Thursday Presentations - HIV Research For Prevention

Transcription

Thursday, 30 October
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
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
HIV Research for Prevention 2014 | HIV R4P
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
Institute of Human Virology, University of Maryland School of
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
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.
www.hivr4p.org
97
ORAL ABSTRACT SESSIONS
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
Oral Abstract Sessions
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
Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United
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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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
Imperial College, Infectious Diseases, London, United Kingdom,
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
Research, Silver Spring, MD, United States
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,
Durham, NC, United States
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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109
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
Institute of Human Virology, University of Maryland School of
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.
www.hivr4p.org
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
Queen’s University Belfast, School of Pharmacy, Belfast, United
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,
Boston, MA, United States
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 ´CD8 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
Oral Abstract Session 30: Antibody Functions and Protection
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.
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
for Behavioral and Preventive Medicine, Providence, RI, United States,
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),
Johannesburg, South Africa
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
Instituto de Investigaciones Biomédicas en Retrovirus y Sida,
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
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.
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145
Posters
Posters 27: Adjuvants
P27.01
P27.02
Monocyte-derived DC Electroporated with
mRNAs Encoding Both Specific HIV Antigens
and DC Adjuvants Are Able to Improve T-cell
Functionality
Intradermal Vaccination against SIV Induces
the Activation and Migration of Langerhans
Cells in Non-human Primates
Alberto C. Guardo1, Laia Miralles1, Joeri L. Aerts2, Kris
Thielemans2,3, Beatriz Mothe4, Javier Martinez-Picado4,
Christian Brander4, Felipe Garcia1, Montserrat Plana1, iHIVARNA
Consortium
IDIBAPS, Retrovirology and Viral Immunopathology Lab. Hospital
Clinic, University of Barcelona, Barcelona, Spain, 2Laboratory of
Molecular and Cellular Therapy, Medical School of the Vrije Universiteit
Brussels, Department of Physiology-Immunology, Brussels, Belgium,
3
eTheRNA, Kortenberg, Belgium, 4AIDS Research Institute IrsiCaixaHIVACAT, Hospital Universitari, Germans Trias i Pujol, Universitat
Autònoma de Barcelona, Badalona, Spain
Lucille Adam1,2,3, Biliana Todorova1,2,3, Roger Le Grand1,2,3,
Catherine Chapon1,2,3, Frederic Martinon1,2,3
CEA, Division of Immuno-Virology, IDMIT Center, Fontenay aux Roses,
France, 2Université Paris-Sud, UMR E1, Orsay, France, 3Vaccine Research
Institute (VRI), Créteil, France
1
1
POSTERS
Background: In the context of therapeutic vaccination of HIV-infected
patients, we have tested in vitro a combination of mRNA sequences that
fulfil two main objectives. On the one hand, a specific T cell activation
immunogen mRNA that focuses the response onto the most vulnerable
targets in the HIV viral proteome and on the other hand, a previously
tested stimulus (TriMix: a mixture of CD70+CD40L+caTLR4 mRNAs) for
appropriate activation of antigen presenting cells (DCs).
Methods: DCs were generated from peripheral blood monocytes
(MDDC) from chronically HIV infected patients by incubation with GMCSF and IL-4. These cells were electroporated with TriMix (15 µg) and/
or HIVACAT (20 µg) mRNA, with their respective controls. After that, DCs
were cocultured with autologous PBMCs for up to 6 days. In addition, the
maturation profile of MDDCs (CD80, CD83, CD86, CCR7) was analyzed
by FACS 24h after electroporation.
Functional analysis was performed using different techniques:
25-multiplex Luminex assay, T cell proliferation by CFSE and IFN-γ
ELISPOT at different time points.
Results: Increased expression of CD80, CD83 and CCR7 was observed
on MDDCs upon electroporation with TriMix mRNA. Functionally, mRNA
electroporated MDDCs were able to stimulate T cells from HIV-infected
individuals on cART in vitro. In fact, MDDCs electroporated with both HIV
antigens and TriMix, induced higher T-cell activation than their respective
separated components or whole AT2-inactivated virus in terms of both
IFNγ secretion and proliferation. Other Th1, Th2 and proinflammatory
cytokines showed a similar profile secretion pattern. Finally, a higher
proportion of stimulated CD8+ T cells, than of CD4+ T cells, was detected.
Conclusions: mRNA electroporation of MDDCs improved their
maturation status and was able to enhance HIV specific T cells responses.
Our results suggest that this mRNA combination could be considered for
a HIV therapeutic vaccination approach.
284
Background: Modern approaches of vaccination aim at targeting
dendritic cells (DC) with specific ligands and adjuvants. Electroporation
(EP), which was used to increase the expression of anti-SIV DNA vaccine
encoded antigens delivered in the skin, strongly improved the specific
immune response in non-human primates (NHP). The aim of this work
was to study the role of local DC, in the induction of this particular
immune response.
Methods: auxoGTU-multiSIV DNA plasmid or PBS was injected
intradermally and associated with EP at vaccinated sites in NHP. Skin
biopsies of injection sites were performed at 24h, 72h and 8 days after
treatment for histology or cell extraction and flow cytometry analysis.
Skin DC, stained with an AlexaFluor 488 labeled anti-HLA-DR antibody,
were imaged with noninvasive in vivo fibered confocal fluorescence
microscopy or monitored continuously during 24h by time-laps confocal
videomicroscopy on skin explants.
Results: EP associated to intradermal injection of the DNA vaccine
induced a recruitment of granulocytes and inflammatory monocytes/
macrophages in epidermis and dermis, as well as a population of
inflammatory dendritic epithelial cells. In epidermis, 24h after treatment,
we observed an initial increase of Langerhans cells (LC) with an upregulation of HLA-DR, CD86 and CD83, demonstrating their maturation,
followed by a decrease of LC number, suggesting a migration to
draining lymph node. In vivo monitoring confirmed the cell mobilization
and showed an increase of DC velocity and displacement after EP. The
skin microenvironment analysis revealed a release of pro-inflammatory
soluble factors (MCP-1, IL-18, IL-15, IL-8) and anti-inflammatory
mediators (IL1RA and sCD40L) by 24h, all considerably enhanced in the
presence of the anti-SIV vaccine.
Conclusions: This study highlights new elements of cell activation
mechanisms in the skin and shows that EP induced a local inflammation
that led to the activation and mobilization of LC. This opens up new
possibilities for vaccine strategies.
P27.03
P27.04
Combinations of TLR4 and TLR7/8 Adjuvants
Administered via the ID or IN Routes
Generate Different Vaccine Antigen-specific
Immune Outcomes in Minipigs
Chimeric Nod2/TLR2 Ligand Amplifies HIV1 Gag p24-specific Mucosal and Systemic
Immune Responses after Sub Cutaneous
Immunization in Mice
Paul F. McKay1, Deborah F. L King1, Jamie F. S Mann1, Guillermo
Barinaga1, Darrick Carter2, Robin J. Shattock1
Capucine Phelip1, Vincent Pavot1, Nicolas Rochereau2, Eric
Perouzel3, Thierry Lioux3, Gérard Tiraby2, Charlotte Primard4,
Stéphane Paul2, Bernard Verrier1
Background: Previous studies have demonstrated synergy between
TLR4 and TLR7/8 stimulation with enhanced CD4 T cell cytokine
production that may augment CD4 dependent Ab generation. In
minipigs we assessed the immune outcome using optimal agonist
TLR4/7/8 ligand adjuvant dosage or combination in a mixed ID and IN
route vaccination regimen.
Methods: Groups of pigs received CN54gp140 via the ID or IN route
with optimally titrated R848 or GLA adjuvants. A schedule of 3xIN+2xID
was compared to 3xID+2xIN, each inoculation given 3 wks apart.
Control animals were unadjuvanted. Pigs were sampled weekly and Agspecific humoral responses assessed by ELISA, avidity and neut assays.
Results: Combinations of R848 and GLA had an additive enhancing
effect when used ID but surprisingly GLA abrogated R848-induced
immunity after IN inoculation. Therefore, ID injections included both
R848 and GLA, IN only R848. Optimized adjuvant dose/combinations
significantly enhanced Ag-specific responses when administered via
either route. At the end of each regimen both groups exhibited similar
quantities but qualitatively different serum Ag-specific Ab. The ID route
vaccinations elicited sera of significantly higher avidity (p=0.042, week
5 - 2 weeks post 2nd vaccination), viral neutralising capacity and mucosal
IgG (p=0.006, week 5). Conversely, serum and mucosal IgA responses,
although low, were significantly higher in the 3xIN primed group (p<
0.05, week 5).
Conclusions: These data begin to address important issues relating to
adjuvant combinations and routes of administration. TLR4 and TLR7/8
agonists combined to enhance Ag-driven immune responses but only
after ID vaccination. The inhibitory effect of GLA on R848-driven
responses after IN inoculation suggests an active contribution by GLA
rather than a simple formulation issue, as adjuvant co-formulation did
not inhibit R848 responses after ID vaccination. Further investigation
of adjuvant combinations in responsive animal models will aid
development of effective vaccines.
Institut de Biologie et Chimie des Protéines, LBTI, UMR 5305 CNRS/
Université de Lyon, Lyon, France, 2Groupe Immunité des Muqueuses
et Agents Pathogènes - INSERM CIC 1408 Vaccinologie, Faculté de
Médecine de Saint-Etienne, Saint-Etienne, France, 3CAYLA - InvivoGen,
Toulouse, France, 4ADJUVATIS, Lyon, France
1
Background: Toll-like receptor (TLR) ligands are critical activators
of innate immunity and are being developed as vaccine adjuvants.
However, their potential synergistic effect in conjunction with other
Pattern Recognition Receptor (PRR), like Nod-like receptor (NLR) agonists
remains poorly studied. In this study, we evaluated both in vitro and
in vivo the adjuvant role of a chimeric Nod2/TLR2 ligand when co
delivered with poly(lactic acid) (PLA) biodegradable nanocarriers, loaded
with HIV-1 Gag p24.
Methods: Chimeric Nod2/TLR2 ligands have been chemically
synthesized and compared with their separate counterparts, Nod2 and
TLR2 with regards to their capacities to induce maturation and proinflammatory cytokines of Human Monocytes derived Dendritic cells.
After sub cutaneous administration of each ligand, co-delivered with
PLA-p24 particles, either alone or in combination, p24-specific Th1/Th2
responses have been evaluated in balb-C mice both at systemic and
mucosal levels (IgG, IgA, Elispot analysis).
Results: In vitro chimeric Nod2/TLR2 ligand induced synergic and strong
up-regulation of maturation markers (CD80, CD83, CD86, DC-LAMP), costimulatory molecules at the DCs surface and proinflammatory cytokines
secretions (IL-1 beta, IL-6, IFN alpha, IFN gamma, TNF alpha) compared
to separate ligands. Interestingly enough, sub-cutaneous administration
of PLA nanoparticles carrying Gag p24 co-administered with Nod2/TLR2
chimera induced strong Ag-speciﬁc IgA and IgG antibody titers both at
systemic and mucosal sites as strong as Cholera Toxin positive controls.
Conclusions: These data demonstrate that targeting both TLR and Nod
pathways with a chimeric ligand such as Nod2/TLR2 could induce strong
mucosal immune responses after sub cutaneous administration when
co-delivered with PLA biodegradable particulate vector. These results
point out the interest of this new class of chimeric molecules as HIV
vaccine adjuvants to induce both strong systemic and mucosal immune
responses after parenteral administration.
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285
POSTERS
Imperial College, Department of Infectious Diseases, Division of
Medicine, London, United Kingdom, 2Infectious Disease Research
Institute, Seattle, WA, United States
1
Posters
P27.05
P27.06
Antigen Formulations and Routes of
Immunization Drive the Magnitude and
Quality of HIV-specific Mucosal Immune
Responses in Mice
Alarmin IL-33 Acts as an Immunoadjuvant to
Enhance Antigen-specific anti-viral Immunity
Thomas Vazquez1, Léa Torrieri-Dramard1, Fabien Pitoiset1, James
Vigneron1, David Klatzmann1, Bertrand Bellier1
I3 Laboratory - INSERM U 959 - GHPS Paris, Paris, France
1
POSTERS
Background: Despite considerable efforts, the development of a
successful HIV vaccine remains challenging. Current attention is mainly
focused on developing strategies to enhance the immunogenicity of
vaccine formulations and the induction of mucosal immunity.
Methods: Here, we investigated the influence of particulate form
of antigen and the route of administration in the induction of HIVspecific immune responses and mucosal immunity. We evaluated
the immunogenicity of HIV-gp140 antigen expressed as proteins or
recombinant Virus-Like Particles (VLPs) and compared different mucosal
(nasal (IN), rectal (IRec), vaginal (IVag)) and parenteral (intradermal (ID),
subcutaneous (SC)) routes of immunization, or their association in primeboost strategies.
Results: We show that the antigen formulation and route of immunization
are critical factors governing the vaccine efficacy. We observed that
DNA vaccine that express HIV-gp140 on VLPs induced higher levels
of immune responses than standard DNA expressing non-particulate
antigens, especially at mucosal level after IN DNA administration.
Similarly, we observed that VLP boost, when administrated by IN or IVag
routes, improved T cell multifunctionality at systemic and mucosal levels
as compared to protein boost.
When mucosal and parenteral immunization were compared, we show
that IgA and IgA-secreting cells were only induced in mucosal tissues
when DNA priming was done by IN route, but not by ID. We also compared
IVag and IRec VLP boosts after IN DNA primes and demonstrated that
both increased antigen-specific T- and B-cell responses in systemic and
local compartments, inducing HIV-specific IgA-secreting cells at proximal
mucosa to the site of immunization.
Conclusions: Taken together, our results showed the importance of the
particulate formulation of the antigen, such as VLPs, and the role and
the interest of mucosal routes of immunization to improve local immune
responses.
286
Daniel Villarreal1, Megan Wise1, Jewel Walters1, Emma Reushel1,
Min Joung Choi2, Nyamekye Obeng-Adjei1, Jian Yan3, Matthew
Marrow3, Niranjan Sardesai3, David Weiner1
University of Pennsylvania, Philadelphia, PA, United States, 2Korea
Food and Drug Administration, Osong-eup, Korea, Democratic People`s
Republic of, 3Inovio Pharmaceuticals, Inc., Blue Bell, PA, United States
1
Background: DNA vaccines are an important approach for the
generation of humoral and cellular immune responses in vivo. We
have recently reported that combining an IL-12 expression vector along
with HIV DNA encoded antigens can induce robust T cell responses in
the clinic (HVTN 080). Identification of novel adjuvants at the innate
to adaptive immune border will likely provide further improvements
in DNA vaccine potency. In this regard we have recently developed
a novel alarmin IL-33 as a new immunoadjuvant in the synthetic EP
delivered DNA vaccine platform. We hypothesized that this endothelial
cell alarmin would be in a particularly effective position to transition the
adaptive immune response after viral infection.
Methods: Mice were immunized with immune plasmid adjuvant IL33 in
combination with DNA followed by EP two to three times at three week
intervals.One week post final vaccination, spenocytes were harvested to
investigate cellular responses.
Results: We report that IL-33 is capable of enhancing potent antigen
(Ag)-specific effector and has a similar robust effect on enhancing
memory T cell immunity in vivo in a DNA vaccine setting. Additionally,
IL-33 augmented vaccine-induced Ag-specific polyfunctional CD4+ and
CD8+ T cell responses. Importantly a large proportion of CD8+ T cells
undergo cytolytic plurifunctional degranulation that is improved over
DNA vaccine alone. We show that IL-33 can particularly expand the
magnitude of Ag-specific CD8+ T cell responses and elicit stronger and
longer lived effector-memory CD8+ T cells. For HIV antigens, both the
magnitude as well as the phenotype of T cells induced by the IL-33 DNA
vaccine are improved.
Conclusions: We have now developed a Rhesus IL-33 genetic adjuvant
for studies in NHP which allow for more detailed studies including
challenges to be performed. These studies will be informative for HIV
vaccine development and lay the groundwork for this novel genetic
adjuvant for study in the HIV vaccine setting.
Posters 28: Behavioral and Social Sciences
P28.01
Does a Weighted Analysis Make any
Difference in the Estimates from a Sample
Survey?
Rajatashuvra Adhikary1, Prabudhyagopal Goswami2, Mandar
Mainkar3
FHI 360, Washington DC, DC, United States, 2FHI 360, New Delhi,
India, 3National AIDS Research Institute, Pune, India
1
POSTERS
Background: Self-weighted sampling designs are not always feasible
in sample surveys. Sample weights are calculated and applied to have
unbiased estimates. However, it is important to understand the extent of
difference between weighted and un-weighted estimates. We examined
these differences among men who have sex with men (MSM) participants
using data from an integrated bio-behavioral survey conducted in 2009
in four districts of Andhra Pradesh, India.
Methods: Two-stage time location cluster sampling approach was used
to recruit 1,608 (around 400 from each district) men aged >18 years
who had sex with another male in one month prior to survey. Sample
size was calculated based on 95% confidence level, 90% power, and
design effect =1.7. Consented participants provided behavioural data in
structured questionnaires and blood and urine specimens. Specimens
were tested for HIV and syphilis. Multivariate analysis was performed
to determine correlates of CCU and HIV prevalence adjusted for sociodemographic variables.
Results: Weighted and un-weighted estimates of consistent condom
use (CCU) with regular male partners, paying male partners, and casual
male partners, HIV prevalence, and syphilis prevalence varied across
the four districts. The absolute difference between the weighted and
un-weighted estimates ranged from 0.7% to 13.2% for CCU and from
0.2% to 5.3% for HIV and syphilis. Z-scores show a significant difference
between weighted and un-weighted estimates in many cases. Weighted
and un-weighted multivariate analysis shows opposite trends in the
adjusted odds ratio (AOR) in few cases and relatively large differences in
the significance level (p-value).
Conclusions: Weighted analysis for a probability-based sample survey
provides unbiased population estimates whereas un-weighted estimates
are representative of the sample. If possible, a self-weighted sampling
design should be adopted, allowing for unbiased estimation for the
sample and population.
www.hivr4p.org
287
Posters
Posters 29: Circumcision and Acceptability
P29.01
P29.02
A Qualitative Exploration of Medical Male
Circumcision among Young Men from
Whizzkids United, Edendale, Pietermaritzburg
Assessing the PrePex™ Device for Voluntary
Medical Male Circumcision in South Africa
Rusha Govender , Janan Dietrich , Jenny Coetzee , Marcus Mc
Gilvray1, Douglas Wilson1
1
2
2
Edendale Hospital, Pietermaritzburg, South Africa, PHRU,
1
2
POSTERS
Background: In Africa male medical circumcision (MMC) has been
shown to reduce HIV acquisition by up to 60%. WHO/UNAIDS
recommendations emphasize that male circumcision should be
considered an effective intervention for HIV prevention in countries
and regions with heterosexual epidemics, high HIV and low male
circumcision prevalence. Mathematical models based on the results of
randomized controlled trials suggest that medical circumcision could
avert two million HIV infections over the next 10 years; however it
is unclear whether medical circumcision would decrease the overall
prevalence of HIV infection when delivered outside the context of the
controlled environment of a randomized trial.
This study was conducted at WhizzKids United (WKU) based at Edendale
Hospital, Kwa-Zulu Natal. WKU is a grassroots South African nonprofit organization that aims to empower and inform high risk youth
about health care and HIV/AIDS prevention. The MMC service facility
at Edendale Hospital performs around 200 circumcisions a month.
WhizzKids United has made the uptake of MMC services more accessible
for adolescents.
Methods: Thirty-one in-depth Interviews and eight focus group
discussions (FGDs) were conducted with young males aged 14-24 years.
Fifty percent of the young males were uncircumcised and fifty percent
reported being circumcised. There were two focus group discussions per
age group were held. One of these consisted of circumcised individuals
and the other consisted of uncircumcised individuals. Thematic analysis
was used for data analysis.
Results: The global themes identified from the uncircumcised
participants included,
(1) medical,
(2) peer pressure,
(3) decision-making,
(4) sexual risk-taking behaviours.
The global themes identified for circumcised participants included,
(1) medical,
(2) traditional,
(3) Becoming a man and
(4) sexual risk- taking behaviours.
Conclusions: The overall findings suggest that young men in Kwa-zulu
Natal are accepting of medical male circumcision.
288
Limakatso Lebina1, Noah Taraburekera2, Minja Milovanovic1,
Nkeko Constance Tshabangu1, Neil Martinson1
Perinatal and HIV Research Unit, Johannesburg, South Africa,
Population Services International, Represented in South Africa by
Society for Family Health, Johannesburg, South Africa
1
2
Background: Voluntary medical male circumcision has been proven
to reduce the risk of acquiring HIV by at least 60% in men who are
circumcised. South Africa is scaling-up medical male circumcision, and
the number of circumcised men is increasing. However, if circumcision
could be safely simplified without compromising efficacy, it could
potentially allow for more circumcisions to be done. The PrePex device
was developed to simplify the circumcision procedure by making sutures,
diathermy, scalpels, and local anesthesia unnecessary. However, there
is no data available on the assessment of the PrePex device in South
Africa.
Methods: A phased, multisite, non-randomised study in which
adolescents (14-17 years) and adult (18-45) men underwent circumcision
using the PrePex device. Data were collected from 9 visits; application,
2 telephonic follow-ups, removal and 5 follow-up visits. Outcome
measures include adverse events, pain and discomfort, procedure time,
sexual resumption, healing time, and PrePex sizes. Data presented is of
the first 318 participants.
Results: A total of 264 adult men and 54 adolescents were recruited for
the PrePex study across the three sites. The overall moderate and severe
adverse events (AE) rate was 2.5% with none of the adverse events
requiring hospitalisation. Mild AE were predominantly obstructed urine
flow. The most commonly used PrePex sizes were size B and C (63%,
197/312). The median resumption for sexual activity was 44 days (IQR:
36.5-53). The device application procedure was quick and caused minor
to no pain however the removal procedure was more painful but the
pain was temporary.
Conclusions: The preliminary results indicate that the PrePex procedure
was easily learnt by the staff at the three sites. The device application
is quick, safe and less painful however removal causes temporary
pain. The AEs encountered were similar in severity to the blade based
circumcision with the exception of urinary obstruction.
P29.03
P29.04 LB
Assessing the Acceptability of the PrePex™
Device for Voluntary Medical Male
Circumcision in South Africa
Does Circumcision Offer Some Protection
from HIV for Men who Have Sex with Men? A
Cross-sectional Study in China
Minja Milovanovic1, Noah Tarabureka2, Nkeko Constance
Tshabangu1, Mmatsie Manentsa1, Neil Martinson1, Limakatso
Lebina1
Han-Zhu Qian1, Yuhua Ruan2, Yu Liu1, Douglas F. Milam1,
Hans M.L. Spiegel3, Lu Yin1, Dongliang Li4, Yiming Shao2,
Sten H. Vermund1
Perinatal and HIV Research Unit, Johannesburg, South Africa,
Population Services International, Represented in South Africa by
Society for Family Health, Johannesburg, South Africa
Vanderbilt University, Institute for Global Health, Nashville, TN,
United States, 2China CDC, State Key Laboratory for Infectious Disease
Prevention and Control, NCAIDS, Beijing, China, 3NIAID, NIH, Henry M.
Jackson Foundation - Division of AIDS, Bethesda, MD, United States,
4
Chaoyang District CDC, Beijing, China
2
Background: Following the recommendation by the World Health
Organisation, medical male circumcision is being scaling-up in South
Africa. Circumcision reduces the risk of acquiring HIV by 60%. Most
circumcision facilities in South Africa use the surgical procedure with
very little use of devices. Previous studies have shown that PrePex is a
quick, easy and acceptable method of circumcision. However, there is
no information on the acceptability of the PrePex device in South Africa.
Methods: A phased, multisite, non-randomised study in which
different individuals across 11 sites in South Africa, were approached
to answer a structured acceptability questionnaire: PrePex users, people
accompanying men coming for circumcision, men refusing PrePex, and
healthcare workers who worked with PrePex and those that would work
with it if it were rolled out. A total of 6 questionnaires were used to
explore: cultural acceptability, trust, recommendation of PrePex and
perceived advantages and disadvantages.
Results: A total of 439 participants, across the different groups, were
recruited for the PrePex acceptability study. The PrePex device was
considered culturally acceptable by 86% of PrePex users, 60% of the
men who refused PrePex and 67% of accompanying. The device was
also highly trusted by PrePex users (99%), accompanying persons (95%)
and 55% of refusals indicated they trusted it. When asked to explain
majority of the responses include perceived safety compared to the
blade procedure. Overall, there was acceptance of the PrePex device and
procedure by healthcare workers, with many respondents expressing
that it is an alternative to traditional circumcision. Furthermore, there
was a strong expression that if nurses were to do PrePex there should be
a certificate from a relevant body.
Conclusions: The results indicate that the PrePex device and procedure
is considered relatively acceptable and there appear to be no major
social barrier to implementation however, cultural differences were
noted.
1
Background: The observational literature on the association of male
circumcision and odds of HIV among men who have sex with men
(MSM) is often limited by a failure to definitively ascertain circumcision
status and/or by a failure to carefully assess sexual position preferences
(insertive, mixed/versatile, or receptive). No randomized clinical trials of
circumcision have been conducted among MSM. We conducted a crosssectional study of MSM in Beijing, China to assess associations, if any, of
circumcision and HIV, controlling for anal sexual positioning role.
Methods: MSM were recruited both from the community and from HIV
clinics. Circumcision status was evaluated by genital examination by
staff trained by an expert urologist. Anal sexual role was assessed by
questionnaire interview. Associations of circumcision status with HIV
infection were assessed through multivariate logistic regression analysis.
Results: The odds of HIV infection by circumcision status and preferred
anal sexual role in 1053 MSM were as follows:
• Receptive/versatile/uncircumcised (n=541, 177 HIV+/ 364HIV-)
REFERENCE GROUP
• Receptive/versatile/circumcised (n=47, 9 HIV+/ 38HIV-) aOR=0.48
(0.22, 1.02)
• Insertive/uncircumcised (n=434, 76 HIV+/ 358HIV-) REFERENCE
GROUP
• Insertive/circumcised (n=31, 2 HIV+/ 29HIV-) aOR=0.35 (0.13, 1.10)
The OR was adjusted for age, ethnicity, marriage, education, occupation,
and Beijing vs. non-Beijing legal residency.
Conclusions: Circumcised MSM were less likely to have acquired
HIV, especially among men practicing predominantly insertive anal
intercourse, with effect sizes comparable to those seen for heterosexual
men in Africa. However, findings may have been due to chance, given
upper bound 95% CI for aOR > 1.0. Circumcision needs clinical trials
evaluation as a potential global HIV prevention strategy for MSM. The
HIV Prevention Trials Network is developing a feasibility study (HPTN
079) for such a trial.
www.hivr4p.org
289
POSTERS
1
Posters
P29.05 LB
P29.06 LB
HIV Is the Primary Exclusion Criterion
in a PrePex™ Male Circumcision Device
Introductory Study in Mozambique
Women’s (Mis) Understanding of Male
Circumcision: A Mixed Methods Study in
Zambia
Mehebub Mahomed1, Beverley Cummings2, Jotamo Come3,
Bossemeyer Debora1, Thais Ferreira1, Leonel Nhambi1, Edgar
Necochea4, Humberto Muquingue1, Kelly Curran4
Nicole A. Haberland1, Christine A. Kelly1, Drosin M. Mulenga2,
Paul C. Hewett2, Barbara S. Mensch1
Jhpiego, Maputo, Mozambique, 2Centers of Disease Control and
Prevention/Mozambique, Maputo, Mozambique, 3Ministry of Health,
National Programme for VMMC Expansion, Maputo, Mozambique,
4
Jhpiego, Baltimore, MD, United States
1
POSTERS
Background: Voluntary medical male circumcision (VMMC) reduces
female to male HIV transmission by approximately 60% and is
recommended by WHO and UNAIDS as a priority intervention in high
HIV prevalence settings. In Mozambique, VMMC for HIV prevention
started in 2009; more than 300,000 males were surgically circumcised
by March 2014; the goal is 2 million by 2016. PrePex™ could potentially
reduce procedure time and increase acceptability of VMMC because
it does not require injectable anesthesia or suturing. In 2013 an
introductory study of the PrePex™ device was conducted in Maputo,
Mozambique to assess the acceptability among providers and clients.
Methods: Adult clients presenting for VMMC at the study site were
offered surgical or PrePex™ circumcision. Those who preferred PrePex™
were screened for inclusion criteria. Exclusion criteria were recorded. The
current WHO guidelines exclude HIV+ men from device circumcision.
