functions in vaccine-induced protection. These studies suggest
that early innate antiviral functions and Fc-mediated functions of PMN
may be important mediators of protection. Consequently, protective
immunity may rely on kinetics of PMN mobilization, activation, and
recruitment during acute HIV/SIV infection.
Methods: We assessed kinetic changes in PMNs in peripheral and
mucosal tissues during acute SIV infection in six rhesus macaques
challenged i.r. with 100,000 TCID50 of SIVmac239X. Flow cytometry,
CBC, and luminex were used to assess PMN and cytokine levels and
related PMN functional markers. Samples were collected pre-SIV and
days 3, 7, 14, 21, 28, 42, and 63 post-SIV.
Results: We observed a significant decrease in systemic IL-17 (p=.0313)
and a trending decrease in G-CSF (p=.0625) early after SIV. Surprisingly,
blood PMN concentrations steadily decreased after infection, and no
significant increase of PMN was detected in gut tissues. Blood PMN
numbers and rectal PMN percentages significantly correlated (p=.0032),
and HLA-DR (p=.0313), CD86 (p=.0156), and FcγRI (p=.0313) were
significantly upregulated on PMN during acute SIV infection.
Conclusions: In contrast to other acute viral infection models, PMNsupporting cytokines are decreased or not induced during acute SIV,
potentially contributing to lack of PMN mobilization from the bone
marrow and recruitment to the tissues. Further, blood and rectum PMN
levels correlate post-SIV, suggesting that blood PMN concentration
may directly impact recruitment to gut tissues. Lastly, upregulation of
markers involved in antigen presentation and Fc-mediated functions
highlight the potential diverse functional roles of neutrophils during
acute SIV infection and the potential importance of inducing neutrophils
for effective prevention strategies.
192
University of Manitoba, Winnipeg, MB, Canada
1
Background: Studies on HIV-resistant women (exposed uninfected)
from the Punwami Sex Worker cohort have shed light on putative
protective mechanisms, suggesting that mucosal immunological factors,
such as serpins, could be mediating HIV-resistance. This project aims
to assess the activity of a panel of serpins against HIV-1 in a preclinical
mucosal tissue explant model.
Methods: Antiviral efficacy of nine blinded serpins was assessed in TZMbl cells, ecto-cervical tissue explants and migratory cells. Incubation of
cells or tissue with serpins for 1 h was followed by addition of an R5tropic virus, BaL. Tissue was exposed to virus for 2 h and then washed.
Following overnight incubation of tissue, migratory cells were harvested
and co-cultured with PM-1 CD4+ T cells without drug. Infection was
determined by measurement of luciferase expression (in TZM-bl cells) or
p24 viral antigen in culture supernatants.
Results: Following screening in TZM-bl cells where dose-response
curves were measured for two serpins, antiviral activity in ecto-cervical
explants was detected for five serpins allowing us to establish an order
of inhibitory potency when mimicking pre-coital use of a serpin-based
microbicide. In addition, the migration of cells out of the explants was
blocked by certain serpins, indicating potential prevention of viral
dissemination following amplification of the founder population.
Conclusions: These results constitute the base for further development
of these mucosal proteins as microbicides. Serpin combinations and
sustained exposure of explants to serpins mimicking repeated dosing
strategies will inform the dosing and formulation of these antiproteases
which have shown potential as effective microbicides able to inhibit HIV1 transmission in pre-clinical assays.
P12.07
P12.08
Restoration of the NK Cells Ability to Mediate
ADCC in HIV-1 Positives after Six Months of
HAART Can be Explained by Normalization of
their Phenotype
Skin Migratory APCs Fine-tune Lymph Node
Microenvironment for the Generation of T
Follicular Helper Cells and Mucosal Immunity
Statens Serum Institut, Department of Microbial Diagnostic and
Virology, Copenhagen, Denmark, 2Odense University Hospital,
Department of Infectious Diseases, Odense, Denmark, 3Copenhagen
University Hospital, Department of Infectious Diseases, Copenhagen,
Denmark
1
Background: Natural killer (NK) cell phenotype and function have
recently gained much attention as playing crucial roles in antibodydependent cellular cytotoxicity (ADCC). Within the context of HIV
infection, the ability of NK cells to mediate ADCC is critical for protection
from disease acquisition and progression.
Methods: We investigated NK cell function, as measured by ADCC,
in HIV-1 positive individuals before and six months after highly active
antiretroviral therapy (HAART) initiation. The ability of antibodies and NK
cells to mediate ADCC was investigated separately and in combination
in an autologous model. The NK cell subset distribution and NK cell
phenotype were analyzed. The ADCC-GranToxiLux assay was used
to evaluate the ability of NK cells and antibodies to mediate ADCC.
The frequency of NK cells expressing receptors was determined by
phenotypic labeling followed by flow cytometry.
Results: The ability of NK cells to mediate ADCC was significantly
increased after only six months of HAART and was not explained by a
normalization of NK cell subsets but rather by a normalization of the NK
cell phenotype. The frequency of NK cells expressing CCR7 and CD27
significantly decreased after six months of HAART. For individuals with
no increase in ADCC after six months of HAART, the frequency of NK
cells expressing NKp46 was down-regulated. The ability of antibodies
to mediate ADCC alone and in combination in an autologous setup was
not improved.
Conclusions: HAART improves the ability of NK cells to mediate ADCC
after six months. This improvement does not correlate with general
immune restoration, as measured by CD4+ T cell counts, but rather
to a normalization of the NK cell phenotype. The investigation of the
immune function during HAART is important in order to gain knowledge
about who may respond to a therapeutic vaccine.
Sorbonne Universités, UPMC Univ Paris 06, Unité Mixte de Recherche
de Santé (UMR S) CR7, Centre d’Immunologie et de Maladies
Infectieuses, Paris, France, 2INSERM U 1135 CIMI, Paris, France, 3Institut
de Biologie et Chimie des Protéines UMR 5305 CNRS/UCBL, Lyon,
France, 4Centre de Physiopathologie de Toulouse-Purpan, UMR 1043,
CHU Purpan, Toulouse, France
1
Background: The understanding of mechanisms of induction of
humoral responses need to be further studied to define vaccination
strategies against HIV. We previously showed that intradermal (i.d)
immunization of mice with PLA-HIV-P24 nanoparticles induced mucosal
immunity in the vagina. The skin is rich in specialized dendritic cells
(DCs) subsets such as Langerhans cells (LCs) and dermal DCs, as well
as other myeloid DC populations. We therefore questioned the role of
skin antigen-presenting cells in the induction of T follicular helper cells
(TFH), which play a pivotal role in B cell help and isotypic class switch for
shaping IgG and IgA-secreting B cells.
Methods: We used Langerin-DTR mice model or ear ablation after ear
i.d vaccination to study the role of skin cells and Langerin+ cells in TFH
and IgG and IgA-secreting B cells induction. We used fluorescent PLA to
track antigen uptake and migration by skin resident and inflammatory
cells. Finally, gene array analysis was performed to reveal key markers
of DLN inflammation after immunization.
Results: Conditional depletion of Langerin+ cells or ear ablation
resulted in partial to full abortion of TFH polarization and p24-specific IgG
and IgA-secreting B cells expansion, respectively. Tracking of injected
PLA nanoparticles showed high ability of LCs and Langerin+ dermal
DCs to capture antigen in the skin and migrate to the T/B interface of
DLNs. Gene array analysis of the whole DLNs of control and ear ablated
mice after i.d immunization revealed distinct molecular signatures of
inflammatory molecules critical for generation of immune responses.
Conclusions: Work is currently in progress to validate the signature
of innate immunity brought by skin APCs to the lymph nodes for TFH
induction and mucosal immunity. Our work emphasizes on the role of
migratory DCs from the skin in fine-tuning the DLN microenvironment.
It will provide insights into mechanisms by which i.d immunization can
induce superior humoral immune responses for the prevention of HIV
infection.
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193
POSTERS
Sanne Skov Jensen1,2, Hans Jakob Hartling3, Jeanette Linnea
Tingstedt1, Tine Kochendorf Larsen1, Susanne Dam Nielsen3,
Court Pedersen2, Anders Fomsgaard1,2, Ingrid Karlsson1
Clement Levin1,2, Charles Nuttens1,2, Olivia Bonduelle1,2,
Helene Perrin1,2, Bernard Verrier3, Nicolas Fazilleau4, Behazine
Combadiere1,2
Posters
P12.09
P12.10
Acute SIV Infection Rapidly Depletes CD4+
pDCs, but Induces Gut-Homing pDCs from
Bone Marrow
Stimulation of Toll Like Receptor 7 and 8
(TLR7/8) Inhibits HIV-1 Replication in the
Pregnant Uterine Mucosa
Haiying Li1, Tristan I. Evans1, Michelle Connole1, Jacqueline Gillis1,
Fay E. Wong1, Yi Yu1, Roger Keith Reeves1
Hicham El Costa1, Heloise Quillay1,2, Marion Duriez1, Claire De
Truchis3, Anne Le Breton3, Mona Rahmati4, Julien Ighil4, Françoise
Barré-Sinoussi1, Marie-Thérèse Nugeyre1, Elisabeth Menu1
New England Primate Research Center, Harvard Medical School,
Southborough, MA, United States
1
POSTERS
Background: Given their potent antiviral functional role, plasmacytoid
dendritic cells (pDCs), the major IFN-α-producing cells, remain a target
of interest for HIV/SIV disease. pDCs accumulate in the gastrointestinal
mucosae during chronic disease, but the kinetics of mobilization and
trafficking are poorly understood. Furthermore, pDCs in multiple tissues
show evidence of dysfunction through unclear mechanisms.
Methods: This planned sacrifice study included 12 rhesus macaques
— six naïve and six intravenously infected with SIVmac239. Animals
had colorectal biopsies prior to infection, followed by longitudinal blood
draws, and tissue collection at day 14 post infection. Phenotypic and
functional analyses were performed using polychromatic flow cytometry.
Results: During acute SIV infection pDCs were rapidly depleted from
blood, spleen and liver, but accumulated in the gastrointestinal tract. The
gut-homing receptor, a4β7, increased significantly on circulating and
splenic pDCs by day 14 following infection, suggesting that trafficking
was a primary mechanism for the loss of pDCs from blood and lymphoid
organs. Interestingly, compared with naïve controls, bone marrow
pDCs in acutely infected rhesus macaques had increased proliferation
and significantly higher levels of a4β7 expression, suggesting that
mobilization and gut-homing of pDCs may be imprinted in the bone
marrow. In multiple tissues CD4+ pDCs were identified as the major
producers of IFN-α and TNF-α production compared to CD4- pDCs, but
were specifically lost during acute SIV infection. It was not, however,
apparent whether loss of CD4+ pDCs was associated with specific
depletion or downmodulation of CD4.
Conclusions: We demonstrate SIV rapidly induces mobilization of bone
marrow and circulating pDCs to traffic to the gut mucosae, but impairs
the pDC functional response by depleting the CD4+ cytokine-producing
subpopulation.
194
1
Institut Pasteur, Paris, France, 2Univ. Paris Diderot, Sorbonne Paris Cité,
Cellule Pasteur, Paris, France, 3A. Béclère Hospital, AP-HP, Clamart,
France, 4Pitié Salpétrière Hospital AP-HP, Paris, France
Background: Elucidating the mechanisms underlying the natural
control of HIV-1 mother to child transmission during the first trimester of
pregnancy may help determine biologic correlates of protection against
HIV-1 transmission in human mucosa. The decidua basalis (uterine
mucosa) is the main materno-fetal interface.
Decidual macrophages (dM) and natural killer (dNK) express functionnal
TLR7/8. Since HIV-1 ssRNA encodes for TLR 7/8 ligands that can mediate
direct activation of the immune system, we investigated the potent role
of TLR7/8 in the control of HIV-1 infection in the decidua.
Methods: Deciduas were obtained from HIV-1 negative women
undergoing elective abortions (8-12 weeks of amenorrhea). Decidual
explants or purified dM were stimulated with R848 (TLR7/8 ligand). The
culture supernatants were collected for soluble factor quantification.
The explants and dM were then used for flow cytometry experiments
or infected with R5 HIV-1 or HIV-1/VSV-G-luciferase pseudotype. HIV-1
entry was assessed by supernatant transfer experiment. Postentry steps
were investigated by PCR quantification. The role of R848 on the dNK
cytotoxic potential was measured by flow cytometry.
Results: HIV-1 replication was blocked efficiently in decidual explants
or purified dM when R848 was added prior to infection and to a lesser
extent when added several days after infection. Stimulated explants and
dM produced high levels of MIP-1α, MIP-1β and Rantes and decreased
significantly the expression of HIV-1 coreceptors. HIV-1 entry was
inhibited by decidual soluble factors. The number of proviral copies was
decreased in stimulated dM. TLR7/8 stimulated dM activated indirectly
dNK cytotoxicity.
Conclusions: Our findings provide evidence that TLR7/8 exerts a
durable and long lasting anti-HIV-1 effect by targeting several steps
of HIV-1 replication cycle and by activating the immune system.These
findings could provide strategies for blocking HIV-1 infection of mucosa.
P12.11
P12.12
Phenotypes of Monocytes and Monocyte
Derived Dendritic Cells (MODC) in
Antiretroviral Naïve Chronically Infected HIV
Patients
Monocytes in Chronic HIV-1 Infection:
Putative Gut Homing Markers and their
Relationship with Immune Activation
Karmistha Poovan1, Hayley Ipp2, Richard H. Glashoff1
Chantal Biya International Reference Center for Research on
the Prevention and Management of HIV/AIDS, Microbiology and
Immunology Laboratory, Yaounde, Cameroon, 2Chantal Biya
International Reference Center for Research on the Prevention and
Management of HIV/AIDS, Infirmary, Yaounde, Cameroon, 3Chantal
Biya International Reference Center for Research on the Prevention
and Management of HIV/AIDS, Medical Analysis Laboratory, Yaounde,
Cameroon, 4Faculty of Health Sciences, University of Buea, Medcine,
Buea, Cameroon
1
Background: Monocytes are immune cells which differentiate into
antigen presenting cells such as dendritic cells (DC). Monocytes have
been sub divided into three subclasses, based on CD14 and CD16
surface markers; namely classical (CD14++CD16+), intermediate
(CD14++CD16+) and non-classical (CD14+CD16+, CD14+CD16+)
monocytes. There is a variation in the phenotypes of these subpopulations
in HIV-1 chronically. Monocytes, are used to obtain MoDC in vitro, and
in vivo for several immunological studies. It is thus imperative to verify if
the variation between the different subclasses of monocytes can impact
their ability to be converted to MoDCs.
Methods: PBMCs purified from peripheral blood of participants, were
used for surface staining with fluorochrome-conjugated antibodies
characteristic of monocytes; and also to enrich for monocytes using
CD14 magnetic beads and magnetic separation columns. The enriched
monocytes were cultured in MoDC differentiation medium to obtain
immature MoDCs, in which maturation signal was added (polyICLC) and
cultured. Surface staining using antibodies was done at each stage to
determine the phenotypic characteristics of the cell populations.
Results: A fourth class of monocytes based on their expression of CD14
and CD16, (CD14++CD16++), which had a direct correlation with CD4+
absolute counts and an indirect correlation with viral load was noticed.
There was progressive down regulation of CD14 on the monocytes as
they converted to MoDCs and up regulation of maturation markers of
DCs (CD80, CD86, CD11c, CD1a and CD83).
Conclusions: Despite a variation in the different monocyte populations
observed between HIV infected people and HIV negative donors, the
monocytes still convert to MoDC similarly.
Stellenbosch University, Division of Medical Virology, Department of
Pathology, Cape Town, South Africa, 2Stellenbosch University, Division
of Haematology, Department of Pathology, Cape Town, South Africa
1
Background: The GALT is a site of rapid replication of HIV-1 during
early infection resulting in GIT damage and microbial translocation into
systemic circulation. This fuels on-going inflammation and immune
activation, which can persist despite the use of ARVs. Monocytes are
key role players in innate immunity, and due to GIT damage in HIV-1
infection, homing of monocytes to this site is envisaged as key in the
pathogenesis process.
Methods: Whole blood samples were collected from HIV + (n = 39)
and HIV- (n=26) volunteers from clinics across Cape Town. Cells were
stimulated with 1mg/µl LPS for 4 hours and then stained with antibodies
prior to analysis of 8 colour panels on a BD FACS CANTO II.
Results: There was a significant increase in the percentage of
inflammatory monocytes (CD14+16+), in the HIV+ group. There was
a higher overall monocyte expression of several chemokine receptors
such as CCR5 (53.4% vs. 10.01%), CCR1 (6.47% vs. 2.94%), CCR7
(86% vs. 76.95%) and CCR2 (52% vs. 31.7%). Other markers that were
increased include TRAIL-R1 (74.52% vs. 55.57%), CD62L (29.15% vs.