Results: During the study, 752 clients aged 18 or older presented for
VMMC and were offered the choice of PrePex™ or conventional surgery;
116 (15.4%) preferred surgical VMMC. Of the 636 clients who chose
PrePex™, 132 (20.8%) were ineligible. HIV infection was the primary
reason for exclusion, restricting 85 (64%) interested HIV+ clients.
Phimosis or narrow foreskin was present in 17 (13%) of the ineligible
clients. Sixteen clients (12%) were unable to communicate in Portuguese
and 8 (6%) lacked communication means (cell phone); both were study
requirements. The remaining 6 (5%) were excluded due to active STI,
sexual dysfunction or previous penile surgery.
Conclusions: One third of adult clients offered PrePex™ either did not
want device circumcision or were ineligible under current guidelines
that exclude HIV+ men. An integrated program offering both device and
surgical VMMC remains the best service delivery option. However, there
is need to assess the safety of PrePex™ among HIV positive clients, as
HIV testing in Mozambique is recommended but not required in routine
VMMC service delivery.
290
Population Council, New York, NY, United States, 2Population Council,
Lusaka, Zambia
1
Background: Randomized controlled trials have demonstrated that
male circumcision (MC) reduces men’s risk of heterosexual acquisition of
HIV, but there is no evidence that individual women benefit directly from
MC. Women’s understanding of the protection afforded - or not - by MC
against HIV and STIs has important implications for risk compensation
and demand.
Methods: This mixed methods study explores the prevalence, depth,
and correlates of women’s understanding of four dimensions of MC
knowledge: awareness of MC, knowledge that MC partially protects
males against HIV, knowledge that MC reduces males’ STI risk, and
knowledge that MC has no effect on females’ HIV risk if she has sex with
a man who is circumcised. We combine data from the first two rounds
of a longitudinal study of Zambian women aged 15-29 (n=933) with indepth interviews conducted among a subsample of respondents (n=45).
Results: Although awareness of MC was high - 77% of women at
baseline reported having heard of MC before it was described in the
interview - women lacked more nuanced knowledge of MC’s protective
effects. In both the quantitative and qualitative surveys, a disconcerting
proportion of women incorrectly believed that MC fully protects men
from HIV and STIs, and that MC similarly offers women partial - or even
complete - protection. Multivariable analysis showed that more highly
educated and wealthier women were better informed about MC than
their more disadvantaged peers.
Conclusions: Efforts are needed to increase understanding about
the limits of MC protection, particularly among more marginalized
women who may have difficulty accessing accurate information about
MC or harbor misperceptions about its protective effects. Messaging
surrounding MC should also more explicitly address women’s needs.
Health professionals should emphasize that, because MC provides men
with only partial protection against HIV and some STIs, women are
still at risk regardless of their partner’s circumcision status. Condoms
remain critical.
Posters 30: Condoms: Attitudes, Use and How to Increase
P30.01
P30.02
Behavioral Attitudes towards the Use of Male
Condoms as a Means of Preventing the HIV/
AIDS Spread
Why Young MSM Do Not Use Condoms
Consistently: A Qualitative Exploration
, Denis Sinzinkayo
3
Yowli Burundi, Bujumbura, Burundi, 2University of Burundi, Medicine,
Bujumbura, Burundi, 3Health Healing Network Burundi, Bujumbura,
Burundi
1
Background:
Assess knowledge of condom use among adolescents of secondary
school,
• Evaluate the cultural and/or religious impact on the use of
condoms in adolescents and Improve the use of condoms as a
means to fight against HIV/AIDS, STIs and undesired pregnancies.
Methods: The first round consisted of conducting a direct interview
towards adolescents with data collection on a questionnaire. The second
time, we gave a survey form to fill out and we compared the results of
the interview with those of the survey form. Close monitoring helped to
yield tangible results.
Results: Of 256 adolescents, 123 were Catholics, 61 were Protestants,
and 38 were Muslims while 34 were pagans. The extremes of age were
13 and 22 with an average age of 13.27. The sex ratio M/F was 4/3. Of
256 adolescents, 88 were from poor settings with a precarious level of
cultural belief. Of them, 41 agreed that they use condoms regardless
of religious and cultural barriers; 48 agreed that they can never use
condoms due to the religious Barriers; 167 admitted that the condom
does not have the same sexual pleasure and so do not use it. Of 256, 17
have already had genital problems and have consulted the doctor. The
study was able to show that most Protestants deny the condoms and
preach abstinence; Muslims willingly use condoms while the Catholics
are conservative.
Conclusions: Religious beliefs affect condom use among adolescents
while culture has no great impact. It is necessary to improve sexual
practices in the adolescents´ environment to prevent the spread of
HIV/AIDS and other STIs, unwanted pregnancies, etc. and promote
educational achievement.
•
Tareerat Chemnasiri1, Anchalee Varangrat1, Supaporn
Chaikummao1, Anupong Chitwarakorn2, Timothy H. Holtz1,3
Thailand MOPH - U.S. CDC Collaboration, Nonthaburi, Thailand,
Thailand Ministry of Public Health, Department of Disease Control,
Nonthaburi, Thailand, 3Centers for Disease Control and Prevention,
Division of HIV/AIDS Prevention, Atlanta, GA, United States
1
2
Background: High HIV incidence and inconsistent condom use were
found among young (18-24 year-old) men who have sex with men
(YMSM) in a recent Bangkok cohort study (8/100 person-years). We
explored reasons behind inconsistent condom use among YMSM using
qualitative methods.
Methods: Eight focus group discussions and ten key informant
interviews were conducted between June 2012 and June 2013.
Individuals were randomly selected by age and history of risk behaviors
from MSM participating in a five-year cohort study in Bangkok, Thailand.
We collected socio-demographic and behavioral data using a short
questionnaire. Group discussions and interviews were audio-recorded,
transcribed, and analyzed using Atlas.ti version 7.17.
Results: All 47 participants were Thai MSM, 18 to 24 years of age,
who were asked to discuss issues related to inconsistent condom
use. Participants expressed feeling an unnatural sensation while
using a condom, having a latex allergy, and feeling pain from sexual
penetration during anal intercourse which reduced sexual pleasure and
sensitivity of penile erection and orgasm. Some participants reported
difficulty finding extra-large or extra-small condoms. Fear of rejection by
partners and lack of condom negotiation skills were noted by receptive
sexual role participants. Individual sexual preference, sensation seeking
behavior, and overwhelming sexual desire were perceived to promote
non-condom use. Being out of condoms and not carrying a condom at
all times were also expressed as promoting inconsistent condom use.
Participants believed not using condoms with a steady partner to be
a sign of mutual trust. Lack of awareness and misconceptions of HIV
transmission caused YMSM to overlook using safe sex practices.
Conclusions: HIV awareness, condom negotiation skills, attitude
change about condom use, and same-sex education are needed for
YMSM in Bangkok. Condom use with steady partners should be strongly
emphasized. Variety of condom size and allergy-free condoms are
needed for this population.
www.hivr4p.org
291
POSTERS
Patrick Bitangumutwenzi
1,2,3
Posters
P30.03
P30.04
Exploring the Impact of Social Marketing
of Female Condoms in the City of Kumba,
Cameroon. 2010-2011
Changes in Sexual Behaviour and HIV
Prevalence among Married Fishermen along
Lake Victoria at two Time Points: A Scorecard
for Prevention Efforts
Alemju Fontu1,2
Association Camerouniase pour Marketing Social, Kumba, Cameroon,
Forestry Conservation and Environmental Society, Kumba, Cameroon
1
2
Zachary Arochi Kwena1, Isaac Mwanzo2, Chris Shisanya3, Lilian
Achiro4, Norton Sang4, Elizabeth Bukusi4
Kenya Medical Research Institute, Center for Microbiology Research,
Kisumu, Kenya, 2Kenyatta University, Community Health, Nairobi,
Kenya, 3Kenyatta University, Geography, Nairobi, Kenya, 4Kenya
Medical Research Institute, Center for Microbiology Research, Nairobi,
Kenya
1
POSTERS
Background: Female condoms are barrier contraceptive methods that
provide protection against conception and acquisition of STIs including
HIV. These barriers which are used and controlled largely by females are
said to provide autonomy and empowerment over other contraceptive
choices.
Methods: The project implemented the following interventions.
Conducted community sensitization campaigns to increase awareness
and benefits of using female condoms.Recruited and trained volunteer
peer educators on interpersonal communication skills, benefits and use
of female condoms.Peer educators go out twice a week to strategic
locations such as the street sides, market places and other public hubs
to educate people on the benefits and correct usage of female condoms
Results: At the end of these interventions conducted from January 2010
to December 2011, 158, 650 persons were reached (Youths 60,000
(37.8%), Women 88,400 (55.7%) and Men 10,250 (.6.5%).
Correct use of the female condoms (FC2) was demonstrated to an
audience of 9,230 [Men 2200 (23%), Women 3,680 (39.8%) and
Youths 3,350 (36.2%)].
Eighty sale points were established at strategic locations in each
community and 172,000 (FC2) were sold by peer educators. 120,400
and 51,600 females and males respectively bought the (FC2).
90% of the persons interviewed disclose seeing a female condom for
the first time during our campaigns and all females who bought the
(FC2)promise to adopt the (FC2) as their contraceptive choice.
Conclusions: Outcome of these interventions reveal that most females
prefer (FC2)to male condoms. The government and other international
donors should support the distribution of (FC2)in order to scale up
prevention of HIV/AIDS in sub-Saharan Africa
292
Background: There are considerable efforts towards reducing new HIV
infections in key affected populations such as fishermen. Assessing
changes in sexual behaviour and HIV prevalence is an important
feedback to these prevention efforts. We evaluated changes in sexual
behaviour and HIV prevalence among married fishermen in fish-landing
beaches in Kisumu County, Kenya.
Methods: We analyzed data from two surveys conducted in 2005/6
with 164, and 2011/2 with 545 married fishermen to evaluate changes
in their sexual behaviour and HIV prevalence at two time points. The
participating fishermen in both surveys were randomly sampled from all
33 fish-landing beaches in Kisumu County. The numbers sampled from
each beach were proportional to the population size of the beaches. In
both surveys, we collected data on socio-economic, sexual behaviour
and HIV sero-status.
Results: A higher proportion of fishermen in 2011/2 survey compared
to 2005/6 survey reported drinking alcohol before sex with extramarital partners (27.7% versus 11.4%; p=0.05) and being involved
in transactional sex (65.8% versus 25.0%; p< 0.01). However, more
fishermen in 2011/2 compared to 2005/6 survey used condoms with
extra-marital partners (34.2% versus 5.4%). Overall HIV prevalence
in 2011/2 survey was marginally lower compared to 2005/6 (21.0%
versus 28.0%; p=0.07). However, there was significant 15 percentage
point drop in HIV prevalence among fishermen below 25 years old that
represent recent infections.
Conclusions: Despite increases in other high risk sexual behaviours,
condom use with extra-marital partners in this HIV key affected
population increased explaining significant drop in HIV prevalence
among youth who represent recent infections.
P30.05
P30.06
Repetitive Risk Reduction Counseling on
Condom Use among HIV Exposed Seronegative (HESN) Persons in Jos, Nigeria
Acceptability and Ease of Use of New Female
Condom Designs among Women Attending
an Urban Reproductive Health Clinic in
Institute of Human Virology, Abuja, Nigeria, 2Plateau State Human
Virology Research Centre, Jos, Nigeria, 3Institute of Human Virology,
University of Maryland, School of Medicine, Baltimore, MD, United
States
1
Background: Condom is the commonest HIV preventive strategy. Over
time, the successes of this strategy have been challenged with several
socio-cultural factors such as male dominance over sexual negotiations;
feeling of mistrust among spouses over condom use and wanting to
achieve pregnancy especially in the African continent. We document
here condom use during a 2-year follow up among HIV exposed seronegatives in a discordant relationship after consecutive risk reduction
counseling in preparation for a future HIV prevention trial in Nigeria
Methods: We conducted a prospective cohort study and followed up
534 HESN partners in established sero-discordant relationship (i.e. at
least 3 months). Relevant ethical approvals and informed consent were
obtained. We provided risk reduction counseling for 10-12 minutes with
emphasis on: importance and proper use of condoms along with free
condoms; the need to watch for symptom of STIs and request immediate
treatment; and the benefit of their HIV+ partner attaining viral
suppression and elevated CD4 count before achieving pregnancy and
thereafter administered standardized questionnaires on risk behavior.
Clinical examinations were done and samples collected for rapid HIV
test and safety labs
Results: 534 enrollees were eligible for 10 follow-up visits with a mean
age of 36 years (19-65years). 257 (48.1%) were female and 277 (51.9%)
males. A total of 7 individual based risk reduction counseling sessions
were provided. About 60% of our female participants are within the
child bearing age (i.e. 19-35years) which explains why these are aspiring
to achieve pregnancy. More so, this group accounts for the 52% who
inconsistently use condom. Nonetheless condom use increased from
40% at baseline to 48% at visit 7 with only 5.8% of females achieving
pregnancy
Conclusions: Among HESN in a marital relationship, repetitive risk
reduction counseling improved the use of condom which highlights the
need for combine HIV preventive strategies considering their significant
exposure to HIV
Mags E. Beksinska1, Jennifer A. Smit1, Greener Ross1, Busi V.
Maphumulo1, Nonhlanhla Mphili1, Sthe Chonco1
MatCH Research, Obstetrics and Gynaecology, Durban, South Africa
1
Background: The availability of new female condom (FC) designs,
which may improve FC acceptability and affordability, will increase
options for couples who choose FCs as their contraceptive and/or
disease prevention method. The acceptability of 2 new products: HLL
FC and Cupid 2 (smaller version of the existing Cupid FC) compared to
the currently available FC2 was evaluated as part of a trial assessing
the functional performance of the 2 new FCs. Here we present the
acceptability results of the trial.
Methods: This randomized, comparative cross-over clinical trial of
3 FCs was conducted among 300 women in an urban reproductive
health clinic in Durban, South Africa. Interviewer- assisted surveys
were employed during 3 follow-up visits to gather data on comparative
acceptability. Numbers and percentages of women in each category of
acceptability were calculated.
Results: In total, 277 women (92.3%) have completed the study to date
using all 3 FC types. Mean age was 27.3 years and 22% had previously
used FCs. Of the total sample entered and analysed to date (n=113), over
80% of women liked all FC types ‘very much’ or liked them ‘somewhat’
and over 90% said each type was comfortable to use For individual
features there were minimal differences between the condoms however
two-thirds of women (66.0%) liked “very much” the pink colour of
Cupid2 compared to 53% of the FC2 and HLL FCs. Although over a third
of women 38.9% found any FC type “difficult to insert but improved with
practice” at first follow-up, by the last follow-up visit only a fifth (19.5%)
reported this, with “very easy to insert” any FC increasing from 9.7% to
37.2% by last visit.
Conclusions: Results from this study show that regardless of FC type,
ease of use increased across each follow-up visit. Although women
expressed preferences for different FC features, acceptability of all FCs
overall was high. A greater range of FCs will provide women with more
choice of protection. Data for the full sample of 300 women completing
the study will be presented.
www.hivr4p.org
293
POSTERS
Evaezi Okpokoro1, Sophia Osawe1,2, Ruth Daitiri2, Grace
Choji2, Stephen Umaru2, Felicia Okolo2, Pam Datong2, Alash’le
Abimiku1,2,3
Posters
Posters 31: Drug Transporters
P31.01
P31.02
Characterisation of Drug Transporter Gene
Expression in Colorectal Tissue and Cell Lines:
Induction with Anti-retrovirals for Microbicide
Optimization
Expression of Drug Transporters in
Cervicovaginal Cell Lines and Modulatory
Effect of Candidate Anti-retroviral
Microbicides
Indrani Mukhopadhya1, Susan Berry1, Emad M. El-Omar1, John
Thomson1, Georgina L. Hold1, Karolin Hijazi1
Kieron A. Smith1, Indrani Mukhopadhya1, Susan Berry1, Georgina
L. Hold1, Karolin Hijazi1
University of Aberdeen, Aberdeen, United Kingdom
1
POSTERS
Background: Drug transporter expression in the colorectal epithelium is
likely to play a role in the mucosal disposition of anti-retrovirals (ARVs) in
rectal microbicidal preparations and impact their efficacy in prevention
of HIV-1 infection. However, there is limited information on expression
levels available. This study assessed expression of 84 drug transporter
genes in human colorectal tissue and representative cell lines pre and
post ARV exposure.
Methods: Drug transporter mRNA expression was quantified from
colorectal biopsies (n=12) and 6 colorectal cell lines using real time PCR.
Relative mRNA expression was quantified in CaCo-2 cells after induction
with tenofovir (TFV; 1000 µM) and dapivirine (DPV; 10 µM) for up to
3 days. Data was analysed using Pearson’s correlation (r), hierarchical
clustering and principal component analysis.
Results: Fifty-eight of the 84 transporters were expressed in colorectal
tissue. SLC28A2/CNT2 was the most expressed uptake transporter (>25
fold increase) followed by efflux transporters ABCB1/P-gp and ABCC3/
MRP3 (4-5 fold increase). No difference was noted between individual
patients, either sexes or biopsy sites (rectum or sigmoid) (r=0.95-0.99).
Similarities between tissue and cell lines were low (r values 0.67-0.81).
Principal component analysis showed distinct clustering of colorectal
tissue and cell lines. TFV stimulation of CaCo-2 cells resulted in >5 fold
increase in expression of SLC28A3, SLC7A8 and TAP1. DPV stimulation
resulted in >6 fold increase in expression of SLC7A11 (26 fold), TAP1 (12
fold) SLC3A2, SLC38A2, SLC16A3,and ABCA3.
Conclusions: This study has enumerated drug transporter mRNA
expression in colorectal tissue and cell lines. There was partial
correlation between cell lines and tissue limiting cell line use as in vivo
surrogate. TFV and DPV did not induce efflux transporters in Caco-2
cells, which could result in enhanced drug delivery to submucosal CD4
T cells. Further studies will elucidate whether combination of ARVs will
be similarly effective.
294
University of Aberdeen, School of Medicine and Dentistry, Aberdeen,
United Kingdom
1
Background: Drug transporters expressed in the cervicovaginal (CV)
epithelium are likely to influence delivery of antiretroviral (ARV)-vaginal
microbicides to subepithelial target cells for HIV-1. This study aims
to characterise drug transporters in CV cell lines and investigate the
impact of dapivirine (DPV) and darunavir (DRV) on gene expression for
development of in vitro assays for testing transport of candidate ARVmicrobicides across the CV epithelium.
Methods: Expression of 84 human drug transporter genes was
investigated in HEC1A, End1E6E7, Ect1E6E7 and VK2E6E7 using RTqPCR. The impact of DPV and DRV on HEC1A and VK2E6E7 drug
transporters expression was analysed over 72 hours.
Results: End1E6E7, Ect1E6E7 and VK2E6E7 cell lines showed similar
baseline expression profiles distinct from HEC-1A. ARV-associated
uptake transporters ENT1, ENT2, OATPD, and OATPE were expressed
in all cell lines. OATP8 was expressed in HEC1A only, and CNT3
expressed in all but HEC1A. Expression of ARV-efflux transporters P-gp
and BCRP was low across all cell lines. Upon stimulation of HEC-1A
with DPV (10uM) upregulation of efflux transporters MRP2, MRP5 and
downregulation of uptake transporters OATP8, OATPE was observed.
VK2E6E7 stimulation with DPV showed downregulation of OATPE and
CNT3. DRV (250uM) induced expression in HEC-1A of MRP2, MRP5,
MRP7 and P-gp. VK2E6E7 stimulation with DRV resulted in upregulation
of OATPD, CNT3, MRP3 and MRP5 and downregulation of MRP4, MRP7
and OATPE. All reported expression changes following ARV stimulation
were > 2 fold.
Conclusions: In the absence of ARVs the cell lines investigated express
uptake transporters reported in CV tissue, but minimal levels of ARVefflux transporters P-gp and BCRP. Cell lines transfected to overexpress
P-gp and BCRP may be suitable for use in transport studies. The
downregulation of uptake transporters and upregulation of efflux
transporters induced by both ARVs, seen in HEC1A, may suggest
reduced drug delivery to subepithelial target cells in the CV tract.
Posters 31: Drug Transporters
P31.03
Transport Characteristics of Antiretroviral
Drugs - Single Agents, Double and Triple
Combinations in Caco-2 Cells
Magda Swedrowska1, Abhinav Kumar1, Charles Kelly2, Ben
Forbes1
King’s College London, Institute of Pharmaceutical Science, London,
United Kingdom, 2King’s College London, Oral Immunology, London,
United Kingdom
1
POSTERS
Background: Combinations of antiretroviral (ARV) drugs, Tenofovir
(TFV), Dapivirine (DPV) and Darunavir (DRV), provide enhanced antiviral
activity compared to single agents when tested in vitro. However, the
impact of co-formulation on drug absorption after delivery to the colorectal mucosa is unclear, and methods are required to screen drug-drug
interactions. The aim of this study is to investigate the permeability
of TFV, DPV and DRV across the colo-rectal epithelium, any role of
transporters in regulating absorption and assess the effect of coadministering the microbicidal agents on their transport.
Methods: Permeability of TFV, DPV and DRV was investigated in vitro
using the Caco-2 epithelial cell model grown on Transwell® inserts for
21- 28 days. The permeability of ARV drugs was measured, interactions
with transporter and effect of co-administration of these ARV drugs, were
evaluated.
Results: The respective absorptive and secretory permeability of
TFV across Caco-2 cell monolayer was 0.12±0.04×10-6 cm/s and
0.13±0.05×10-6 cm/s. DPV demonstrated high permeability coefficient,
36±2.9×10-6 cm/s and 27±2.5×10-6 cm/s in the absorptive and
secretory directions, respectively. TFV and DPV flux was not influenced
by the presence of transporter inhibitor verapamil or co-administration
with other ARV. Transepithelial transport of DRV was 6-fold greater in
the secretory direction compared to absorptive direction. Modulation of
the transport of DRV at 10 µM by verapamil was shown (ER decreased
from 6 to 1). Co-administration of DRV with TFV, DPV and DPV/TFV in
combinations resulted in equivalent transport as single agent.
Conclusions: TFV and DPV were passively transported across Caco-2
cells (non-vectorial and concentration independent). Transport of DRV
was vectorial, concentration dependent and affected by transporter
inhibitors, suggesting that DRV is a substrate of P-glycoprotein.
Tested ARV drugs were unaffected by the presence of double or triple
combinations.
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295
Posters
Posters 32: Ethics and the Law
P32.01
P32.02
Ethical Considerations in Implementing a
Biometric Co-enrolment Prevention System in
Clinical Trials in South Africa
Fair Subject Selection and HIV Vaccine
Efficacy Trials: Canada’s Global and Domestic
Responsibilities
Jayajothi Moodley1, Vaneshree Govender1, Sarita Naidoo1,
Dhevium Govender1, Gita Ramjee1,2, Patrick Charls3
Rika Moorhouse1
South African Medical Research Council, HIV Prevention Research
Unit, Durban, South Africa, 2London School of Hygiene & Tropical
Medicine, Department of Epidemiology and Population Health, London,
United Kingdom, 3South African Medical Research Council, National
Information Technology Department, Durban, South Africa
Ottawa Hospital Research Institute (OHRI), Ottawa, ON, Canada
1
1
POSTERS
Background: Preventing co-enrolment of participants in clinical trials
ensures participant safety and data integrity. To facilitate co-enrolment
checks, a Biometric Co-enrolment Prevention System (BCEPS) was
implemented in 2010 by the SAMRC National Information technology
(IT) Division, in collaboration with the HIV Prevention Research Unit
(HPRU). This is a web based system capturing participant’s identification
details in real time. Here, we report on the ethical considerations in
implementing BCEPS across clinical research sites (CRSs).
Methods: Ethical approval for use of this system was obtained from
MRC Ethics Committee. All participating research organisations signed
a memorandum of agreement, prior to use of BCEPS. Participants who
screened at the CRSs had their name, South African identity number
and fingerprints captured onto the system. This information was verified
and updated at all study visits. If a participant attempted to co-enrol at
another site, the system flagged this as a potential co-enrolment.
Results: Ethical considerations for implementation included ensuring
confidentiality and data security, through password protection and
fingerprint access by designated staff. Participant fingerprints are stored
as encrypted codes which cannot be copied. A participant information
sheet is used to ensure the participant’s understanding and right to
refuse, or to withdraw from BCEPS at any time. Investigator discretion
is used to decide if it is safe for the participant to enrol into the study;
or continue with study product use without co-enrolment checks. The
period of storage of data within the database is up to 15 years as per
South African good clinical practice.
Conclusions: BCEPS is a novel approach to prevent co-enrolment in
clinical trials. By addressing the ethical aspects and compliance with
good clinical practice, we are able to ensure that the system protects
participant rights and safety, and ensures data integrity.
296
Background: HIV vaccine research is a shared global enterprise, with
the potential to yield great benefits for populations at risk. To this end,
Canada’s contributions to domestic and global efforts are yielding
important new knowledge and scaling up capacity for biomedical HIV
prevention trials in priority settings. However, there remains a gap in
bioethics scholarship on Canada’s global and domestic obligations to
enroll late phase trials of preventive HIV candidate vaccines. Specifically,
there is a dearth of literature on how to operationalize ethical standards
for biomedical HIV prevention trials to ensure fair subject selection in
Canadian settings. During the STEP Study, community gatekeepers in
Canada expressed concerns about the moral defensibility of recruitment
in communities of marginalized populations including sex workers,
Aboriginal people, and people who use illicit drugs. This perception of
moral transgression suggests the need for more detailed and documented
moral analysis on fair subject selection specific to Canadian populations,
and in anticipation of future preventive HIV vaccine trials (HVTs).
Methods: I present key ethical considerations for stakeholders to weigh
and balance in order to develop moral accounts of fair selection of HVT
participants in Canada, with a focus on the principle of “justice”.
Results: There are at least six key ethical considerations for Canadian
stakeholders to consider in the fair selection of subjects for HVTs: duty
to include; duty to exclude; respect for communities; risk-benefit profile
and net risk; public health research; and global distributive justice.
Conclusions: This exploration encourages an ethical analysis of
fair subject selection that accounts for both domestic and global
responsibilities, and that uses a “reflective equilibrium” approach. More
debate, discussion and commentary among HVT stakeholders in Canada
is necessary to ensure that key moral tensions have been described,
addressed and, where possible, consensus reached.
Posters 32: Ethics and the Law
P32.03
HIV and the Law: The Impact of the Law on
HIV Research and the Role of Researchers
Michael Ulrich1
Henry M. Jackson Foundation - Division of AIDS, National Institutes of
Health, National Institute of Allergies and Infectious Diseases, Division
of AIDS, Rockville, MD, United States
1
POSTERS
There is a growing need to fully explore the connection between the
law and HIV, and the negative impact the law can have. Internationally,
there is an alarming growth in the popularity of anti-gay laws, which are
likely to have a significant impact on the spread of HIV. While the myth
that HIV is a disease that plagues homosexuals alone has long been
proven false, nevertheless, men having sex with men (MSMs) are a high
risk population that suffers from stigma and discrimination that often
results in them avoiding the health care system or not receiving the
treatment they need. These laws only stand to exacerbate this problem.
Furthermore, the stereotype that a man who has contracted HIV is gay
has unfortunately not been completely eliminated. As such, these laws
may cause heterosexual, as well as homosexual, HIV-positive men to
avoid the health care system, fearing criminalization and prejudice. In
addition to anti-gay laws, there are HIV criminalization statutes that
still not been eliminated, with recent legal cases exhibiting the impact
that the legal system can have on public health, and the HIV epidemic
specifically. The purpose of this presentation is to analyze and educate
how these laws and the enforcement and interpretation of them can
impact the HIV community and, in fact, hinder the goal of slowing the
spread of the disease. By increasing stigma and misconceptions, while
reducing education and understanding, the effect of criminalization
laws needs to be fully explored and understood to aid in their eventual
elimination. Moreover, there is a growing need to explore the roles
of researchers and potential obligations they may have in relation to
participants and the legal threats they face.
www.hivr4p.org
297
Posters
Posters 33: Evaluation of Novel Compounds in Cell-Based Systems
P33.01
P33.02
HIV-1 Shows Increased Sensitivity to
Griffithsin Derivatives
Antiviral Activity and Mode of Action
of Griffithsin against HSV-2 and HPV:
Preliminary Studies of a Potential non-ARV
Combination Microbicide
Kabamba Bankoledi Alexandre1, Karen W. Buckheit2, Lauren
Haugh-Krumpe3, Brian Constantine3, Robert W. Buckheit2, Barry
R. O’Keefe3
Council for Scientific and Industrial Researches, Bioscience, Emerging
Health Technology, Pretoria, South Africa, 2ImQuest BioSciences,
Frederick, MD, United States, 3National Cancer Institute, Molecular
Target Laboratory, Frederick, MD, United States
1
POSTERS
Background: The lectin griffithsin (GRFT) is a homodimer isolated from
the red alga griffithsia sp. GRFT has demonstrated potent and broad
anti-HIV-1 activity across subtypes and is one of the leading HIV-1
microbicide candidates. The GRFT Derivatives 2MG, 2MG3, 3MG and
4MG are made of arrays of two, three and four monomeric GRFT units,
respectively.
Methods: We evaluated HIV-1 subtype A, B and C against 2MG,
2MG3, 3MG, 4MG and GRFT using the TZM-bl neutralization assay.
GRFT derivatives were also tested for their inhibition of the cell-to-cell
transmission of HIV-1. The 234 and 295 glycans, shown to be important
in GRFT binding to HIV-1, were introduced in the virus by site directed
mutagenesis, and their effects on 2MG, 2MG3, 3MG and 4MG binding
studied. GRFT resistant viruses were generated by culturing HIV-1 under
escalating concentrations of the lectin. These resistant viruses were then
tested for sensitivity to 2MG, 2MG3, 3MG and 4MG.
Results: In general 2MG and 2MG3 were as potent as GRFT against all
the viruses tested while 3MG and 4MG were more potent against HIV-1
subtype A and C. GRFT was also less potent than these two derivatives
in the inhibition of cell-to-cell transmission of HIV-1. Similar to GRFT,
the introduction of the 234 and 295 glycans affected HIV-1 sensitivity
to 2MG and 2MG3; while it did not affect 3MG and 4MG neutralization
of the virus. Lastly, GRFT resistant viruses showed sensitive to 3MG and
4MG.
Conclusions: The 3MG and 4MG derivatives were more potent than
GRFT in inhibiting HIV-1 infection. Also viruses that showed resistance
to GRFT remained sensitive to these compounds. It is possible that 3MG
and 4MG binding site on the viral envelope is different from that of GRFT
given that the 234 and 295 glycans do affect their neutralization of the
virus. The data generated from these studies suggests that linking GRFT
into arrays of more than two monomeric units increases its potency
against HIV-1.
298
Keith Levendosky1, Olga Mizenina1, Kyle Kleinbeck1, Larisa
Kizima1, Aixa Rodríguez1, Ninochka Jean-Pierre1, Melissa
Robbiani1, Barry R. O’Keefe2, Thomas Zydowsky1, José A.
Fernández-Romero1
Population Council, New York, NY, United States, 2Molecular Targets
Laboratory, Center for Cancer Research, NCI at Frederick, Frederick,
MD, United States
1
Background: Griffithsin (GRFT) is a promising HIV microbicide candidate.
Nixon et. al. have shown that GRFT blocks herpes simplex 2 (HSV-2)
infection in a mouse model, proposing inhibition of cell-to-cell spread as
the mode of action (MOA). Using in vitro studies we further investigated
the MOA of GRFT against HSV-2 and studied its antiviral activity against
human papillomavirus (HPV). We also combined GRFT with zinc acetate
(ZA) and/or carrageenan (CG) to render a more potent microbicide.
Methods: We used XTT assay to define non-cytotoxic concentrations of
GRFT, ZA, CG or their combinations. Assays for anti-HIV, anti-HPV and
anti-HSV-2 activities were performed in TZM-bl cells or PBMCs using
MAGI and p24 ELISA; in HeLa cells using a luciferase assay; and in
Vero cells using plaque forming units (pfu) assay. We performed timeof-addition and temperature dependence experiments to differentiate
inhibition of viral adsorption from entry. Surface plasmon resonance
(SPR) was used to assess GRFT binding to viral glycoproteins and
immunohistochemistry was used to determine the specific glycoprotein
involved. Antiviral activities of prototype GRFT/CG (GC) and GRFT/ZA/CG
(GZC) gels in a vaginal HSV-2 mouse model were evaluated.
Results: GRFT shows modest in vitro antiviral activity against HSV-2
G (IC50=5.8µg/ml) and HPV 6, 16, 18, 45 PsVs (IC50=10.8-26.3µg/ml),
compared to potent anti-HIV activity (IC50=0.7-1.4ng/ml). As with HIV,
GRFT blocks the entry but not the adsorption of HSV-2 and HPV to
target cells. The combined analyses of SPR and immunohistochemistry
for HSV-2 gD, suggest that GRFT binds to HSV-2 gD. GC and GZ had
synergistic in vitro antiviral activity against HIV and HPV (CI < 1). GC
and GZC gels significantly reduced (p< 0.05) HSV-2 vaginal infection in
vivo when administered up to 2h before challenge with 106pfu/mouse.
Conclusions: GRFT blocks HSV-2 and HPV entry to target cells and
combination with CG and/or ZA may result in a potent/broad-spectrum
non-ARV microbicide.
P33.03
P33.04
Humic Acids (HA) Strongly Potentiate Anti-HIV
Effects of AZT, Griffithsin, and Cyanovirin
Polyanionic Functionalized Carbosilane
Dendrimers as Potential Microbicides to
Prevent HIV-1 Sexual Transmission
Ivanovsky Institute of Virology, Moscow, Russian Federation,
Immunomica LLC, Moscow, Russian Federation, 3Center for Cancer
Research, National Cancer Institute, Frederick, MD, United States,
4
Strasbourg University, Strasbourg, France, 5University of Pittsburgh
Medical Center and Magee-Women Research Institute, Pittsburgh, PA,
United States, 6Eastern Virginia Medical School, CONRAD, Norfolk, VA,
United States
1
2
Background: The objective of this study was to assess the anti-HIV
activity of HA and the synergistic potential of their combinations with
NRTI or lectin proteins (LP).
Methods: Anti-HIV efficacy of HA was evaluated in PBMC, MDM, DC,
Caco-2, and HEC-1A cells (using R5 HIV-1 BaL); syncytium formation
(using CEM SS) and TZM-bl assays; and in cell-free model systems
with recombinant HIV enzymes. Virus replication was detected by p24
HIV-1 antigen (intracellular and/or released) ELISA. Cytotoxicity was
determined using the MTT assay.
Results: HA suppressed HIV-1 Bal replication in MDM with IC90 = 1.5 µg/
ml and IC50 = 0.4 µg/ml. The activity in PBMC and DC was less pronounced
(IC90 values were 6.0 and 20 µg/ml, respectively). CD4-independent
entry of HIV-1 into endometrial HEC-1A cells was suppressed with IC100
= 100 µg/ml and IC50 = 10 µg/ml, regardless of the dose of the virus
(at 103 and 104 TCID50/ml). HA cytotoxicity in this system was low (CC50
not attainable; ATP and TEER levels remained unaffected up to 100 µg/
ml). Similar results were obtained with colorectal Caco-2 cells. Selectivity
indices in the cell-virus systems studied were in excess of 2000. HA had
no spermicidal activity up to 3 mg/ml (370C, 30 min).
HA augmented the antiviral effects of NRTI (AZT) and LP (griffithsin or
cyanovirin). Synergistic effects were determined by concentration-matrix
antiviral assays. Combination indices (CIs) showed synergy between HA
and either AZT (CI = 0.14) or LP (CIs between 0.3-0.7).
HA affected at least two phases in the life cycle of HIV: (1) virus
entry (inhibition of syncytium formation) and (2) reverse transcription
(experiments with recombinant reverse transcriptase).
Conclusions: HA hold significant promise as safe and efficacious drugs
for the treatment of HIV infection. The observed synergistic effects
may have a utility in HIV prevention strategies (HA may be used as a
component of vaginal and/or rectal microbicides).
Enrique Vacas-Córdoba1, Francisco J. De la Mata2, Rafael
Gómez2, Marjorie Pion1, Mª Ángeles Muñoz-Fernández1
Hospital General Universitario Gregorio Marañón, Laboratorio
InmunoBiología Molecular, Madrid, Spain, 2Universidad de Alcalá,
Departamento de Química Inorgánica, Alcalá de Henares, Spain
1
Background: Topical microbicides are researched as potential tools
in order to stop the HIV sexual spreading in all risk groups. Since the
majority of clinical trials in HIV-1 patients have failed, nanotechnology
offers novel suitable approaches to develop new microbicidal
compounds, such as dendrimers.
Methods: In vitro and in vivo studies were performed to evaluate
the safety, biocompatibility, anti-HIV ability and mechanism of two
polyanionic carbosilane dendrimers. Moreover, the antiviral activity of
carbosilane dendrimer/ARV combinations against R5, X4 and dual tropic
HIV-1 isolates was evaluated in human primary cells and TZM.bl cell line
using Calcusyn software.
Results: Sulphated and naphthylsulfonated functionalized carbosilane
dendrimers G3-S16 and G2-NF16 are shown as safety and effective
compounds against HIV. They impede laboratory and clinical primary
HIV-1 isolates infection in activated PBMC and inhibit HSV-2 infection
in vitro. Dendrimers are able to inhibit viral infection at fusion and thus
at the entry step. They impede the binding of viral particles to target
cells surface and membrane fusion, through the blockage of gp120/
CD4 interaction. In addition, dendrimers can inhibit cell-to-cell HIV
transmission and difficult infectious synapse formation.
G3-S16 or G2-NF16 did not produce changes in proinflammatory
cytokines profile in treated epithelial cells, in PBMC proliferation,
microbiota or sperm survival. Moreover, no irritation, inflammation or
vaginal lesions were detected in female mice after dendrimers vaginal
administration.
As well, G3-S16 and G2-NF16 showed a synergistic activity profile with
AZT, efavirenz, maraviroc and tenofovir in the majority of combinations
tested against X4 and R5 tropic HIV-1 in cell lines as well as in primary
human cells.
Conclusions: Carbosilane dendrimers and their combinations with ARV
can be effective antiviral agents, supporting further clinical research on
these as potential microbicides in the context of blocking HIV-1 sexual
transmission.
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299
POSTERS
Eduard Karamov1,2, Galina Kornilaeva1, Kabamba Alexandre3,
Barry O’Keefe3, Christiane Moog4, Ian McGowan5, Gustavo
Doncel6, Irina Zalenskaya6, Ali Turgiev2, Alexander Tatarintsev2
Posters
P33.05
P33.06
Antiviral Action of Sulfonate Anionic
Carbosilane Dendrimer as a Topical
Microbicide against HIV Infection
Identification of a Novel Acylguanidine-based
Inhibitor of HIV-1 Replication
Daniel Sepúlveda-Crepo , Javier Sánchez-Rodriguez , María Jesús
Serramia1, Ana López1, Esther Alonso1, Rafael Gomez2, Francisco
Javier De La Mata2, Jos Luis Jiménez1, Mª Ángeles MuñozFernández1
1
1
Hospital General Universitario Gregorio Marañón, IISGM, Networking
Research Center of Bioengineering, Madrid, Spain, 2Universidad de
Alcalá, Campus Universitario, Alcalá de Henares, Networking Research
Center of Bioengineering, Biomaterials and Nanomedicine (CIBERBBN), Madrid, Spain
Philip M. Mwimanzi1, Ian Tietjen2, Aniqa Shahid1, Scott C. Miller2,
David Fedida2, Zabrin Brummer1,3, Mark Brockman1,3
Simon Fraser University, Faculty of Health Sciences, Burnaby, BC,
Canada, 2University of British Columbia, Vancouver, BC, Canada, 3British
Columbia Centre for Excellence in HIV/AIDS, Vancouver, BC, Canada
1
1
POSTERS
Background: Microbicides include moderately specific macromolecular
anionic polymers that block HIV and other STIs, and antiretrovirals
(ARVs) that inhibit HIV entry and reverse transcription. Based on
nanotechnology, we show a novel water-soluble anionic carbosilane
dendrimer, 2G-S16, as an advantageous molecule against HIV-infection.
Methods: 2G-S16 was synthesized containing sulfonate peripheral
groups. Cellular in vitro or in vivo models were used to study the safety,
biocompatibility and anti-HIV ability of 2G-S16. Study of dendrimer/
ARVs effects and the EC50 were performed using Calcusyn software.
Results: 2G-S16 shows as safety and effective compound against HIV1 and HIV-2 with great potential as topical microbicides. 2G-S16 has
a great capacity to block HIV entry inside epithelia cells derived from
uterus and vagina due to 2G-S16 protect the epithelial monolayer
from cell disruption. Also impede laboratory and clinical primary X4,
R5 and X4/R5 HIV isolates infection in activated PBMCs. 2G-S16 does
not change the proinflammatory cytokines profile in treated epithelial
cells, in PBMC proliferation, on microbiota or sperm survival. We
research the mechanism of action and shown that 2G-S16 inhibits
HIV infection at fusion and thus and entry step. 2G-S16 impedes the
binding of HIV particles to target cell surface and membrane fusion,
through the blockage of gp120/CD4 interaction. Interestingly, 2G-S16
in combination with tenofovir or maraviroc obtains 100% HIV-inhibition
and displayed a synergistic profile at low micromolar doses against a
broad-spectrum of HIV strains in TZM.bl. Also, no irritation or vaginal
lesions were detected in female rabbit genital tracts and in CD1 (ICR)
mice after daily different concentrations of 2G-S16 for 7 days. The proof
of concept is been performed by using 2% (W/V) hydroxylethyl cellulose
gel (HEC) with 3% of 2G-S16 in humanize mice.
Conclusions: 2G-S16 is effective to inhibit HIV infection and transmission
within genital mucosa.
300
Background: Increased access to therapy has reduced HIV-1 morbidity
and mortality, but new drug classes would further enhance treatment
options and counter resistance. Acylguanidine-based molecules are
active against diverse viruses. One member of this group, BIT225, is
reported to inhibit HIV-1 by blocking the viroporin function of Vpu,
however its clinical utility is limited by toxicity. We investigated the antiHIV-1 activity of a novel acylguanidine compound, SM111.
Methods: We used a GFP-reporter T cell assay to test SM111’s ability
to inhibit replication of NL4-3 and four recombinant strains encoding
major NRTI- and/or NNRTI-resistance mutations in Pol (e.g. D67N and/or
K103N, respectively). Viruses were cultured in the presence of SM111 (0100µM), AZT (NRTI; 100nM) or EFV (NNRTI; 100nM) and infected GFP+
cells were monitored by flow cytometry. Drug activity was assessed on
day 6 compared to media controls. Cytotoxicity was evaluated using
ViaCount (Millipore). NL4-3 was also passaged in 100µM SM111; three
independent drug-resistant strains were isolated and sequenced.
Results: SM111 inhibited NL4-3 in a dose-dependent manner between
10µM and 100µM. HIV-infected cells were reduced >98% at 100µM
(44.3% [42.8-46.4] in absence vs. 0.64 % [0.56-0.76] in presence of
drug). Similar activity was observed against NRTI and NNRTI resistance
strains (>95% reduction in all cases). In contrast to BIT225, SM111 was
not toxic at any dose tested. Notably, SM111-resistant strains encoded
mutations in the transmembrane of Vpu, including a 5AA deletion, a
substitution or insertion of a stop codon at highly conserved W22.
Conclusions: SM111 is a novel compound that can inhibit wild type as
well as NRTI- and NNRTI-resistant HIV-1 strains, indicating that it has a
different mechanism of action than current drugs. Resistance patterns
suggest that SM111’s target is Vpu, but additional studies are necessary
to explore the mechanism of this promising prototype.
Funded by CIHR and the Michael Smith Foundation for Health Research
P33.07
P33.08
Microbicide, SsALF-24 Prevents HIV Infection
through the Blockade of gp120 Binding to
CD4 Receptor
Design and Discovery of Pyrazole and
Pyrimidine as Novel Class of Potent Nonnucleoside Reverse Transcriptase Inhibitors
(NNRTIs)
Kvr Reddy1, Ankit Shroff1
Background: Thirty years after its first case, HIV/AIDS has become
a pandemic and is raging in many parts of the world. UNAIDS report
of 2011 states that there were 1.7 million deaths through AIDS and
2.5 million were newly infected by the virus. Inspiring responses by
researchers and doctors have resulted in survival of marginal populations
of patients on potent antiretroviral therapy. Thus, preventing entry of the
virus into host is the best way to tackle this infection. A major route of
infection is the sexual route. Hence, developing a safe vaginal product
that can block HIV entry into the host is of utmost importance.
Methods: Anti-HIV1 activity of SsALF-24 was analyzed by p24 ELISA,
cell-cell fusion and Luciferase assays. Binding efficiency of SsALF-24
to gp120 was determined using Surface Plasmon Resonance (SPR)
spectroscopy. Toxicity of SsALF-24 on vaginal epithelial cells was
assayed by Trans-Epithelial Electrical Resistance (TEER). Cytokine
profile of human vaginal epithelial cells (VK2/E6E7) on interaction with
SsALF-24 was determined by estimating the levels of Interleukin-6 (IL6), Interleukin-8 (IL-8), Monocyte Chemotactic Protein -1 (MCP-1) and
Interkeukin-1α (IL-1 α) using ELISA, Western blot, Flow cytometry, q-PCR.
Results: SsALF-24 is derived from SsALF (Scyalla serrata AntiLipopolysaccharide factor). p24 ELISA, cell-cell fusion and Luciferase
assays show that SsALF-24 binds to gp120 and thereby prevents the
binding of the latter to CD4 receptor. This blocks the downstream
events of infection. SsALF-24 is not toxic to vaginal epithelial cells as
demonstrated by the results of TEER and microsphere experiments.
Cytokine profile of vaginal epithelial cells on exposure to SsALF-24
indicates that it may have anti-inflammatory activity.
Conclusions: SsALF-24 binds of gp120 and prevents the latter’s binding
to CD4 receptor and subsequent viral entry. Hence, SsALF-24 may be
developed as a microbicide that prevents viral entry.
Udaya Pratap Singh1, Hans Raj Bhat1
Sam Higginbottom Institute of Agriculture, Technology & Sciences,
Deemed University, Drug Design & Discovery Laboratory, Department
of Pharmaceutical Sciences, Allahabad, India
1
Background: Non-Nucleoside Reverse Transcriptase Inhibitors (NNRTIs)
considered as structurally diverse group of compounds, act by inhibiting
the reverse-transcriptase (RT) enzyme in an allosteric mode by binding to
the polymerase active site causing a distortion of the catalytic aspartate
triad in a non-competitive fashion. Consequently, a wide chemical
opportunity exists, due to the flexibility of the NNRTI binding pocket
(NNIBP) in the RT. In continuation of our ongoing efforts in the discovery
of economic NNRTI agents, herein, we wish to report a novel class of
pyrazoles and pyrimidines in order to obtain more effective candidates.
Methods: A novel series of pyrazole and pyrimidine compounds has
been developed via cyclocondensation reaction. These molecules have
been subsequently tested for anti-HIV activity using TZM-bl cell lines
along with Luciferase expression profile of the TZM-bl cells after infecting
with NL4.3 virus and MTT assay for the cytotoxicity determination.
Results: In Anti-HIV assay, molecules having pyrazole amine with distant
position of electron withdrawing group showed 91-98 % inhibition.
Further these compounds in Luciferase assay showed considerable
inhibition of infection. While in cytotoxicity assay, it was observed that an
increase in the concentration of most active compounds from 25 mg/mL
to 125 mg/mL did not appreciably lower the percentage of cell viability.
A close inspection of the best docked pose of most active compound
clearly establish that it attained a ‘’horseshoe-like’’ conformation and
interaction with the Tyr181 and Tyr188 of the p66 subunit in the NNIBP.
Conclusions: As a concluding remark, we have developed a novel
series of compounds with potent anti-HIV activity. It was confirmed that
the designed molecules have the possibility of introducing chemical
diversity around the core skeleton to generate new, potent molecules.
www.hivr4p.org
301
POSTERS
National Institute for Research in Reproductive Health, Molecular
Immunology & Microbiology, Mumbai, India
1
Posters
P33.09
P33.10
Developing an Effective Rectal Microbicide:
Inhibiting HIV Transmission in Human
Colorectal Tissue and Humanized Mice with
CD4 Aptamer-siRNA Chimeras
Glycolysis Inhibitors as Potential Anti-HIV
Compounds
Lee Adam Wheeler1, Judy Lieberman1
1
Harvard Medical School, Boston, MA, United States
1
Background: The continued spread of the HIV epidemic underscores
the need to interrupt transmission. One attractive strategy is a topical
microbicide. We previously showed that intravaginal application of
siRNAs targeted to CD4+ cells protected humanized mice from sexual
transmission, but to date no microbicide candidates have proven
effective for the prevention of the rectal transmission of HIV.
Methods: Here, we use chimeric RNAs composed of a CD4-specific
aptamer fused to siRNAs targeting the HIV coreceptor CCR5, gag,
and vif to knockdown target gene expression in CD4+ cells in human
colorectal tissue and in vivo, by employing a small animal model of HIV
transmission.
Results: Here, we demonstrate that CD4 aptamer-siRNA chimeras
(CD4-AsiCs) maintain stability for over 24h in a colorectal lavage, and
over 36h when suspended in an FDA-approved hydroexthylcellulose
(HEC) gel formulation. We provide evidence that CD4-AsiCs effectively
deliver siRNAs to CD4+ cells in human colorectal tissue explants, and
in the colorectal tissue of humanized mice without stimulation of an
inflammatory response after administration. When applied intra-rectally
to humanized mice, CD4-AsiCs in solution provide substantial but
incomplete protection from HIV transmission. When administered in an
HEC gel formulation however, all treated mice are protected against HIV
transmission for up to 8 weeks.
Conclusions: From these data, we conclude that topical CD4-AsiCs
administered in a HEC gel formulation could be used as the active
ingredient of a microbicide to prevent the rectal transmission of HIV.
POSTERS
302
Elijah Songok1, Benjamin Nzau1, Mark Wainberg2, Frank
Plummer3, Solomon Mpoke1
Kenya Medical Research Institute, Centre for Virus Research, Nairobi,
Kenya, 2McGill University, Montreal, QC, Canada, 3University of
Manitoba, Medical Microbiology, Winnipeg, MB, Canada
Background: In a bid to determine correlates of HIV protection, a
whole blood microarray study revealed that HIV exposed seronegative
(HESN ) sex workers have a significantly lowered glycolyis rate. This
suggested that reduction of cell glycolytic activity may be protective
or beneficial against HIV Infection. This phenomenon has been noted
among cancer cells, where compounds that lower glycolysis rate have
been developed as potential therapeutics against malignant carcinoma
. Our observation among HESN suggests that similar to cancer cells,
HIV maybe utilizing the glycolytic pathway for its energy needs. We
have carried a preliminary in vitro analysis to determine if the glycolytic
inhibitor,s lonidamine LND and 2 Deoxyglucose (2DG), has an anti-HIV
effect. and a potential therapeutic against HIV.
Methods: Viral Reverse Transcriptase (RT) was measured from culture
supernatant using RT assay on pretreatment (4hr drug exposed followed
by 2hr virus infection) and co treatment. Cell cytotoxity on mock
infected C8166 cells was done in parallel with RT assay using Trypan
blue exclusion test and Vi-cell (Beckmancounter) cell viability analyser.
Results obtained were normalized and presented as percentages.
Results: Lonidamine co treatment at 100 µM resulted in more than 76%
HIV inhibition while 2-DG concentrations at 5mM had HIV inhibition
percentage above 90% inhibition. Cell toxicity assessment at similar
concentrations resulted in 75% reduction in actual cell count with LND
co treatment. The 2-DG component maintained a cell proliferation of
more than 90%.
Conclusions: Glycolytic inhibitors 2-DG and LND have potential anti-HIV
activity. However it suggests a better safety profile of 2-DG at effective
inhibitory concentrations.
P33.11 LB
Targeting the Glucocorticoid Receptor with
Selective Modulators for Prevention against
HIV-1 Infection
Chanel Avenant1, Michele Tomasicchio1, Roslyn Michelle Ray1,
Andrea Du Toit1, Yashini Govender1, Hazel Hunt2, Janet Patricia
Hapgood1
1
University of Cape Town, Cape Town, South Africa, 2Corcept
Therapeutics, Storrington, United Kingdom
POSTERS
Background: Due to their anti-inflammatory properties, glucocorticoids
(GCs) are widely used therapeutically. In the female genital tract,
inflammation correlates with increased HIV infection, suggesting the
use of GCs in multipurpose prevention therapy. However, the effects
of different GCs and the role of the glucocorticoid receptor (GR) in HIV
infection and the mechanisms involved are unknown, and are likely to
be relevant to HIV-1 prevention strategies.
Methods: PBMCs or End1/E6E7 endocervical epithelial cells were
incubated with different GCs. PBMCs were infected with virus prepared
from HIV-1BAL-LUC IMCs. Inflammatory markers were measured by qRTPCR or FACs. GR function was assessed by western blotting, ChIP and
siRNA experiments.
Results: Infection assays in PBMCs showed that while Dex and
CORT113176 increased infection, pre-treatment with the GR antagonist
RU486 and a selective GR modulator CORT108297 did not increase,
but potentially had a protective effect, consistent with a role for the GR
in HIV-1 infection. Differential regulation of GILZ, IL6 and RANTES genes
by different GCs indicated a mechanism whereby some GCs increase,
while others protect against HIV infection. Furthermore, Vpr was found
to activate the GR, resulting in repression of both basal and induced
cytokine genes in the absence of GCs.
Conclusions: HIV exploits the host GR to favor viral infection via several
strategies. In the absence of GCs, the viral protein Vpr modulates GR
activity to change expression of basal and induced cytokine genes,
suggesting that GR levels play a critical role in HIV pathogenesis. GR
agonists repress immune function, while at the same time increase HIV1 infection, suggesting their use should be avoided in high risk areas.
Selective GR modulators exhibit variable effects on HIV-1 replication
and gene-specific effects on expression of cytokine genes. Some show
potential application for combination therapy in the female genital tract
by both repression of RANTES and repression of HIV-1 infection.
www.hivr4p.org
303
Posters
Posters 34: Glycans and Antibody Effector Functions
P34.01
P34.02
Identification of Specificities of Broadly
Neutralizing Plasma Antibodies Obtained
from HIV-1 Clade C Infected Indian Donors
HIV-1 gp120 Impairs B Cell Proliferation
by Inducing TGF-β1 Production and FcRL4
Expression
Jayanta Bhattacharya1, Shilpa Patil1, Sweety Samal1, Saikat
Boliar1, Tripti Srivastava1, Manish Bansal1, C Ritcher King2, K G.
Murugavel3, Suniti Solomon3, Bimal K. Chakrabarti1
Claudia Cicala1, Katija Jelicic1, Raffaello Cimbro1, Fatima Nawaz1,
Xin Zheng2, Jun Yang2, Richard Lempicki2, Donald Van Ryk1,
Jocelyn Ray1, Joseph Hiatt1, Catherine Schwing1, Danlan Wei1,
Massimiliano Pascuccio1, John Kehrl1, James Arthos1, Anthony S.
Fauci1
HIV Vaccine Translational Research Laboratory, THSTI-IAVI HIV
Vaccine Design Program, Translational Health Science & Technology
Institute, Gurgaon, India, 2International AIDS Vaccine Initiative, New
York, NY, United States, 3YRG Care, Chennai, India
1
POSTERS
Background: A number of broad and potent neutralizing human
monoclonal antibodies (mAbs) against diverse regions of HIV-1 Env
have been reported. However, it is not known whether HIV-1 clade C
infections in India mount a neutralizing antibody response against any
of those known epitopes. In the present study, we examined
(1) plasma samples obtained from anti-retroviral (ART) naïve chronically
infected Indian donors for their capacity to neutralize a diverse virus
panel representing different subtypes and
(2) we mapped the specificity of the epitopes targeted.
Methods: Two hundred plasma samples were assessed for their
capacity to neutralize Env-pseudotyped viruses in a TZM-bl cell assay.
CD4 binding site (CD4bs), N160, N332-glycan and MPER directed
neutralizing activity of the analyzed plasma samples were determined
by using TriMut core protein, N160A/N332A mutant and HIV2/HIV-1
chimeric viruses respectively.
Results: 10/200 (5.5%) plasma antibodies were found to neutralize
>60% of the 50 panel viruses of distinct geographical origin and
belonging to clades A, B. C, B/C,A/E and A/G with a median ID50>200.
Amongst these ten broadly neutralizing plasma (BNP) samples, two (1%)
were found to be highly potent (median ID50 > 400). Interestingly,
none of the 200 plasma samples showed any specificity to CD4 binding
site (CD4bs), indicating a possible inability of clade C strains circulating
in India to elicit CD4bs directed antibodies. In addition, none of the
neutralizing activity by the BNP samples was dependent on N160 and
N332 glycans. Three of the BNP samples showed specificity to the gp41
membrane proximal external region (MPER). This MPER reactivity was
different than that of the known MPER neutralizing mAbs.
Conclusions: Our data indicates that the BNP samples obtained from
Indian donors likely target novel epitopes and would pave way towards
identification of new vulnerable site/s on HIV-1 envelope.