15.56%) and IL-10 (3.67% vs. 0.48%). Upon stimulation monocytes
appeared to respond similarly in both study groups, but the HIV+ group
did display unique and significant increases in IL-10 (2.53%), CD69
(1.41%), CD116 (4.09%) and HLA-DR (6.75%). The inflammatory subset
showed an increased expression of CD69 at baseline.
Conclusions: In HIV+ the total monocyte population shows a trend of
increased migratory markers including gut specific markers CCR7 and
CCR2, indicating they are primed to exit circulation to travel towards sites
of infection. The increased inflammatory subset seen in HIV+ individuals
is known to be associated with the persistence of the inflammatory state
and with the release of pro-inflammatory cytokines. Although chronic
HIV-1 infection does not seem to functionally impair monocytes, it does
lead to up-regulated expression of key activation and homing markers,
providing potential intervention biomarker targets.
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195
POSTERS
Nadesh Ngechae Nji1, Jules C. Tchadji1, Achile Nangue Nangue1,
Georgia Ambada1, Carol Stephanie Ngane Sake1, Abel Lissom1,
Edith Temgoua2, Betrand Sagnia1, Samuel Sosso3, Rachel
Kamgaing3, Jule Clement Assob Nguedia Assob4, Godwin
Wapimawah Nchinda1
Posters
P12.13
P12.14
Activation of Toll-like Receptor 2 (TLR2)
Heterodimers by HIV-1 Proteins Significantly
Increases HIV Infection and Inflammation
Phenotypic Characterization of Natural Killer
(NK) Cells in Antiretroviral Naïve Chronically
Infected HIV-1 Patients (the CIRCB Afrodec
cohort)
Bethany M. Henrick1,2, Xiao-Dan Yao1,2, Kenneth L. Rosenthal1,2
McMaster University, Department of Pathology & Molecular Medicine,
Hamilton, ON, Canada, 2McMaster University, McMaster Immunology
Research Centre, Hamilton, ON, Canada
1
Carole Stéphanie Sake Ngane1,2, Jules C. Tchadji1,3, Achille
Nangue1, Nadesh Nji1, Georgia Ambada1,3, Edith Temgoua1,
Samuel Sosso1, Rachel Kamgaing1, François-Xavier Etoa2, Godwin
Nchinda1
1-CIRCB/Microbiology and Immunology Lab, Yaoundé, Cameroon,
University of Yaoundé I, Department of Biochemistry, Yaoundé,
Cameroon, 3University of Yaoundé I, Yaoundé, Cameroon
1
POSTERS
Background: Immune activation is a critical driver of HIV-1 infection
and pathogenesis, however our understanding of HIV innate immune
activation remains incomplete. Recently, we showed that soluble TLR2
directly interacted with HIV proteins. Here, we investigated TLR2 and its
heterodimers as innate pattern recognition receptors for HIV-1 structural
proteins.
Methods: Using cells stably expressing TLR2, primary human T cells
and cell lines expressing selected TLRs, we determined the ability of HIV
structural proteins to activate IL-8 production through a NFκB signalling
pathway and the TLR2-dependent effect on HIV infection. Knockdown
of TLR expression with siRNA and anti-TLR2 antibodies were used to
confirm our results. Co-immunoprecipiatation was used to determine
direct interaction of TLR2 with HIV proteins.
Results: Cells expressing TLR2 had significantly increased HIV-1
integration and NFκB-dependent IL-8 production compared to cells
lacking TLR2. Primary T cells exposed to TLR2 ligand, Pam3CSK4, or
HIV-1 p17, p24 or gp41, but not to gp120, produced significantly more
IL-8 compared to cells incubated with anti-TLR2 antibodies. HIV p17
and gp41 were recognized by TLR2/1, while p24 activated cells through
TLR2/6. These results were confirmed by TLR2/1 siRNA knockdown,
as well as studies of HEK293 cells expressing selected TLRs. HIV
gp41, p17 and p24 were shown to co-immunoprecipitate with TLR2.
Interestingly, in competition assays, p24 blocked p17 and gp41-induced
TLR2 activation, thus providing a novel mechanism by which HIV can
modulate innate sensing.
Conclusions: Our results demonstrate that HIV-1 structural proteins p17,
p24 and gp41 act as pathogen-associated molecular patterns (PAMPs)
and signal through TLR2 and its heterodimers leading to significantly
increased HIV infection,NFκB-dependent activation and proinflammatory
cytokine production. These findings have important implications for our
understanding of HIV immune activation, pathogenesis and vaccine
development.
196
2
Background: Natural Killer (NK) cells are effectors cells of the innate
immune system which play a critical role as a first line of defense against
viral infection.They are characterized as CD3-CD56+CD16+/- cells and
effect cytotoxic activity against target cells as well as cytokines and
chemokines production. NK cytotoxicity is modulated by inhibitory and
activating receptors. HIV-1 infection maintains immune system in a
sustained state of activation and causes immune dysfunction.However
the impact of NK cell phenotype modulation has not been acessed in
the context of Antiretroviral naïve HIV-1 infection.
We shall verify the existence of HIV-specific NK cells in antiretroviral
naïve chronically infected HIV-1 patients.
Methods: 35 treatment naïveHIV-1 patients, CD4 greater than 350
cell/µl and 30 healthy donors were recruited. Age ranged from 21 to
65 years old. Patients group was composed of 05 patients with Viral
load(VL)< 2Log,23 VL between 2-4,5 Log and 07 VL>4,5 Log.PBMCs
were obtained from whole blood using ficoll-paque density grandient.
NK cells are analyzed by multiparametric flowcytometry in bulk PBMCs
. samples were aquired using BD FACScanto II machine and data
analysed by flowjo 7.4
Results: There was a differential modulation of NK cell phenotype as we
observed a decreased expression of activating markers NKP30, NKP44,
NKP46 and a significant increase (P< 0,05) in the expression of HLADR
, CD38 and NKG2D in treatment naïve HIV-1 infected patients relative
to healthy donors. Down regulation of NKG2A expression was also
observed in the patients group.
Conclusions: This study shows phenotypic pertubations on NK cells
in treatment naive HIV-1 infected patients. Observation of increased
expression of activating receptors NKG2D, CD38 and HLA-DR is a
pathological signature of HIV-1 infection however a decreased NKG2A
in expression could be necessary in increase cytolytic function of NK
cells.
P12.15
P12.16
Human Beta Defensins and RNases: Antiviral
Effect during Sexual Exposure to HIV-1
Sex Differences in TNFAIP3 Levels in pDC
upon TLR7 Stimulation
Wildeman Zapata1,2, Wbeimar Aguilar-Jimenez1, Zhimin Feng3,
Aaron Weinberg3, Aniello Russo4, Nicoletta Potenza4, Hernando
Estrada5, Maria Teresa Rugeles1
Susanne M. Ziegler1, Morgane Griesbeck2,3, Judy Chang4, Marcus
Altfeld1,2
Background: Individuals demonstrating natural resistance to HIV
infection, despite repeated unprotected sexual exposure, are referred
to as HIV-1-exposed seronegative (HESN), and are considered a model
to determine mechanisms of protection. Innate immune factors, such as
human beta defensins (HBD), and RNases, previously known for their
antiviral potential, are present at mucosal barriers, main ports of HIV
entry, suggesting their involvement in avoiding the establishment of HIV
infection.
Methods: To evaluate the role of these factors in the natural resistance
to HIV-1, we performed a study, including 60 HESN, and 61 healthy
controls (HC). Vaginal, endocervical and oral mucosal samples were
taken from these individuals; RNA was extracted, and mRNA transcripts
of HBD-2, -3, and the Ribonucleases, eosinophil-derived neurotoxin
(EDN), RNase-1 and angiogenin were quantified by qPCR. In addition,
we explored the step(s) of the viral cycle that was inhibited by these
soluble factors, using an in vitro single-round, recombinant-based, viral
infectivity assay.
Results: Oral mucosa of HESN expressed significantly higher levels of
hBD-2 (p=0.04) and hBD-3 (p=0.04) mRNA compared to HC. Likewise,
vaginal mucosa of HESN showed significantly higher mRNA levels of
RNase-1 compared to HC (p=0.036); in addition, endocervical mucosa
showed higher mRNA levels of EDN (p=0.04) and Angiogenin (p=0.02)
compared to HC. Finally, HBDs and RNases inhibited HIV-1 replication, in
a dose dependent manner at the following steps of the viral replication
cycle: entry, reverse transcription and nuclear import.
Conclusions: Taken together, these results suggest that innate immune
factors, such as HBD-2 and -3, and RNases inhibit HIV-1 infection during
either oral or genital exposure of HIV. In fact, HBDs and RNases inhibited
in vitro replication of HIV-1, pointing their potential role as antiretroviral
molecules to be explored in the near future.
Heinrich Pette Institute, Leibniz Institute for Experimental Virology,
Viral Immunology, Hamburg, Germany, 2Ragon Institute of MIT, MGH
and Harvard, Boston, MA, United States, 3CIMI/ U1135, Paris, France,
4
Monash University, Department of Infectious Diseases, Melbourne,
Australia
1
Background: The outcomes of many diseases differ between women
and men. It is well documented that women have a higher incidence
as well as more severe pathogenesis of autoimmune diseases.
Furthermore, accumulating evidence indicates a role of sex-based
divergence of infectious diseases, including influenza and HIV-1. In HIV1 infection, clinical studies have shown faster disease progression and
stronger immune activation in females compared to males for the same
level of viral replication, as well as better control of initial viremia in
women during primary infection (Meier et al 2009 Nat Med, Sterling
et al 2001 NEJM, Farzadegan et al 1998 Lancet). It has been suggested
that this is mainly due to Toll-like receptor (TLR)-mediated responses of
plasmacytoid dendritic cells (pDCs), the main producers of IFN-α. We
investigated the role of the TNF-α induced protein (TNFAIP) 3 (A20),
an estrogen-regulated early NF-κB-responsive gene that encodes a
ubiquitin-editing protein involved in negative feedback regulation of NFκB signaling. Thus, TNFAIP3 is known to be a potent anti-inflammatory
signaling molecule that restricts multiple intracellular signaling cascades
including TLR signaling.
Methods: 100 pDC were sorted and stimulated for 2h with a TLR7
ligand. Using fluidigm technology the expression level of TNFAIP3
mRNA was determined.
Results: We detected a significant difference in TNFAIP3 mRNA
upregulation in pDCs after 2h TLR7 stimulation between males and
females (p< 0.05). pDCs from males showed a 2-log higher expression
level of TNFAIP3 mRNA compared to females, associated with lower
production of IFN-α in pDCs from males in response to HIV-1-derived
TLR7 ligands and HIV-1.
Conclusions: Taken together, these data demonstrate that sex differences
described in response to HIV-1 are associated with the enhanced
expression of the sex hormone-dependent inhibitory regulator of TLR
signaling TNFAIP3 in pDCs of men compared to women, providing a
novel target to modulate the inflammatory IFN-α response to HIV-1.
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197
POSTERS
Universidad de Antioquia UdeA, Facultad de Medicina, Grupo
Inmunovirología, Medellin, Colombia, 2Universidad Cooperativa de
Colombia, Facultad de Medicina, Grupo Infettare, Medellin, Colombia,
3
Case Western Reserve University, School of Dental Medicine,
Department of Biological Sciences, Cleveland, OH, United States,
4
Second University of Naples, Department of Life Sciences, Naples,
Italy, 5HERES Health, Lending Institution of Health, Santa Marta,
Colombia
1
Posters
P12.17
Phenotypic and Functional Characteristics of
Siglec-7, NKG2A and NKG2C Expressing NK
Cells in Chronic HIV-1 Clade C Infection
Michael Z. Zulu1, Kewreshini Naidoo1, Zenele Mncube1,
Manjeetha Jaggernath1, Philip Goulder1,2, Thumbi Ndung’u1,3,
Marcus Altfeld1,3,4, Christina Thobakgale1
Doris Duke Medical Research Institute, University of KwaZulu-Natal,
HIV Pathogenesis Programme, Durban, South Africa, 2Peter Medawar
Building for Pathogen Research, University of Oxford, Department of
Paediatrics, London, United Kingdom, 3Ragon Institute of MIT, MGH
and Harvard, Boston, MA, United States, 4Heinrich-Pette Institute,
Department of Viral Immunology, Hamburg, Germany
1
POSTERS
Background: Natural Killer (NK) cells play a critical role in the control
of HIV-1 infection. HIV viremia has been shown to induce several
phenotypic and functional abnormalities in NK cells such as a decrease
in NKG2A+ and expansion of NKG2C+ NK cell population. Also, Siglec-7
was identified a cellular marker associated with NK cell abnormalities
in HIV-1 infection. However, there is still a paucity of data on factors
mediating NK cell receptor changes and identification of dysfunctional
NK cell subsets during different stages of clade C HIV-1 infection.
To assess mediators of NK cell dysfunction, our study examined the
phenotype and function of NK cells based on Siglec-7, NKG2A, NKG2C
and CD57 expression in HIV-1 clade C chronically infected individuals
versus healthy donors.
Methods: NK cell phenotypic profiles were characterized by assessing
Siglec-7, NKG2A, NKG2C and CD57 expression on PBMCs of combination
antiretroviral therapy (cART)-naïve HIV-1 chronically infected individuals
(n=15), HIV-1 chronically infected individuals who have been on cART
for at least 12 months (n=15) versus healthy individuals (n=15). The
cytolytic potential of individual NK cell subsets between the three study
groups was determined by flow cytometry assessment of CD107a and
IFN-γ following overnight stimulation with K562 target cells.
Results: Healthy individuals had the highest frequency of NK cells
expressing Siglec-7 when compared to HIV-infected subjects (p=0.002).
Interestingly, CD57 expression on total NK cell population was the
highest in healthy donors compared to HIV-1 infected subjects (p=0.01).
Overall, NKG2C expression in total NK cells was higher compared
to NKG2A; however, there were no significant differences in NK cell
degranulation and IFN-γ secretion across the three study groups.
Conclusions: HIV-1 viremia alters Siglec-7 and CD57 expression on NK
cells of infected subjects. Phenotypic changes induced by viremia may
lead to functional dysregulation on NK cell subsets during chronic HIV-1
clade C infection.
198
Posters 13: Key Populations
P13.01
P13.02
Awareness, Knowledge of HIV and Utilisation
of HIV Preventive Services among Young
People in Nigeria
Prior HIV Testing and ARV Use among HIV
Positive Female Sex Workers in Kigali Rwanda
2
University of Ibadan, Health Promotion and Education, Faculty of
Public Health, Ibadan, Nigeria, 2Stellenbosch University, Division of
Community Health, Cape Town, South Africa
1
Background: Human Immunodeficiency Virus (HIV) infections remain a
public health challenge in Nigeria. Despite the widespread knowledge
of HIV coupled with the expanded access to HIV-related services, risky
behavioural practices are prevalent among young people and the
utilization of HIV preventive services remains low. The aim of this study
is to examine the trend in the knowledge of HIV and utilisation of HIV
preventive services among young people in Nigeria between 2003 and
2008.
Methods: Data was obtained from Nigeria Demographic Health Survey
conducted in 2003 and 2008. The data for indicators on comprehensive
knowledge of HIV, comprehensive knowledge of prevention of motherto-child transmission, percentage who had ever had HIV test and
collected result, percentage of pregnant women who were counselled,
tested for HIV and received results and male circumcision for the 1524 years age group were extracted. The statistical significance of the
observed trend was tested.
Results: In 2008, level of comprehensive knowledge of HIV among
female youth was 22.2% while in 2003, it was 32.6%. Comprehensive
knowledge of prevention of mother-to-child transmission among women
increased from 5.1% to 23.2% and from 4.4% to 29.3% among men
in 2003 and 2008 respectively. In 2003, only 4.6% women had ever
had HIV test and collected result and in 2008 this increased to 9.2%
while among men, the percentage increased from 6.4% to 7.4%. Only
45.4% of the women interviewed knew where to get tested for HIV
in contrast to 59.3% men. In 2008, only 8.4% pregnant women were
counseled, tested for HIV and received results and 97.7% male has had
circumcision done.
Conclusions: The level of awareness and utilisation of HIV preventive
services is very low in Nigeria among the youth. Male circumcision is an
exceptional case because it is a culturally and religious norm for almost
every male to be circumcised in Nigeria. There is a need to change
the strategy for HIV youth awareness and utilisation of HIV services in
Nigeria.