304
1
NIH/NIAID, LIR, Bethesda, MD, United States, 2NIH/NIAID, Laboratory
of Immunopathogenesis and Bioinformatics, SAIC, Frederick, MD,
United States
Background: During the early stages of HIV infection the immune system
of the infected individual is impaired. Among the defects described is
the impairment of normal B cell function, that includes a significant
delay in the development of the anti-HIV humoral immune response.
The mechanisms underlying this delay are not fully understood. In the
present study, we asked whether gp120-induced signaling through α4β7
on B cells could disrupt their function. We show that gp120 binds and
signals through α4β7 on naïve B cells, resulting in an abortive proliferative
response.
Methods: We performed microarray analysis, RT-PCR, CFSE proliferation
assay, ELISA, FACS analysis, cell cycle analysis on freshly isolated primary
human B cells. B cells were stimulated with CpG, anti-human IgM and
anti-CD40 mAb, in presence or not of recombinant HIV-1 gp120.
Results: gp120 directly impaired the activation and proliferation of
naive B cells by releasing TGF-β1, which in turn up-regulated expres
sion of the inhibitory IgA receptor FcRL4. Co-culture of B cells with
HIV-infected autologous CD4+ T cells also increased B cell FcRL4
expression. Treatment of stimulated B cells with gp120 decreased the
expression of the co-stimulatory receptor CD80 but not CD86. gp120
impaired proliferation of naïve B cells stimulated with both a TI and a
TD stimulation. gp120 affected the responses of memory B cells to TI
and TD stimulation in a relatively modest way compared with the sup
pressive effect we observed for naïve B cells.
Conclusions: These findings indicate that, in addition to mediating
chronic immune activation, viral proteins can contribute directly to
HIV-associated B cell dysfunction. These studies have implications for
understanding the immune-pathogenic mechanisms of HIV-1 infection,
particularly the ability of high levels of viremia, observed during acute
HIV infection, to blunt the early antibody response to HIV.
P34.03
P34.04
Shielding of the HIV-1 Envelope Membrane
Proximal External Region (MPER) from
Antibody
The Dynamics of HIV-1 gp120 N-linked
Glycans in the Context of Broadly Crossneutralizing Antibodies
Rajesh Abraham Jacob1,2, Thandeka Moyo1,2, Fatima Abrahams1,
Berta Grau Pujol1, Jeffrey R. Dorfman1,2
Ereshia Gabier1, Natasha Wood1, Simon Travers1
International Centre for Genetic Engineering and Biotechnology, Cape
Town, South Africa, 2University of Cape Town, Division of Immunology,
Cape Town, South Africa
University of the Western Cape, SANBI, Cape Town, South Africa
1
1
POSTERS
Background: The Membrane Proximal External Region (MPER) of HIV1 gp41 envelope is an attractive vaccine target. We have previously
identified 253-11 (CRF_02AG) as unusually neutralization-resistant and
studied its sensitivity to anti-MPER antibodies.
Methods: Antibodies reactive against the 253-11 MPER were identified
using a chimeric HIV-2 virus displaying the 253-11 MPER sequence
in place of its own. The presence of dominant anti-MPER neutralizing
antibodies was tested by assessing the reduction in neutralization after
depleting MPER-specific antibodies. We also studied MPER swap viruses
between 253-11 and 928-28, a subtype-matched virus that is sensitive
to anti-MPER antibodies in cohort sera.
Results: We found that 253-11 is rarely neutralized by anti-MPER
antibodies in sera from individuals HIV-infected >1yr. However,
recognition of that MPER in the HIV-2 chimera was common: 19/177
(9%) sera recognized 253-11’s MPER in the chimeric virus but not the
original 253-11 virus. At least 13/19 of these sera neutralized other HIV1 isolates via MPER, indicating that these anti-MPER antibodies were not
generally defective for neutralization of HIV-1 and that many viruses do
not share this pattern of resistance to anti-MPER antibodies. Importantly,
sensitivity of 928-28/253-11 MPER swap viruses to these 19 sera
and to MPER-specific mAb Z13e1 is primarily controlled by sequences
outside the MPER region. Several epitopes were recognized by the
sera, suggesting that conformational differences of MPER between the
native virus and the HIV-2 chimera are unlikely to explain 253-11’s
neutralization resistance.
Conclusions: We consider several explanations for our data and
propose that the most parsimonious explanation is obstruction of access
to MPER prevents anti-MPER neutralization of 253-11. In this case, only
the limited proportion of anti-MPER antibodies that can penetrate similar
obstructions would be able to provide the protection against the large
number of HIV-1 variants that would be desirable in a vaccine.
Background: HIV-1 prevention through vaccination remains challenging
due to the high mutation rate and variability amongst viral strains
meaning that the identification of epitopes that elicit a truly broadly
cross clade neutralizing virus is an immensely complex task. Identifying
vaccine targets is further complicated by the presence of N-linked glycans
on the surface of gp120 that protects the virion from recognition by
the host immune system. Recently, however, broadly cross-neutralizing
(BCN) antibodies have been identified that recognize specific glycans on
the surface of gp120 and result in the neutralization of a broad panel
of HIV viruses. Studies have identified that the glycans bound at either
N332 or N334 in the C3 region at the base of the V3 loop are critical for
susceptibility to, or evasion of neutralisation. The structural mechanism
of this critical process is not yet fully understood.
Methods: Using the full gp120 structure, containing both the V1/
V2 loops and V3 loop, we performed a molecular dynamics study to
model the effect of the movement of glycosylation from 332 to 334
on both the ‘glycan shield’ and the underlying protein and, thus, on the
susceptibility to neutralization.
Results: We have identified features of the glycan-glycan, as well
as glycan-protein, interaction that may appear to play a role in the
recognition of these epitopes by BCN antibodies as well as in the
evasion of neutralization.
Conclusions: Our current research therefore contributes to the
understanding of the role of N-linked glycosylation in the context of
BCN antibodies and their associated epitopes.
www.hivr4p.org
305
Posters
P34.05
P34.06
Post-attachment Neutralization by Singlechain Variable Fragment (scFv) from an
Anti-V3 Monoclonal Antibody with Crossreactivity
HIV-1 Neutralisation Breadth Is Positively
Associated with Presence of Anti-MPER
Antibodies and Not of Anti-PG9/16-site
Antibodies
Yasuhiro Maruta1, Takeo Kuwata1, Kazuki Tanaka1, Yusuke
Nakahara2, Kristel Ramirez1, Muntasir Alam1, Yoshika Egami1,
Izumi Hirata1, Yoshiaki Suwa2, Hiroshi Morioka2, Shuzo
Matsushita1
Thandeka Moyo1,2, Rajesh A. Jacob1,2, Michael Schomaker3, Berta
Grau4, Fatima Abrahams4, Jeffrey R. Dorfman1,4
Matsushita Project Laboratory, Center for AIDS Research, Kumamoto
University, Kumamoto, Japan, 2Department of Analytical and
Biophysical Chemistry, Graduate School of Pharmaceutical Sciences,
Kumamoto University, Kumamoto, Japan
Division of Immunology, University of Cape Town, Cape Town,
South Africa, 2International Centre for Genetic Engineering and
Biotechnology, Cape Town, South Africa, 3Centre for Infectious Disease
Epidemiology & Research, Cape Town, South Africa, 4International
Centre for Genetic Enginnering and Biotechnology, Cape Town, South
Africa
Background: The neutralization antibody response against HIV-1 is
crucial for controlling HIV-1 infection. Such is the case of the epitope
in the V3 loop that is exposed after binding of gp120 to CD4. However,
it is difficult for anti-V3 IgGs to bind their epitope following CD4-gp120
interaction because IgG is too large to access the close physical proximity
of gp120 and the cellular membrane.
Methods: We constructed scFv from 1C10, an anti-V3 monoclonal
antibody with cross-reactivity. The 1C10 scFv was produced in E.coli,
purified and refolded by “on column refolding” process. In addition to
the usual single cycle neutralization assay, we employed temperature
regulated neutralization assay (TRN assay) and neutralization assay using
SOSJR-FL virus (SOS assay) to evaluate post-attachment neutralization
using TZM-bl cells as a target. TRN assay and SOS assay allows
determining whether scFv can access efficiently to the narrow space
between HIV-1 and the V3 epitope of the envelope after attachment of
the virus to CD4.
Results: Neutralization activity of 1C10 scFv was greater than that
of IgG counterpart against JR-FL, YU2 and 89.6 strains of HIV-1 (IC50
values of scFv were about 5, 3 and 3 times better than values with
IgG, respectively). In addition, 1C10 scFv neutralized viruses resistant
for neutralization by 1C10 IgG, such as SVPB8 (IC50 of IgG; >50 µg/ml,
scFv; 5.23 µg/ml). TRN assay results showed that neutralization capacity
of IgG was considerably reduced after the virus bound to CD4. On the
other hand, neutralization activity of scFv at TRN assay was equivalent
to that at usual single cycle neutralization assay. In SOS assay, 1C10
scFv neutralized SOSJR-FL, but 1C10 IgG did not.
Conclusions: These results suggest that the anti-V3 scFv can neutralize
HIV-1 even after attachment of the virus to the target cells. For this
reason, the use of scFv may be one of the promising strategies to
overcome neutralization resistance of HIV-1.
Background: The Membrane Proximal External Region (MPER) and the
PG9/16-site are both potential targets for anti-HIV-1 antibody-based
vaccines. The MPER of the gp41 subunit of the envelope glycoprotein is
relatively conserved and plays a key role in viral fusion with target cell
membranes. The PG9/16-site is a quaternary epitope which is expressed
on trimeric envelope protein spanning conserved regions of variable
loops of the gp120 subunit. The PGT/2G12 antibodies recognise an
epitope made primarily from a glycosylation at sites 301 and/or 332.
Methods: We explored associations between neutralisation breadth
and presence of antibodies directed against these sites in sera from
177 antiretroviral-naïve HIV-1-infected (>1yr) individuals. Anti-MPER
antibodies were detected using chimeric 7312A HIV-2 viruses engrafted
with three different HIV-1 MPER sequences: a consensus subtype
C sequence or the MPER sequence from Yu2 (B) or 253-11 (CRF02_
AG) viruses. Dominant anti-PG9/16-site antibodies were detected by
determining what proportion of neutralisation of any of 3 viruses was
abrogated by mutations at positions 160 or 169 within the PG9/16 site.
Results: Anti-MPER activity and activity directed against epitopes
overlapping the PG9/PG16 site were common. Neutralisation breadth of
the MPER-recognising sera was significantly higher than that of the nonMPER recognising samples. We found no evidence for an equivalent
association for sera containing anti-PG9/16 site antibodies. Variability
within the MPER, measured by analysing 3829 envelope sequences,
was substantially lower than that in the PG9/16-site and other broadly
neutralising antibody targets. A similar comparison for the PGT/2G12
specificity will also be presented.
Conclusions: Our data suggest that inducing anti-MPER antibodies
by vaccination is more likely to be productive than inducing broadly
neutralising anti-PG9/16 site antibodies, even if PG9/16-site immunogen
models can be engineered.
1
POSTERS
306
1
P34.07
P34.08
Identification of Key Determinants for the
Unusual Neutralization Sensitivity of the
MW965.26 Env
Strain Specific Anti-HIV Antibody Evolution
during Acute Infection and Viral Escape
1
2
1
Rutgers Biomedical and Health Sciences, Public Health Research
Institute, Newark, NJ, United States, 2Tulane University, Health Sciences
Center, New Orleans, LA, United States
1
Background: The V3 domain of gp120 is the most immunogenic region
of the functional Envelope (Env) spikes found on the surface of a HIV
virion. However, in most cases high affinity antibodies (Abs) developed
to this region are unable to neutralize the majority of circulating isolates
as the V3 loop is usually effectively masked. ConC, the virus encoded by
the clade C consensus sequence, is extremely resistant to neutralization
by V3 Abs. On the other hand, the clade C tier 1a virus MW965, is the
most neutralization sensitive viral isolate described to date and can be
neutralized by a wide range of these Abs. Compared to ConC, MW965
has a longer more highly glycosylated V1/V2 region, which usually
correlates with greater masking. It is thus unclear why this isolate is so
sensitive.
Methods: To identify key neutralization determinants in the MW965
Env, chimeric Env constructs were produced between this Env and ConC
using various cloning techniques. Neutralization sensitivity to several
α-V3 and α-quaternary neutralizing epitope (QNE) Abs was determined
using pseudovirus produced from Env expressing plasmids and pNL4-3.
luc.R-E-. Reversal of neutralization phenotype indicated the identification
of a determinant(s).
Results: The construction of a chimeric Env using the SF162 (unmasked)
backbone and MW965 V1/V2 revealed that this V1/V2 is in fact
capable of masking the V3. This suggests that another Env region(s)
is responsible for this neutralization sensitivity despite the masking
properties of the V1/V2 domain. Neutralization assays using additional
ConC and MW965 chimeric Env constructs, ruled out the C1-V3 and
gp41 Env regions and narrowed several key determinants to the C3 and
C5 Env regions.
Conclusions: The identification of specific residues in these domains
defines a novel mechanism for regulating neutralization sensitivity in a
circulating isolate. Additionally, several of the chimeric Env constructs
may provide additional insights for designing immunogens capable of
eliciting effective antibody responses.
Cathrine Scheepers1,2, Dshanta Naicker1, Chaim Schramm3,
Zhizhang Sheng3, Arshad Ismail1, Salim S. Abdool Karim4,
Bronwen Lambson1, Penny Moore1,2,4, Lawrence Shapiro3, Lynn
Morris1,2,4
Center for HIV and STIs, National Institute for Communicable Diseases
(NICD), Sandringham, South Africa, 2University of the Witwatersrand,
Virology, Sandringham, South Africa, 3Columbia University,
Biochemistry and Molecular Biophysics and Department of Systems
Biology, New York, NY, United States, 4Center for the AIDS Programme
of Research in South Africa (CAPRISA), Durban, South Africa
1
Background: We previously identified an individual (CAP88), from
the CAPRISA 002 cohort, with potent strain-specific neutralization.
This response mapped to the C3 region of gp120 and was detected
at 11 weeks post-infection (wpi) and then later waned co-incident with
viral escape. A monoclonal antibody (CAP88-CH06) was isolated at
34 wpi which utilized the IGHV4-39*01, D3-3*01 and J4*02 genes,
had 5.9% divergence from germline, and CDRH3 length of 17 amino
acids. This study examined the evolution of the CAP88-CH06 heavy
chain immunoglobulin genes over 121 weeks, starting from 11 weeks
of infection.
Methods: RNA and DNA were extracted from PBMCs from donor
CAP88 at four time-points (11, 17, 38 and 121 wpi). The heavy chain
VDJ regions of IGHV4-39 gene were PCR amplified and sequenced by
Illumina MiSeq. The resulting sequences were blasted against a database
of germline IGHV and IGHJ sequences from IMGT and compared to the
CAP88-CH06 sequence.
Results: We detected ~3,500 sequences that were highly related to
CAP88-CH06. The majority (75%) of these were from RNA at 11 wpi
(1,051 sequences, 39%), 17 wpi (1,625, 61%) and 1 sequence each
in 38 wpi and 121 wpi. This corresponded to the antibody response
which first appeared at 11 wpi, peaked at 26 wpi and by 54 weeks
had declined. Most of the DNA sequences were from 11 wpi (n=761)
followed by 38 wpi (n=113) and fewer than 10 from 17 wpi and 121
wpi. The 38 wpi DNA sequences were closely related to the 11 wpi
sequences of both RNA and DNA.
Conclusions: We have identified clonally related antibody sequences
from 4 different time-points from CAP88 in both RNA and DNA. The
frequency of sequences in RNA corresponded with plasma neutralizing
antibody titres. DNA-derived sequences from later time-points clustered
in phylogenetic trees with RNA-derived sequences from earlier timepoints, suggesting that they were from the memory B cell compartment.
www.hivr4p.org
307
POSTERS
Zakiya Qualls , James Theis , James Robinson , Abraham Pinter
1
Posters
P34.09
P34.10
Engineering Antibodies to Enhance Activity
and Increase Half-life
Antibody-based PrEP and Cross-reactivity
Kevin J. Whaley1, Steve Hume2, Larry Zeitlin3
Stuart A. Sievers , Sonal N. Patel , Kathleen Bennett , Florian
Klein2, Michel C. Nussenzweig2, Pamela J. Bjorkman1
1
1
1
California Institute of Technology, Biology and Biological Engineering,
Pasadena, CA, United States, 2Rockefeller University, Laboratory of
Molecular Immunology, New York, NY, United States
1
POSTERS
Background: HIV/AIDS remains one of the most serious current threats
to global public health. Although anti-HIV drugs have been effective
among the wealthiest populations, a vaccine and/or new methods to
prevent infections are needed to control HIV globally. Strategies to
combat HIV-1 require structural knowledge of how antibodies recognize
HIV envelope proteins and how the immune system eliminates viruses.
Until recently, only a small number of broadly neutralizing antibodies
against HIV-1 had been characterized, and the immunological basis
for their breadth and potency remains poorly understood. However,
it was recently demonstrated that antibodies could be engineered to
greatly enhance their breadth and potency (Diskin et al., Science 2011).
Unfortunately, this and other engineering efforts have resulted in a
decrease in antibody half-life in mouse and non-human primate models.
This decrease in half-life correlates with an increase in reactivity to a
variety of antigens, termed polyreactivity.
Methods: In order to make better targets for passive delivery
therapies, we are working to increase the half-life of antibodies while
maintaining their breadth and potency using a variety of computational
and structure-based techniques. One technique involves reducing the
spatial aggregation propensity, in which an algorithm finds dynamically
exposed hydrophobic patches on the surface of proteins (Chennamsetty
et al., PNAS 2009). To this end, we have constructed several mutations in
regions that have been predicted to have high aggregation propensities,
and have tested them for polyreactivity and potency in neutralization
assays.
Results: Initial results show that these novel reagents have reduced
polyreactivity, yet they still maintain their potency in in vitro
neutralization assays.
Conclusions: We are currently pursuing in vivo experiments in mice
to further understand the relationship between antibody potency,
polyreactivity, and half-life.
308
1
Mapp Biopharmaceutical, San Diego, CA, United States, 2Kentucky
Bioprocessing, Inc., Owensboro, KY, United States, 3Mapp
Biopharmaceutical, Inc., San Diego, CA, United States
Background: Broadly neutralizing antibodies (bNAbs) are being
developed for topical and systemic pre-exposure prophylaxis. Since
some bNAbs (e.g. 4E10) have been reported to interact with non-viral
epitopes, the cross-reactivity of bNAbs is an important safety parameter
to be documented in regulatory submissions. The objective of this study
was to determine the cross‑reactivity of Nicotiana (-N) manufactured
anti-HIV bNAbs 4E10-N and VRC01-N, and anti-HSV glycoprotein D
bNAb HSV8-N with cryosections of human tissues.
Methods: In order to detect binding, the antibodies were biotinylated
and applied to cryosections of normal human tissues (3 donors per
tissue) at two concentrations (2-20 µg/ml). Commercially available
Synagis was biotinylated and used as a control. The study was GLP
compliant.
Results: 4E10-N variably stained a variety of tissue elements in the
human tissue panel. VRC01-N also produced staining of tissue elements;
however, the staining with VRC01-N was generally present in fewer
tissues and with reduced intensity and frequency. No staining with
HSV8-N or Synagis was observed in the human tissue panel examined.
Conclusions: The majority of observed staining was cytoplasmic
in nature, which is of little toxicologic concern since the cytoplasmic
compartment is generally considered to be inaccessible to antibodies
administered in vivo. The toxicologic concern for the observed staining
of extracellular elements in selected tissues with 4E10-N and VRC01-N
is unknown. Since no staining was observed with HSV8-N the binding
of VRC01-N cannot be attributed to the Nictotiana-based manufacturing
system.
P34.11 LB
P34.12 LB
Superinfected Patient Pseudovirus Exhibits
Resistance to Broadly Neutralizing Antibodies,
but Sensitivity to Autologous Plasma Postsuperinfection
Acute HIV-1 Subtype C Infection Is Associated
with Rapid Increase of Tissue-like Memory
and Decrease in Resting Memory B-cells
New York University - School of Medicine, Department of Pathology,
New York, NY, United States, 2Medical Diagnostic Center, Yaoundé,
Cameroon, 3Manhattan Veterans Affairs Harbor Healthcare Systems,
New York, NY, United States
1
Background: Superinfected HIV patients provide the unique
opportunity to investigate the immune response after challenge with
diverse HIV antigens and remain the major source for recombinant
strain production. The competition and/or coexistence of two or more
viral strains drive viral evolution and diversity, eventually triggering
the generation of more broad and potent neutralizing antibodies
(Abs) as compared to singly infected patients. The objective of this
study was to analyze the genetic evolution of the viruses found
within a superinfected Cameroonian patient, and to determine the
neutralization sensitivity of the recombinant viruses to both autologous
and heterologous neutralizing Abs.
Methods: Longitudinal plasma samples from an HIV-1 superinfected
Cameroonian patient were analyzed for their Env diversity, evolution,
and recombination events. Pseudoviruses were generated from
timepoints before, during, and after superinfection, and were then
tested for sensitivity to homologous plasma and to heterologous
neutralizing mAbs.
Results: Env sequence analysis indicated that the CRF02_AG infected
subject became superinfected with an F2 strain resulting in a massive
increase in viral diversity. Later, the patient´s viral repertoire was
narrowed to quasispecies closely clustering with the initial strain. The
highest neutralization was observed with autologous plasma from
timepoints after superinfection but before the intiation of ART. Of
interest, the superinfecting F2 strain was resistant to neutralization with
most known broadly neutralizing antibodies (bnAbs) including PG09
and VRC01 while the original infecting CRF02_AG strain was sensitive
to these bnAbs.
Conclusions: The resistance of the F2 strain to known bnAbs urges
the need to generate nAbs from such patients and to monitor the
emerging recombinant strains. HIV-1 superinfected patients may serve
for the generation of new bnAbs covering multiple subtypes and deliver
important knowledge for future vaccine design.
KwaZulu Natal Research Institute for TB & HIV, University of KwaZulu
Natal, Durban, South Africa, 2HIV Pathogenesis Programme, Durban,
South Africa, 3Ragon Institute of MGH, MIT and Harvard, Massachusetts
General Hospital and Harvard Medical School, Boston, MA, United
States, 4Massachusetts General Hospital, Infectious Diseases, Boston,
MA, United States
1
Background: HIV chronic infection (CI) is characterized by perturbations
in B cell homeostasis, phenotype and function. There are limited data
describing B cell dynamics during HIV acute infection (AI). Characterizing
B cell subsets during AI might help define signatures that shape the
humoral response in HIV infection.
Methods: Eleven women who became HIV-1 infected during a
longitudinal follow-up study in Durban, South Africa were analyzed.
Samples were analyzed at baseline (pre-infection), at 1 week, 2 week, 1
month and 3 months post detection of plasma viremia. Multicolor flow
cytometry was used to identify B cell subsets based on expression of
CD21CD27 on live CD19+ lymphocytes and defined as follows; activated
memory (AM) CD27+CD21-, resting memory (RM) CD27+ CD21+, naïve
cells (N) CD27-CD21+, tissue-like memory (TLM) CD27-CD21- and
plasmablasts (PBs) CD27+CD38+ cells. HIV-specific antibodies against
subtype C gp120, gp41 and p24 antigens were determined by ELISA.
Results: Compared to a baseline negative sample, we observed rapid
and significant expansion of TLM cells post HIV infection (PI); 1week (p
= 0.0003), 2 weeks (p = 0.0018), 1 month (p = 0.046) and 3 months
(p= 0.043). In contrast, RM cells were significantly lower throughout
AI compared to baseline; 1week (p = 0.0007), 2 weeks (p = 0.016), 1
month (p = 0.019) and 3 months (p= 0.018). We observed significant
expansion of AM cells at 2 weeks and 1 month (p = 0.008, p= 0.009
respectively) followed by contraction by 3 months (p = 0.260) PI.
Peripheral blood PBs peaked by a median of 18 days (range 8-56 days)
PI. There was a temporal relationship between peak viral load, PB peak
and detection of HIV specific antibodies.
Conclusions: Perturbations in B cell subsets occurs immediately
following HIV-1 infection and this may therefore determine the
subsequent development of anti-HIV antibodies.
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309
POSTERS
Ralf Duerr1, Sonal Soni1, Colleen Courtney1, Josephine Meli2,
Johnson Ngai2, Luzia Mayr1, Phillipe Nyambi1,3
Jennifer K. Mabuka1,2, Anne-Sophie Dugast3, Zelda Euler3,
Yathisha Ramlakhan2, Krista Dong3, Bruce D Walker2,3,4, Thumbi
Ndung’u1,2,3, Galit Alter3
Posters
Posters Posters 34: Glycans and Antibody Effector Functions
P34.13 LB
Potent SIV-specific Antibodies Targeting the
Cyanovirin Binding Site
Rosemarie Mason1, Cameron Adams1, Carole Bewley2, John
Mascola1, Mario Roederer1
Vaccine Research Center, National Institute of Allergy and Infectious
Diseases, National Institutes of Health, Bethesda, MD, United States,
2
Laboratory of Bioorganic Chemistry and Laboratory of Molecular
Biology, The National Institute of Diabetes and Digestive and Kidney
Diseases, National Institutes of Health, Bethesda, MD, United States
1
POSTERS
Background: The SIV model of HIV infection is useful for studying
vaccine mediated and immune correlates of protection but knowledge
of the specificity, function and efficacy of protective SIV antibody
responses is limited. Binding of cyanovirin-N (CVN), a potent inhibitor of
HIV and SIV, occludes the CD4bs and 2G12 epitope - both major sites
of HIV Env vulnerability. We used CVN with SIVgp140 to isolate CVNbsspecific mAbs.
Methods: Trimeric SIVgp140 and CVN proteins were used to generate
2 distinct probes with differential binding for CVNbs-specific B cells.
Sequential staining with these probes was used to isolate CVNbs-specific
B cells by indexed single cell sorting. Individual mAbs were cloned and
expressed in vitro and tested for SIV-specific binding and neutralization.
Results: We sorted CVNbs-specific B cells from SIV-infected rhesus
macaques. Of 15 mAbs screened, 11 bound to monomeric SIVgp120
and trimeric SIVgp140 but not SIV Env 15mer peptides. Pre-adsorption
of SIVgp140 with CVN blocked binding of 4 out of 11 SIV-specific mAbs
confirming their specificity for the CVNbs and validating our probe
strategy. Interestingly, only the CVNbs-specific mAbs (ITS50, ITS51, ITS52
& ITS53) neutralized SIVmac251.30 (Tier 2) with ITS50 and ITS51 also
cross-neutralizing primary isolate HIV-2 (7312A). Of note, ITS51 showed
detectable albeit low level neutralization of SIVmac239. The remaining
7 mAbs were binding but non-neutralizing. SIV CD4bs- and V1V2specific mAbs did not compete with ITS50, ITS51 or ITS52 for binding
to SIVgp140 trimer suggesting distinct and non-overlapping binding to
SIV Env. Additional mapping with glycan-deficient SIVgp140 protein
and individual glycan-deletion mutant viruses will determine binding
specificity and glycan-dependence of these CVNbs-specific mAbs.
Conclusions: We isolated and characterized SIV CVNbs-specific mAbs
by a novel competitive binding probe strategy that may be adapted for
isolating additional SIV- and HIV-specific mAbs in order to optimize HIV
vaccine development.
310
Posters 35: HIV Drug Resistance in vitro
P35.01
P35.02
Resistance Profile of the Diaryltriazine Nonnucleoside Reverse Transcriptase Inhibitor
and Candidate Microbicide UAMC01398
Investigating HIV-1 Resistance to CCR5
Antagonist Maraviroc for the Design of New
Prevention Strategies
Kevin K. Ariën1, Muthusamy Venkatraj2, Johan Michiels1, Katleen
Vereecken1, Jurgen Joossens2, Pieter Van der Veken2, Leo
Heyndrickx1, Jan Heeres2, Koen Augustyns2, Guido Vanham1
Jacqueline K. Flynn1,2, Michael Roche1,2, Geza Paukovics1,
Hamid Salimi1,2, Renee C. Duncan1, Miranda S. Moore1, Anne
Ellet1, Lachlan R. Gray1,2, Becky Jubb3, Mike Westby3, Damian
F. J. Purcell4, Sharon R. Lewin1,2, Benhur Lee5, Richard J Payne6,
Melissa J. Churchill1,2, Paul R. Gorry1,2
Institute of Tropical Medicine, Antwerpen, Belgium, 2University of
Antwerp, Antwerpen, Belgium
1
Burnet Institute, Melbourne, Australia, 2Monash University, Melbourne,
Australia, 3Pfizer Global Research and Development, Kent, United
Kingdom, 4University of Melbourne, Melbourne, Australia, 5UCLA, Los
Angeles, CA, United States, 6University of Sydney, Sydney, Australia
Background: We previously selected UAMC01398 as a lead compound
from a screen with 60 diaryltriazine analogues, in the context of a multipartner program on microbicide development (FP7-CHAARM). This new
NNRTI has a superior toxicity profile compared to Dapivirine (DPV) and
it retains nM activity against DPV-resistant viruses. We now report on the
resistance profile of this new candidate microbicide.
Methods: Resistance was induced in dose-escalation studies and in
single high dose experiments. Mutations were identified by sequencing
and subsequently confirmed in IC50 experiments with site-directed
mutagenesis in a pNL4.3 molecular clone. Cross-resistance to other
clinical and experimental NNRTIs was studied. Finally, the replication
capacity of the UAMC01398-resistant viruses was assessed.
Results: Dose-escalation studies revealed the following mutations in the
RT gene: V90I, V106A, E138K, V179M, H221Y, F227C and M230I. Full
blown resistance was selected only after 150 days. At least 4 of these
mutations are required in concert for resistance against UAMC01398.
Cross-resistance was assessed against DPV, Etravirine, Rilpivirine (RPV),
Lersivirine, MIV170, Efavirenz and Nevirapine. Only Etravirine and
RPV retained partial activity (sub µM). Single high dose exposure to
UAMC01398 did not select for resistant HIV. Finally, we clearly show
that UAMC01398-resistant viruses are significantly less fit than wild type
virus.
Conclusions: UAMC01398 is a strong new candidate microbicide with
superior toxicity, activity against DPV-resistant HIV, and a complex
resistance profile that is not easily selected but similar to RPV.
Background: Maraviroc (MVC) is a CCR5 antagonist currently used for
the treatment of HIV-1 and is being tested as a Pre-exposure prophylaxis
prevention strategy. MVC resistance can occur, however the mechanisms
behind its development are unclear. Elucidating these mechanisms will
assist in the design of improved CCR5 antagonists for use in prevention
and therapy.
Methods: Envs were cloned from plasma from two phase III MOTIVATE
clinical trial participants, who developed phenotypically verified MVC
resistance in vivo. Both participants were male with a similar duration
of infection (14yrs). MVC resistance was characterised by measuring the
maximal percent inhibition (MPI) and the ability of the Envs to recognise
the MVC-bound confirmation of CCR5 (Affinofile assay). Tropism
alterations for the infectivity of CD4+ T cell subsets and macrophages
were characterised and CCR5 engagement of resistant Envs was tested
for neutralization by sulfated peptide fragments of the CCR5 N-terminus.
Results: The MPI values and affinity profiling showed that these Envs
displayed a divergent ability to recognise MVC-bound CCR5, characterised
by either a relatively efficient (MPI ~10%, less CCR5-dependant) or
inefficient (MPI ~90%, more CCR5-dependant) recognition. Only the
MVC-resistant Env with efficient recognition of MVC-bound CCR5 (MPI
~
10%) displayed a tropism shift for CD4+ T cells characterized by a
significant expansion of infected CM T cells, potentially affecting the HIV
reservoir size and no change in macrophage infectivity. Interestingly,
both resistant Envs were susceptible to neutralization by a sulphated
peptide fragment of the CCR5 N-terminus.
Conclusions: Our results suggest that the pattern of HIV tropism
alterations for susceptible cells can vary depending on the magnitude of
MVC resistance. Despite divergent phenotypes of resistance, both Envs
showed an increase reliance on sulfated CCR5 N-terminus residues,
which could provide a new avenue to block HIV-1 entry through CCR5
N-terminus sulfopeptidomimetic drugs.
www.hivr4p.org
311
POSTERS
1
Posters
Posters 35: HIV Drug Resistance in vitro
P35.03
P35.04
Resistance Profile of CD4 Mimic Small
Compounds (CD4MCs) and the Structure
Analysis by Molecular Dynamic (MD)
Simulation
Combinations of Entry and Reverse
Transcriptase Inhibitors as Candidate
Microbicides
Shigeyoshi Harada1, Masaru Yokoyama2, Samatchaya
Boonchawalit1,3, Hironori Sato2, Shuzo Matsushita3, Kazuhisa
Yoshimura1,3
National Institute of Infectious Diseases, AIDS Research Center,
Shinjuku-ku, Japan, 2National Institute of Infectious Diseases, Pathogen
Genomics Center, Musashimurayama, Japan, 3Kumamoto University,
Center for AIDS Research, Kumamoto, Japan
1
POSTERS
Background: CD4MCs inhibit the gp120-CD4 interaction and can
also expose masked epitopes of neutralizing antibodies on the gp120
protein. In this study, we investigated the phenotypic change in the
CD4MCs resistant isolates against CD4MCs, other entry inhibitors and
anti-Env neutralizing monoclonal antibodies (nMAbs).
Methods: Resistant variants were induced by five CD4MCs using
the primary KP-5P virus (subtype B, R5) in PM1 cells. We constructed
infectious clones with CD4MC-resistant mutation following in vitro
selection. The susceptibility of the infectious clones to the inhibitors
was evaluated using TZM-bl cells. We also simulated the gp120 3D
structures by MD simulation model.
Results: Resistance against CD4MCs was associated with V255M,
T375N/I, or M426I substitutions. We examined susceptibilities of these
mutated clones to the CD4MCs, maraviroc (MVC), an entry inhibitor
IC9564, CD4bs nMAb 3D6, and CD4i nMAb 4E9C. V255M, T375I, and
M426I were associated with high level of resistance to all CD4MCs
tested, while there was no substantial difference between the wild type
and the mutated clones in sensitivity of MVC and IC9564. The V255M
and M426I clones became resistant to 4E9C, whereas the clone with
T375I showed low sensitivity to both 3D6 and 4E9C. MD simulations
of KP-5P gp120 in complex with NBD-556 showed that (i) V255M
mutation abolished the interaction of NBD-556 and gp120, and (ii)
M426I mutation disconnected a hydrogen bond between Lys130 and
Glu429, thus the NBD-556 binding site shifted different from the usual.
Conclusions: These data may give important knowledge for combination
of NBD and other entry inhibitors or nMAbs.
312
Carolina Herrera1, Natalia Olejniczak1, Javier García Pérez2,
José Alcamí2, Loïc Martin3, Oliver Hartley4, Charles Kelly5, Robin
Shattock1
Instituto de Salud Carlos III, Madrid, Spain, 3Comissariat à l’Energie
Atomique, Giff-sur-Yvette, France, 4University of Geneva, Geneva,
Switzerland, 5King’s College London, London, United Kingdom
1
2
Background: Multiple drug combinations as microbicides have been
shown to be highly effective in preclinical studies against wild type
HIV-1 isolates. This study aims to assess the activity of entry inhibitors
(EIs) combinations with a nucleotide reverse transcriptase (RT) inhibitor
(NRTI), against resistant HIV-1 and SIV isolates
Methods: Antiviral efficacy of dual combinations of an NRTI, tenofovir
(PMPA), and EIs, a CD4 mimetic miniprotein, M48-U1, or CCR5 inhibitors,
5P12-RANTES or maraviroc; was evaluated. The combinations were
assessed in cellular (TZM-bl cells and activated PBMCs) and colorectal
explant models. Preincubation of cells or tissue with the drugs
individually or in combination, for one hour was followed by addition
of virus. NRTI-escape mutants with point mutations K65R +/- M184V in
HIV-1YU.2 and SIVmac32H RT were used. Infection was determined by
measurement of luciferase expression (in TZM-bl cells) or p24/p27 viral
antigen in culture supernatants
Results: All PMPA-EI dual combinations inhibited the NRTI-resistant
clones in all cellular and explant models tested. The dose-response
curves of combinations including M48-U1 or 5P12-RANTES reflected
the activity of the EI with no increase of potency of these drugs when
combined with PMPA. The same result was observed with the gelformulated version of M48-U1. Interestingly, an increase of activity was
observed for maraviroc and PMPA when used in combination against all
resistant isolates tested
Conclusions: The positive results obtained against clade B NRTI-resistant
HIV-1 isolates in this pre-clinical evaluation indicate that combinations
of EIs with PMPA are good candidate microbicides able to block wildtype viruses and, importantly, NRTI-resistant isolates, which have been
shown to be in increasing prevalence
Posters 36: HIV Incidence and Prevalence
P36.01
P36.02
Participation in Clinical Research Could
Modify Background Risk for Trial Outcome
Measures
Trends of Reported HIV Sexual Risk
Behaviour and HIV Incidence among
Fisherfolk in Uganda Receiving Clinic-based
Routine HIV Counseling and Testing
MRC/UVRI, Uganda Research Unit on AIDS, Entebbe, Uganda,
International AIDS Vaccine Initiative, New York, NY, United States
1
2
Background: Data on HIV incidence and retention are needed to
inform study design of efficacy trials. However, the selection criteria and
interventions during an actual clinical trial could reduce HIV incidence
and thus affect the statistical power. We investigated the effect of
inclusion and participation in a simulated vaccine efficacy trial (SiVET)
on HIV and pregnancy incidence in a fisherfolk cohort in SW Uganda.
Methods: High-risk vounteers aged 18-49 years from fishing
communities 30-40km from the MRC/UVRI research centre were
recruited in HIV open cohort. High risk was defined as history of multiple
sex partners, unprotected sex, STI presence and absence from home
for ≥ 2 days in the preceding 3 months. Consenting volunteers with at
least 3 months of follow-up, no contraindications for hepatitis B vaccine
and willing to use contraception were administered a licensed Hepatitis
B vaccine at 0, 1 and 6 months to mimic a candidate vaccine. The
cohort was followed quarterly for a year. HIV incidence, pregnancy and
retention rates were compared.
Results: Of 853 (55% men) individuals screened from Jan 2012-Feb
2014, 575 (60% men, mean age 28) were enrolled into the open cohort,
282 (73% men) of whom enrolled into the SiVET between Jul 2012-Feb
2013. In both groups there was reduction of risky behaviours, (p< 0.05).
A total of 13 HIV incident cases occurred in 93.0 PYO [brackets 95% CI];
incidence 13.9/100 PYO [8.1-24.1] and 10 cases in 311.6 PYO; incidence
3.2 [1.7-6.0] in the open cohort and SiVET respectively. A total of 26
pregnancies were observed in 42.7 Women Years of Observation (WYO);
incidence 60.9 [41.5-89.5], and 4 pregnancies (71.4WYO); incidence 5.6
[2.1-14.8] in the open cohort and SiVET respectively.
Conclusions: Although reduction in risky sexual behaviours was
observed in the open cohort and SiVET, lower HIV and pregnancy
incidence rates were observed in the SiVET. The low HIV incidence could
impact on sample size estimates for a prevention trial.
Ubaldo Mushabe Bahemuka1, Andrew Abaasa1, Eugene
Ruzagira1, Freddie Mukasa Kibengo1, Juliet Ndibazza1, Gershim
Asiki1, Jerry Mulondo1, Matthew Andrew Price2, Patricia Fast2,
Anatoli Kamali1
Medical Research Council/Uganda Virus Research Institute Unit on
AIDS, Entebbe, Uganda, 2International AIDS Vaccine Initiative (IAVI),
1
Background: HIV counseling and testing (HCT) has been shown to
reduce HIV risk behaviour and is central to HIV prevention programs.
We investigated risk behaviour and HIV incidence trends in a fisherfolk
cohort on Lake Victoria, Uganda.
Methods: HIV negative volunteers aged 18-49 years, at high risk of HIV
infection and willing to undergo HCT were enrolled. At every quarterly
visit, they received HCT. Condoms and STI treatment were also provided.
Risk behaviour data on alcohol consumption before sex, multiple or new
sex partners, condom use and exchange of gifts for sex in the past 3
months were collected at baseline and every 6 months for 2 years. We
fitted multilevel logistic regression models to investigate the trends.
Results: A total of 428 (63% men) volunteers, mean age 28 years were
enrolled. There were significant reductions in reported risk behaviours
over the 2-year follow-up. The proportion reporting ≥2 partners
decreased from 80% at baseline to 45% at month 6 and to 43% at
month 24 for males; for females the decrease was from 42% at baseline
to 13% at month 6 and to 6% at month 24; P< 0.01). Similarly there
were significant reductions among men (P=0.01) reporting new partners
but of borderline statistical significance among females (P=0.09). In
both sexes there were significant decreases in reported non-condom
use, transactional sex and in having sex when drunk. HIV incidence (in
brackets 95% CI) reduced from 8.2/100 person years (5.1-13.5), to 7.3
(5.0-10.6), 6.5 (4.6-9.1) and 6.0 (4.3-8.3) at 6, 12, 18 and 24 months
respectively (p=0.21).
Conclusions: In this study there was a substantial reduction in selfreported risk behaviour in the first 6 months and marginal reduction
in the later period. However, a modest HIV incidence reduction was
observed. This calls for an urgent need for combination prevention
strategies in this population.
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313
POSTERS
Andrew M. Abaasa1, Gershim Asiki1, Jonathan Levin1, Ubaldo
Bahemuka1, Eugene Ruzagira1, Freddie M. Kibengo1, Jerry
Mulondo1, Juliet Ndibazza1, Matthew A. Price2, Pat Fast2, Anatoli
Kamali1
Posters
P36.03
P36.04
Development of a Risk Scoring Tool to Predict
HIV-1 Acquisition in African Women
Age-disparate Partnerships and Risk of HIV1 Acquisition among South African Women
Participating in the VOICE Trial
Jennifer E. Balkus1,2, Jingyang Zhang1, Gonasagrie Nair3, Thesla
Palanee4, Gita Ramjee5, Clemensia Nakabiito6, Marthinette
Taljaard7, Baningi Mkhize8, Zvavahera Mike Chirenje9, Jeanne M.
Marrazzo2, Elizabeth R. Brown1,2, Barbra A. Richardson1,2
Jennifer E. Balkus1,2, Gonasagrie Nair3, Elizabeth Montgomery4,
Anu Mishra2, Thesla Palanee5, Gita Ramjee6, Ravindre Panchia7,
Pearl Selepe8, Barbra A. Richardson1,2, Zvavahera Mike Chirenje9,
Jeanne M. Marrazzo2
Fred Hutchinson Cancer Research Center, Vaccine and Infectious
Disease Division, Seattle, WA, United States, 2University of Washington,
Seattle, WA, United States, 3CAPRISA/University of Kwa Zulu Natal,
Durban, South Africa, 4Wits Reproductive Health and HIV Institute,
University of the Witwatersrand, Johannesburg, South Africa, 5South
African Medical Research Council, Durban, South Africa, 6Makerere
University - Johns Hopkins University Research Collaboration, Kampala,
Uganda, 7The Aurem Institute, Klerksdorp, South Africa, 8Chris Hani
Baragwanath Hospital, Johannesburg, South Africa, 9UZ - UCSF, Harare,
Zimbabwe
Fred Hutchinson Cancer Research Center, Vaccine and Infectious
Disease Division, Seattle, WA, United States, 2University of Washington,
Seattle, WA, United States, 3CAPRISA/University of Kwa Zulu Natal,
Durban, South Africa, 4RTI International, San Francisco, CA, United
States, 5Wits Reproductive Health and HIV Institute, University of the
Witwatersrand, Johannesburg, South Africa, 6South African Medical
Research Council, Durban, South Africa, 7Chris Hani Baragwanath
Hospital, Johannesburg, South Africa, 8The Aurem Institute, Klerksdorp,
South Africa, 9UZ - UCSF, Harare, Zimbabwe
Background: In many African countries, women account for more than
half of all new HIV-1 infections; however, not all women are at equal
risk of acquiring HIV-1. A risk prediction tool that can identify women
at highest risk for HIV-1 acquisition could improve prevention research
efficiency and inform HIV-1 prevention activities in policy and clinical
settings.
Methods: Using baseline data from VOICE (MTN-003), a randomized,
double-blinded, placebo-controlled trial conducted in South Africa,
Uganda and ZImbabwe that assessed safety and effectiveness of daily
oral and vaginal chemoprophylaxis for HIV-1 prevention, we used
standard methods for the development of clinical prediction rules to
generate a risk scoring tool to predict HIV-1 acquisition over the course of
one year. The predictive ability of the score was assessed by calculating
area under the curve (AUC) and the score was internally validated using
10-fold cross-validation.
Results: Among 5,029 women enrolled in VOICE, 4,834 women had
complete data for factors of interest and were included in the analysis;
of these, 248 acquired HIV-1 within one year after enrollment (HIV
incidence=6.05% [248/4,093 person-years]). The final risk score
resulting from multivariable modeling included the following baseline
factors: participant age, married/living with a partner, financial or
material support from partner, partner has other partners, curable STI,
HSV-2 status and alcohol use. The maximum possible score was 12;
36% of participants had a score > 6 and accounted for 66% of HIV-1
infections. The AUC for the score was 0.72 and mean AUC from 10-fold
cross validation was 0.70, indicating good predictive ability.
Conclusions: A discrete set of characteristics which can be easily
assessed were highly predictive of HIV-1 acquisition over one year.
External validation of the risk score is required to evaluate the tool’s
performance when applied to different populations of women at risk for
HIV-1 infection in Africa.
Background: Age-disparate relationships where the male partner is
older than the female partner have been associated with increased HIV
acquisition risk in women. A recent analysis of data from South Africa
failed to observe an association between age-disparate partnerships and
HIV acquisition. We assessed the association between partner age and
HIV acquisition among South African women in VOICE.
Methods: VOICE was a randomized, double-blinded, placebo-controlled
trial conducted at 11 sites in South Africa that assessed the safety
and effectiveness of daily oral and vaginal chemoprophylaxis for HIV
prevention in women. Cox proportional hazards models stratified by site
were used to assess participant-reported male partner age at enrollment
and HIV acquisition risk in the first year of follow-up.
Results: Among 4077 South African women enrolled, 3789 had
complete data for this analysis. Of these, 26% and 5% reported having
a male partner >5 and >10 years older at enrollment, respectively.
There were 230 HIV infections within 1 year of follow-up (3181 personyears). Reporting a male partner >5 years older was not associated with
HIV acquisition (HR 1.00; 95% CI 0.74, 1.35). Findings were similar
for reporting a male partner >10 years older (HR 0.92; 95% CI 0.49,
1.74). Results for both male partner age categories were similar among
younger and older women (women < 25 years: male >5 years, HR 1.01
[95% CI 0.71, 1.44]; male >10 years, HR 1.24; [95% CI 0.58, 2.66]
versus women ≥25 years: male >5 years, HR 0.95 [95% CI 0.55, 1.65];
male >10 years, HR 0.68 [95% CI 0.21, 2.17]). Results were consistent
after adjusting for known baseline risk factors for HIV acquisition in
VOICE.
Conclusions: These data corroborate recent reports and may suggest
a shift in local epidemiology of heterosexual HIV transmission. Given
the limitations of these analyses (use of enrollment data and inclusion
of only one partner in analysis), regular assessment of characteristics to
identify women at greatest risk of HIV acquisition is needed.
1
POSTERS
314
1
P36.05
P36.06
Estimating HIV Incidence for Identification
of Microbicide Trial Sites in India: A Crosssectional Study
Zambia FSW-risks Descriptors: HIV, Retention,
Condom Use and Trichomonas Vaginalis/
Sperm Trends over Time
Nomita Chandhiok1, Ramesh S. Paranjape2, Sanjay M.
Mehendale3, Archana Beri2, Sanjay Chauhan4, R Hari Kumar5,
Seema Sahay2, Reynold Washington6, Marianne Callahan7
Linda J. Kimaru1, Tyronza Sharkey2, Marydale Oppert3, Kathleen
Wu2, Rachel Parker3, William Kilembe2, Mubiana Inambao1,
Amanda Tichacek3, Susan Allen3
Indian Council of Medical Research, Division of Reproductive and
Child Health, New Delhi, India, 2National AIDS Research Institute, Pune,
India, 3National Institute of Epidemiology, Chennai, India, 4National
Institute for Research in Reproductive Health, Mumbai, India, 5National
Institute for Nutrition, Hyderabad, India, 6Karnataka Health Promotion
Trust, Bengaluru, India, 7CONRAD, Eastern Virginia Medical School,
Arlington, VA, United States
1
Rwanda Zambia HIV Research Group, Ndola, Zambia, 2Rwanda Zambia
HIV Research Group, Lusaka, Zambia, 3Rwanda Zambia HIV Research
Group, Atlanta, GA, United States
1
Background: India has the second largest burden of HIV in the world
with estimated 2.1 million infections. Of these, 39.3% are estimated
to be women. The present study was designed to characterize HIV-1
incidence in select clinical site(s) suitable for conducting HIV prevention
trials in India.
Methods: This cross-sectional study was conducted in Female Sex
Workers (FSWs) between Jan 2012 and June 2013 in six districts with
historical evidence of concentrated HIV epidemic, from three high HIV
prevalence states of India, namely Maharashtra, Andhra Pradesh and
Karnataka. A total of 9138 FSWs were enrolled. HIV incidence was
estimated on sero-positive samples using serological assays such as
BED-Capture Enzyme Immuno-assay (CEIA) and two avidity assays:
modified GS HIV ½ (Biorad) and LAg HIV-1 (Sedia). In addition,
molecular estimation was carried out on sero-negative samples using
pooled PCR. Furthermore, HIV Incidence was calculated using Recent
Infection Testing Algorithm (RITA) which was developed by including
BED-CEIA, an avidity assay, and CD4 count in series.
Results: While the HIV prevalence in this population was estimated at
9.26%, the incidence by serological assays ranged from 0.144 to 2.4.
The False Recent Rates (FRR) for the two avidity assays were different;
however, the incidence estimates were comparable. Similar HIV
incidence was also obtained using RITA. HIV incidence by Pooled PCR
was significantly different from serological assays.
Conclusions: The incidence estimates varied with the test applied.
Although there is no gold standard test available, two avidity based
assays gave comparable results. Adding CD4 counts to RITA did not
improve the outcome. The low HIV incidence estimates, obtained using
serological assays in a cross-sectional study of FSWs in high prevalence
states, suggest that efficacy trials for vaginal microbicides would require
large sample sizes in India.
Background: Female Sex Workers (FSWs) in Zambia are at high risk
for HIV and may be eligible for HIV prevention clinical trials. HIVFSWs were invited to enroll into a prospective cohort to determine the
incidence and risk factors for HIV.
Methods: From 2012 to 2014 FSWs in Lusaka and Ndola were invited
for VCT/STI services from known FSWs hotspots through direct and peer
FSW outreach activities. FSWs received HIV/STI testing and counseling
and were offered Long Acting Reversible Contraceptives (LARC, IUD and
implant). HIV- FSWs were invited for enrollment and follow-up visits
quarterly for HIV/STI testing and counseling. Demographic and risk
assessment questionnaires were done at each visit.
Results: Among 733 FSWs screened, 391 (53%) were HIV-. Of the HIV-,
9% were positive for T. Vaginalis, 3% were positive for syphilis, 3% had
presence of semen on their vaginal swabs and 19% were on LARC. The
297 (75%) HIV- FSW who returned for enrollment reported an average
of 21 clients a month including and 2 repeat/regular clients. 17%
reported condom use always/most of the time and 83% sometimes/
never. 48% reported non-condom use due to client request/refusal. 187
(64% of those enrolled) FSWs came for at least 1 follow up visit, with
a total of 803 months of follow up. 11 initiated LARC. 32% reported
disclosing their HIV status by client request and 60% had voluntarily
disclosed. 64% had requested the HIV status of their clients. At followup 7% were positive for T. Vaginalis, 3% had presence of semen, and
70% reported not using a condom within the last month. Annual HIV
incidence was 6%.
Conclusions: HIV- FSW are at high risk for HIV/ STI infection and should
be targeted for prevention trials. Though HIV- FSW received counseling,
STI and LARC services, they maintained high-risk behaviors including
unprotected sex. Disclosure of HIV- status may be dangerous for FSW
if clients use this as an excuse to refuse condom use. Asking clients for
their HIV status shows awareness but may not yield truthful information.
www.hivr4p.org
315
POSTERS
Posters
P36.07
P36.08
Hematological Profiles of HIV-infected Adults
Initiating Highly Active Antiretroviral Therapy
(HAART) in Uganda
Characteristics of Clients Undergoing Repeat
HIV Counseling and Testing Compared to
Clients Newly Tested for HIV in Nyanza
Province, Kenya
Rachel Kyeyune1, Elmar Saathoff2, Amara Ezeamama3, Wafaie
Fawzi4, Thomas Loescher2, David Guwatudde5
Infectious Diseases Institute, Research, Kampala, Uganda, Medical
Center of the University of Munich, Division of Infectious Diseases
and Tropical Medicine, Munich, Germany, 3University of Georgia,
Epidemiology and Biostatistics, Athens, GA, United States, 4Harvard
School of Public Health, Nutrition, Boston, MA, United States, 5Makerere
University College of Health Sciences, School of Public Health,
Epidemiology and Biostatistics, Kampala, Uganda
1
2
POSTERS
Background: Cytopenias are the most common HIV-associated
hematological abnormality. Cytopenias become more prevalent as
HIV progresses and are often fatal. Sex, race, geographical location
and comorbidities such as tuberculosis have been associated with
cytopenias. Data from resource-limited settings about the prevalence,
correlates and trends in cytopenia are limited. This analysis assessed
the prevalence and correlates of cytopenia at initiation of HAART and
the trend in cytopenias among HAART-treated AIDS patients in Uganda.
Methods: This is a secondary analysis of hematological data of 400
adults enrolled into the Multivitamins, HAART and HIV/AIDS Trial
(NCT01228578). Anemia was defined according to WHO guidelines
and leucopenia and thrombocytopenia were defined using study site
laboratory reference ranges for lack of generally accepted standardized
definitions for these 2 cell lines. Univariate and bivariate analyses were
done to describe the patient population and log-binomial regression was
used to quantify the correlates of cytopenia. Multilevel Mixed-effects
linear regression was used to examine the change in the 3 cell lines over
18 months of HAART
Results: Sixty five percent had at least one form of cytopenia and the
prevalence was higher in females(PR 1.21 CI 1.01-1.43) and higher with
decreasing CD4 count and decreasing body mass index. Anemia was
the most common occurring in 47.8%. Adjusted models showed that
hemoglobin values were 0.03g/dl higher with each month of HAART
(p< 0.001) while white blood cell counts and platelets were lower by
0.01(p=0.009) and 0.15 (p=0.522) units respectively with each month
of HAART.
Conclusions: Cytopenias are a frequent complication in HIV-infected
adults at initiation of HAART in Uganda. Females, a decreasing CD4
count and decreasing body mass index were associated with having
a cytopenia. This data shows that HAART improves hemoglobin status
and alters the white blood cell and platelet counts independent of sex,
immunological and nutritional parameters.
316
Patrick O. Owiti1, Kevin Owuor2, Hillary Ng’eno1, Nicollate
Awuor1, Patricia Ong’wen1, Starley B Shade3, Jayne Lewis-Kulzer3,
Elizabeth A Bukusi1, Craig R. Cohen3
Kenya Medical Research Institute, Family AIDS Care and Education
Services, Kisumu, Kenya, 2University of Reading, Statistical Services
Center, Reading, United Kingdom, 3University of California San
Francisco, Obstetrics, Gynecology and Reproductive Sciences,
Pediatrics, Medicine, San Francisco, CA, United States
1
Background: According to Kenya AIDS Indicator Survey 2007, only
35.6% of Kenyan adults had ever tested for HIV. Since then, routine
HIV counseling and testing (HTC) has increased in health facilities. We
compared characteristics of new and repeat clients tested for HIV to
inform efforts to improve testing uptake.
Methods: This retrospective study included a proportional stratified
random sample of adult clients (≥18 years) tested at the outpatient
department from October-December, 2011 at 9 high patient volume
facilities in Nyanza Province (collectively testing >12,000 clients in
one quarter). Routine data were abstracted from health facility registers
through systematic selection of every 51st adult patient. Variables
included age, gender, HIV status, individual vs. couples testing, test
type (new/repeat) and interval to repeat test. Descriptive statistics were
presented as medians or proportions. Logistic regression was used to
assess differences between new and repeat HIV testers.
Results: Among the 555clients sampled, the median age was 27 years
(IQR22-35), 365(66%) were female, 397(71%)were repeat testers and
521 (94%) tested as individuals. Median time to repeat test was 4 months
(IQR3-7).New testers were older (aOR=1.36 per 10 year age increase;
95%CI1.17-1.58). HIV prevalence among new testers was higher than
among repeat testers (27% vs. 13%, respectively) (aOR2.64; 95%
CI1.65-4.21).No significant gender differences were found between new
and repeat testers (OR1.31; 95 % CI0.89-1.92) or individual vs. couple
visit (OR1.21; 95% CI0.58-2.56).