Nurilign Ahmed1, Etienne Karita1, Rachel Parker2, Rosine
Ingabire1, Julian Nyombayire1, Robertine Sinabamenye1, Jean
Nduwamungu3, Jean Bizimana3, Gisele Umviligihozo4, Honoree
Uwamahoro3, Amanda Tichacek2, Eric Hunter2, Susan Allen2
Project San Francisco, Kigali, Rwanda, 2Emory University, School of
Medicine, Department of Pathology and Laboratory Medicine, Atlanta,
GA, United States, 3Projet San Francisco, Rwanda Zambia HIV Research
Group, Kigali, Rwanda, 4Profectus Biosciences, Kigali, Rwanda
1
Background: In Rwanda, an estimated 3% of the adult population is
infected with HIV (3.6% of adult women and 2.3% of adult men). This
prevalence is higher in urban setting 7.1% compared to rural setting
2.3%, and HIV risk is highest in urban FSW. Urgent access to ART
for FSWs is vital to reduce new infections in clients. Rwanda national
guidelines for ART for FSWs affirm that HIV+ FSWs should initiate ART
regardless of CD4 cell count.
Methods: FSWs were invited for services at Project San Francisco in
Kigali Rwanda from September 2012- April 2014. Prior HIV testing,
use of ARVs, and risk factors including average number of clients, and
disclosure of HIV status to/from clients were assessed.
Results: In this period, 677 FSWs were invited and tested for HIV and
372 (55%) were HIV+. Of these, 265 (71%) knew their HIV status
beforehand and 186 (70%) FSWs were using ARVs. On average, FSW
had 16 clients/month, one fifth of whom were regular/repeat clients.
51% of HIV+ FSWs disclosed their HIV status to their clients (73%
willingly, 27% by client request). 25% of HIV+ FSWs reported requesting
the HIV status of their clients and 11% of HIV+ FSWs were willing to get
tested with their clients. FSW charged a median of $8 for services (range
$2-$50) and reported a variety of successful strategies to minimize risk
of violence from clients. Oral sex was reported by 10% and anal sex by
15% of FSW; 31% reported using condoms all the time and 44% most
of the time. One in three were illiterate in the local language and one in
eight could understand French or English.
Conclusions: HIV testing should be promoted among FSW and
counseling should emphasize Rwanda guidelines regarding ART.
Research is needed to better understand obstacles to testing and ART
in HIV+ FSW. It is encouraging that many HIV+ FSW disclose their HIV
status to clients and are willing to be jointly tested with regular/repeat
clients; this may be an avenue for prevention research. Inconsistent
condom use remains a concern.
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199
POSTERS
Mojisola Morenike Oluwasanu , Olatunji O. Adetokunboh
1
Posters
P13.03
P13.04
Knowing Whom we Are trying to Protect:
An Assessment of HIV Risk in South African
Adolescent Females
Expanded Use of Antiretrovirals (ART) for
Treatment and Prevention for Female Sex
Workers in South Africa
Shaun L. Barnabas1,2, Heather B. Jaspan3,4, Smritee Dabee1,
Shameem Z. Jaumdally1, Hoyam Gamieldien1, David Lewis5,
Anna-Lise Williamson1,6, Thola Bennie2, Angel Phuti2, Martin van
der Watt7, Janan Dietrich7, Nicola Mulder8, Clive Gray9, Thomas
J. Hope10, Francesca Chiodi11, Robin Shattock12, Lynn Morris5,13,
Nonhlanhla N. Mkhize5, Glenda Gray7,14, Linda-Gail Bekker2,15, JoAnn S. Passmore1,6, Women’s Initiative in Sexual Health (WISH)
Robyn Eakle1,2, Gabriela Gomez3, W.D. Francois Venter1, Helen
Rees1
University of Cape Town, Medical Virology, Cape Town, South Africa,
Desmond Tutu HIV Foundation, Cape Town, South Africa, 3University
of Cape Town, Department of Immunology, Cape Town, South Africa,
4
Seattle Biomedical Research Institute, Seattle, WA, United States,
5
National Institute for Communicable Diseases, Johannesburg,
South Africa, 6National Health Laboratory Service, Cape Town, South
Africa, 7Perinatal and HIV Research Unit, Johannesburg, South Africa,
8
University of Cape Town, Computational Biology, Cape Town, South
Africa, 9University of Cape Town, Immunology, Cape Town, South
Africa, 10Northwestern University, Department of Cell and Molecular
Biology, Feinberg School of Medicine, Chicago, IL, United States,
11
Karolinska Institutet, Department of Microbiology, Tumor and Cell
Biology, Stockholm, Sweden, 12Imperial College, Department of
Infectious Diseases, Division of Medicine, London, United Kingdom,
13
National Health Laboratory Service, Johannesburg, South Africa,
14
HIV Vaccine Trials Network, Johannesburg, South Africa, 15Infectious
Disease and Molecular Medicine, University of Cape Town, Cape Town,
South Africa
Wits Reproductive Health & HIV Institute, Johannesburg, South Africa,
London School of Hygiene & Tropical Medicine, Infectious Disease
Epidemiology, London, United Kingdom, 3Amsterdam Institute for
Global Health and Development, Amsterdam, Netherlands
1
2
1
2
POSTERS
Background: South African adolescent females have been
underrepresented in HIV prevention trials though they have a
disproportionately high risk for HIV acquisition. We hypothesized that
changes associated with puberty may influence this susceptibility.
Methods: The Women’s Initiative in Sexual Health (WISH) study is
investigating factors associated with HIV risk in young South African women,
by enrolling HIV-negative adolescents between 16-22 years. This study plans
to enroll 150 participants from Masiphumelele, Cape Town and 100 from
Soweto, Johannesburg. As part of WISH, information on vaginal pH and the
prevalence of STIs (C. trachomatis, N. gonorrhoea, T. vaginalis, M. genitalium,
HSV-2, syphilis, candida) and bacterial vaginosis (BV) was collected.
Participants were surveyed on sexual risk and hormonal contraceptive use.
Results: Preliminary results in 48 enrolled participants show the majority
of adolescents used injectable hormone contraception (80% Nur-Isterate
and 16% Depo-Provera), with only 4% choosing oral contraceptives.
Almost all, 98%, of the participants self-identified as heterosexual. Their
median age of sexual debut was reported to be 16 years (IQR 16-17);
with their median lifetime number of sexual partners being 2 (range 1-5).
Reported condom use with their last sex act was 70%. Of the participants,
20% had been pregnant and all carried to term. Although only 25%
reported having had a previous symptomatic STI, approximately half of
the women were infected with an STI at enrolment; with C. trachomatis
being the most common by far. Furthermore, 40% had BV (Nugent >7),
with a median vaginal pH of 4.9. While STI prevalence at follow-up visits
declined following treatment, BV did not.
Conclusions: These data highlight the vulnerability of South African
adolescent females to STIs, BV and potentially HIV despite self reported
condom use. This study will provide insight into the reproductive health
of South African adolescents and may influence the design of future
preventative strategies.
200
Background: Recent trials demonstrated the efficacy of ART in reducing
the risk of transmission in HIV-positive patients and of pre-exposure
prophylaxis (PrEP) in reducing the risk of HIV acquisition. Adherence and
implementation challenges relating to service delivery may hinder the
ability of these interventions to make a significant impact. Sex workers
have been identified as a critical community to access HIV prevention
technologies.
Methods: This demonstration project seeks to test the ‘real-world’
viability of offering oral PrEP (for those testing HIV negative) and
immediate HIV treatment (for those testing HIV positive) to female sex
workers (FSWs) to prevent acquisition and transmission as part of a
routine package of clinical care in two urban sites in South Africa. The
main outcome studied will be retention at 12 months. The study will
have two arms:
1) PrEP arm for those testing negative; and
2) immediate treatment arm where women will be started on treatment
for CD4 counts of >350 cells/mm3.
We will investigate process and other health indicators (i.e. uptake,
safety, rates of pregnancy, sexual behaviour, rates of co-infections, and
use of SMS reminders). A qualitative research component will aim to
better understand the motivations and barriers to uptake of PrEP and
immediate treatment from perspectives of participants and providers.
Finally, an economic evaluation will inform a cost-effectiveness analysis
combined with estimates of impact through epidemiological modelling.
Results: In early 2017, after following FSWs for up to 24 months, we
will have finalised the analyses of all outcomes, as well as clinical data,
qualitative research, cost data, and impact.
Conclusions: This is one of two demonstration projects integrating
immediate treatment and PrEP service delivery in a real world setting.
Our results will aim to inform national and global policy making of the
viability of PrEP and immediate treatment as interventions prioritised for
high risk populations.
P13.05
P13.06
Tenofovir Gel Use in Women at High Risk of
HIV Infection: A Retrospective Analysis of the
Sex Worker Sub-group within the CAPRISA
004 Cohort
‘Facing our Fears’: Facilitated Film Viewings as
a Community Engagement Tool in Research
Involving MSM in Kenya
of Oxford, Headington, United Kingdom, 3University of Washington,
Seattle, WA, United States
CAPRISA/University of Kwa Zulu Natal, Durban, South Africa,
Columbia University Mailman School of Public Health, New York, NY,
United States
1
Background: Sex workers in Durban, South Africa are at high risk for
HIV infection and would benefit from effective prevention technologies.
A small urban sex worker cohort participated in the CAPRISA 004 trial
and their demographics, trial outcomes and adherence estimates have
not been exclusively described previously.
Methods: We conducted a retrospective analysis of demographic
data, sexual frequency, gel applicator use and assessed adherence
using applicator returns in conjunction with behavioural data from the
CAPRISA 004 trial sex worker cohort.
Results: In total 18 eligible female sex workers (self-identified) were
enrolled (two ineligible women were excluded from this analysis).
At baseline median (IQR) age was 22 years (IQR: 20-27) and 27.8%
completed high school. At enrolment, approximately 77.8% reported
condom use at last sex act, and 61.1% reported having a stable partner.
Eleven participants were randomized to the placebo arm and 7 to the
active arm. Overall, median trial participation duration was 21 months
(IQR: 11-28 months). Median sex acts reported per month was 10 (IQR:
7 to 13.5) and median number of applicators dispensed per month
was 13 (IQR: 10-20). Of these 60% (IQR: 40-93%) of applicators were
returned as used. Adherence, based on returned used applicators and
self -reported sex, was 50% (IQR: 45-100) overall. HIV incidence was
7.53 per 100PY (95% CI 0.9 - 27.2). However, there were only two HIV
infections, both being in the placebo arm and HIV incidence in that arm
was 13.8 per 100PY (95%CI 1.7 - 49.8). This is in contrast to the HIV
incidence rate in the parent trial which was 9.1 per100PY (95%CI 6.911.7) in the placebo arm.
Conclusions: Despite their high risk for HIV infection the small CAPRISA
004 sex worker cohort demonstrated moderate adherence to study gel.
However, it is noteworthy that the active arm had 100% protection and
that the sex workers had higher adherence to this coital gel regimen
compared to women prescribed daily gel in the VOICE trial in South
Africa
Background: Research with men who have sex with men (MSM) in Africa
requires ongoing community engagement to ensure safety and access
to care of participants. We assessed views of stakeholders regarding a
community engagement film more than 3 years after a research clinic
was attacked in Coastal Kenya.
Methods: We made a 16-minute video ‘Facing Our Fears’ capturing
footage of the attack in 2010, interviews with attack leaders and victims
in 2010 and 2013, and various reflections of community members
on MSM research. We conducted 10 facilitated film viewings (FFV)
followed by focus group discussions (FGD) with 8-20 participants each,
comprising of religious leaders, LGBT members, health care providers,
policy makers and media representatives to assess the film’s usefulness
as a communication tool, perceived security risks if the film would be
released publicly, and if it supported community ownership of MSM
research. FGD were tape-recorded or noted by hand. Transcripts were
analysed thematically by two of the authors.
Results: FFV presented a platform for discussion about involvement of
MSM in medical research, importance of stakeholder collaboration, and
LGBT rights in general. Visual representations of community engagement
efforts were seen to be effective in highlighting the importance of
including MSM in health care and research. FGD participants identified
concerns over the possible risks to LGTB communities and those
working with them following public release, due to homophobic and
discriminatory atmosphere in much of Africa.
Conclusions: FFV were seen as an empowering tool for raising awareness
of MSM research and same-sex relations in general. Facilitated viewings
allowed the film to be used as a training tool for successful LGBT
research with community engagement, and may mitigate the possible
risks of further stigma following open release.
1
2
www.hivr4p.org
201
POSTERS
Tanuja N. Gengiah1, Lise Werner1, Quarraisha Abdool Karim1,2,
Salim S. Abdool Karim1,2
Evans Gichuru1, Salla Sariola2, Elisabeth M. Van der Elst1,
Peter Mugo1, Murugi Micheni1, Susan M. Graham1,3, Catherine
Molyneux1,2, Eduard J. Sanders1,2
Posters
P13.07
P13.08
Are Nigerian Healthcare Providers (HCP)
Prepared for Men who Have Sex with Men
(MSM): Lessons from the Mystery Client
Survey in Nigeria?
The Anti-same Sex Marriage Law Implications
on HIV Interventions for Men who Have Sex
with Men in Nigeria
Chiedu Chike Ifekandu1,2
Population Council, HIV Division, Abuja, Nigeria, 2Ahmadu Bello
University, Development Communications/Theatre Arts, Samaru-Zaria,
Nigeria
1
POSTERS
Background: Services in the most healthcare facilities in Nigeria are
targeted at the general population; the services they provide tend not
to be friendly towards the health needs of men who have sex with men
(MSM). The need to address the situation informed the mystery client
survey to health facilities to evaluate the quality of treatment, care and
services rendered to MSM in Nigeria.
Objectives: To evaluate the quality of HIV prevention services for Men
who have sex with men in Nigeria.
Methods: The study was conducted in 12 Nigerian states. Mystery Client
Survey methodology was used for the study. Pre-recruited MSM visited
healthcare Facilities anonymously, posing as a regular customer/client.
After leaving the facility, a standardized reporting form (questionnaire)
was discretely completed by the mystery client immediately after each
visit to capture the facility with regard to their assessment of provision of
quality services. The mystery client in this survey sought one of the two
services: HCT or STI consultation services. Quantitative data was entered
and analyzed using SPSS version 20.0 Descriptive statistics, including
95% confidence intervals (CI) and Chi square tests of comparison for
differences between categorical variables were conducted.
Results: The mean age of the respondents is 22years +/-SD. The study
size is 104 MSM. More than 76% reported that information at the clinic
was neither inclusive of their specific sexual needs nor address stigma
against MSM. About 40% reported that their sexual practices and
behaviours were not explored during the pre and post-test counselling.
More than 50% reported that the HCP exhibited a judgmental attitude
towards MSM, while 47% reported that follow-up, care and support as
well as referral were not made after the pre and post-test counselling
sessions.
Conclusions: From this study, sensitizing HCP on sexual diversity can
help in changing their attitudes and believe towards.
202
Chiedu Chike Ifekandu1, Bala Abullahi1, Ibrahim Suleiman1, Hafiz
Abdullahi1, Jean Njab1
Population Council, HIV Division, Abuja, Nigeria
1
Background: HIV prevalence in Nigeria among men who have sex with
other men is at 17.2% (IBBSS 2010). Despite the alarming prevalence
figures, Nigerian president signed the anti-same-sex bill into an Act. This
law does not criminalize MSM alone, but incurs 10 years jail sentence
for institutions and healthcare providers working with MSM as well.
Objectives: To evaluate how the law will impact on HIV interventions and
possibly come up with a more effective strategy for field implementation.
Methods: There are 48 MSM peer educators on the Strengthening HIV
prevention Services (SHiPS) project in four northern Nigerian states.
These 48 peer educators reach a total of 1200 MSM with six month
timeline. The strategies being utilized before the Act was enacted was
the combined prevention packaging of intervention (CMPPI); using the
cohort formation approach.
A well-structured questionnaire were administered to the 48 peer
educators on their background knowledge of the MSM community
within their area of coverage, their knowledge of the content of the antisame-sex Act and the impact on peer participation in the SHiPS Project
and brainstorm on new strategies in reaching the MSM peers without
exposing them to danger.
Results: More than one-quarter of the peer educators reported than
most of their peers are declining participation for SHiPS project. Over
half of the peer educator have read the same-sex marriage prohibition
Act and believes that they are vulnerable to harassments from the law
enforcement agencies or the general public while having their peer
sessions. Almost all the peer educators suggested on the change of the
approach from peer education sessions to inter-personal Communication
(IPC).
Conclusions: Advocacy led the National Agency for the control of AIDS
should be paid to the federal ministry of health and the presidency using
informed evidence from national survey to convince the government
from public health angle while MSM should not be criminalized.