Conclusions: The majority of adult patients seeking HIV testing have
been tested previously. Those newly tested for HIV have a higher HIV
prevalence than repeat testers. This may indicate that initial testing
is reaching higher risk individuals and that HTC along with other
preventive interventions may have led to lower HIV incidence amongst
repeat testers. Further attention to behavioural differences between new
and repeat testers should be examined.
P36.09
P36.10
ABSTRACT WITHDRAWN
HIV and STI Incidence and the Association
with Number of Lifetime Sexual Partners Case for Combination HIV/STI Prevention
Strategies
Renee A. Street1, Neetha Morar1, Handan Wand2, Gita Ramjee1
South African Medical Research Council, Durban, South Africa, 2Kirby
Institute, University of New South Wales, Sydney, Australia
Background: South Africa has a generalised HIV epidemic driven
largely by heterosexual transmission. Multiple sexual partnerships is
believed to be an important driver of the HIV epidemic. The aim of this
study is to describe socio-demographic characteristics of women by the
number of lifetime sexual partners and its association with incident HIV
and sexually transmitted infections (STI).
Methods: The Methods for Improving Reproductive Health in Africa
(MIRA) clinical trial was conducted between 2003 and 2006. In Durban
(KwaZulu-Natal Province), a total of 1485 women were enrolled from
two sights (a peri-urban clinic in Umkomaas and a rural clinic in Botha’s
Hill) and were followed up for a total of 24 months. The chi squared
test was used to compare categorical parameters. Kaplan-Meier survival
analyses were carried out to estimate the crude HIV and STI incidence
rates over time. All analyses were performed using Stata V.10.0 (College
Station) and SAS V. 9.2
Results: Women with a greater number of lifetime sexual partners (5+)
were older (35+ years of age), had early sexual debut (< 16 years of
age) and were unmarried (p< 0.001). Cohabitation status and level of
education were not significantly associated with lifetime number of
sexual partners. Women with 5+ lifetime sexual partners had a crude
HIV incidence reported at 10.5 per 100 person years (PY). Women with
3 lifetime sexual partners were at higher risk of STI acquisition (25 per
100 PY) when compared with women with 1 lifetime sexual partner (14
per 100 PY).
Conclusions: This study supports existing evidence that reducing
partner turnover is key in HIV prevention. However combination HIV/
STI prevention strategies are essential to achieve maximum impact
on HIV prevention. Women remain critical participants in investigative
biomedical and translational HIV prevention efforts.
www.hivr4p.org
317
POSTERS
1
Posters
P36.11
High HIV Incidence in Young Men who Have
Sex with Men Engaged in Sex Parties, Factors
Associated with Sex Party, Bangkok MSM
Cohort Study, 2006-2014
Warunee Thienkrua1, Sarika Pattanasin1, Tareerat Chemnasiri1,
Anchalee Varangrat1, Wichuda Sukwicha1, Supaporn
Chaikummao1, Sumetha Hengprasert1, Anupong Chitwarakorn2,
Timothy H. Holtz1,3
Thailand-MoPH U.S. CDC Collaboration, Nonthaburi, Thailand,
Department of Diseases Control, Ministry of Public Health, Nonthaburi,
Thailand, 3Division of HIV/AIDS Prevention, U.S. Centers for Disease
Control and Prevention, Atlanta, GA, United States
1
2
POSTERS
Background: Evidence concerning sex party participation and
recreational drug use along with continuing high rates of HIV and
STIs among MSM in Thailand has been described in several studies.
We investigated HIV incidence among young MSM (YMSM) and
characteristics associated with sex party participation.
Methods: Thai men, ≥18 years from the Bangkok metropolitan area
who reported sex with another man in the past 6 months were enrolled
and followed-up every four months for HIV testing and audio computerassisted self-interview behavioral questionnaire completion. We defined
engaging in sex parties as having group sex and using recreational or
erectile dysfunction drugs in the past four months preceding enrollment.
We calculated HIV incidence among YMSM 18-24 years of age from
2006-2014 using survival analysis. Factors associated with sex party
participation were analyzed using logistic regression.
Results: Of 1744 men enrolled, 712 (40.8%) were YMSM, 77/712 (11%)
reported engaging in sex parties, 428/712 (60%) reported unprotected
anal intercourse in the past 4 months. HIV incidence among YMSM
engaged in sex parties was 8 per 100 Person-Years. Factors associated
with sex parties were: sex with casual partners at pub/disco [Adjusted
Odd Ratio (AOR) 5.8, 95% CI 2.0-16.9], paid for sex (AOR 4.1, 95% CI
1.9-8.8), sex with foreigner (AOR 3.3, 95% CI 1.9-5.8), self-report of STIs
(AOR 2.8, 95% CI 1.5-5.1), had HIV infection (AOR 2.6, 95% CI 1.5-4.7),
reporting Internet use to find casual partners (AOR 2.0, 95% CI 1.1-3.5),
ever had suicidal idea (AOR 2.1, 95% CI 1.2-3.7), and experience of
coercive sex (AOR 1.9, 95% CI 1.03-3.5).
Conclusions: Engaging in sex parties was common among Bangkok
YMSM in our cohort. Sex at entertainment venues and commercial
sex are strongly associated with sex parties, both high-risk behaviors
for incident HIV. Innovative and creative HIV interventions directed at
MSM engaging in high-risk behaviors is needed to promote safer sex
behaviors.
318
Posters 37: Immunogenetics
P37.01
P37.02
Variants in Vitamin D Pathway and Antiviral
Response Genes Interact to Modulate the
Natural Resistance to HIV-1 Infection
The Expression Analysis of Hexokinase
1 Gene in the Pumwani Commercial Sex
Worker Cohort, Nairobi, Kenya
Wbeimar Aguilar-Jiménez1, Wildeman Zapata1,2, Antonio Caruz3,
Joan Fibla4, Marina Laplana4, Antonio Rivero5, Juan A. Pineda6,
Maria T. Rugeles1
Winnie Apidi1, Ruey Chi Su1, Joshua Kimani1,2, Frank Plummer1,2,3,
Blake Ball1,2,3
Background: The immunomodulatory functions of vitamin D (VitD)
may orchestrate anti-HIV-1 responses and influence resistance to HIV-1infection exhibited by HIV-1-exposed seronegative (HESN) individuals.
Methods: We performed a nested case-control study, involving HIV1-exposed seropositives (cases) and seronegatives (controls), from two
cohorts: sexually-exposed from Colombia, and parenterally-exposed
from Spain. The association of 140 variants in 9 genes of the VitD
pathway, and in 13 genes of the antiviral response with resistance/
susceptibility (R/S) to HIV-1 was evaluated.
Results: Twenty-four variants were associated with R/S to HIV-1 infection
at p< 0.05 (including 6 variants at false discovery rate [FDR] ≤ 20%).
Similarly, 12 haplotypes in both pathways were also associated with
R/S to HIV-1 after Bonferroni correction. Some variants in VitD pathway
genes displayed epistatic interactions with those in antiviral genes and
such interactions were associated with R/S to HIV-1 infection at p< 0.01,
but not after Bonferroni correction. Furthermore, most of the HESNs
exhibited the same combinations of the variants in VitD pathway and
antiviral genes (p< 0.003 after Bonferroni correction). Remarkably, we
observed that HESN individuals carrying resistance-associated variants
in genes coding for the vitamin D receptor (VDR), 27-OH hydroxylase
(CYP27A1), and Toll-like receptor 2 (TLR2), had higher levels of VitD in
plasma, of VDR mRNA in blood leucocytes and mRNA of beta-defensins
in mucosa, suggesting that the expression of these molecules could be
genetically determined. Interestingly, variants in antimicrobial peptides
that exert their function on mucosal surfaces were associated with R/S
in sexually- but not in parenterally-exposed individuals, highlighting
differences in mechanisms underlying resistance to HIV-1, depending
on the route of exposure.
Conclusions: These results suggest that the VitD pathway may act
in concert with antiviral genes modulating the resistance phenotype
observed in HESN individuals.
University of Manitoba, Medical Microbiology & Infectious Diseases,
Winnipeg, MB, Canada, 2University of Nairobi, Nairobi, Kenya, 3Public
Health Agency of Canada, National Microbiology Laboratory, Winnipeg,
MB, Canada
1
Background: Altered susceptibility to HIV-1 infection has been observed
in a commercial sex worker (CSW) cohort in Nairobi, Kenya, where
a subset of women are classified as HIV-1 exposed yet seronegative
(HESN). A gene expression analysis conducted showed differential
regulation of the glycolysis/gluconeogenesis pathway in HESN CSWs.
The first and potentially critical regulatory step in glucose metabolism
is its entry into lymphocytes, where glucose binds to Hexokinase-1 that
prevents it from leaking out. Together with the Glucose Transporter 1,
Hexokinase-1 regulates the first rate-limiting steps of the entire glucose
metabolism by phosphorylating glucose into glucose-6-phosphate,
which is the starting material for glycolysis.
Methods: The study population was randomly selected from the
Pumwani Sex Worker Cohort, Nairobi including: HIV highly exposed yet
seronegative (HESNs) CSWs (>7 years); newly enrolled HIV-uninfected
(< 3 years) CSWs, 85% of whom would likely seroconvert to HIV-1
positive; and lowly-exposed HIV negative antenatal clinic attendees with
low exposure to HIV. Total RNA was extracted from PBMCs using Trizol;
cellular Hexokinase-1 mRNA levels were quantified by quantitative real
time PCR using SYBR Green. Statistical analysis was performed using
Mann-Whitney U Test. Differences were considered to be significant if P<
0.05. Each assay was normalized using 18s rRNA gene.
Results: We observed significantly lower level of Hexokinase-1 mRNA
expression in HESNs when compared to that in newly enrolled HIV
uninfected CSWs. (Hexokinase-1 p=0.0323)
Furthermore, the levels of Hexokinase-1 mRNA in HESN and the HIV
negative antenatal clinic attendees were quite similar. (Hexokinase-1
p=0.6448)
Conclusions: Lower expression of Hexokinase-1 in HESN might suggest
lower regulation of glucose uptake. Hexokinase is a rate-limiting
enzyme in the glycolysis pathway. Following studies of expression and
uptake are underway to understand its role in glucose metabolism in
HIV resistance.
www.hivr4p.org
319
POSTERS
Grupo Inmunovirología, Universidad de Antioquia UdeA, Medellin,
Colombia, 2Grupo Infettare. Universidad Cooperativa de Colombia,
Medellin, Colombia, 3Unidad de Inmunogenética, Facultad de Ciencias
Experimentales, Universidad de Jaén, Jaen, Spain, 4Unitat de Genètica
Humana, Departament de Ciències Mèdiques Bàsiques, IRBLleida,
Universitat de Lleida, Lleida, Spain, 5Maimonides Institute for Research
in Biomedicine of Cordoba (IMIBIC)/Reina Sofia University Hospital,
Cordoba, Spain, 6Unidad Clínica de Enfermedades Infecciosas y
Microbiología. Hospital Universitario de Valme, Sevilla, Spain
1
Posters
P37.03
P37.04
Subtype-Specific HIV-1 Adaptation to Host
HLA
Characterization of the 3’ Untranslated
Region of HLA-G in HIV-1 Infected Black
South African Mothers and their Infants
Guinevere Q. Lee1, Jonathan Carlson2, Chanson J. Brumme1,
Helen Byakwaga3,4, Conrad Muzoora3, Daniel MacMillan5, Natalie
Kinloch5, Kyle Cobarrubias5, Mark A. Brockman1,5, Peter W. Hunt4,
Jeff N. Martin4, Mary Carrington6, David R. Bangsberg7, P. Richard
Harrigan1,8, Zabrina L. Brumme1,5
BC Centre for Excellence in HIV/AIDS, Vancouver, BC, Canada,
Microsoft Research, Seattle, WA, United States, 3Mbarara University
of Science and Technology, Mbarara, Uganda, 4University of California
San Francisco, San Francisco, CA, United States, 5Simon Fraser
University, Faculty of Health Sciences, Burnaby, BC, Canada, 6Frederick
National Laboratory for Cancer Research, Cancer and Inflammation
Program, Laboratory of Experimental Immunology, SAIC Frederick,
Inc., Frederick, MD, United States, 7Massachusetts General Hospital and
Harvard University, Boston, MA, United States, 8University of British
Columbia, Department of Medicine, Vancouver, BC, Canada
1
2
POSTERS
Background: HIV-1 adapts to HLA alleles expressed by infected hosts
and host populations. However, the extent to which HIV’s genetic context
influences population-level adaptation to HLA remains incompletely
understood. The Ugandan HIV epidemic, where subtypes A and D
co-circulate, provides a unique opportunity to distinguish universal
pathways of HIV adaptation from those that are subtype-specific.
Methods: High-resolution HLA class I and HIV RNA gag genotyping
were performed for 513 antiretroviral-naïve patients from Kampala and
Mbarara, Uganda. Recombinant and non-A/D sequences were excluded,
leaving 200 subtype A and 135 subtype D gag sequences for analysis.
HLA-associated polymorphisms were identified in each subtype via
statistical association with phylogenetic correction, after which a logistic
regression approach was used to determine cases where the same HLA
allele drove significantly different escape pathways between subtypes.
Results: Of 103 unique HLA alleles observed in the study cohort, only
3 (B*58:01, C*16:02, A*26:12) differed in frequency between subtypes
A and D (p< 0.05), consistent with a single host population where HIV
subtypes co-circulate. A total of 55 HLA-associated polymorphisms at
25 Gag codons were identified at the population level in subtype A;
36 HLA-associated polymorphisms at 25 Gag codons were identified
in subtype D (p< 3x10-4, q< 0.2). Comparative analysis revealed that
>35% of these adaptations differed significantly between subtypes. For
example, B*57:03 drove the selection of T242N in subtype D (Odds
Ratio~250, p=2x10-10) but not in subtype A (inter-subtype comparison
p=8x10-6), whereas Y79F selection by A*01:01 was six-fold stronger in
subtype A (OR~20) versus D (p=2x10-3).
Conclusions: HIV’s genetic context exerts a substantial influence on
its ability to adapt to HLA. Establishing whether this is attributable to
differential epitope presentation, mutational constraints or other factors
is relevant to vaccine design.
320
Heather A. Hong1,2, Maria Paximadis1,2, Glenda E. Gray3, Louise
Kuhn4, Caroline T. Tiemessen1,2
Centre for HIV & STIs, National Institute for Communicable Diseases,
NHLS, Johannesburg, South Africa, 2Faculty of Health Sciences,
University of the Witwatersrand, Johannesburg, South Africa, 3Perinatal
HIV Research Unit, Chris Hani Baragwanath Hospital, Soweto, South
Africa, 4Gertrude H. Sergievsky Centre, College of Physicians and
Surgeons, Mailman School of Public Health, Columbia University,
Department of Epidemiology, New York, NY, United States
1
Background: HLA-G is a tolerogenic molecule capable of inhibiting
the cytolytic activity of natural killer cells and cytotoxic CD8 T-cells. A
14bp insertion-deletion (indel) within the 3’ untranslated region (3’UTR)
has been found to alter HLA-G expression, wherein high expression
was associated with the Del/Del genotype. Other single-nucleotide
polymorphisms (SNPs) within the 3’UTR have also been reported to
influence HLA-G expression by binding several microRNAs. Recently,
the 14bp indel has been associated with susceptibility to vertical
transmission of HIV-1; however the roles of the other 3’UTR SNPs have
not been extensively investigated.
Methods: We sequenced the 3’UTR of HLA-G in a total of 216 Black
South African HIV-1 infected mother-infant pairs as well as 71 HIVnegative controls. Mother-infant pairs were classified as HIV-1 nontransmitting (NT, n=144) or HIV-1 transmitting (TR, n=72).
Results: We identified 8 previously reported polymorphisms arranged in
8 distinct haplotypes. Heterozygosity at three of the polymorphisms (the
14bp indel, +3010C/G and +3142G/C SNP) were significantly underrepresented in HIV-positive mothers compared to HIV-negative controls
(P=0.019, P=0.001 and P=0.001, respectively). SNPs +3010C/G and
+3142G/C were in complete linkage disequilibrium, and the +3142G/C
SNP was predicted to bind to three miRNAs (miR-148a, miR-148b, and
miR-152). There were no significant differences in variants of the 3’UTR
between NT and TR mothers, or their exposed-uninfected and HIVinfected infants.
Conclusions: This is the first description of HLA-G 3’UTR variability
in a Black South African population. The data suggest that, in our
cohort, variations within the 3’UTR of HLA-G did not influence vertical
transmission. However, significantly higher heterozygous representation
for the 14bp indel and +3142G/C SNP in HIV-negative controls as
compared to HIV-infected individuals, suggests a fine balance in HLA-G
expression may be necessary for protection against sexual acquisition
of HIV-1.
P37.05
P37.06
Fcγ Receptor Variability in the South African
Population - Will this Impact on HVTN097
Vaccine Efficacy?
A Potential Role for CXCR6 in Long-term
Nonprogression of HIV-1 Infected Black South
African Individuals
Ria Lassauniere1,2, Caroline T. Tiemessen1,2
Anabela C. P. Picton1, Maria Paximadis1, Neil Martinson2, Caroline
T. Tiemessen1
Background: Vaccine-induced IgG binding antibodies interacting
with Fcγ receptors (FcγR) may reduce infection risk through antibodydependent cellular cytotoxicity, triggering of soluble antiviral factors,
or phagocytosis. The potential role of FcγR genetic variation in vaccine
efficacy (VE) was previously demonstrated by the significant association
of the FcγRIIc-T118I variant with VE in RV144 vaccinees. With the
RV144 follow up trial (HVTN097) commencing in Southern Africa, we
considered it important to establish the FcγR genetic variability in South
Africans, given that this may influence VE.
Methods: We genotyped FcγR functional variants and gene copy
number in 131 South African Black individuals. The following variants
were assessed: 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. The FcγR
data from South African Black individuals were compared with that of
the 1000 Genomes Project for Asians, European Caucasians and other
African populations.
Results: The FcγRIIc-118I allele previously associated with VE was not
detected in the 131 South African Black individuals. Furthermore, none
were predicted to express the FcγRIIc protein. Genotype frequencies
were significantly different when comparing South African Black
individuals with Asians (FcγRIIa-p.H131R, P < 0.0001; and FcγRIIIa-p.
F158V, P < 0.01), or with European Caucasians (FcγRIIb-p.I232T, P <
0.0001; and FcγRIIIa-p.F158V, P ≤ 0.001). Of note were the significantly
different genotype frequencies between African populations (South
African, Kenyan, Nigerian) for FcγRIIa-p.H131R, FcγRIIb-p.I232T, and
FcγRIIIa-p.F158V, suggesting differing FcγR-mediated immune function
capabilities.
Conclusions: The HVTN097 trial will commence in different African
populations, who are characterized by substantial inter-population
genetic diversity, which may differentially influence VE in the populations
most in need of an effective HIV vaccine.
Centre for HIV and STIs, National Institute for Communicable Diseases,
NHLS and Faculty of Health Sciences, University of the Witwatersrand,
Johannesburg, South Africa, 2Perinatal HIV Research Unit, Chris Hani
Baragwanath Hospital, Soweto, South Africa
1
Background: A role for the HIV secondary coreceptor, CXCR6, in
long-term nonprogression (LTNP) to AIDS has been reported. A single
nucleotide polymorphism (SNP), rs2234358, located 42bp downstream
from the CXCR6 termination codon, significantly associates with LTNP
but not elite control (EC) in cohorts of European descent. In this study
we investigated the role of rs2234358 in HIV-1 disease progression in
populations of sub-Saharan descent.
Methods: Study participants included HIV-1-infected Black South African
individuals with differing disease phenotypes (progressors, n=109;
LTNPs, n=47; ECs, n=11) and healthy HIV-uninfected Caucasian (n=28)
and Black (n=36) individuals. A continuous region encompassing the
CXCR6 open reading frame (ORF) and the untranslated regions (7.0kb)
was amplified in 4 overlapping sections. Amplified fragments were
sequenced and analysed for the presence of SNPs, indels and intragenic
haplotypes to identify potential rs2234358-associated haplotypes. Realtime PCR assays were developed to detect the rs2234358 SNP and
another SNP, CXCR6-E3K, also shown to associate with clinical outcome
of HIV-1 infected individuals.
Results: The rs2234358 SNP frequency did not differ between LTNPs
(51.1%) and progressing (56.9%) Black individuals (P=0.386). CXCR6
gene sequencing failed to detect linkage disequilibrium between
rs2234358 and other SNPs within CXCR6. CXCR6-E3K, located in the
ORF, was found to be present at high allelic frequencies within control
Black individuals (44.4%) and absent in the Caucasian individuals
genotyped. In LTNPs heterozygosity for the E3K SNP was overrepresented
(72.3%; 85.3% of whom also had rs2234358) compared to progressing
(34.9%) individuals (P< 0.001, OR=0.17).
Conclusions: The E3K SNP is likely to affect both CXCR6 cell surface
expression and/or binding of its ligand. Thus, we hypothesize that the
high prevalence and interplay of both these mutations in Black South
African individuals may be masking the effect of the rs2234358 SNP on
LTNP within this population.
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321
POSTERS
National Institute for Communicable Diseases, Centre for HIV and
STI’s, Johannesburg, South Africa, 2University of the Witwatersrand,
Faculty of Health Sciences, Johannesburg, South Africa
1
Posters
P37.07
P37.08
Cytokines Genes Polymorphisms in Ukrainian
HIV-1 Infected Individuals
Multiple T-cell Epitopes of HIV-1 Nef
Containing Positively Selected Mutations
Associated with Different Disease Outcome
Anna Ivanivna Piddubna1
Sumy State University, Infectious Diseases and Epidemiology
Department, Sumy, Ukraine
1
Elnaz Shadabi1, Raghavan Sampathkumar1, John Ho2, David La2,
Rupert Capina2, Binhu Liang2, Jeff Tuff2, Joshua Kimani3, T. Blake
Ball1,2, Francis Plummer1,2, Ma Luo1,2
University of Manitoba, Department of Medical Microbiology,
Winnipeg, MB, Canada, 2National Microbiology Laboratory, Public
Health Agency of Canada, Winnipeg, MB, Canada, 3University of
Nairobi, Nairobi, Kenya
1
POSTERS
Background: The objective of the research was to study distribution
character of the allelic variants of cytokines genes in HIV-1 infected
Ukrainians.
Methods: Data for the study were DNA samples, received from 200
inhabitants of Ukraine: 78 HIV-infected, 22 - HIV-negative individuals
from the group of high risk of contamination, 100 healthy blood
donors. IL-4 (-590C/T), IL-10 (-592C/A) and TNF-α (-308G/A) genes
polymorphisms detection was made with PCR-RLFP method.
Results: By analysis of frequency of IL-4 gene allelic variants it has
been discovered that homozygotes by the main allel were the dominant
variant. Among people with HIV T/T minor gene carriers were 4.5 more
often met in comparison with control group (p< 0.05) that can prove
the tendency to association of the mentioned genotype with infection.
Distribution of allelic variants of IL-10 gene promoter region in position
-592 is characterized by homozygote dominance by the main gene.
Among the individuals with HIV A/A minor allel carriers were 3.4 more
often met in comparison with control group (p< 0.05). Individuals with
A/A genotype were not identified in group of high risk of virus infection.
The abovementioned proves the tendency to association of minor allel
carrier state with HIV infection.
The occurrence of the homozygous combination of the allelic variant G/G
of the promoter of TNF-α has been shown to prevail almost twofold over
the occurrence of the variant G/A among all groups. High frequency of
heterozygote by the main allel has been recorded among the individuals
with HIV. Thus, G/A genotype frequency in group of HIV-infected people
2 and 1.5 exceeded the appropriate indices of group of high risk of
infection and comparison group correspondingly (p< 0.05) that points
to the tendency to association of the mentioned variant with infection.
Conclusions: Cytokines genes variations may contribute to the
acquisition of HIV infection and encourages carrying out of further
populations studies in this sphere of HIV-infection immunogenetics.
322
Background: HIV-1 Nef plays a major role in enhancing the
pathogenicity of the virus through various mechanisms such as downregulation of CD4 and HLA class I surface expression and interfering
with cell signaling pathways. Identifying and characterizing CD8+ T
cell epitopes in Nef that are under host immune selection can help in
selecting targets for an effective vaccine.
Methods: 326 subtype A Nef sequences from treatment naïve
patients of a Kenyan sex-worker cohort were generated using 454
pyrosequencing. Positively selected (PS) mutations were determined
using a bioinformatics approach, quasi analysis. Peptides were designed
with mutation placed in anchor position 2, 5, 8, 9 of epitopes of HLA
class I alleles for validation with ELISpot assay using patient PBMCs.
Results: E70D, I109V and I176M were associated with rapid CD4
decline (p=0.010, 0.015, 0.025 respectively). H124N and K190M were
associated with slow CD4 decline (p=0.001 and 0.029). The five PS
mutations were significantly associated with HLA class I alleles including
A*23:01 (E70D, p=0.002; I176M, p=0.003), A*02:01 (I109V, p=0.028;
H124N, p=0.021), B*58:01 (I109V, p=0.048), A*3002, B*57:03 and
C*02:01 (H124N, p=0.026, 0.0004, and 0.011 respectively) and
C*06:02 (K190M, p=0.037). ELISpot analysis identified 27 novel
epitopes containing either the consensus or the PS mutations. Six new
epitopes contained E70D, five epitopes contained K190M, and I109V
and H124N were each contained by eight new epitopes. No epitopes
containing I176M was confirmed by ELISpot. It is possible that I176M
represents compensatory mutations due to functional requirements
under host immune selective pressure.
Conclusions: Identification and characterization of epitopes containing
beneficial and detrimental PS mutations can provide important insight
for selecting immunogens for an effective HIV vaccine. More detailed
investigation of T-cell responses, such as poly-functionality and
proliferation to these mutations will be conducted to further characterize
these Nef epitopes.
Posters 38: Innovations in Vaccine and Microbicides Studies in Lab and Monitoring
P38.01
P38.02
Operational Challenges for the Set-up of
Gram Stain Analysis for Diagnosing Bacterial
Vaginosis in a Local Laboratory in Durban,
South Africa
Impact of Tenofovir 1% Gel on Hepatitis B
Virus Resistance in CAPRISA 004
Medical Research Council, HIV Prevention Research Unit, Durban,
South Africa
1
Background: Studies have demonstrated the association between
BV and HIV acquisition in woman. The prevalence of BV reported in
clinical trials conducted by the Medical Research Council HIV Prevention
Research Unit (MRC HPRU) is between 5-9%. In an attempt to reduce
turnaround time for results, enable direct contact with research clinics,
facilitate staff capacitation and reduce costs, the HPRU was selected
by the Protocol Reference Laboratory to perform in-house testing for
diagnosing BV. We report here on the operational challenges associated
with the set-up of the laboratory to perform the analysis.
Methods: To perform the testing, the laboratory underwent
infrastructural changes in order to implement a staining workbench.
Correct waste management disposal was also part of the set-up process
as well as procurement for staining reagents, consumables and internal
quality control (IQC) slides. Staff that were involved in the testing had to
undergo rigorous training.
Results: Operational challenges included: training and evaluating only
staff who specialize in clinical pathology and microbiology, quality of
gram stain reagents, QC processes, waste management and manual
IQC slides which were compared to commercially available ones.
Commercial gram stain reagents and IQC slides were preferred as less
labour intensive with enhanced staining results. Waste receptacles
proved futile and a local municipal waste discharge permit was obtained
instead. To date, 1605 slides have been analyzed. The following errors
were noted during the QC process: incorrect slide preparation (47%) and
inadequate sample material for diagnosis (0.3%), however, retraining of
clinical staff resulted in a significant reduction (50%) of errors.
Conclusions: Despite operational challenges HPRU has been successful
in the set-up of gram stain analysis for BV diagnosis as a Protocol
Reference Laboratory. This enables BV to be diagnosed in real-time
rather than shipping to an international laboratory where slides are
analyzed at study end.
Centre for the AIDS Programme of Research in South Africa, Durban,
South Africa, 2College of Medicine, University of Cincinnati, Division of
Digestive Diseases, Department of Internal Medicine, Cincinnati, OH,
United States, 3Columbia University Mailman School of Public Health,
Department of Epidemiology, New York, NY, United States
1
Background: The intermittent, before-and-after sex, dosing regimen
used in the CAPRISA 004 trial, where women were only required to use
the gel on days that they had sex, meant that women in the trial who
were infected with Hepatitis B Virus (HBV) could be exposed to variable
levels of a single antiretroviral agent in the presence of HBV, potentially
increasing the risk for the development of antiviral resistance. Here we
assessed the impact of intermittent tenofovir gel use on HBV resistance.
Methods: Hepatitis B virus DNA was extracted from stored samples
of women identified as being HBV surface antigen positive during
the trial. Extracted DNA was amplified by polymerase chain reaction
(PCR) and positive samples were gel purified and sequenced using the
Applied Biosystems 3130 x; automated sequencer. Five of the samples
were cloned into the pGEM-T Easy vector. Sequences were submitted
to the Stanford University HBV drug resistance database for analysis
of drug resistance, resistance loci and resistance-associated mutations.
To confirm conventional PCR results, the same HBV DNA was then
amplified by nested PCR and tested for resistance using the commercial
INNO-LiPA HBV DR V3 kit.
Results: A 3.2 kb PCR product corresponding to the pol region of HBV was
successfully amplified in 19/37 (51%) of the β-globin positive samples,
13 from women assigned to tenofovir and 6 from women assigned
to placebo. All samples clustered phylogenetically with HBV subtype
A. There was no difference in the frequency of amino acid variation
between the tenofovir and placebo arms. None of the known tenofovir
resistance mutations (M240V/I, L180M, A194T, V214A, N238T) were
identified using both the commercial kit and in-house PCR assay.
Conclusions: No resistance mutations associated with tenofovir were
detected in the 19 sequences examined. These data provide further
support for the safety of intermittent 1% tenofovir gel use as HIV preexposure prophylaxis for HBV-infected individuals.
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323
POSTERS
Natasha Gounden1, Nathlee Abbai1, Rashika Maharaj1
Cheryl Baxter1, Sinaye Ngcapu1, Jason T. Blackard2, Eleanor A.
Powell2, Patricia K. Penton2, Quarraisha Abdool Karim1,3, Salim
Abdool Karim1,3
Posters
P38.03
P38.04
Setting up and Oversight of Local Clinical Trial
Site Laboratories with Limited Resources: A
Case Study of South Africa’s FACTS 001 Study
Clinical Research Site Preparedness for
Clinical Emergencies - Implications for Preexposure Prophylaxis, Microbicide and
Vaccine Trials
Sarah S. Cohen1, Ishana Naidoo1, Romeo Martin2, Nomfundo
Maduna2, Maletsatsi Moloelang3, Nwabisa Ndzamela3, Christian
Kasango4, Andrew Tlagadi5, Gail Stockenstrom6, Nomzamo P.
Tabata6, Thuleleni Dlungwana7, Siphiwe Gumede7, Manyabeane
A. Phaahla8, Zaheda Ismail9, Elizabeth Rammutla10, Sinead
Delany-Moretlwe1, Glenda Gray11, Helen Rees1, Lindiwe
Nhlangulela4
Wits Reproductive Health and HIV Institute, University of the
Witwatersrand, Research, Johannesburg, South Africa, 2Wits
Reproductive Health and HIV Institute (WRHI), Research, Johannesburg,
South Africa, 3Perinatal HIV Research Unit, Kliptown - FACTS, Soweto,
South Africa, 4Aurum Institute for Health Research, Laboratory,
Rustenburg, South Africa, 5Aurum Institute for Health Research,
Laboratory, Tembisa, South Africa, 6Desmond Tutu HIV Foundation,
Laboratory, Cape Town, South Africa, 7MatCH Research (Maternal,
Adolescent and Child Health Research), Laboratory, Pietermaritzburg,
South Africa, 8Medunsa Clinical Research Unit, Laboratory, Garankuwa,
South Africa, 9Qhakaza Mbokodo Research Clinic, Laboratory,
Ladysmith, South Africa, 10Setshaba Research Centre, Laboratory,
Soshanguve, South Africa, 11Perinatal HIV Research Unit, Chris Hani
Baragwanath Hospital, Research, Soweto, South Africa
1
POSTERS
Background: Ensuring Good Clinical Laboratory Practice (GCLP) in
low-resource field laboratories sites during clinical trials is challenging.
We describe processes for ensuring GCLP compliance at all nine South
African sites in the FACTS 001 trial.
Methods: All nine sites established laboratories for collection,
processing and shipping of samples to a central laboratory; three sites
also processed and stored onsite. A coordinating team developed a
study-wide lab analytical manual (LAM), checklist for lab assessments,
and a laboratory working group that met monthly by phone to discuss
issues. Regular onsite assessments of laboratories were completed at
six-weekly intervals to ensure conformance with protocol and GCLP.
All visit findings were recorded in reports which were reviewed and
assessed for trends in findings over this period.
Results: An average of 10 assessments per site was completed in
the period July 2012 to May 2014. Monthly lab working group calls
provide opportunities for sites to resolve findings, and discuss common
challenges and retraining needs.Interventions for improvement included
retraining of personnel, vertical audits of sample records, and provision
of guidance in documenting laboratory incidents and deviations.
Following this, improvement was noted in general, and fewer deviations
were identified suggesting that the interventions were successful at
improving overall laboratory quality assurance.
Conclusions: While audits help ensure compliance with protocol
requirements, regular supportive supervision to sites helps identify areas
of deviation sooner and allows for rapid intervention and correction of
potential problems.
324
Tashni Nayager1, Vaneshree Govender1
South African Medical Research Council, HIV Prevention Research Unit,
1
Background: The frequency of clinical emergencies is far lower in HIV
prevention trial sites versus medical healthcare facilities. Despite this,
patients with “urgent” and “emergent” conditions such as cardiac arrest,
acute asthma, seizures, anaphylaxis, hypoglycaemia and shock, must be
provided with immediate care to decrease morbidity and mortality. Our
aim was to evaluate if emergency trolleys were appropriately stocked, if
staff were adequately trained in clinical emergency management and if
systems for monitoring adequacy were sufficient.
Methods: At the HIV Prevention Research Unit (HPRU), the primary
study population over the last decade has been healthy 18-45 year old
women, with a smaller population of infants and HIV sero-converters.
The stringent screening eligibility process excludes individuals with
serious comorbidities, such as unstable cardiovascular disease.
Emergency trolley equipment, supplies and resuscitation medication
were evaluated for availability, expiry dates and functional acceptability
with monitoring checklists.
Results: An internal audit of 6 clinical research sites identified a need for
a consistent schedule for trolley checks, timeous ordering of new stock,
regular maintenance of oxygen cylinders, review of required medication,
easy access to medication in a temperature controlled environment,
easy access to resuscitation algorithms and a study operational
procedure. Due to the infrequent occurrence of medical emergencies,
staff competency was assured with refresher training. Appropriate
systems were in place for immediate referral to a Department of Health
care facility based on a Memorandum of Understanding.
Conclusions: A clinical emergency management plan is a necessity in
a research site conducting HIV prevention trials. The level of emergency
care provision should be tailored to the study and site requirements, and
be relevant to the study population. Safety of clinical trial participants is
of the utmost importance.
P38.05
P38.06
Introduction of a Novel Monitoring Tool to
Reduce Specimen Archive Errors
Comparison of 2 Different BD PROBETEC™
ET Extraction Methods for Detecting of
Chlamydia trachomatis and Neisseria
gonorrhoeae in HIV Prevention Trials
South Africa, 2London School of Hygiene & Tropical Medicine,
Epidemiology and Population Health, London, United Kingdom
1
Background: The Medical Research Council (MRC), HIV Prevention
Research Unit (HPRU) has been involved in the conduct of multiple HIV
prevention clinical trials. In order to efficiently track clinical samples from
the 6 Clinical Research Sites (CRSs), the laboratory used the Laboratory
Data Management System (LDMS) for specimen archive. However,
previous specimen verification quality error reports had identified
gaps from the collection to archive process within the LDMS system.
Therefore, the laboratory developed a novel monitoring tool as part of
the quality specimen management programme.
Methods: The monitoring tool was set up for: quality control checks
of LDMS documentation against physical specimen verification and
monitoring of monthly quality trends. The tool captured the following
information: type of errors identified per CRS; participant identifiers
(PTIDS); staff responsible for errors, Corrective Actions and Preventative
Actions (CAPA) and re-trains performed.
Results: For the period of September 2012-December 2013, out of
369867 samples archived, there were 180 errors: LDMS typographical
errors (26/180), incorrect collection dates (19/180), incorrect visit codes
(47/180), incorrect Case Report Form (CRF) completion (27/180), incorrect
tracking sheet completion (3/180) and CRF and LDMS discrepancies
(58/180). The number of errors on average per month ranged from 7 to
15. The new monitoring tool enabled stringent monitoring of specimen
management with a significant decrease in errors to a total of 4 errors
in December 2013.
Conclusions: Since the introduction of the award winning novel
monitoring tool an improvement to MRC specimen archive quality has
been noted. We recommend that the tool can be adapted by other
organizations to improve their long term and high volume specimen
archive processes.
Rashika Maharaj1, Nathlee Abbai1, Gita Ramjee1,2, Lakshmi
Jagesur1
Department of Epidemiology and Population Health, London, United
Kingdom
1
Background: Nucleic acid amplification tests for the detection of C.
trachomatis (CT) and N. gonorrhoeae (NG) in genital tract specimens
has become the standard diagnostic method used in most laboratories
for HIV prevention trials. The BD ProbeTec ET System, use Strand
Displacement Amplification (SDA) technology for the direct, qualitative
detection of CT and NG from DNA extracted from endocervical swabs,
male urethral swabs, and urine specimens. The aim of this study is to
compare two DNA extraction methods (M) (M1 as per package insert;
M2 an international developed in house method) for the detection of CT
and NG using the BDProbeTec ET instrument.
Methods: Our sample size included 60 vaginal swabs. The first set of
extractions (method 1) was performed by conventional lysing, priming
and amplification. The second method (2) included additional washing
and centrifugation steps prior to lysis, priming and amplification. During
the amplification process, external quality controls [College of American
Pathologists: CAP ] were included in the runs. As part of the validation
process, samples were also processed at an external/reference laboratory.
Results: There was 100% concordance for CT/GC between the results
obtained by the external/reference laboratory and MRC HPRU Central
Laboratory using method 1, 100 % sensitvity and specificity on GC/CT
for all 60 swab samples. Method 2 achieved a 78 % concordance; CT
had 6 false negatives =70% sensitivity; 85 % specificity; while GC had 4
false negatives=80% sensitivity; 90 % specificity. CAP panels achieved
100% pass on both methods.
Conclusions: Method 1 as per package insert was superior to method
2 achieving a higher sensitivity and specificity. It is vital in populations
with high HIV risk with co-infections with CT and NG that methods used
for detection are accurate, specific and in keeping with the package
insert which this study illustrates.
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325
POSTERS
Lakshmi Jagesur1, Rashika Maharaj1, Avika Haridutt1, Nathlee
Abbai1, Duduzile Ndwandwe1, Gita Ramjee1,2
Posters
P38.07
P38.08
Hematologic and Chemistry Normal
Laboratory Values among Healthy Ugandan
Women Screened for a Pre-exposure
Prophylaxis Trial: the MTN-003(VOICE) Study
Strategies to Optimise Data Quality Metrics
in the ASPIRE Trial at the Wits Reproductive
Health and HIV Institute in Johannesburg
Flavia Matovu Kiweewa1, Holly M. Gundacker2, Mike Mubiru1,
Betty Kamira1, Brenda Mirembe Gati1, David Ojok3, Samuel
Kabwigu1, Philippa Musoke1,4, Clemensia Nakabiito1, Mary Glenn
Fowler1,5
MU-JHU Research Collaboration, Kampala, Uganda, 2SCHARP-FHCRC,
Seattle, WA, United States, 3MU-JHU Core Laboratory, Kampala,
Uganda, 4Makerere University College of Health Sciences, Department
of Pediatrics and Child Health, Kampala, Uganda, 5Johns Hopkins
University, Baltimore, MD, United States
Pranitha Ramchuran1, Krishnaveni Reddy1, Helen Rees1, Thesla
Palanee1
Wits Reproductive Health & HIV Institute, School of Clinical Medicine,
University of the Witwatersrand, Johannesburg, South Africa
1
1
POSTERS
Background: Universal laboratory toxicity grading tables are used to
screen and monitor adverse event (AE)s in clinical trials for HIV prevention
and treatment; use of these tables excludes otherwise eligible volunteers
and can make AE assessment and product management challenging in
specific populations. We computed selected hematologic and chemistry
normal ranges specific to healthy non-HIV infected women screened
for an HIV prevention trial (MTN-003) and compared findings to U.S.
established intervals.
Methods: Excluding women with hepatitis, syphilis, and pregnancy, we
analyzed data from 538 women aged 18-45 screened in Kampala from
Nov 2009-Dec 2011. We calculated 95% normal intervals as the 2.5%
and 97.5% limits for the population, and compared data against U.S.derived lab intervals from Massachusetts General Hospital and Division
of AIDS (DAIDS), Dec 2004 toxicity tables (clarification dated Aug 2009)
to determine the number of women with any AE per DAIDS grading
criteria.
Results: Compared to intervals from the U.S., we found slightly lower
2.5th centiles for red cell indices (hemoglobin (Hgb), hematocrit, MCV,
red blood cell counts), lower white blood cells and neutrophils, but
higher eosinophils. Chemistry parameters were comparable with U.S.based ranges except for a lower 2.5th centile for serum phosphorus and
higher 97.5th centile for ALT and AST. When graded against U.S.-derived
DAIDS criteria, we observed 96 AEs from 87 (16%) volunteers with
grade ≥ 1 results including decreased neutrophils and phosphorus in
48 (9%) and 23 (4%) women respectively. There were 8 (1%) grade 3
and no grade 4 AEs.
Conclusions: Similar to existing local intervals, we found differences in
upper and lower ranges for some hematologic and chemistry indices
among healthy Ugandan women compared to US norms. About 1 in
6 women were described as having grade ≥1 toxicity for either Hgb,
white cell indices, or phosphorus. Local laboratory ranges should be
considered for toxicity grading in international research settings.
326
Background: Quality data collection and their timeous reporting are
critical in clinical trials. In ASPIRE, a phase 3 safety and effectiveness
study of the Dapivirine Vaginal Ring in HIV prevention, data quality
is assessed by the quality control (QC) error rate calculated by the
number of errors per 100 case report forms (CRFs) submitted to the Data
Management Centre (DMC). Data timeliness is defined as the percentage
of CRFs received at the Data Management Centre (DMC) within 7 days
of the visit.
Methods: To ensure good quality data and timeliness at Wits RHI,
several strategies were implemented that include the formation of a
multi-disciplinary quality management (QM) team comprising Clinical
Quality Improvement Mentors (CQIMs), Quality Assurance (QA)/QC
officers and a datafax team. The CQIMs conduct clinical review and
immediate retraining of the clinical team, QA/QC officers conduct
error trend analyses and the site datafax team conducts a final QC
prior to datafax. Other strategies include regular review and revision
of QM processes within the Clinical Quality Management Plan; timely
re-training of the team on errors trends identified and collective team
review and ownership of monitoring and audit findings. An additional
tool assisting this process is iDataFax; a DMC data system that provides
early access to errors identified on datafaxed CRFs. Its use facilitates
identification of errors trends more frequently to inform improved CRF
completion and shorter QC resolution times.
Results: Implementation of these strategies has led to the development
of a more structured and focused QM team and instilled a culture of
accountability among the study team who consider quality in all aspects
of trial implementation.
Conclusions: These strategies are continuously reviewed and amended
to suit the needs of the study and the dynamic nature of data collection
and with consistent implementation have and continue to assist the
team to strive to optimize data quality and timeliness metrics in ASPIRE.
Posters 39: Molecular Epidemiology
P39.01
P39.02
Using Viral Dynamics to Connect Clinical
Markers of Disease Progression to Sequence
Evolution during HIV Infection
Adaptation of HIV-1 Envelope Glycoprotein
gp120 to Humoral Immunity over the Course
of the Epidemic
Andrew E. Adams1, Zabrina L. Brumme2, Alexander R.
Rutherford1, Ralf W. Wittenberg1
Melanie Bouvin-Pley1, Marion Morgand1, Alain Moreau1,
Laurence Meyer2,3, Cécile Goujard2,3, Hugo Mouquet4, Michel C.
Nussenzweig5, Craig S. Pace6, David D. Ho6, Pamela J. Bjorkman7,
Daniel Baty8, Patrick Chames8, Marie Pancera9, Peter D. Kwong9,
Pascal Poignard10, Francis Barin1,11, Martine Braibant1
Simon Fraser University, Mathematics, Burnaby, BC, Canada, 2Simon
Fraser University, Faculty of Health Sciences, Burnaby, BC, Canada
1
INSERM U 966, Tours, France, 2CESP INSERM U 1018, Paris, France,
AP-HP, Hôpital de Bicêtre, Le Kremlin-Bicêtre, France, 4Institut Pasteur,
Paris, France, 5Howard Hughes Medical Institute (HHMI), Rockefeller
University, New York, NY, United States, 6Aaron Diamond AIDS
Research Center, Rockefeller University, New York, NY, United States,
7
California Institute of Technology, Pasadena, CA, United States, 8CRCM
INSERM U 1068, Marseille, France, 9Vaccine Research Center, NIH,
Bethesda, MD, United States, 10International AIDS Vaccine Initiative
(IAVI), Neutralizing Antibody Center, Scripps Research Institute,
Immunology and Microbial Science, La Jolla, CA, United States,
11
Laboratoire de Bactériologie-Virologie, CHU Bretonneau, Tours, France
1
Background: Since 2009, a large panel of broad and potent
monoclonal neutralizing antibodies (MoNAbs) against HIV-1 have been
isolated. These MoNAbs can protect from HIV-1 infection and suppress
established infection in animal models. Because their efficacy should
be evaluated in human clinical trials, it is of importance to define the
sensitivity of the most contemporary transmitted variants to these
MoNAbs. We, and others previously, reported that HIV-1 has become
more resistant to neutralization over the course of the epidemic (Bunnik
et al, Nature Med 2010, Bouvin-Pley et al, PloS Pathog 2013).
Methods: Here we extended the analyses to the most potent MoNAbs
described since then, either more recently isolated or improved by
structure-based gene modifications.
Results: We fully confirmed the first observations showing an increasing
resistance of HIV-1 clade B over time to MoNAbs targeting the major
gp120 epitopes but not to MoNAbs targeting the gp41 MPER. Despite
this evolution, some MoNAbs still were able to neutralize efficiently the
most recently transmitted HIV-1 variants (2006-2010). The most potent
MoNAbs were the bi-specific PG9- and PG16-iMab that alone were able
to neutralize all variants at less than 0.4 mg/mL. The sensitivity to iMAb
remained similar over time, suggesting that the trend of increasing
resistance to PG9-/PG16-iMAb may be attributed only to the antigen
binding domain of PG9/PG16. NIH45-46m2 (and -m7), 10-1074 and
10E8 were also highly potent and, if combined, reached the potency
of PG9-/PG16-iMAb. We also observed that 3BNC117 was almost as
potent as the modified NIH45-46 antibodies, and that the lama-derived
JM4IgG2b was the most potent Ab among those that do not target the
major gp120 neutralizing epitopes.
Conclusions: These data clearly suggest a continuous drift of the env
gene of HIV-1 clade B over the epidemic, and that not a single epitope
is concerned but the entire gp120 as a whole. The consequences of this
adaptation on the envelope functionality are being explored.
www.hivr4p.org
327
POSTERS
Background: Understanding how HIV establishes infection and, if
left untreated, eventually overcomes the immune system is crucial to
the development of a vaccine or cure. The high rates of turnover and
evolutionary adaptability exhibited by HIV pose a particular challenge
to HIV vaccine development. Our focus is to understand the dynamics of
two of the most commonly tracked clinical markers of an HIV infection:
CD4+ T cells/mm3 (CD4 count) and HIV RNA/ml (viral load).
Methods: We developed a stochastic system of differential equations to
model HIV infection that uses equilibration, adaptation, and inheritance
to model the initial infection as well as successive generations of
viral lineages. The model allows viruses to generate new lineages in
proportion to their viral load with an inherited fitness. These lineages
compete for immune cells to infect and drive decline in CD4 count
through a series of small adaptations. We use this model to demonstrate
how viruses with a sufficiently high mutation rate could overcome
the immune system, even when most changes are expected to be
detrimental to viral fitness.
Results: We have calibrated our model to match viral load set
points and rates of CD4 decline from 91 HIV-infected individuals
studied longitudinally during early stages of the disease. Our model
demonstrates how a genetically diverse population of viruses could be
sustained in an environment with high rates of competition, turnover,
and the development of an immune response. The underlying stochastic
process also generates a phylogenetic structure which can be used to
explore different hereditary patterns in the underlying viral lineages.
Conclusions: Our model demonstrates that the high rate of mutation
and recombination in the HIV genome can contribute to slow disease
progression. It suggests the diversity of HIV lineages is a consequence
of lineages having similar fitness and the high levels of competition
which create a balance in the expansion of existing lineages and their
replacement by new lineages.
3
Posters
P39.03
P39.04
Characterisation of Transmitted and
Non-transmitted HIV in Index-recipient
Transmission Pairs
Significance of HIV-1 Western Blot Bands
Appearance in Clinical Trials - Point of
Seroconversion and Window Period in Rural
Kwazulu-Natal; South Africa
Lotte Bracke1, Elisabeth Willems1, Astrid Gall2, Paul Kellam2,
Sandra Coppens1, Conor Meehan3, Georgios Pollakis4,5, Ben
Berkhout4, Guido Vanham1, Marion Cornelissen4, Leo Heyndrickx1,
Kevin Ariën1
Institute of Tropical Medicine, Department of Biomedical Sciences,
Virology Unit, Antwerp, Belgium, 2Wellcome Trust Sanger Institute,
Cambridge, United Kingdom, 3Institute of Tropical Medicine,
Department of Biomedical Sciences, Mycobacteriology Unit, Antwerp,
Belgium, 4Center for Infection and Immunity Amsterdam (CINIMA),
Academic Medical Center, University of Amsterdam, Amsterdam,
Netherlands, 5Institute of Infection and Global Health/CIMI, University
of Liverpool, Liverpool, United Kingdom
1
POSTERS
Background: Many of the viral and host factors associated with HIV
transmission are still poorly understood. In 60-80% of the mucosal
infections, a single transmitted/founder virus is responsible for the
establishment of a productive infection, indicating a strong genetic
bottleneck upon transmission. We aim to better understand the viral
factors involved during transmission by studying the genetic variability
and replicative characteristics of viruses isolated from transmission pairs.
Methods: We had access to blood samples from 5 index-recipient
transmission pairs of MSM. All samples were obtained shortly after
transmission. Plasma was used for full genome sequencing and PBMC
were cocultured with HIV negative donor PBMC by limiting dilution to
obtain biological clones.
Results: We isolated a total of 270 biological clones from the 5
transmission pairs. Nearly the complete gp120 from 7-18 clones was
sequenced for each of the 10 individuals. As expected, these sequences
group nicely with the plasma sequences. In only one transmission
pair studied, an identical clone was found in both index and recipient.
Phylogenetic analysis showed a low genetic diversity in the recipients, in
contrast to a greater genetic diversity among the clones from the index
patients. Sequences from recipients also showed shorter V1, V2 and V4
loops, indicating a more compact envelope compared to viruses from
the index patients. We also found fewer potential N-linked glycosylation
sites in three recipients compared to their indexes.
Conclusions: Our sequences and phylogenetic analysis confirm
observations from others. Currently the replication capacity of the
biological clones is assessed in dual infection/competition assays. This
will allow us to rank the fitness of the clones obtained from all the
transmission pairs. A selection of clones will also be tested against
neutralizing antibodies and entry inhibitors. These experiments should
give us an indication of which virus characteristics favour transmission.
328
Rashika Maharaj1, Nathlee Abbai1, Gita Ramjee1,2, Lakshmi
Jagesur1
Kingdom
1
Background: Information on acutely HIV-1 infected individuals is very
useful for treatment, pathogenesis and disease progression. Besides
accurately predicting HIV-1 infection, sequential appearance of specific
bands of the western blot offers a window of opportunity to develop
a less subjective tool for monitoring disease progression. The aim of
this analysis is to investigate the significance and order of HIV-1 band
appearance at two time points during seroconversion in HIV Prevention
trials.
Methods: Between July 2007 and December 2012 the Medical Research
Council (MRC) HIV Prevention Research Unit (HPRU) Central Laboratory
did a retrospective analysis on Biorad Genetics HIV-1 Western Blot
test performed on 652 plasma samples collected at two time points
i.e. point of sero-conversion (T1, n=326) and 1-2 post sero-conversion
(T2, n=326). All testing was performed according to the manufactures’
instructions.
Results: Of the 326 at T1/T2: presence of HIV-1 Gp 160 (#/%)
=303(93%) /324(99%), Gp 120 =182 (56%) /255(78%), Gp 65
=159(49%)/ 251(77%), p55/51=306(94%)/ 324(99%), Gp 41=
168(52%)/ 250(77%), p40=304 (93%)/ 323(99%), p31=195(60%)/
276(85%), p24=312(96%)/ 323(99%), p18=132(41%)/ 189(58%). p24
antigen was the first marker to be detected within 2 weeks of infection
evident in 99% of the samples tested. Antibodies to the envelope
glycoproteins (gp 160 and gp 120) and the trans membrane glycoprotein
(gp 41) appeared within 2 to 4 weeks of infection. Samples at both time
points showed that Gp160, p55/51, p40 and p24 were predominantly
evident in > 93-99% of all acute sero-convertor cases with majority
displaying at least four of the nine bands characteristic of the virus by
Western blotting, with the lowest number of three bands characteristic
of the virus displayed by any sample.
Conclusions: The use of WB banding patterns during early infection
will prove useful as there is a renewed interest for new surveillance
technologies in this area and may assist in future vaccine development
P39.05
P39.06
Changes in Viral Population Kinetics
Following HIV-1 Superinfection
The Role of N-glycosylation in DC-SIGN
Interactions with Transmitted Founder
Variants of HIV-1 Subtype C Envelope
University of Cape Town, Clinical Laboratory Sciences, Cape Town,
South Africa, 2CAPRISA/University of Kwa Zulu Natal, Durban, South
Africa
1
Background: Elucidation of factors influencing HIV superinfection
with a second viral strain, despite pre-existing immune responses to
the primary infecting strain, may provide insights into correlates of
protection. Superinfection is frequently associated with a spike in viral
load which could be due to loss of control of the primary infecting virus,
or the superinfecting virus, or both. To evaluate if pre-existing responses
to primary infection differentially controlled viral populations following
superinfection, we estimated viral population kinetics at time points
before and after superinfection.
Methods: We performed 454 deep sequencing of two genomic regions,
gag p17 (332bp) and env C2C3 (403bp), on three participants who were
known to be superinfected. To enable quantification of the input cDNA,
and control for PCR and sequencing errors, the cDNA was labelled with
a unique primer ID.
Results: In all three participants the temporary increase in viral
load associated with superinfection was predominantly due to the
superinfecting viral strain, which became the major circulating variant
in two of the three participants. Within the regions examined, we did
not detect recombination in the gag region, however in one of three
participants recombination was found within the env C2C3 region in
~20% of viral sequences 15 weeks after superinfection.
Conclusions: The superinfecting virus was not controlled following
infection within two participants, suggesting that responses elicited to
the primary strain were not cross-protective against the second strain
at the time of superinfection. In one participant however, there was
subsequent control of the superinfecting strain following its introduction,
this could be indicative of a broader CTL response and warrants further
investigation.
Evelyn N. Lumngwena1, Liliwe Shuping1, Netanya Bertniz1,
Claudia Cicala2, James Arthos2, Zenda Woodman1
University of Cape Town, Molecular and Cell Biology, Cape
Town, South Africa, 2National Institutes of Health, Laboratory of
Immunoregulation, NIAID, Bethesda, MD, United States
1
Background: The design of effective vaccines and microbicides
requires understanding early steps in mucosal transmission. As the
genetic bottleneck at transmission favors variants with fewer potential
N-glycosylation sites (PNGs) and shorter variable loops, we investigated
whether Envelope (Env) PNGs could influence the ability of variants to
cross the genital mucosa by altering gp120 interactions with DC-SIGN,
which could favour transfer to CD4+ cells, and also alter localized
immune responses in the genital epithelium.
Methods: We initially determined whether pseudovirus-Envs from
transmitted founders (TF) had enhanced DC-SIGN binding, transinfection and increased IL-10 secretion over matched chronic Envs. As
DC-SIGN interactions with Env favors high mannose N-glycans, we also
deleted gp120 PNGs bearing high mannose glycans either singly, or in
combination, in matched CAP239 Env T/F and chronic clones.
Results: T/F Envs induced more IL-10 secretion than chronic controls.
When PNGs were deleted from the CAP239 envs, the effect on
pseudovirion entry, DC-SIGN binding and trans-infection was clone
specific, suggesting that specific N-glycans affect Env function differently
in different clones. For example deletion of PNG 448 reduced entry
efficiency, DC-SIGN binding and trans-infection of TF by ~50% when
compared to wild type, while either enhancing or maintaining these
phenotypes in the chronic infection clone. Only deletion of the PNG 241
reduced IL-10 induction for T/F clones.