P13.09
P13.10
Need to Raise the Level of Knowledge of PrEP
and Rectal Microbicides (RM) among Men
who Have Sex with Men (MSM)
HIV and Sexually Transmitted Infection (STI)
Testing among Female Sex Workers (FSWs) in
Urban Zambia
Rewa Kohli1, Soumyashree Mohanty1, Praveen Jadhav1, Ramesh
Paranjape2
Jennifer A. Kotlewski1, Linda Kimaru1, Tyronza Sharkey1, Marydale
A. Oppert2, William Kilembe3, Mubiana Inambao1, Hervette
Nkwihoreze1, Amanda Tichacek2, Nurelign Ahmed4, Rachel
Parker2, Susan Allen2
Background: Very little information is available about acceptability of
new biological prevention tools such as PrEP and RM among MSM in
whom the HIV risk is high. We explored knowledge and willingness
about these tools among MSM community in India.
Methods: Qualitative study using repeated in-depth interviews among
39 consenting MSM was conducted in Pune. “Expert peer” and social
networking helped recruitment. Interview guide aided discussion. Data
collection and analysis occurred concurrently.
Results: MSM were between 20-55 yrs of age, mostly educated till
10th grade, 12 were highly educated, and 4 MSM were HIV positive,
9 MSM engaged in sex work regularly and 10 occasionally. Few MSM
had correct knowledge about HIV. Despite using condoms 23 MSM
perceived the risk of HIV infection because of fear of condoms tear/
nonuse owing to liquor consumption. Majority lacked knowledge about
other prevention options while 2 mentioned PEP. PrEP was acceptable
as it provided added protection in cases of condom nonuse and failure,
multiple partners and for a stress free sex life. Majority was fine with daily
intake but others wanted it before sex as sex did not happen frequently.
Most would continue intake till risk behavior lasts. Concerns about sideeffects and adherence were expressed. Side-effects affecting physical
appearance and functioning of internal organs were not acceptable Free/
cost below Rs 500/m was fine. Efficacy should be nearly 100 percent. All
MSM were willing to use the RM as it would provide HIV protection. Most
MSM were habituated to using lubricants and favored gel formulation.
If RM enhances libido it would sell very well. Availability preferred at
pharmacy with/without prescription; or NGOs/public hospital at nominal
cost/free.
Conclusions: It is critical to raise awareness of community about new
prevention tools such as PrEP and RM, for successful positioning when
efficacious options become available. Tools should be effective, easily
available at low cost and be without major side-effects.
1
Rwanda Zambia HIV Research Group, Ndola, Zambia, 2Rwanda Zambia
HIV Research Group, Atlanta, GA, United States, 3Rwanda Zambia
HIV Research Group, Lusaka, Zambia, 4Rwanda Zambia HIV Research
Group, Kigali, Rwanda
Background: FSWs in Sub-Saharan Africa are a vulnerable group in
the AIDS epidemic. Due to barriers in access to testing and counseling,
numerous sexual partners, and little control over safe sex practices with
clients, FSWs are at particularly high risk for HIV/STIs. In this study, FSWs
in urban Zambia were surveyed and tested for HIV, T. Vaginalis (TV), and
syphilis.
Methods: FSWs were recruited from known activity hubs through
peer outreach in Lusaka and Ndola, Zambia. Interested FSWs received
Voluntary HIV Counseling and Testing, RPR serologies for syphilis,
and self-administered vaginal swab for TV. A risk assessment survey
was administered, including questions on previous testing and use of
antiretrovirals (ARV).
Results: 733 FSWs were screened over an 18-month period. At baseline,
323 (44%) were HIV positive (HIV+), 391 were HIV negative (HIV-), 9
were indeterminate, and 10 declined testing. 45% had some level
of spoken and/or written illiteracy. Among those testing HIV+, 40%
reported previous HIV testing and of those 7% were on ARVs. Of HIV+
FSW, 10% were positive for TV and 25% were positive for syphilis. In the
HIV- group, 9% were positive for TV and 3% were positive for syphilis.
14% of HIV+ participants reported treatment for STI symptoms in the
12 months before this screening. None of the participants could name
health or social services, whether provided by NGO or government, that
cater to FSW.
Conclusions: Less than half of FSW had been previously tested for HIV.
Among HIV+ women who had been previously tested fewer than one
in ten were on ART. Low literacy may contribute to poor comprehension
of referral services, and fear of stigma may deter FSW from seeking
care. Reported treatment for past STI was low, though screening for
asymptomatic STI uncovered many cases of syphilis and T.vaginalis. The
type of FSW-friendly services provided in this study may be helpful in
increasing HIV and STI screening and uptake of ART. Particular attention
to low literacy is critical in this effort.
www.hivr4p.org
203
POSTERS
National AIDS Research Institute, Socio-behavioral Research
Department, Pune, India, 2National AIDS Research Institute,
Immunology Department, Pune, India
1
Posters
P13.11
P13.12
Trends and Factors Associated with
Unprotected Anal Intercourse among Young
Men who Have Sex with Men in Bangkok,
Thailand, 2006-2013
Serodiscordant Couples’ Cohort: An Important
Cohort to Explore Correlates of Protection to
HIV Infections
Sirirat Lertpruek1, Wipas Wimonsate1, Sarika Pattanasin1, Kesinee
Satumay1, Wichuda Sukwicha1, Jaray Tongtoyai1, Kanokpan
Pancharoen1, Anupong Chitwarakorn2, Timothy H. Holtz1,3
Thailand-MoPH U.S. CDC Collaboration, Nonthaburi, Thailand,
Ministry of Public Health, Department of Diseases Control, Nonthaburi,
Thailand, 3Division of HIV/AIDS Prevention, U.S. Centers for Disease
Control and Prevention, Atlanta, GA, United States
1
2
POSTERS
Background: Risk factors for HIV acquisition among young men who
have sex with men (YMSM, aged 18-24 years) in Bangkok have been
described. We investigated behavior associated with unprotected anal
intercourse (UAI) at baseline and during follow-up visits among YMSM
enrolled in the Bangkok MSM Cohort Study (BMCS).
Methods: Thai men from the Bangkok metropolitan area, aged ≥18 years
who had engaged in sex with another man in the past six months were
enrolled and followed up every four months. At enrollment, participants
received HIV testing, and were asked about HIV-risk behaviors during the
preceding four months using audio computer-assisted self-interviews.
We analyzed the factors associated with UAI at baseline and over time
using logistic regression and Generalized Estimating Equations (GEE),
respectively.
Results: We enrolled 712 YMSM during 2006-2010, 621 (87%)
contributed at least one-follow-up visit through 2013. HIV prevalence
was 21%. The overall UAI at baseline was 60% and declined to less than
20% at the 36-month visit, thereafter remaining stable. For each visit
increase, we found a 2% decrease in odds for reported UAI regardless
of HIV infection.
A significant association with UAI at baseline was found among YMSM
living with a partner [Adjusted Odd Ratio (AOR) 3.1, 95% Confidence
Interval (CI) 1.7-5.6] compared with YMSM living with family. Factors
associated with UAI over time were: ≥ 2 sexual partners (AOR 3.6, 95%
CI 3.0-4.4), current HIV infection (AOR 0.5, 95% CI 0.4-0.6), club drug
use (AOR 1.5, 95% CI 1.1-1.9), erectile dysfunction drug use
(AOR 1.6, 95% CI 1.1-2.1), participation in group sex (AOR 1.8, 95% CI
1.5-2.2).
Conclusions: YMSM who enrolled in the BMCS exhibited a high level
of UAI. The trend over time suggests that intensive HIV-risk reduction
counseling and HIV testing campaigns may be associated with a
significant reduction in self-reported UAI in this population. However, a
longitudinal effect may limit the generalizability.
204
Sophia Osawe1,2, Evaezi Okpokoro3, Ruth Datiri2, Grace
Choji2, Felicia Okolo2, Stephen Umaru2, Pam Datong2, Alash’le
Abimiku2,3,4
1
Institute of Human Virology, Jos, Nigeria, 2Plateau State Human
Virology Research Centre, Jos, Nigeria, 3Institute of Human Virology,
Abuja, Nigeria, 4Institute of Human Virology, University of Maryland
School of Medicine, Baltimore, MD, United States
Background: Seronegative partners of a discordant relationship are
exposed to HIV and other STIs. Despite availability of ARVs to HIV+
partners, they had varying levels of detectable virus in our cohort and yet
their partners remain HIV negative. Studying this more ‘natural’ model
of HIV exposure, which is slightly different from the HESN described by
other investigators, can help understand mechanisms of protection to
HIV infection. Data from a cohort of discordant couples can explore risk
factors in this group and correlates of protection to HIV.
Methods: A cohort was developed in October 2011 and has been
followed up for 2 years and 6 months. Behavioral data collection,
medical history, examinations and safety labs were conducted at each
of the 11 follow up visits. HIV positive partners were offered CD4 counts
and viral loads as well as risk reduction and couples counseling with
their partners.
Results: Out of 683 screened, 534 were eligible for the study with 257
(48.1%) females and 277 (51.9%) males. Condom use was collected at
all follow up visits with baseline data showing only 40% of enrollees
reporting consistent use of male condoms. Partner viral loads and CD4
counts done revealed that 1/3 had viral loads above >1,000 copies/ml
despite being on ARVs and almost 50% had CD4 counts < 350 cells/
µl. Condom use increased slightly during the follow period up to 47%.
A total of 22 women (8.6%) became pregnant so far. Recorded HIV and
Hepatitis C incidence were 1.1% and 3.7% respectively.
Conclusions: Our cohort was made up of couples in a stable home with
99% married. A high number of enrollees expressed their desires to have
children explaining the low use of condoms which did not significantly
increase through the follow up period despite consistent risk reduction
counseling and free condoms. Exposure to HIV is relatively high in the
cohort. With documented inconsistent condom use and detectable viral
loads this cohorts make an interesting group to look into correlates of
protection which is planned.
P13.13
P13.14
Testing for HIV as Individuals or Couples:
Preference of MSM in Lagos State Nigeria
Use of Virtual Cohorts in Zambia’s Couples’
Voluntary Counseling and Testing (CVCT) as a
Screening Mechanism for Clinical Trials
Adekemi Sekoni1, Sikeade Alagbe1, Olukemi Odukoya1,
Kofoworola Odeyemi1
University of Lagos, Lagos, Nigeria
1
Tyronza Sharkey1,2, Rachel Parker1,3, William Kilembe1,2, Mubiana
Inambao1,4, Frances Priddy5, Pat Fast5, Matt Price5, Amanda
Tichacek1,3, Eric Hunter3, Susan Allen1,3
Rwanda Zambia HIV Research Group, Atlanta, GA, United States,
Zambia Emory HIV Research Project, Lusaka, Zambia, 3Emory
University, Atlanta, GA, United States, 4Zambia Emory HIV Research
Project, Ndola, Zambia, 5International AIDS Vaccine Initiative (IAVI)-New
York, New York City, NY, United States
1
Background: CVCT, recommended by WHO, is used to identify virtual
cohorts (VC) for Zambia Emory HIV Research Project (ZEHRP). VC are at
risk groups (HIV concordant negative (M-F-) and discordant couples (DC))
that are followed up and offered various services as standard of care
within government and stand-alone clinics. Costs to maintain VC are
leveraged by program activities offering these services. Data acquired
from these programs are used to calculate seroincidence, follow-up rates
and pre-screen potential volunteers for clinical trials.
Methods: VC are verbally consented, identified and followed in > 70
centers from 2008-2014. Couple HIV status and basic demographics
are collected using ZEHRP data collection tools that capture Zambian
health indicators, which are reported to the districts. Seroincidence by
couple HIV status, % of index partners on/off ARV at seroconversion,
and follow-up rates are calculated.
Results: Of 148,839 couples tested, 129,181 were identified for VC
enrolment (109,677 or 74% M-F- and 17,619 or 12% discordant). 73%
of VC were ages 18-40 with 96% cohabiting >3 months. Follow-up
rates within VC are 6241PY overall with 5083PY in M-F- and 1158PY in
discordants. In 112 seroconverted couples, 62 were initially discordant
and 50 were M-F-. Incidence rates were highest in DC with 5.36/100PY
(95% CI 4.11-6.87) and lowest in M-F- couples with 0.98/100PY (95% CI
0.73-1.30). In DC with linked transmissions (n=48, 77.4%), 27/48 (56%)
of index partners had not initiated ARV and 21/48 (44%) were on ARV
before seroconversion.
Conclusions: VC allow for calculating seroincidence, follow-up rates and
is used for pre-screening of volunteers for clinical trials. In discordant
couples, VC has illustrated the continued need for inclusion of this high
risk group in clinical trials as one size does not fit all. Future plans are
to use VC to follow-up other HIV high risk groups in Zambia. VC has the
potential to be applied to other diseases/infections for clinical trials and
epidemiologic research.
www.hivr4p.org
205
POSTERS
Background: Testing as couples rather than as individuals has been
shown to confer added benefits because it allows individuals to also
know their partners status and therefore avoid the challenges associated
with disclosure of status.
Methods: This cross-sectional descriptive study was carried out to
assess testing for HIV as individuals and as couples among MSM in eight
local government areas of Lagos State, Nigeria using respondent driven
sampling to reach 320 men. An interviewer-administered questionnaire
was used to collect information after informed written consent was
obtained from the participants. Univariate, bivariate and multivariate
data analysis was carried out using IBM SPSS 20 software, association
was established at p< 0.05.
Results: Majority of the respondents was within the age range 20-29
years, Christians, self reported bisexual, had post secondary school
education and was unemployed. Half of the respondents were single
and not in a steady relationship. Majority of the respondents (85.3%)
had been tested for HIV, and out of those that have not been tested for
HIV, 48.9% are willing to do the test. About half (52.8%) of those who
tested as individuals had disclosed their results to their main sexual
partners. Less than a quarter of respondents that had been tested did
so with their main sexual partner Willingness to test for HIV as a couple
(CVCT) among respondents who had previously tested as individuals
was poor as only about a third (33.5%) was willing to go for CVCT.
Statistically significant association was observed between the age of
the respondents, employment status, graded knowledge of HIV/AIDS,
knowledge of at least an individual who was tested for HIV or is HIV
positive, sexual behavior of respondents and being tested for HIV as
individuals but not as couples on bivariate analysis.
Conclusions: Couple HIV testing is not popular among our study
population, it is necessary to promote this practice among key population
in other to achieve zero new infections.
2
Posters
P13.15
P13.16 LB
Getting a Foot in the Door: Adolescent
Retention in a Mock HIV Prevention Clinical
Trial
Facilitators and Challenges to ART Adherence
among Men who Have Sex with Men (MSM) in
Coastal Kenya
Elizabeth E. Tolley1, Joy N. Baumgartner2, Sam Field3, Jennifer
Headley1, Anna Kaale4, Anna Minja4, Sylvia Kaaya5
Murugi Micheni1, Andrew Secor2, Elisabeth van der Elst1,
Bernadette Kombo1, Jane Simoni2, Don Operario3, Eduard
Sanders1, Susan Graham2
FHI 360, Social & Behavioral Health Sciences, Durham, NC, United
States, 2FHI 360, Social & Behavioral Health Sciences, Washington,
D.C., DC, United States, 3FHI 360, Quantitative Sciences, Durham, NC,
United States, 4Muhimbili University of Health and Allied Sciences,
Dar es Salaam, Tanzania, United Republic of, 5Muhimbili University
of Health and Allied Sciences, Psychiatry and Mental Health, Dar es
Salaam, Tanzania, United Republic of
1
POSTERS
Background: Young women below age 18 are underrepresented in HIV
prevention trials, despite being among the most at risk groups in Africa.
Their non-inclusion is due in part to ethical concerns. But, adolescents
may also face challenges with trial follow-up due to mobility, lower HIV
risk perception and less access to sexual and reproductive health (SRH)
services. We examine whether retention differs among adolescents aged
15-17 and young women aged 18-21, and what factors might account
for this difference.
Methods: We enrolled 135 sexually-active, non-pregnant, HIV-negative
women, aged 15-21, into a 6-month mock clinical trial in Dar es Salaam,
Tanzania, with visits at baseline and months 2, 4, and 6. A mediation
analysis assessed direct and indirect effects of age and potential
mediators on number of missed visits. Mediators included theoreticallyderived barriers and facilitators to retention.
Results: Adolescents held significantly lower risk perception and were
less likely to have used SRH services compared to young adults. Both
of these factors were positively associated with missed visits, suggesting
that adolescents should have missed more visits than adult women.
However, controlling for mediators, adolescents were less likely
[trending towards significance] to miss visits than adult women. These
results suggest that HIV risk perception and previous SRH service use
may act as suppressor variables in the relationship between age and
retention in a clinical study.
Conclusions: Adolescents were no more likely to miss visits than
adult women, despite having lower risk perception and less exposure
to SRH services. Interventions that realistically increase adolescent risk
perception and link them to services - getting their feet in the door - may
support their retention in future HIV prevention trials and in SRH services
more generally. Also, these analyses suggest that doing so may lead to
improvements in retention beyond what is seen in an older cohort of
study participants.
206
of Washington, Seattle, WA, United States, 3Brown University,
Providence, RI, United States
1
Background: In coastal Kenya, 20% of HIV infections occur among
MSM, a highly stigmatized group facing many barriers to care
engagement and ART adherence. We aimed to identify key facilitators
and challenges among HIV-positive MSM, with the goal of developing
an intervention to improve clinical outcomes
Methods: We conducted individual in-depth interviews (IDI) with
purposively sampled HIV-positive MSM and focus group discussions
(FGD) with their health providers using semi-structured, open-ended
topic guides based on an access-information-motivation-behavioural
skills model of adherence, with added focus on provider trust, stigma
and discrimination. Detailed interviewer notes and transcriptions were
translated into English and reviewed for common factors influencing
ART adherence.
Results: 30 IDI and 4 FGD were carried out and analysed. Conformity
was observed between individual-level factors proposed by the
conceptual framework and those reported. Barriers included poverty,
limited access to MSM-friendly information and services, poor HIV
and ART knowledge, lack of psychosocial support, nondisclosure of
HIV status, medication side effects, mental health challenges and
substance abuse. Facilitators included economic empowerment, tailored
knowledge, peer support, selective disclosure, self-acceptance and pilltaking skills. Resiliency, not identified in the original model, emerged
as an important protective factor against stigma and discrimination. A
trained ART-experienced peer working with trusted care providers was
the favoured means for adherence support.
Conclusions: Our final conceptual model comprises factors at the
individual, community and policy levels, with access to MSM-friendly
services and information, disclosure, self-acceptance and support being
key facilitators despite challenging stigma and discrimination. Trust in
providers and peers appeared to mediate a hostile social environment
and should be fostered through training and structural support.
P13.17 LB
P13.18 LB
Sexual Health, Alcohol Use, Childhood
Sexual Abuse, and Mental Health Outcomes
Among Spanish-speaking Latino MSM in the
Northeastern United States.
Effectiveness of the Community PROMISE
and Enhanced Community PROMISE
Interventions among Female Sex Workers in
the Dakar Region, Senegal
Omar Martinez1, Elwin Wu2, Joseph Spadafino3, Theo Sandfort2,
Andrew Z Shultz1, Javier Lopez Rios1, Hugo Ovejero4, Scott D
Rhodes5, Eva Moya6, Silvia Chavez Baray6, Brian Dodge7, Alex
Carballo-Dieguez2
Moussa Sarr1, Julie Pulerwitz2, Ibou Thior2, Marièma Soumaré3,
Ibrahima Traoré4, Carlos Suarez1, Elizabeth King2, Lindsay
Hugues2, Daouda Gueye3, Amadou Sougou3, Aminata Mboup4,
Souleymane Mboup4
HIV Center for Clinical and Behavioral Studies at Columbia University,
Division of Gender, Sexuality, and Health, New York, NY, United
States, 2Columbia University, New York, NY, United States, 3Arizona
State University, Phoenix, AZ, United States, 4Latino Commission on
AIDS, New York, NY, United States, 5Wake Forest School of Medicine,
Winston-Salem, NC, United States, 6University of Texas, El Paso, TX,
United States, 7Indiana University Bloomington, Bloomington, IN,
United States
Westat, Rockville, MD, United States, 2PATH, Washington, DC, United
States, 3AWA, Dakar, Senegal, 4Cheikh Anta Diop University, Dakar,
Senegal
Background: Little is known about the health status of predominantly
Spanish-speaking Latino men who have sex with men (MSM).
Methods: Between January and March of 2014 a cohort of Latino
MSM (N=176) was recruited to participate in Latinos en Pareja, an HIV/
STI prevention intervention adaptation study. A multinomial logistic
regression model predicting problematic alcohol consumption was
carried out; demographic characteristics, sexual risk factors, childhood
sexual abuse experiences, and mental health outcomes were included
in the model.
Results: Prevalence estimates of problematic alcohol consumption in
the past 30 days and clinically significant depressive symptoms (CES-D
score ≥ 10) were 47% and 68%, respectively. Internal consistency
reliability coefficients of the CES-D scale were satisfactory (Cronbach
α=0.86). Among participants who reported sexual activity before the age
of 17 (n=130, 74%), 39 participants (30%) reported childhood sexual
abuse. In univariable analyses, characteristics and covariates associated
with problematic alcohol consumption included having more than one
sexual partner in the past 3 months, engaging in risky sexual behavior
(operationalized as condom nonuse in the past 3 months), being in a
relationship, reporting intimate partner violence, screening for clinically
significant depressive symptoms, and having experienced childhood
sexual abuse. In the multinomial logistic regression model, problematic
alcohol consumption was predicted by having more than one sexual
partner in the past 3 months, engaging in risky sexual behavior, being
in a relationship, and reporting intimate partner violence.
Conclusions: Further work is needed to develop effective prevention
intervention approaches for problematic alcohol consumption among
Latino MSM. Given the gap in research on Latino MSM and the high
prevalence estimate of childhood sexual abuse among this subpopulation,
there is a need to steer effective preventive and treatment interventions
to meet the particular needs of this community.
1
Background: In Senegal, the HIV prevalence is ≤ 1% in the general
population, but very high among female sex workers (FSWs) with
rates up to 30%. The objective of this PATH funded study through a
Canadian International Development agency (CIDA) grant is to assess
the effectiveness of the Community PROMISE intervention, an effective
community-level STD/HIV prevention intervention that relies on role
model stories and peers from the target community (Community Promise
Only). A structural component was also added to the Community
PROMISE activities by targeting bars, bar owners/staff and clients
(Enhanced Community Promise).
Methods: After adapting and implementing the proposed interventions,
we used data from pre and post test surveys of FSWs conducted in
the Dakar region to assess differences in consistent condom use with
clients and with regular partners. Comparisons were also done between
districts of Dakar randomly assigned to Community Promise Only versus
Enhanced Community Promise Interventions. Univariate and multivariate
logistic regressions were used to assess effectiveness.
Results: The average age of the FSW population was 36.33 years
(STD=9.62). Most FSWs were Senegalese (96.7%), and from the Pulaar
(31.5%), Sereer (27.4%) and Wolof (21.6%) ethnic groups. Approximately
half of FSW (46.9%) never attended school.
Overall, we found evidence of increased condom use with clients at
post-test versus pre-test (p = 0.04). Consistent condom use increased
from 79% at pre-test to 85% at post-test (non-significant) in Community
Promise only districts; but reached 93% in Enhanced Community
Premise districts (p = 0.006). We did not find evidence of increased
condom use with regular partners.
Conclusions: Our results showed evidence of increased consistent
condom use with clients when both FSWs and clients are targeted by
our intervention activities. We see a great potential in diffusing the
adapted intervention throughout FSWs communities in Senegal and in
other countries in Africa.
www.hivr4p.org
207
POSTERS
1
Posters
Posters 14: MPT Development
P14.01
P14.02
Drug-drug Interaction Studies Investigating
the Impact of Levonorgestrel on Antiviral
Potency of Dapivirine
MZC and 1% TFV Gel: Multipurpose
Prevention Approaches
Abbey B. Evans , Jonathon Holt , Jeremy Nuttall , Robin Shattock
1
2
2
1
Imperial College London, Group of Mucosal Infection & Immunity,
London, United Kingdom, 2International Partnership for Microbicides,
Silver Spring, MD, United States
1
POSTERS
Background: Intravaginal rings represent an ideal platform to
deliver both anti-viral microbicides and hormonal contraception. An
intravaginal ring releasing dapivirine (DPV), a potent NNRTI, is currently
undergoing late stage clinical efficacy testing. Levonorgestrel (LNG), a
synthetic progestogen and hormonal contraceptive, could be used in
conjunction with DPV offering the potential of dual protection against
HIV infection and pregnancy. Initial drug-drug interaction studies have
been performed to determine the potential impact of LNG on the
antiviral potency of DPV.
Methods: Initially studies were performed to determine the
biocompatibility of LNG with common cell lines used to evaluate antiviral potency of candidate drugs. Having determined the maximal
compatible dose of LNG in the different cell models, the anti-viral
activity of DPV alone or in combination with LNG was determined using
TZM-bl, PM-1 CD4+ T cells and PBMC. Viral replication was assessed by
quantitation of luciferase or p24.
Results: A concentration of 10µg/ml was selected as the maximal
compatible dose for use in all assays. In TZM-bl cells LNG alone showed
significant inhibitory activity against HIV-1 at concentrations >1µg/ml.
This is likely attributable to the progestogen-driven anti-proliferative
effect observed in epithelial cell lines, from which TZM-bl are derived.
However LNG had no direct activity against HIV-1 infection when using
PM-1 CD4+ T cell lines and PBMC, which do not express progesterone
receptors. Titration of DPV alone or in the presence of LNG displayed
superimposable curves indicating no impact of LNG on the anti-viral
activity of DPV.
Conclusions: LNG does not alter the anti-viral potency of DPV against
HIV-1BaL in PM-1 CD4+ T cells or human PBMC and is therefore unlikely
to influence the anti-viral potency of DPV when co-formulated in an
intravaginal ring.
208
Ninochka Jean-Pierre1, Patrick Barnable1, Larisa Kizima1, Aixa
Rodríguez1, Samantha Seidor1, Meredith Clark2, Gustavo Doncel2,
Melissa Robbiani1, Thomas Zydowsky1, Natalia Teleshova1, José
A. Fernández-Romero1
Population Council, New York, NY, United States, 2CONRAD Eastern
Virginia Medical School, Arlington, VA, United States
1
Background: Next generation microbicides may be broad-spectrum
multipurpose prevention technologies with improved efficacy and
safety. CAPRISA-004 showed that 1% TFV vaginal gel prevented HIV-1
and HSV-2. Here we compare the in vitro and in vivo safety and efficacy
profile of 1% TFV gel versus a microbicide gel (MZC) containing 50 mM
MIV-150 (M), 14 mM zinc acetate (Z) and 3% carrageenan (CG).
Methods: Increased HSV-2 susceptibility in mice (n=20), TEER in Caco2 cells, and macaque vaginal explant histology were used to estimate
damage to epithelia. XTT and Cyquant were used to evaluate in vitro
cytotoxicity in TZM-bls and PBMCs. Anti-HIV activity was tested against
HIV-1 lab strains (n=5), primary isolates and multidrug resistant strains/
clones (n=28) in TZM-bls or PBMCs. CC50 and IC50 values were used to
estimate therapeutic indexes (TI=CC50/IC50). Anti-SHIV-RT activity
(104 TCID50/explant) of diluted gels was assessed in macaque vaginal
explants. In vivo anti-HSV-2 activity (5x103 pfu/mouse, n=20/treatment)
was evaluated in mice (gel applied 1 h before and after HSV-2 vaginal
challenge). Levels of TFV and TFV-DP in mouse cervicovaginal tissues
were quantified by LC-MS/MS. Fisher’s exact test (P< 0.05) was used for
statistical comparison in all HSV-2 murine models. Kruskal-Wallis and
Dunn’s Multiple Comparison test (P< 0.05) in the explant model.
Results: Both gels were safe. However, diluted 1% TFV reduced TEER
values in Caco-2 monolayers. MZC showed greater in vitro antiviral
activity (2 to 80-fold) than 1% TFV gel against HIV-1 in TZM-bls. Both
gels showed good TI (>25-800) in PBMCs, except for one and two MDR
HIVs with 1%TFV and MZC, respectively. MZC fully protected explants
from SHIV-RT (p< 0.009) with greater anti-SHIV-RT activity than 1%
TFV gel. MZC fully protected mice from HSV-2 (p< 0.0001), but the 1%
TFV gel did not. The lack of protection could be associated with subtherapeutic levels of TFV and TFV-DP in mouse tissues.
Conclusions: MZC is a promising microbicide candidate for clinical use.
P14.03
P14.04
Formulation Development of a Combination
Silicone Elastomer Ring for Vaginal Delivery
of Dapivirine and Levonorgestrel
Matrix Vaginal Ring Formulations that
Maintain Target in-vitro Release Rates of
Dapivirine and Levonorgestrel (Alone or in
Combination) over 90 Days
University of Ulster, School of Pharmacy and Pharmaceutical Sciences,
Coleraine, United Kingdom, 2Queen’s University Belfast, School of
Pharmacy, Belfast, United Kingdom, 3International Partnership for
Jonathon Holt1, Andrew Brimer1, Susan Fetherston2, Peter Boyd3,
Brid Devlin1, Karl Malcolm3
1
1
Background: IPM is currently evaluating a 25mg dapivirine (DPV) vaginal
ring (VR) in Phase III clinical trials as a 30-day HIV microbicide product.
A second generation VR delivering both DPV and levonorgestrel (LNG)
is being developed as a multi-purpose prevention technology (MPT)
offering simultaneous HIV prevention and hormonal contraception for
at least 60 days. In order to maintain DPV release rates for 60 days at
levels likely to provide protection, it will be necessary to increase the
DPV loading beyond 25mg.
Methods: Eleven prototype, silicone elastomer (Nusil® MED-4870),
matrix VRs containing various loadings of DPV (0, 100, 150 and 200mg)
and/or LNG (0, 16 and 32mg) were manufactured by injection molding
at 160°C for 60 sec. In vitro release testing under sink conditions was
performed for at least 60 days. Release of DPV and LNG from the VRs
was quantified by HPLC-UV.
Results: In vitro release of DPV and LNG obeyed root time kinetics for
all VRs tested, as expected for matrix-type devices and indicative of a
permeation-controlled release mechanism. Increasing the DPV or LNG
loading significantly increased the release rate for the same active (P <
0.001). The addition of LNG (16 or 32 mg) in the VRs did not significantly
influence DPV release. LNG release was significantly increased by the
presence of DPV, as compared to LNG only rings with equivalent LNG
loading. Each formulation tested exceeded the in-vitro release target for
DPV (200 µg at d60). Day 60 DPV release was 284-285 µg, 363-376
µg, and 437-454 µg for the 100, 150 and 200 mg DPV VRs respectively.
Conclusions: The data confirm the potential for development of a MPT
VR strategy based on a matrix ring containing DPV and LNG. Dapivirine
load of 200 mg should maintain requisite levels of dapivirine for 60
days in-vitro. In-vitro release of LNG is enhanced by the presence of DPV.
In-vivo testing of DPV-LNG rings will be necessary in order to test drugdrug release effects under physiological conditions
Background: IPM’s silicone-based Dapivirine Vaginal Ring is in Phase
III trials as a vaginal microbicide for 1 month continuous use. New
formulations have been assessed with in-vitro drug release of greater
than 90 days. By extending the use duration per ring, the overall
production cost per patient per month may be decreased to nearly one
third that of the monthly dapivirine vaginal ring.
Methods: Silicone vaginal rings manufactured at Queen’s University
Belfast using similar manufacturing processes as IPM’s monthly
dapivirine vaginal ring were produced as either dapivirine (DPV) only
(25 and 200mg) or a combination of DPV (200mg) & LNG (32 mg).
Nusil Med-4870® (a Pt-catalyzed, addition cured silicone polymer) was
used to produce rings with an overall diameter of 5.67cm, cross-section
diameter of 7.82mm and weighing 8g (160°C cure for 60 sec).
Daily in-vitro release (IVR) of DVP and/or LNG was assessed up to 92
days in 50mL of 1:1 IPA: water. Media was assayed by HPLC for DPV
and LNG.
Results: The DPV IVR target was 105 µg, which is the day 28 (25 mg)
Dapivirine Vaginal Ring release rate. Formulations containing 200 mg
DPV exceeded the target (d30 = 628 µg; d60 = 437 µg; d92 = 301 µg).
In addition, when formulated in combination with LNG, DPV IVR (µg/
day) remained above the target (d30 = 634; d60 = 454; d92 = 299).
LNG IVR was also above target (35 µg/day) for all 92 days tested (d30
= 155; d60 = 91; d92 = 46).
The day 1 release for a 200mg DPV ring (4132 µg DPV alone; 6038 µg
DPV with LNG) and for a 32 mg LNG ring (684 µg) remained at levels
below those established as safe in nonclinical toxicity studies.
Conclusions: The IVR data presented indicate that it may be possible
to produce a DPV-only microbicide ring and a DPV -LNG combination
microbicide and contraceptive ring for use over 90 days using a silicone
matrix ring. Furthermore, within the given range, elevation of DPV
loading (or co-loading with LNG) does not increase in-vitro release to
unsafe levels.
IPM, Silver Spring, MD, United States, 2University of Ulster, Coleraine,
United Kingdom, 3Queen’s University Belfast, Belfast, United Kingdom
www.hivr4p.org
209
POSTERS
Susan Fetherston1,2, Clare McCoy2, Diarmaid Murphy2, Peter
Boyd2, Andrew Brimer3, Jonathon Holt3, Brid Devlin3, Wendy
Blanda3, Jeremy Nuttall3, Chris Gimour3, Karl Malcolm2
Posters
P14.05
P14.06
In Vitro Design and Drug Delivery of
Combination Dapivirine and Contraceptive
Hormone Ethylene Vinyl Acetate (EVA) Vaginal
Ring Prototypes
Evaluation of the Factors Contributing to
Levonorgestrel Binding in Addition Cure
Silicone Elastomer Vaginal Rings
Jonathon Holt1, Tiffany Derrick1, Andrew Brimer1, Andrew Loxley2,
Brett Breaker2, Matthew Bigert2, Michael Grieco2, Bruce Frank2,
Brid Devlin1
IPM, Silver Spring, MD, United States, 2Particle Sciences Inc.,
Bethlehem, PA, United States
1
POSTERS
Background: Dapivirine (DPV) is currently formulated in a silicone
ring for Phase III clinical trials as a vaginal microbicide. NuvaRing® (a
marketed vaginal contraceptive ring) is EVA-based and has many of the
characteristics desired for a DPV/hormone combination ring. This work
describes the design and in vitro testing of EVA vaginal ring prototypes
containing DPV and various contraceptive hormones.
Methods: Two contraceptive hormone options were co-delivered with
DPV from either matrix or core-sheath designs: Etonogestrel (ETO)
combined with Ethinyl Estradiol (EE); Levonorgestrel (LNG). In vitro
elution (IVE) targets were based on the DPV ring currently in Phase III
trials, NuvaRing® (ETO and EE) and literature (LNG). The 60d IVE profile
was determined for matrix ring designs for all hormones and for coresheath rings with various loads of LNG and DPV in the core, and several
sheath thicknesses (blank or with 10mg of DPV).
Results: Matrix ring prototypes released DPV and all hormones with first
order kinetics, however, the day 1 bursts for ETO and LNG were high
(>1000 µg). Rings loaded with 25 mg DPV released drug at target rates
on day 60 and the day 1 burst was within expected limits. Core sheath
designs had a blunted initial release compared to the matrix rings but
the anticipated zero order kinetics was not achieved with a loaded core
and blank sheath. DPV loaded sheaths further increased the overall DPV
release across 60 days but only the lower LNG target (35 ug/day) was
achieved with any configuration of LNG load. Stability tests indicated a
need for refrigerated storage conditions to prevent migration of LNG to
the ring surface and transfer to the packaging.
Conclusions: These studies demonstrate that EVA would be a suitable
polymer for the dual delivery of dapivirine and contraceptive hormones
from a vaginal ring under refrigerated conditions. These conditions were
deemed unsuitable for use in developing countries due to cost and
therefore alternative polymers are being evaluated.
210
Karl Malcolm1, Diarmaid Murphy1, Clare McCoy1, Peter Boyd1,
Susan Fetherston2, Andrew Brimer3, Jonathon Holt3, Wendy
Blanda3, Brid Devlin3, Jeremy Nuttall3, Chris Gimour3, Tiffany
Derrick3
1
Background: Following progress of the dapivirine (DPV)-releasing
silicone elastomer (SE) vaginal ring (VR) into Phase III clinical studies,
there is now interest in developing next-generation rings that
additionally provide contraception. Levonorgestrel (LNG) is a safe and
effective progestin that is being widely considered for use as a hormonal
contraceptive agent in future multipurpose prevention technology (MPT)
products. Although LNG has previously been incorporated into various
controlled release SE devices, minimal attention has focused on its
propensity to irreversibly react with addition cure SE systems. Here, for
the first time, we investigate this LNG binding phenomenon and outline
strategies for overcoming it.
Methods: VRs containing various loadings of DPV and LNG were
manufactured and in vitro release assessed. Different LNG-only SE
samples were also prepared to assess the following parameters:
(i) addition cure vs. condensation cure SEs;
(ii) different types of addition cure SEs;
(iii) mixing time,
(iv) cure temperature,
(v) cure time; and
(vi) LNG particle size.
After manufacture, the LNG-only samples were assayed for total drug
content using a solvent extraction method. The SE curing reaction and
the LNG binding reaction was probed using nuclear magnetic resonance
(NMR) spectroscopy.
Results:
Under certain drug/formulation/processing conditions, LNG was
not recoverable from VRs. Further studies using non-ring samples
showed that:
(a) the phenomenon was only observed with addition cure SEs (and not
condensation cure SEs);
(b) the extent of binding was dependent upon the type of addition cure SE;
(c) micronised LNG showed significantly greater binding than nonmicronised LNG;
(d) the extent of binding correlated with increased mixing time, cure
time and cure temperature.
Conclusions: Careful control of the API characteristics, the SE
composition, and the manufacturing conditions will be necessary to
establish a practical VR formulation for controlled release of LNG.
P14.07
P14.08
Thermal Properties and Eutectic Behaviour
of Dapivirine in Combination with Steroid
Hormones and Other Antiretrovirals
Selection of a Hormonal Contraceptive Agent
for Inclusion with Dapivirine in a Silicone
Elastomer Vaginal Ring
Karl Malcolm1, Clare McCoy1, Diarmaid Murphy1, Peter Boyd1,
Andrew Brimer2, Jonathon Holt2, Brid Devlin2, Wendy Blanda1,
Jeremy Nuttall2, Chris Gimour2
Clare McCoy1, Peter Boyd1, Susan Fetherston2, Ian Major1,
Diarmaid Murphy1, Brid Devlin3, Andrew Brimer3, Jonathon Holt3,
Jeremy Nuttall3, Chris Gimour3, Wendy Blanda3, Karl Malcolm1
Kingdom, 2International Partnership for Microbicides, Silver Spring, MD,
United States
Background: Combination drug products can display thermal behaviour
that is more complex than for the corresponding single drug products.
For example, the contraceptive vaginal ring (VR) Nuvaring contains a
eutectic (lowest melting) composition of etonogestrel (ETN) and ethinyl
estradiol. Here we report the predisposition of dapivirine (DPV) to
form reduced melting/eutectic mixtures when combined with other
contraceptive hormones and antiretrovirals, and discuss the implications
for development of combination microbicide and multipurpose
prevention technology (MPT) products.
Methods: Binary mixtures of DPV with darunavir (DRV), levonorgestrel
(LNG), ETN or maraviroc (MVC) were prepared either by physical mixing or
by solvent evaporation. Selected binary mixtures were also incorporated
into silicone elastomer (SE) VR devices. Thermal behavior of the mixtures
was analyzed using differential scanning calorimetry (DSC) operating
in standard heating ramp mode (10 °C/min). DSC data were used to
construct two component phase diagrams for each binary system.
Results: Drug mixtures typically showed reduced melting transitions
for both drug components, with clear evidence for a eutectic mixture
at a well-defined intermediate composition. Eutectic temperatures and
compositions for the various mixtures were: 40% DPV / 60% ETN 170°C; 25% DPV / 75% MVC - 172°C; 65% DPV / 35% LNG - 192°C.
In each case, the eutectic composition was also detected when the drug
mixtures were incorporated into SE VRs. For the DPV/DRV system, the
thermal behaviour is complicated by desolvation from the darunavir
ethanolate polymorph.
Conclusions: When DPV is combined with small molecular weight
hydrophobic drugs, the melting temperature for both drugs is typically
reduced to a degree dependent on the composition of the mixture.
At specified compositions, a low melting eutectic system results. The
formation of eutectic behavior in binary drug systems needs to be
carefully characterised in order to define product performance and
drug release.
1
Background: A vaginal ring (VR) that can prevent both unintended
pregnancy and the spread of sexually transmitted infections has the
potential to significantly improve women’s sexual and reproductive
health. Here, we report in vitro evaluation of four hormonal contraceptive
compounds - levonorgestrel (LNG), ethinylestradiol (EE), etonogestrel
(ET) and nestorone (NES) - for co-formulation with dapivirine (DPV) in a
silicone elastomer (SE) vaginal ring device.
Methods: The influence of each steroid hormone on the SE cure
characteristics was assessed using flow rheology. The thermal stability of
each API and the potential for drug-polymer and drug-drug interactions
were assessed using thermal gravimetric analysis (TGA) and differential
scanning calorimetry (DSC). Matrix-type and reservoir-type VRs
containing various loadings of each steroid were manufactured using
both a condensation-cure and an addition-cure SE systems and in vitro
release rates assessed.
Results: TGA confirmed all steroids were stable in the temperature
range required for DPV ring manufacture. DSC analysis confirmed
that all steroids showed eutectic behaviour when combined with DPV.
Rheological analysis demonstrated that EE and ET, but not LNG or
NES, inhibited curing of addition-type SE system. All steroids could be
effectively released from the SE VRs. However, only LNG release could
be sustained for the target 30 day period.
Conclusions: Based on the results of these studies, LNG was selected as
the most suitable contraceptive agent for further development with DPV
in a dual-purpose vaginal ring.
www.hivr4p.org
211
POSTERS
1
Posters
P14.09
P14.10
The Need for Multi-product Technologies for
Women Participating in Vaginal Microbicide
Trials
Multipurpose Prevention of HIV and
Pregnancy: Perspectives of Users, Providers
and Community Advisors in Zimbabwe and
Malawi
Duduzile E. Ndwandwe1, Nathlee S. Abbai1, Handan Wand2, Gita
Ramjee1
Unit, Durban, South Africa, 2University of New South Wales, The Kirby
Institute, Sydney, Australia
1
POSTERS
Background: Sub-Saharan Africa continues to bear a towering share
of the global HIV burden with 70% new HIV infections. HIV prevalence
among young women remains more than twice as high as among young
men throughout sub-Saharan Africa Therefore, it is important to perform
a needs assessment for women enrolled in HIV prevention trials as a
strategy to increase adherence to investigational products. The aim of this
study is to assess commitment to prevention interventions by measuring
pregnancy and HIV incidence in relation to product adherence.
Methods: This study included data collected from women (n=1456)
enrolled in the Carraguard vaginal microbicide gel trial. Associations
between gel adherence and pregnancy and HIV incidence in women
during the study follow-up were described using Kaplan-Meier and
Cox regression analysis. Statistical analysis was performed using STATA
release 12.0 (College Station, Texas, TX, USA).
Results: The highest crude incidence rates for pregnancy (6.7 per 100
person year (py), 95% Confidence interval (CI) 5.1, 8.9) and HIV (8.5 per
100/py, 95% CI: 6.2, 11.6) were reported for women that were in the
2nd quartile of % gel used with condoms (35-< 55%). After adjusting
for age, number of sexual partners in the last 3 months, number of
sex acts in the last week and marital status; women that had very low
gel adherence with condom use were at increased risk for incident
pregnancy (1st quartile < 35%, Hazard Ratio (HR) 2.4, 95% CI: and
HIV infections (2nd quartile 35-< 55%, HR: 3.5, 95% CI: 1.8, 7.0) when
compared to high adherers (>75% gel adherence with condom use, HR:
1 per 100 p/y respectively).
Conclusions: These findings suggest that adherence to study product
and condom promotion is critical to prevent infection with HIV as well
as reduce pregnancy incidence.
212
Cynthia Woodsong1, Petina Musara2, Adlight Chandipwisa2,
Elizabeth Montgomery3, Patty Alleman4, Zvavahera Mike
Chirenje2, Tsungai Chipato2, Francis Martinson5, Irving Hoffman6
International Partnership for Microbicides, Silver Spring, MD, United
States, 2UZ-UCSF Collaborative Research Programme, Harare,
Zimbabwe, 3RTI International, Women’s Global Health Imperative,
San Francisco, CA, United States, 4USAID, Office of Population and
Reproductive Health, Washington, DC, United States, 5UNC Project,
Lilongwe, Malawi, 6UNC, UNC Hospitals, Chapel Hill, NC, United States
1
Background: Prevention of HIV and pregnancy could be accomplished
with a single “Multipurpose Prevention Technology” (MPT), and a number
of topical vaginal products are currently under development. There is
limited information on the interests of potential MPT users and those
advising on use, particularly in sub-Saharan Africa. Rather, assumptions
about interest in such a product are inferred from what is known about
acceptability and use of vaginal microbicides or contraceptives.
Methods: This paper presents data on concerns and preferences for
multipurpose prevention of HIV and pregnancy. Data were collected
in two on-demand microbicide gel studies in Malawi and Zimbabwe.
Participants were women using candidate vaginal products, their
male partners, health professionals and community stakeholders.
All participants provided an interview which was audio-recorded,
transcribed, coded for content, and analyzed for key themes concerning
interest and concerns about multipurpose prevention.
Results: Participants indicated strong interest in a vaginal HIV prevention
product that could also prevent pregnancy. Key reported advantages
included convenience, the potential to avoid side effects experienced
with current contraceptive methods, concerns about long term effects of
contraceptives, and concerns about the health burdens of HIV infection
during pregnancy. The main disadvantage of an MPT was recognition
that while interest in preventing HIV is constant, contraceptive needs
change over time.
Conclusions: The study provides much-needed primary data on interest
in an MPT to prevent HIV and pregnancy. This interest may be further
strengthened if a product is also available for prevention of only HIV.
Health professionals and community advisors may be more likely to
recommend an MPT, and women and men may be more willing to use
an MPT, if they can be reassured that use will have no long-term effect
on fertility, and they can switch from single to multipurpose prevention
as needs change over time.
Posters 15: Novel Formulations, Agents and Microbicides
P15.01
P15.02
ABSTRACT WITHDRAWN
Acceptability of a Polyurethane Tenofovir
Disoproxil Fumarate (TDF) Intravaginal Ring
for HIV Prevention
Dana L. Watnick1, Marla J. Keller1, Lilia Espinoza1, Betsy C.
Herold1, Laurie J. Bauman1
Albert Einstein College of Medicine, Bronx, NY, United States
Background: There is urgent need for female-controlled HIV prevention
strategies such as topical pre-exposure prophylaxis. Unfortunately,
intravaginal ring (IVR) development has had little input from users.
As part of an ongoing Phase 1 TDF IVR safety and pharmacokinetic
trial among 30 US women, we are qualitatively investigating how HIV
risk perception and ring acceptability, specifically related to partner
concerns, menstruation and timing of ring exchange, may affect user
receptivity to ring use.
Methods: Female participants (18-45 years) are randomized to a
polyurethane TDF or placebo IVR for 14 days of continuous use and
participate in two in-depth interviews. Data are collected and analyzed
using a Grounded Theory approach. We report results for the first 10
women who completed the trial (6 TDF, 4 placebo).
Results: No one perceived herself at risk for HIV, but all conceded
infidelity (“even spouses might cheat”) and 3 identified rape, sex work
and “living in Africa” as HIV risk factors. Trusting one’s partner to “always
be faithful” and absence of a sexual relationship were reasons to forego
ring use. Issues with partner acceptability were male drug exposure,
interference with sex, suspicion of her cheating, and facilitating his
cheating. Most women (n=8) had concerns about ring use during
menses: potential infection from blood on the ring; the body’s need
to clean out toxins (including the drug itself); tampons soaking up
medication; blood blocking the outward flow of drug; and not needing
a ring because of abstinence during menses. Most (n=8) preferred to
remove the ring monthly before menses or to clean it to avoid infection
or “funky odor”. Seven subjects questioned ring effectiveness the longer
it was left inside the vagina.
Conclusions: Lack of perceived HIV risk and partner concerns are
potential barriers to ring uptake. Menstruation and frequency of ring
exchange are inextricably linked around the monthly cycle; half prefer
monthly ring changes.
www.hivr4p.org
213
POSTERS
1
Posters
P15.03
P15.04
Jatropha sp. Extracts Induces CD4
Internalization and Inhibits HIV-1 Entry
Development of a Novel Bioadhesive
Microbicide Gel Formulation for Prophylactic
Protection against HIV and HSV-2
Paola Silveira1, Rodrigo Orlandini2, Rodrigo Cunha1, Thais
Barbizan3, Luis Pianowski3, Luis Da Silva2, Amilcar Tanuri1,
Renato Aguiar1
Federal University of Rio de Janeiro, Rio de Janeiro, Brazil, 2Federal
University of São Paulo, Ribeirao Preto, Brazil, 3Kyolab Laboratories,
Valinhos, Brazil
1
POSTERS
Background: In 2012, were recorded approximately 2.3 million of
new HIV infections. These numbers show the need to develop drugs
that can prevent the infection, since an effective vaccine for HIV is not
available. Microbicides are an alternative for primary prevention and
control the HIV epidemic. Here, we screened extracts from the Brazilian
plant Jatropha sp. to evaluate cytotoxicity and potential antiviral activity
against HIV-1 in TCD4 cells.
Methods: The cytoxicity and antiviral activity of Jatropha sp. extracts
were initially screened in TCD4+ lymphocytes. Cells were pre-exposed
to extracts and the HIV infectivity was measured by luciferase reporter
gene cloned in the HIV genome.The downmodulation of CD4 receptor
was checked by flow cytometry. To elucidate the molecular mechanisms
of CD4 internalization several mutants of CD4 tail were transfected in
HeLa cells treated and the CD4 staining. CD4 degradation was evaluated
by the lysosome inhibitor and immunoblotting. Cellular morphology
after treatment was checked by electron microscopy and the cytokines
produced by treated cells was measured by Bioplex system. The cell
activation was evaluated by flow cytometry with specific antibodies.
Results: The THS fraction decreased HIV-1 infectivity up to 80% in
a dose-dependent manner with no cytoxicity. THS treatment induces
internalization of CD4 receptor in PBMC cells, direction to early
endosomes, followed by lysosomal degradation. The CD4 internalization
promoted for THS treatment was mediated by PKC activation and
consequent phosphorylation of CD4 tail serine residues. Our results of
electron microscopy showed that the THS stimulate macropinocytic.
Finally, it was observed that the compound activated cells with high
expression of CD69 and increased secretion of IL-8 and MIP-1beta.
These data suggests that THS treatment does not causes T cell anergy.
Conclusions: THS induces CD4 internalization and inhibits HIV entry
suggesting that compounds can be potentially used as microbicide to
prevent HIV transmission.
214
Gita N. Shankar1, Gaurav Bhatia1, Carsten Alt2
SRI International, Pharmaceutical Sciences, Menlo Park, CA, United
States, 2Palo Alto Institute for Research and Education, Palo Alto, CA,
United States
1
Background: Over-the-counter access to an inexpensive topical
microbicide could reduce transmission of human immunodeficiency
virus (HIV) and would increase women’s control over their health and
eliminate the need to obtain their partners’ consent for prophylaxis.
Chronic infection with herpes simplex virus-2 (HSV-2) has been shown
to facilitate HIV infection and speed the progression to disease. Our
objective is to develop a drug formulation that adheres to the vaginal
surface for extended time, while protecting against both HIV and HSV-2.
We present here development studies of a novel bioadhesive vaginal
delivery platform “SR-2P” which is composed of two FDA-approved
polymers, acting as a depot for two approved antiviral drugs, acyclovir
and tenofovir.
Methods: Prototype gels were developed and evaluated for pH,
osmolality, buffering capacity and viscosity under simulated vaginal
semen dilutions, and bioadhesivity using minipig vaginal tissues. Release
profile of drug loaded SR-2P was performed through porcine vaginal
mucosa. Vaginal irritation studies were performed in mice and rabbits.
Results: Results show that SR-2P retains its structure and bioadhesivity
upon dilution in presence of simulated fluids under stress conditions.
The in-vitro release profile of SR-2P demonstrated that ~40% tenofovir
and ~38% acyclovir was retained in the porcine vaginal mucosa, in the
presence of vaginal semen fluids even after 6 hours of contact time.
SR-2P caused minimal irritation in a mouse and a rabbit vaginal model.
Our results indicate that SR-2P containing acyclovir and tenofovir
protects Vero cells from infection with HSV-2 in vitro and mice from
HSV-2 in vivo.
Conclusions: This preliminary study demonstrates that SR-2P gel could
be used as a vaginal microbicide for prophylaxis against vaginal HIV and
HSV-2. Research reported in this publication was supported by NIAID/
NIH Award No.AI098658. The content is solely the responsibility of the
authors and does not necessarily represent the official views of the NIH.
P15.05
P15.06
Design of Biodegradable Nanoparticles for
Vaginal Delivery of the anti-HIV Protein
Griffithsin
Nanodelivery of CSIC for Enhanced Solubility
and Rapid Macrophage Uptake
1
2
3
University of Pittsburgh School of Pharmacy, Department of
Pharmaceutical Sciences, Pittsburgh, PA, United States, 2Magee Women
Research Institute, Pittsburgh, PA, United States, 3University of Louisville
School of Medicine, Louisville, KY, United States, 4University of
Pittsburgh School of Medicine, Department of Obstetrics, Gynecology,
and Reproductive Sciences, Pittsburgh, PA, United States
1
Background: Griffithsin (GRFT) is an anti-HIV protein which blocks
gp120 binding to CD4 receptor-expressing cells and binds to mannoserich glycans on viral glycoproteins (gp120, gp41, and gp160). Its
broad-spectrum anti-HIV activity and unique safety profile support its
development as a topical microbicide. However, its proteinous nature
and poor tissue permeability limit its antiviral activity to the vaginal
lumen. That leaves HIV targeted cells within submucosa and deeper
vaginal tissue vulnerable without the protection of GRFT, which will
consequently diminish its potential as a topical microbicide product. The
objective of this study was to use poly (lactic-co-glycolic acid) (PLGA)
nanoparticles (NPs) to enhance the penetration of GRFT leading to more
efficient delivery of this anti-HIV drug.
Methods: A double emulsion-solvent evaporation method was utilized
to manufacture GRFT loaded PLGA NPs. Particle size, size distribution
and zeta potential of NPs were determined utilizing dynamic light
scattering. Images of the nanoparticles were taken by transmission
electron microscopy (TEM). The amount of GRFT encapsulated in the
nanoparticles was determined by HPLC method. The anti-HIV activity
and toxicity of GRFT nanoparticles was studied in a TZM-bl cell based
assay.
Results: The developed GRFT loaded PLGA NPs showed nearly spherical
shape with a particle size of 257-318 nm and narrow size distribution
(PDI < 0.2). Particle size was confirmed using TEM. Particle zeta potential
was -30 mV. Encapsulation efficiency for GRFT was found to be 42%. In
vitro studies showed that the IC50 of GRFT loaded NPs was comparable
with GRFT (»3 nM). In these same studies cells viability of both PLGA NPs
and GRFT loaded NPs was completely maintained.
Conclusions: These studies represent the incorporation of GRFT into
a PLGA NPs system. The encapsulation of GRFT into PLGA NPs did not
impact its anti-HIV activity and safety. NP incorporation of GRFT may
enhance its tissue penetration leading to increased protection.
Tiantian Gong1,2, Michael Parniak3, Phalguni Gupta4, Lisa
Rohan1,2,5
Magee Womens Research Institute, Pittsburgh, PA, United States,
University of Pittsburgh School of Pharmacy, Pittsburgh, PA, United
States, 3University of Pittsburgh, Department of Microbiology &
Molecular Genetics, School of Medicine, Pittsburgh, PA, United States,
4
University of Pittsburgh Graduate School of Public Health, Department
of Infectious Diseases and Microbiology, Pittsburgh, PA, United States,
5
University of Pittsburgh, Department of Obstetrics, Gynecology and
Reproductive Sciences, Pittsburgh, PA, United States
1
2
Background: CSIC is a hydrophobic, tight-binding, non-nucleotide
reverse transcriptase inhibitor with high potency against HIV-1 in
vitro. To enhance its aqueous solubility a three phase nanoparticle
engineering technology was used to manufacture CSIC nanocrystals
(NC) with acceptable properties for intravaginal application.
Methods: Nanocrystals were prepared using co-precipitation, hydration,
and stabilization steps. Particle size distribution, polydispersity index
(PdI), and zeta potential were determined using dynamic light scattering.
NC drug release and stability were assessed. Surface morphology was
investigated by transmission electron microscopy (TEM). Differential
scanning calorimetry (DSC) was used to monitor crystal properties. Drug
content was analyzed by HPLC-UV. Biological attributes including mucin
interaction, bioactivity, and macrophage uptake were studied.
Results: Nanocrystal stabilization was achieved through use of
a combination of Pluronic F98 (0.3mg/mL) and hydroxypropyl
methylcellulose (HPMC) E5 (5mg/ml). NC obtained using this system had
a particle size of 260nm (PdI < 0.4) and surface charge of -7.2mV. Particle
size was maintained for 8h in a liquid state, and for at least 2 month
after lyophilization. An amorphous state following co-precipitation was
confirmed using DSC. Following hydration and stabilization, needleshaped crystals resembling the raw CSIC powder were obtained
(confirmed by DSC and TEM). Aqueous solubility of the NC CSIC form
was increased by ~500 fold. Encapsulation efficiency was >80%
(0.7mg/mL) and 75% of loaded CSIC was released from NC within 4
days. No change in particle size was observed in the presence of mucin
indicating limited mucoadhesive properties. NC formation resulted in no
loss of bioactivity. In vitro studies showed rapid macrophage uptake via
an energy-dependent pathway.
Conclusions: Nanocrystals can be used to improve CSIC aqueous
solubility and drug targeting which are critical for drug localization at
the target site.
www.hivr4p.org
215
POSTERS
Haitao Yang , Jing Li , Charlene S. Dezzutti , Kenneth Palmer ,
Lisa C. Rohan1,2,4
1,2
Posters
P15.07
P15.08
Vaginal and Rectal Use of OTC Lubricants:
Safety in the Macaque Model
NMR Metabonomics in an in vitro Model of
HIV-1 Latency
Dorothy L. Patton1, Yvonne Sweeney1, Sharon Hillier2
Thato P. Nonodi1, Debra Meyer1
University of Washington, Obstetrics & Gynecology, Seattle, WA,
United States, 2Magee Women’s Research Institute, Pittsburgh, PA,
United States
1
1
POSTERS
Background: Over-the-counter (OTC) lubricants are marketed for use
during vaginal and anal receptive intercourse but these products have
not been extensively studied for safety. The macaque model provides a
standardized means for evaluation of vaginal and rectal product safety.
Methods: Four OTC lubricants available in the USA were evaluated
including three aqueous-based personal lubricants (hypo-osmolar
FemGlide, iso-osmolar Pre-Seed, hyper-osmolar KY Jelly), and
hyperosmolar, lipid based Elbow Grease cream. Six macaques
completed each study arm. Animals underwent 4 daily applications of
each lubricant, with measurements collected prior to and 30-minutes
after each use, and 24-hours after the final exposure. The integrity
of the mucosal epithelium at the site of product use was assessed by
colposcopy following vaginal use and assessment of epithelial sloughing
in rectal lavage samples following rectal application. The microflora and
pH was also assessed before and after daily exposure to test agents.
Results: No evidence of product-related epithelial disruption was
noted with vaginal use of any of the four products. Two products were
associated with increased epithelial sloughing in rectal lavage samples
after use (KY Jelly and Pre-Seed). The vaginal and rectal microbiota was
not grossly affected by product use; however repeated vaginal use of
KY Jelly suppressed the presence of beneficial lactobacilli in half of the
animals. pH measures were only slightly and temporarily influenced by
the product pH in all cases.
Conclusions: OTC personal lubricants had variable effects on the
genital and rectal tracts of pigtailed macaques. While FemGlide and
Elbow Grease Cream had acceptable safety profiles with vaginal and
rectal use, some other OTC lubricants adversely affected the vaginal
microbiota or were associated with epithelial sloughing following rectal
application. Given the wide use of personal lubricants during sexual
activity, additional safety studies are warranted.
216
University of Pretoria, Biochemistry, Pretoria, South Africa
Background: HIV latency is viewed as one of the important reasons
why eradication of infection has been elusive thus far. Studies on how
the virus affects immune system cells during its active and latent phases
could provide targets for drug development as well as a means to
monitor the progress and status of infection. Since the implementation
of ‘omics’ studies, whole cells/systems/pathways have been elucidated
by the use of high throughput analytical tools like nuclear magnetic
resonance (NMR) spectroscopy and mass spectrometry (MS).
Methods: In this study metabonomics analysis of U1 cells (a
promonocytic HIV-1 latency and chronic infection model) was used to
investigate and compare HIV-1 latency to active infection. The uninfected
parent cell line (U937) served as a necessary control. Metabolites from
these cells were extracted and analysed by NMR.
Results: Six important metabolites were detected; these metabolites
were identified using relevant databases and found to be lactate, lipid,
diethylthiophosphate, methylsuccinate, choline and methylacetoacetic
acid. Changes in the peak height of lactate peak between the different
viral phases presents as a possible marker of infection status.
Conclusions: Lactate, is a product of glucose metabolism which is
already known to be influenced by active HIV-1 infection. Data presented
here suggests that even in latency the virus appears to have an effect
on glucose metabolism.
P15.09
P15.10
Drug-drug Interactions between the
Dapivirine Vaginal Ring (Ring-004) and
Miconazole Nitrate Vaginal Capsule (GynoDaktarin®)
Broad-spectrum Anti-HIV-1 Activity of
Anionic Carbosilane Dendrimers and Synergy
in Combination with Maraviroc and Tenofovir
as Topical Microbicide
Annalene Nel1, Wouter Haazen2, Marisa Russell1, Jeremy Nuttall3,
Neliette Van Niekerk1, Nicoline Treijtel4
Daniel Sepúlveda-Crespo1,2,3, María Jesús Serramía1,2,3, Javier
Sánchez-Rodríguez1,2,3, Raquel Lorente1,2,3, Rafael Gómez3,4,
Francisco Javier de la Mata3,4, Jose Luis Jiménez2, Mª Ángeles
Muñoz-Fernández1,2,3
Background: The Dapivirine Vaginal Ring-004 (25 mg dapivirine) is a
topical microbicide currently being evaluated in Phase III trials. Given
the potential for interactions with co-administered vaginal products,
pharmacokinetic assessments were performed during co-use of Ring004 and the antifungal miconazole nitrate.
Methods: An open-label, randomised, crossover trial was conducted
among 36 healthy, HIV-negative women, aged 18-40 years. Participants
used dapivirine Ring-004 for 28 days, alone or together with a single
dose of miconazole nitrate (1200 mg vaginal capsule). A single dose
of miconazole nitrate was administered alone in a third treatment
period. Washout periods of 3 weeks were included between treatments.
Dapivirine and miconazole concentrations were determined in plasma
and vaginal fluid (CVF) samples, and residual dapivirine levels were
assessed in used rings.
Results: A single vaginal dose of miconazole nitrate at the time of
dapivirine ring insertion increased the systemic exposure (Cmax and
AUC) of dapivirine by approximately 20%, but decreased dapivirine
CVF levels up to 14 days post-ring insertion (decreases of 26% in Cmax
and 69% in AUC0-24h). No significant difference was observed between
dapivirine ring residual levels with and without concomitant miconazole,
suggesting similar dapivirine release. Dapivirine CVF levels remained at
least 100 times higher than the in vitro IC99 in cervical tissue (3.3 ng/
mL). Local and systemic miconazole exposure was increased after coadministration with Ring-004 (1.4 to 6-fold higher). Concomitant use
of the two products was safe and well tolerated. One product-unrelated
SAE occurred (acetabulum fracture); one Grade 2 event of vulvovaginal
candidiasis (ring alone) was considered product-related.
Conclusions: Concomitant use of Ring-004 and a single vaginal dose
of miconazole nitrate altered the local and systemic levels of both
drugs, but these changes are considered unlikely to affect adversely the
efficacy of either drug.
Hospital General Universitario Gregorio Marañón, Laboratorio
Inmunobiología Molecular, Madrid, Spain, 2Hospital General
Universitario Gregorio Marañón, Plataforma de Laboratorio, Madrid,
Spain, 3Networking Research Center of Bioengineering, Biomaterials
and Nanomedicine (CIBER-BBN), Madrid, Spain, 4Universidad de Alcalá,
Departamento de Química Inorgánica, Alcalá de Henares, Madrid,
Spain
1
Background: Self-administered topical microbicides may be very
helpful tool for women and homosexuals to decrease new HIV
infections. Polyanionic carbosilane dendrimers are considered HIV1 entry inhibitors and antiretrovirals (ARVs) are the most advanced
microbicides. Consequently, the combination approach should be
taken into consideration when designing a new microbicide. In this
regard, double o triple-combination of dendrimers and ARVs acting in
the early stages of HIV-1 replication is an indispensable and beneficial
tool in terms of efficacy and long-term safety in fighting the HIV/AIDS
epidemic. We will show the latest results of the combination of two or
three carbosilane dendrimers with tenofovir (TFV) and/or maraviroc
(MRV) used as microbicides.
Methods: Cytotoxicity in different cell lines in vitro, the anti-HIV-1 in TZM.
bl cell line, vaginal irritation and subsequent histological analysis were
showed. 48h post-infection inhibitory activity profile was determined
by luciferase activity. Study of combined effects and the 50% effective
concentration (EC50) were performed using Calcusyn software.
Results: Two- and three-drug combinations showed a greater broadspectrum anti-HIV-1 activity than the single-drugs, preserved this activity
in acid environment or seminal fluid, and did not activate inflammatory
response. The strongest combinations were with G2-STE16 dendrimer at
constant ratios. They demonstrated strong synergistic activity profile due
to the combination indices varied between 0.06 and 0.74. Additionally,
no irritation was detected in female mice after dendrimer vaginal
administration.
Conclusions: The two- and three-drug combination increases their
antiviral potency and act synergistically as potential microbicide.
Our results deserve especially further clinical research on dendrimer/
dendrimer,
dendrimer/ARVs,
dendrimer/dendrimer/dendrimer,
dendrimer/dendrimer/ARVs, or dendrimer/ARV/ARV as microbicide
against HIV-1.
www.hivr4p.org
217
POSTERS
International Partnership for Microbicides, Clinical Affairs, Paarl,
South Africa, 2SGS Life Science Services, Clinical Pharmacology
Unit Antwerpen, Antwerp, Belgium, 3International Partnership for
Microbicides, Product Development, Silver Spring, MD, United States,
4
Kinesis Pharma BV, Clinical Pharmacokinetics, Breda, Netherlands
1
Posters
POSTERS
P15.11
P15.12
More…? Less…? Just Right…? The Role
of Perceived Volume in Gel and Film
Perceptibility During Intercourse, and its
Impact on Product Preference
Vaginal Film User Evaluations: Developer
Considerations from Initial Impressions and
User Sensory Perceptions and Experiences
during Vaginal Sex
Kathleen M. Morrow1, Rochelle K. Rosen2, Sara Vargas3, David
Katz4, Fava Joseph3, Erna M. Kojic5, David Friend6, Lisa Rohan7,
Anthony Ham8, Robert Buckheit8
Kathleen M. Morrow1, Lisa Rohan2, Rochelle K. Rosen3, Joseph
Fava4, Anthony Ham5, David Friend6, David Katz7, Erna M. Kojic8,
Robert Buckheit5
1
Miriam Hospital & Alpert Medical School of Brown University, Centers
for Behavioral and Preventive Medicine, Providence, RI, United
States, 2Miriam Hospital & Brown University School of Public Health,
Centers for Behavioral and Preventive Medicine, Providence, RI,
United States, 3Miriam Hospital, Centers for Behavioral and Preventive
Medicine, Providence, RI, United States, 4Duke University, Biomedical
Engineering and Ob/Gyn, Durham, NC, United States, 5Miriam Hospital
& Alpert Medical School of Brown University, Division of Immunology,
Providence, RI, United States, 6CONRAD, Arlington, VA, United States,
7
Magee Womens Research Institute, Pittsburg, PA, United States,
8
ImQuest BioSciences, Inc., Frederick, MD, United States
1
Background: We analyzed coital and pericoital vaginal perceptibility in
2 volumes of HEC gel (2 mL, 4 mL) and one 1”x2” film to understand
the role of volume and other properties in user sensory perceptions and
experiences (USPEs) of vaginal formulations.
Methods: 24 monogamous HIV/STI-negative heterosexual couples
enrolled; 100% completed 3 product evaluation visits (random
order). Women inserted product, then had vaginal sex with their male
partner. All completed USPE surveys and 30 (15 couples) completed
in-depth qualitative interviews (IDI) about each product’s properties and
performance. USPE scale scores, indicating ranges of sensations and
experiences perceived by users, as well as qualitative thematic analyses,
contribute to findings.
Results: USPE scale scores showed significant differences in pairwise
comparisons between each of 2 gel volumes and the film. Perceptibility
(USPE) scores were lowest overall for film, and highest overall for 4mL
gel. Volume differences were particularly relevant for:
1) sensations at initial penetration;
2) ease of stroke, perceived wetness, and physical awareness of product
during coitus;
3) perceived leakage and messiness; and
4) perceptions of product being sexually stimulating.
IDIs described ranges of experiences with differences by volume and
formulation. As expected, higher volume gel was generally considered
to be the most lubricating and messiest. 2mL gel was perceived by some
to be less messy and provide less lubrication, but this was not universal.
Film was perceived as not adding volume; most said that it did not add
lubrication. 44% of all participants (both genders) chose the 4mL gel as
their preferred product; 29% chose the 2mL gel; 27% chose the film.
Conclusions: Gel data indicate user-perceived differences by volume.
Film data provides context showing the importance of the role of volume
in product experience. While volume is important, user opinions of
product performance for penetration, pleasure, and covert use are not
determined by volume alone.
Background: Drug delivery in prevention technology is critical, but is
predicated on use adherence. The role of formulation characteristics/
properties of vaginal film (i.e., impressions of size, texture, color) and
film perceptibility (user sensory perceptions/experiences) was explored
during insertion and vaginal sex.
Methods: 24 monogamous HIV-/STI- heterosexual couples enrolled in
a mixed methods study. All
(100% retention) completed 3 formulation evaluations, 1 a placebo film.
Female participants provided qualitative data regarding film attributes
(size: 1x1, 1x2, 1x3, 2x2-inch; texture: textured, smooth; color: clear,
translucent, opaque) then inserted 1”x2” film and had vaginal sex
with their partner. Both partners completed USPE surveys and in-depth
qualitative interviews.
Results: Individual film characteristics were first rated separately, then a
final preference for film design was given. Relatively equal numbers of
women chose each film size. 13 of 23 women chose one smooth/one
textured side. The majority chose translucent or opaque coloring. Data
revealed a range of expectations about product performance. Some
perceived the 1”x1” films as difficult to insert and less efficacious, others
felt the 1”x1” film would insert easier, dissolve quicker, and result in
less leakage. 2”x2” films elicited similar expectation ranges. USPE scale
scores were low; women felt little-to-no lubrication, coating, leakage,
etc. Design parameters called for a film that users would not be aware
of and that would generate no leakage or messiness: these properties
were achieved. However, low lubricity and coating sensations at initial
penetration or during the first coital strokes were problematic, even
causing a few couples to stop sex.
Conclusions: The addition of vaginal film to the formulation parameter
space for USPEs of vaginal products offers important considerations
for developers. Given previous USPE analyses, initial lubrication may
impact use adherence and thus is currently being explored in new film
development.
218
Miriam Hospital & Alpert Medical School of Brown University, Centers
2
Magee Women’s Research Institute, Pittsburg, PA, United States,
3
Miriam Hospital & Brown University School of Public Health, Centers
4
Miriam Hospital, Centers for Behavioral and Preventive Medicine,
Providence, RI, United States, 5ImQuest BioSciences, Inc., Frederick,
MD, United States, 6CONRAD, Arlington, VA, United States, 7Duke
University, Biomedical Engineering and Ob/Gyn, Durham, NC, United
States, 8Miriam Hospital & Alpert Medical School of Brown University,
Division of Immunology, Providence, RI, United States
P15.13
P15.14
Using Molecular Dynamics Techniques to
Investigate the Influence of Glycans on the
HIV-1 gp120 Envelope Protein Trimer
Development of a Temperature-recording
Vaginal Ring for Monitoring User Adherence
1
1
University of the Western Cape, South African National Bioinformatics
Institute, Cape Town, South Africa
1
Background: The gp120 glycoprotein present on the surface of the HIV
virion is crucial for the recognition and binding to receptors on the host
cell surface, as well as facilitation of the fusion of the viral envelope
to the host cell membrane. The surface of gp120 is covered with
N-linked glycans that can influence HIV-1 infectivity as well as affect the
recognition of the virus by the host immune system, acting as a “glycan
shield” from antibody recognition.
Methods: Previous research has shown that the N-linked glycans bound
to the surface of a gp120 monomer have a significant impact on the
underlying dynamics of the protein.
Here, we have expanded on this work by undertaking molecular
dynamic modeling to explore the effect of N-linked glycans on the
dynamics of the full gp120 trimer.
Results: Our results illustrate the significance of the silent and active
faces of the gp120 monomers, as well as showing the trimer-specific
behavior of the glycans and glycan-protein interactions.
Conclusions: Collectively our results provide a better understanding
of the role that the glycan composition and distribution play during
infection of a new cell.
Peter Boyd1, Clare McCoy1, Diarmaid Murphy1, Manjula LustiNarasimhan2, Berglind Helgadóttir3, Karl Malcolm1
Kingdom, 2World Health Organization (WHO), Geneva, Switzerland,
3
Star-Oddi Ltd., Gardabaer, Iceland
1
Background: Vaginal ring devices are being actively developed for
controlled delivery of HIV microbicides and as multi-purpose prevention
technology (MPT) products combining hormonal contraception with
prevention of HIV and other sexually transmitted diseases. Presently,
there is no reliable method for monitoring user adherence in HIV vaginal
ring trials; previous acceptability studies have included some type of
participant self-reporting mechanism, which have often been unreliable.
More objective, quantitative and accurate methods for assessing
adherence are needed.
Methods: A silicone elastomer vaginal ring containing an encapsulated
miniature temperature recording device has been developed that
can capture and store real-time temperature data during the period
of designated use. Devices were tested in both simulated vaginal
environments and following vaginal placement in cynomolgus
macaques. Various use protocols and data sampling rates were tested to
simulate typical patient usage scenarios.
Results: The temperature logging devices accurately recorded
vaginal temperature in macaques, clearly showing the regular diurnal
temperature cycle. When environmental temperature and vaginal
temperature was significantly different, the device was able to accurately
pinpoint the insertion and removal times. Based on the data collected
it was possible to infer removal periods as short as 5 min when the
external environmental temperature was 25 °C. Accuracy increased with
data sampling rate.
Conclusions: This work provides proof-of-concept for monitoring
adherence using a vaginal ring device containing an encapsulated
temperature logger. The addition of one or more active agents into
the ring body is not anticipated to affect the temperature monitoring
function. A clinical study to compare self-reported user adherence data
with that obtained by the device would be highly informative.
www.hivr4p.org
219
POSTERS
Clint Mercuur , Natasha Wood , Simon Travers
1
Posters
P15.15
P15.16
Potential of RNA Aptamers in the Prevention
of HIV-1 subtype C Infections
Overcoming Phase Behavior and Surface
Crystallization in a Tenofovir-levonorgestrel
Intravaginal Ring
Grace Mothepane London1, Makobetsa Khati1,2, Bongani Mayosi2
CSIR, Biosciences, Pretoria, South Africa, 2University of Cape Town,
Medicine, Cape Town, South Africa
1
Meredith Clark1, Justin Clark2, Todd Johnson2, Namdev Shelke2,
David Friend1, Patrick Kiser3
CONRAD, Arlington, VA, United States, 2University of Utah, Salt Lake
City, UT, United States, 3Northwestern University, Evanston, IL, United
States
1
POSTERS
Background: Compounds that have been used to prevent human
immunodeficiency virus type-I (HIV-1) infections include synthetic
chemicals, plant extras and monoclonal antibodies. Although most
of these compounds have potent antiviral activity, they often fail to
progress to later stages of clinical trials due to high toxicity and lack
of specificity. Therefore, as an alternative to circumvent the above
mentioned limitations we used aptamers, which are small nucleic acid
ligands that recognize their target with high specificity and have no
toxicity in clinical applications.
Methods: In this study, we evaluated efficacy of four gp120-aptamers
against Env pseudovirus panel derived from HIV-1 subtype C, using virus
inhibition assay in TZM-bl cells, as well as toxicity. Binding specificity
of one potent aptamer (CSIR1.1) to gp120 was determined by EnzymeLinked-Immunosorbent Assay. Subsequently, a virus inhibition assay
was performed to test whether CSIR1.1 can inhibit HIV-1 pseudotyped
with vesicular stomatitis virus envelope glycoprotein (HIV-VSVG.
Results: All four aptamers inhibited infectivity of 81-84 % of the tested
viruses with mean inhibition concentration (IC50) of 6.4-9 nM. The
specificity results showed that CSIR1.1 only bound to gp120 and did
not bind other tested proteins (HIV-1 gp41, mycobacterium tuberculosis
virulent protein (CFP10), human interferon gamma (IFN-γ) and BSA).
CSIR1.1 also failed to inhibit HIV-1 pseudotyped with VSV-G protein and
showed no toxicity in vitro, even at concentration (500 nM), which was
5 × higher than one used for virus inhibition assays.
Conclusions: Aptamers showed significant efficacy against HIV-1
subtype C isolates and specificity to gp120 without causing cytoxicity
effects. These properties make aptamers attractive candidates for
prevention of HIV-1.
220
Background: We have reported on the design of an intravaginal ring
(IVR) engineered to deliver both tenofovir (TFV) and levonorgestrel
(LNG). Because these drugs have divergent properties and release
rates, sophisticated designs are required to deliver them in a controlled
manner. These designs force us to push the materials science behind
the IVR to the edge of what is possible. While exploring the physical
chemistry of the IVR we discovered and solved a number of stability
challenges that made the clinical development of this IVR possible.
Methods: TFV/LNG IVR as previously described were subjected to
various time-temperature profiles; drug release rates, presence of
surface crystallization by polarized light microscopy, and polyurethane
(PU) phase behavior by DSC were evaluated.
Results: Rings made with 35wt% swelling PU and placed at 40°C for
0 and 60d went from 21±5 to 4±1 mg/d TFV due to changes in the
effective diffusion coefficient of TFV in the PU with time. DSC showed a
change in the water hydration state of the PU with time at 40°C with an
exothermic peak at -50°C and enthalpy of -25 J/g. To compensate for this
change we added a 14d annealing step at 40°C to the manufacturing
process that allows the PU to achieve a more equilibrium conformation
resulting in stable DSC and drug release profiles. Annealed rings made
with a 60% swelling PU achieved steady-state release rates at 9±2 mg/d
and eliminated the hydrated water enthalpy. LNG segments placed on
storage showed a temperature dependent formation of LNG crystals
on the surface due to supersaturation of drug in the PU with crystals
forming after 30d at 40°C but not for at least 1yr at 4°C. Coating the
LNG segment with glycerol or PVP, in which LNG has a low activity,
inhibits LNG crystallization for >1yr at 40°C. No LNG crystallization has
been observed to date (5mo) on finished TFV/LNG IVR stored at 40°C.
Conclusions: Quantitative evaluation of the phase behavior of the IVR
allowed us to engineer an IVR not requiring cold chain storage.
P15.17
P15.18
Mass Transport Theory Improves
Compartmental PK Modeling of Microbicides
and Helps Guide Product Science and
Development
Testing of Nanoparticle-based ARV Drug
Combinations for Inhibiting Cell-free and Cellcell HIV Transmission
University of Washington, Department of Bioengineering, Seattle, WA,
United States, 2Seattle University, Department of Chemistry, Seattle,
WA, United States
1
1
Background: Understanding microbicide functionality, product design
and evaluation derive from synergy of experimental (in vitro, in vivo)
and computational (modeling) analyses. Modeling can deconstruct
and elucidate many elements of cause and effect in this complex,
multivariate process. Its potential has not been fully realized in the
science and development of anti-HIV strategies, e.g. in pharmacokinetic
analysis. Mechanistic modeling of microbicide PK processes - for
different vehicles (e.g. gels, rings) and different types of drugs - can
provide a framework to:
(1) identify salient properties of vehicle, drug, host environment and
dose regimen that govern PK;
(2) input those properties to make quantitative comparisons and
predictions that help design and interpret in vivo studies in humans
and animals, and in vitro studies of drug release/transport and HIV
neutralization.
Methods: Principles of mass transport theory were applied to vaginal
delivery of microbicide drugs by clinical and prototype gels and rings.
Compartments in the models were vehicle, lumenal fluids, epithelium,
stroma and blood stream. Diffusion and convection were primary
mechanisms of drug transport. Systems of coupled partial differential
equations were derived/solved, outputting drug concentrations vs. time/
location in each compartment.
Results: Results include tradeoffs in gel volume (2-4mL) and loading
(0.5-1.5%) for Tenofovir, IQP0528 and other drugs in achieving mucosal
concentrations that reach prophylactic (e.g. EC50) levels, showing how
frequency of dosing can sustain these levels. IVR geometry and release
flux history are related to such target concentrations, including effects
of transient ring removal. Scaling rules for human vs. animal dosing are
given.
Conclusions: Mechanistic modeling fills gaps in microbicide drug
delivery science, helping define and organize effects of the many factors
involved. This knowledge translates to methodology for use in rational
product design and performance evaluation, in vitro and in vivo.
Background: Strategies that enable ARV drugs to be easily combined
and provide sustained antiretroviral activity have the greatest potential to
impact the efficacy of next generation topical microbicides. To overcome
challenges associated with formulating multiple ARV compounds that are
chemically incompatible, nanoparticles were fabricated to encapsulate
individual ARV drugs t

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