Conclusions: As pseudovirion entry efficiencies of most PNG mutants
were reduced for both CAP239 Env clones, it is difficult to determine the
role that each might play in DC-SIGN interactions. However, the TF Envs
induced MDDCs to secrete higher levels of IL-10 compared to matched
chronic infection controls, suggesting that localized anti-inflammatory
responses in the genital epithelium might play a role in HIV transmission.
www.hivr4p.org
329
POSTERS
Murray G. Logan1, Daniel J. Sheward1, Colin Anthony1, Nigel
Garret2, Carolyn Williamson1
Posters
P39.07
P39.08
Real Time Fitness Assay of Two CRF01_A/E
HIV-1 Transmitted Founder Variants
Outgrowth of Subtype C Envelope Viral
Populations in Dual-infected Individuals Is Not
Always Associated with Entry Efficiency
Melanie Merbah1, Gustavo Kijak1, Leigh Anne Eller1, Eric SandersBuell1, Brendan T. Mann1, Devin M. Pillis1, Anne Marie O’Sullivan1,
Meera Bose1, Jenica L. Lee1, Kultida Poltavee1, Nelson Michael1,
Jerome Kim1, Merlin Robb1, Sodsai Tovanabutra1, Agnes-Laurence
Chenine1
MHRP/WRAIR/HJF, Silver Spring, MD, United States
1
POSTERS
Background: RV217/ECHO study presents a unique opportunity to
identify subjects very early in HIV infection, with a median time to last
negative nucleic acid test (NAT) of 4 days. Two transmitted/founder
(T/F) variants were isolated from a Thai volunteer, with a representation
of 99% and 1% for the major (Mj) and the minor (mn) variants
respectively at the peak of viral load. Using next-generation, targeted
deep sequencing, we confirmed the presence of that mn variant on the
day of the first positive NAT. After peak viremia the mn variant grew
exponentially, reaching >30% by day 31. Six months post infection,
the mn variant became the predominant quasispecies. We hypothesized
that viral fitness is responsible for this viral dynamic profile.
Methods: Full-length infectious molecular clones (FLIMC) of the Mj and
mn variants were generated. Each Mj and mn FLIMC was engineered to
express the fluorescent proteins, GFP and mCherry. The four constructs
Mj.C2, Mj.G2, mn.C2 and mn.G2 were tested for infectivity in cell lines
(TZMbl & A3R5) and primary cells (PBMC, macrophages, dendritic cells)
and monitored by flow cytometry, fluorescent microscopy and p24
capture assay; cells were singly infected and co-infected. Cell tropism
was also assessed using CXCR4 and CCR5 Ghost cells.
Results: Both mn and Mj FLIMCs showed productive infection in cell
lines and primary cells and used the co-receptor CCR5 exclusively.
Neither the GFP nor mCherry proteins modified in vitro infection kinetics
of Mj and mn FLIMCs. Cell lines and primary cells were co-infected
with different ratios of Mj/mn. Contrary to their representation at the
time of initial infection, we found that the mn variant was more fit,
as measured by replication kinetics, than the majority transimitted Mj
variant. Interestingly, dual infection of single cells was a very rare event.
Conclusions: We successfully applied a new fitness assay to evaluate
multiple T/F in acute infection and found fitness didn’t account for
dominance of the Mj variant during acute infection.
330
Shatha Sultan Ahmed Omar1, Daniel Sheward2, Carolyn
Williamson2, Zenda Woodman1
University of Cape Town, Molecular and Cell Biology, Cape Town,
South Africa, 2University of Cape Town, IIDMM, Cape Town, South
Africa
1
Background: Dual infection with two phylogenetically distinct HIV
variants has been reported to be associated with rapid disease
progression possibly due to the emergence of early recombinant viruses
with high viral fitness resulting in high viral loads. As it has previously
been shown that Envelope (Env) plays an important role in HIV fitness,
this study aimed to determine the relationship between in vivo viral
outgrowth and Env entry efficiency, and to identify fitness determinants
that can be used as novel targets for drug and vaccine design.
Methods: Highlighter plots of SGA-derived env sequences of four
dual infected individuals (sampled at enrolment and approximately
3, 6, and 12 months post infection (mpi) was used to determine the
relative frequency and fluctuation of invivo viral populations within each
participant. Representative amplicons were cloned and compared using
a pseudivirion entry efficiency assay.
Results: our data indicated the presence of recombinants at enrolment
for 3/4 participants and the outgrowth of these viruses at 12 mpi for
all participants suggesting rapid recombination. However, this rapid
recombination was not always associated with enhanced entry efficiency
as recombinants at 12 mpi had similar entry efficiency to those at the
earliest time point for 2/4. In one participant (CAP84) recombination
occurred within the signal peptide and gp41, with the recombinant
carrying an additional N-glycan site (PNG) at position 339 in gp120.
While the recombinant entered TZM-bl cells as efficiently as the primary
virus, a chimeric Env carrying the gp41 of the recombinant but lacking
the PNG had a 4-fold increase in entry efficiency. When this PNG (N339)
was introduced by site directed mutagenesis, entry efficiency decreased
suggesting that recombination did select for a fitter variant but that the
introduction of an N-glycan lowered fitness.
Conclusions: Therefore, viral outgrowth in HIV-1 dual infected
individuals is likely due to a balance between replicative fitness and
immune escape.
P39.09
P39.10
HIV-2/SIV Vpx Protein Interacts with Human
Nup153 and Regulates Viral Pathogenesis
The Sexually Driven Epidemic in Youngsters
of China’s Southwestern Border Region
Was Caused by Dynamic Emerging Multiple
Recombinant HIV-1 Strains
Satya Prakash Singh1, Pankaj Gupta1, S Mahalingam1
Indian Institute of Technology Madras, Biotechnology, Chennai, India
1
State Key Laboratory for Infectious Disease Prevention and Control,
National Center for AIDS/STD Control and Prevention, Chinese Center
for Disease Control and Prevention, Beijing, China, 2Dehong Center
for Disease Control and Prevention, Dehong, China, 3Guangdong
Provincial Institute of Public Health, Guangdong, China, 4Yunnan Center
for Disease Control and Prevention, Kunming, China, 5Fred Hutchinson
Cancer Research Center, Seattle, WA, United States
1
Background: Dehong prefecture of Yunnan province was the gateway
of China’s AIDS epidemic. Most HIV-1 strains were first found in Dehong
before spreading to other parts of the country. Study on HIV-1 genetic
recombinants will provide key information on virus transmission
dynamics and help to inform both local and national HIV prevention
strategies.
Methods: In Dehong, we surveyed all HIV infected young people
(age ≤25 ) diagnosed in the first quarter of each year in 2009- 2012.
Thepol genes fragment from a totalof 205 HIV-1 infected people were
amplified and sequenced. The HIV-1 subtypes and recombinant patterns
were identified by phylogenetic and breakpoint analyses. Near fulllength genome sequencing was performed to characterize the new
recombinants and potential circulating recombinant forms (pCRFs).Risk
factors for generating various recombinant viral forms were analyzed by
Chi-square tests and logistic regression methods.
Results: Two thirds (131/205) of the HIV-1 infections in young people in
Dehong were caused by new recombinant viruses. About 40% (54/131)
of them were found to form 11transmission clusters, which was termed
pCRFs. The rest (77/131) of them have not yet been able to form any
transmission clusters and belonged to unique recombinant forms
(URFs). The generation of the new HIV-1 recombinants was significantly
associated with people with low education, residents outside the capital
city of Dehong and being Myanmar residents.
Conclusions: By properly sampling the young people, we revealed that
the ongoing HIV epidemic in Dehong was caused by high proportion
of new recombinant viruses, which was not found in sexually driven
epidemic before. Considering Dehong had generated wide-spreading
HIV-1 CRF_07, 08 BC, great efforts should be put on preventing the
existing pCRF from further spreading and containing the URFs evolving
to future CRFs. Prevention strategy should focus on those most at-risk
for multiple exposures, residents with low education, on border region
and migrating Burmese.
www.hivr4p.org
331
POSTERS
Background: Vpx, a virus associated accessory protein is encoded
by Human Immunodeficiency Virus type 2 (HIV-2) and Simian
Immunodeficiency Virus (SIVsm/SIVmac lineage) is known to be
involved in the nuclear import of viral DNA in non-dividing primary
target. Interestingly, Vpx mutant virus fails to replicate in non-dividing
cells. The mechanism by which Vpx helps in the nuclear transport of
viral genome remains unknown.
Methods: Co-immunoprecipitation was performed to study the proteinprotein interaction in transiently transfected HEK cells. Protein colocalization was assessed by immunofluoresence. Using homology
modeling of Vpx and Nup153 amino acid sequences, putative binding
motif for Vpx in Nup153 was predicted. Site directed mutagenesis
was employed to generate mutant viruses which were defective for
interaction with Nup 153. Nuclear import ability of wild type and mutant
virus was analysed by 2-LTR assay.
Results: Molecular transport across nuclear envelope is governed by
nucleo pore complexes (NPCs), composed of 30 nucleoporins (NUPs). We
found that interaction between Vpx and human Nup153 was necessary
for viral DNA import. We mapped the domain of Nup153 critical for
interaction with Vpx and our data suggests the role of a zinc finger
domain (610-869aa) to be critical. Vpx interaction with Nup153 was
impaired by exchange of serine (63,65) and tyrosine (66, 69 and 71)
residues to alanine and resulted in abrogation of nuclear localization.
Interestingly, the SIV PBj1.9 with mutant Vpx was found to have reduced
nuclear import ability in 2-LTR assay.
Conclusions: Our data gives insight into the mechanism of nuclear
import by Vpx interaction with Nup153. Novel treatment methods using
phosphorylation inhibitors specific for Vpx could be devised in future.
Huamian Wei1, Jenny H. Hsi1, Song Duan2, Yi Feng1, Cui He1,
Yuhua Ruan1, Xiang He3, Lingjie Liao1, Yanling Ma4, Yunda
Huang5, Manhong Jia4, Hui Xing1, Yiming Shao1
Posters
P39.11 LB
A New Cost-effective Viral RNA PCR-based
Diagnostic for Detection of Early Subtype C
HIV-1 Infection
Debby Basu1,2, Tyronza Sharkey2, William Kilembe2, Susan
Allen2,3, Eric Hunter1
Emory University, Emory Vaccine Center, Atlanta, GA, United States,
Zambia Emory HIV Research Project, Lusaka, Zambia, 3Emory
University School of Public Health, Department of Global Health,
Atlanta, GA, United States
1
2
POSTERS
Background: Strategies for detecting early HIV-1 infection can be
costly, inefficient and time-consuming, especially in settings where highthroughput screening is needed but viral load (VL) testing instruments
may not be accessible or economical. We have developed a costeffective in-house viral RNA PCR that can empower limited-resource
laboratories to screen for acute HIV-1 infection by amplification of viral
regions in gag, pol and gp41.
Methods: Viral RNA was extracted from plasma, and a multiplexed onestep reverse-transcriptase first round PCR, containing the outer primer
sets for three regions in a single reaction, was performed. Nested second
round PCR specific to each region followed. To validate that this multiplex
assay could be used on batched plasma pools and to determine the limit
of detection, we tested 14 samples of known VL, ranging from 1.2x107
to 1.1x104 copies/mL, by diluting these plasmas from early infection
1:10 with uninfected plasma.
Results: Positive amplification in at least 1 of the 3 diagnostic regions
was observed for all 14 mock-pooled plasma samples, with the majority
(11/14) showing positive amplification in all 3 regions. In the lowest VL
case, input copies as low as an estimated 16 copies per first round PCR
reaction still resulted in positive amplification in gp41.
Conclusions: We have developed a multiplexed RT-PCR assay to
detect early HIV-1 infection in patient plasma in a cost-effective, highthroughput manner. All samples tested were positive in at least one of
three viral regions. The majority resulted in positive amplification in
all three regions, thus showing potential universality of the assay in
detecting viral RNA from Subtype C plasma samples with a range of
VLs. This strategy costs less than $8/sample tested, and thus represents
an economical strategy to screen patients for evidence of early HIV-1
infection in low-resource settings. This research was supported by NIH
FIC R25 TW009337 (DB), FIC 2D43 TW001042 (WK), RO1 MH095503
(SA), R37 AI51231and IAVI (SA).
332
Posters 40: Mucosal Immune Activation and Inflammation
P40.01
P40.02
Vaginal Concentrations of Lactic Acid Potently
Inactivate HIV-1 Compared to Short Chain
Fatty Acids Present During Bacterial Vaginosis
Effects of Endogenous and Exogenous Female
Reproductive Hormones on Gene Expression
and Barrier Function in Female Genital
Epithelia
Ayesha Islam1, Jai G. Marathe2, Jeff Pudney3, Seyoum Ayehunie4,
Robin R. Ingalls5, Deborah J. Anderson6
1
Burnet Institute, Centre for Biomedical Research, Melbourne,
Australia, 2Monash University, Microbiology Department, Melbourne,
Australia, 3University of Melbourne, Department of Microbiology and
Immunology, Melbourne, Australia, 4ReProtect Inc., Baltimore, MD,
United States, 5Johns Hopkins University, Department of Biophysics,
Baltimore, MD, United States
1
Background: Bacterial vaginosis (BV) is caused by an imbalance in
vaginal microflora and is a major risk factor for sexually transmitted
infections, including HIV in women. Microflora composition is likely
influenced by low vaginal pH (~3.8), maintained by racemic DL-lactic
acid (LA) (~110mM). BV alters the pH (>4.5) and the short chain fatty
acid (SCFA) profile to predominantly acetic acid (BV) vs LA (non-BV).
Our previous studies highlight the potent HIV virucidal activity of LA;
however, the virucidal activity of BV-associated SCFAs is unknown.
Methods: Virucidal activity of physiologically relevant non-BV
associated SCFAs at pH 3.8 versus BV-associated SCFAs at pH 5 were
compared against subtype B transmitted/founder (T/F) strains, RHPA and
CH058, subtype C 92BR025 and subtype EA CMU02. Anti-HIV activity
of 100mM of DL-LA (pH3.8) and 100mM of acetic acid (pH 5) was
determined over time. The structure activity relationship (SAR) of SCFAs
and HIV-1 virucidal activity was assessed at the same pH and at equal
concentrations of the active protonated forms.
Results: Non-BV associated SCFAs (pH3.8) rapidly inactivated T/F strains
causing a 1000-fold drop in HIV-1 infectivity while BV associated SCFAs
(pH5) caused little inactivation. This potent virucidal activity could be
attributed to DL-LA (a non-BV SCFA), and not low pH. DL-LA had the
greatest virucidal activity against subtypes B, C and EA over related BVassociated SCFAs at the same pH and concentration of the active form.
SCFA SAR analysis revealed potent virucidal activity is associated with
the presence of hydroxyl groups, especially on the α-carbon; which is
attenuated by the presence of a CH3 group on the carboxylic acid.
Conclusions: We show that LA, a non-BV SCFA, is a more potent HIV
virucide than SCFAs produced during BV, suggesting that BV-associated
SCFAs are not as protective for the female reproductive tract. SAR
analysis reveals chemical elements required for HIV-1 activity that could
inform the design of SCFA analogues.
Background: Little is known about how female reproductive hormones
estradiol-17β (E) and progesterone (P) influence vaginal barrier and
immune function. Furthermore the synthetic progestin contraceptive
Depo-Provera (DMPA) promotes vaginal SIV acquisition in macaques
and may enhance HIV acquisition in women. We have studied the
effects of endogenous and exogenous hormones on vaginal epithelial
barrier function and molecular mechanisms of immune defense.
Methods: We conducted an Affymetrix 1.0 ST microarray study to
examine gene expression in MatTek vaginal (VEC) and endocervical
(VEN) tissues after differentiation in media containing physiologic
E (75nM) or E+P (75 and 700nM, respectively) or 130 nM DMPA for
10 days. To assess barrier function, tissues were seeded apically with
CMFDA-stained macrophages and infiltration was assessed by confocal
microscopy.
Results: Our study confirms the hormonal responsiveness of these
tissues, and identifies several genes that are significantly up and downregulated following exposure to hormones. Pathways identified by
DAVID and Ingenuity Pathway Analysis (IPA) reflect classical hormone
responses and epithelial differentiation, as well as a number of others
that potentially affect HIV and other sexually transmitted infection
acquisition including mediators of innate immunity, cell death, and
tight junction molecules. VEC-DMPA showed increased membrane lipid
storage but decreased steroid (E) responses and retinol metabolism.
VEC-E showed increased lysozyme expression (3x) and decreased
Caspse14 (-8x) expression versus hormone untreated VEC tissue (p<
0.05). Notably, gene expression profiles of VEN were distinct from VEC.
E treatment of VEC prevented infiltration of macrophages by >50%,
providing further evidence of its barrier enhancing effects.
Conclusions: Female reproductive hormones and DMPA have distinct
effects on molecular pathways underlying immune defense in vaginal
and endocervical epithelium. E appears to fortify vaginal barrier function.
Boston University, OB/GYN, Boston, MA, United States, 2Boston
University, School of Medicine, Boston, MA, United States, 3Boston
University, Boston, MA, United States, 4MatTek Corp, Ashland, MA,
United States, 5Boston Medical Center, Infectious Diseases, Boston, MA,
United States, 6Boston University, OB/GYN, Microbiology, Boston, MA,
United States
www.hivr4p.org
333
POSTERS
Muriel Aldunate1,2, David Tyssen1, Catherine Latham1, Paul
Ramsland1,2,3, Patrick Perlmutter2, Thomas Moench4, Richard
Cone5, Gilda Tachedjian1,2,3
Posters
P40.03
P40.04
Softcup Compared to Cervicovaginal Lavage
Sampling: Determining Total and HIVspecific IgGs in the Female Genital Tract - A
Randomized Study
Proteomics Based Methods for Toxicity
Monitoring of Rectal Microbicides
Adam Burgener1, Florian Hladik2, Kenzie Birse3, Ian McGowan4
Public Health Agency of Canada, Winnipeg, MB, Canada, 2University
of Washington, Seattle, WA, United States, 3University of Manitoba,
Winnipeg, MB, Canada, 4Microbicide Trials Network, Pittsburgh, PA,
United States
1
Derseree Archary1, Lenine Julie Liebenberg1, Lise Werner1, Sahil
Tulsi1, Nelisile Majola1, Nivashnee Naicker1, Sarah Dlamini1,
Natasha Samsunder1, Salim S. Abdool-Karim1,2, Jo-Ann S.
Passmore1,3,4, Lynn Morris5, Nigel Garrett1
Centre for AIDS Programme of Research in South Africa (CAPRISA),
Durban, South Africa, 2Mailman School of Public Health, Columbia
University, Department of Epidemiology, New York, NY, United States,
3
University of Cape Town, Cape Town, South Africa, 4National Health
Laboratory Services, Cape Town, South Africa, 5National Institute for
Communicable Diseases, Johannesburg, South Africa
1
POSTERS
Background: Determining optimal methods for genital specimen
collection for detection and quantification of mucosal immune responses
is a priority for the HIV vaccine prevention field. Here we investigated
whether Softcup® (EuroFemPro, Netherlands) sampling yields greater
quantities of HIV-specific immunoglobulins (IgG) and total IgG when
compared to the standard, cervicovaginal lavage (CVL) method in a
randomized study.
Methods: Forty HIV-infected antiretroviral-naïve women from an HIV
cohort study were randomized to undergo either Softcup sampling
followed by CVL, or CVL alone. We measured HIV-specific IgGs against
Gag p24, p66 (RT), gp41MN and gp120 in 19 Softcups, their matching
CVLs and 20 randomized CVLs using Luminex multiplexing assays.
Results: The average time of Softcup insertion was 114 minutes (range
75-143). Eighteen of 19 Softcup samples had HIV-specific IgG to all
four antigens compared to 17/20 randomized CVLs. Detection of IgG
to gp41MN was highest with gp120 the lowest for both Softcup and
CVL. All four HIV-specific IgGs (MFIs) were significantly higher when
Softcup was compared to randomized CVL (all p< 0.001). Higher specific
activity [MFI/total IgG (ng/ml)] for Gag p24, gp41MN and gp120 (all
p< 0.05) in Softcup compared to CVL was found, indicating that even
lowly expressed gp120 IgG was significantly detected from Softcup.
Recovery of total IgG from Softcup was significantly higher than from
randomized CVLs (p< 0.0001). Softcup collection did not compromise
detectable HIV-specific IgGs in the subsequent CVL with no differences
of the IgG titres of the matching CVL and randomized CVL samples; and
total IgG in Softcup and matching CVL were highly correlated (r=0.73;
p=0.0006).
Conclusions: Softcup sampling is an ideal replacement to CVL alone,
and a subsequent lavage after Softcup provides equivalent IgG detection.
These data suggest that Softcup sampling should be considered a
replacement method to assess antibody responses at the female genital
tract.
334
Background: The HIV prevention field requires better tools for studying
mucosal drug toxicity and general inflammation. Standard tools that
study individual components in isolation likely underestimates the
global effects that may underpin complex immunological processes.
Advances in mass spectrometry allow for the measurement of a large
number of immunological parameters simultaneously. Here we evaluate
the utility of this technology for examining rectal mucosal samples, and
the effect of the known mucosal irritant nonoxynol-9 on the host rectal
mucosal proteome.
Methods: Rectal sponge samples were collected from 7 individuals pre
(Visit 1) and after 7 consecutive days (Visit 3) of once-daily doses of
Nonoxynol-9 (N9) or placebo HEC gel controls (MTN-007 phase 1 clinical
trial). Sponge eluates were analyzed by tandem mass spectrometry.
Results: 488 unique proteins were identified, that covered many
functions that may be important for product toxicity including dermatitis
(57, 6.78E-29), skin development (23, 4.57E-18), allergic response
(47, p=3.0E-20), psoriasis (127, p= 5.67E-38), cell death (127, 3.68E16), necrosis (107, 4.82E-13), and cancer (279, 1.1E-10). 46 proteins
were differentially expressed (p< 0.05) between N9 and HEC controls,
after 7 days of exposure (V3) (filtered to non-treated controls (n=8,
p< 0.05). Hierarchical clustering showed a general overexpression of
proteins in the N9 treatment arm. Biofunctional analysis indicated these
factors belong to skin disorders (20) and general inflammation (11),
and apoptotic, carcinogenic, and pro-inflammatory canonical pathways,
indicating a wide range of inflammatory and toxic effects of N9 (p<
0.05, BH corrected).
Conclusions: This indicates that proteomics methods are suitable for
monitoring immune factors in rectal mucosa, and mucosal-irritant
associated responses. This shows the utility of systems biology
techniques and adds to the available toolsets for toxicity monitoring of
future microbicide candidates.
P40.05
P40.06
The Action of Cilia in the Upper Reproductive
Tract of Women Influences the Distribution of
HIV
Functional Assessment of Antibody Activity in
Mucosal Tissue Explant and Cellular Inhibition
Assays
Ann M. Carias1, Daniel Stieh1, Danijela Maric1, Thomas J. Hope1
Hannah M. Cheeseman1, Katja Klein1, Abbey Evans1, Deborah
King1, Robin J. Shattock1
Northwestern University, Chicago, IL, United States
1
Imperial College London, Group of Mucosal Immunology and
Infection, London, United Kingdom
Background: Understanding the precise mechanisms of HIV
transmission across genital barriers is essential for studying vaccine
development and inhibitory compounds. Previously, we illustrated that
HIV can penetrate the intact female epithelium both ex vivo and in vivo,
suggesting a diffusion mechanism for HIV entry; however, these studies
were primarily focused on the lower female reproductive tract (FRT). We
recently found that SIV based vectors can reach the ovary. The distance
to transverse the uterus and fallopian tube of the upper FRT to reach the
ovary is too far to reach by simple diffusion.
Methods: To investigate the possible role of cilia on the diffusion of
HIV in the upper FRT, we examined the localization of cilia by staining
for microtubules. Bundles of microtubules are present in the extended
structure of individual cilia. To assess how ciliated epithelium might
influence the movement of fluorescent beads and virus, we directly
visualized the dynamics of fluorescent particles using time-lapse
imaging.
Results: We find that the movement of cilia has a significant influence
on the diffusion of virus and nanoparticles. First, the beating of the cilia
accelerates and alters the diffusion of particles by causing convection
currents in the regions adjacent to the ciliated epithelia. Second,
the beating of the cilia pushes the particles away from the ciliated
epithelium, essentially preventing particles from interacting directly with
the underlying epithelial barrier.
Conclusions: These data reveal that the ciliated epithelium of the upper
FRT provides protection from potential contacting particles. It seems
likely that this action also drives an acceleration of the diffusion of
the particles. The net effect of the lack of binding to the tissue and
acceleration drives particles to traverse the long distance to the ovary.
These data suggest that the anatomy of the FRT, combined with the
action of cilia, must be taken into account to achieve complete protection
by inhibitory modalities.
Background: The RV144 vaccine trial yielded results of 31.2% efficacy
in protection from HIV-1 infection, despite a lack of broadly neutralising
antibodies (BnAb). Subsequent analysis has demonstrated a correlate
of protection was high ADCC (antibody-derived cellular cytotoxicity)
IgG and low levels of IgA antibodies to the V1V2 region of env. Here,
we investigate the ability of a range of neutralising (nAb) and nonneutralising (nnAb) antibodies to prevent or lower HIV-1 infection in
cellular and tissue explant models.
Methods: A range of monoclonal antibodies with discrete epitope
specificity and structure isolated by the CHAVI consortium from
vaccinated and infected subjects have been screened in cellular and
tissue based models designed to mimic the initial events in transmission.
Results: A range of nnAbs and nAbs were screened for activity against
HIV-1BaL infection of macrophages, dendritic cells, dendritic trans/cis
infection of T cells, and mucosal explants. 21/25 nnAbs displayed no
antiviral activity in these models. Four nnAbs had inhibitory activity
against macrophage infection, however none had any inhibitory activity
against infection of mucosal tissue explants (rectal, penile, cervical).
Only the nAb CH31 displayed inhibitory activity across all models.
Conclusions: To date only nAb have been able to prevent infection
across all models tested. These data suggest that nnAbs may be
insufficient to prevent the initial infection of target cells in mucosal tissue.
Nevertheless, these conclusions come with caveats. First, experiments
were performed with HIV-1BaL, there may be differential activity against
other strains. Second, polyclonal nnAbs may show increased function.
Third, the tissue explant models represent a static system with no influx
of effector cells. Although such explants contain numerous cells capable
of performing effector functions, it cannot be excluded that initial
infection in vivo could result in recruitment of additional effector cells
able to control or eliminate infection.
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335
POSTERS
1
Posters
P40.07
P40.08
Genital IRIS, Immune Activation and
Inflammation in the Female Genital Tract
Influences HIV Shedding in HIV-infected
Women Starting HAART
Effects of Intra-vaginal Drying Agents on
Mucosal Immune Activation and Risk for HIV
Acquisition in South African Women
Smritee Dabee1, Jean-Mari Kriek1, David Lewis2,3, Maseko
Venessa2, Mkhize Nonhlanhla2, Pamela P. Gumbi3, Zizipho
Mbulawa1, Anna-Lise Williamson1,4, Heather B. Jaspan1,4, Jo-Ann
S. Passmore1,4
Institute of Infectious Disease and Molecular Medicine, University
of Cape Town, Department of Clinical Laboratory Sciences, Cape
Town, South Africa, 2National Institute for Communicable Diseases,
Johannesburg, South Africa, 3Institute of Infectious Disease and
Molecular Medicine, University of Cape Town, Cape Town, South Africa,
4
National Health Laboratory Services, Cape Town, South Africa
1
POSTERS
Background: Initiation of HAART has previously been associated with
development of genital IRIS. As more HIV+ individuals are eligible to
start treatment in sub-Saharan Africa, reactivation of subclinical sexually
transmitted infections (STIs) may influence genital HIV shedding and
transmission to uninfected partners. We investigated the impact of
HAART on STI prevalence, genital immune activation, inflammation, and
genital HIV viral loads in HIV+ women initiating HAART.
Methods: Blood and cervical mononuclear cells were obtained from
HIV+ women before, and 1 month, after starting HAART. T-cell activation
and proliferation (CD38, HLA-DR, Ki67) were measured by FACS. IL1β, IL-6, IL-8, IP-10, MIP-1α, MIP-1β, TNF-α, IL-7, G-CSF and IL-10
concentrations were measured by Luminex. Screening for C. trachomatis,
N. gonnorhoea, M. genitalium, T. vaginalis, and HPV was performed.
Results: Although HAART significantly reduced blood T-cell activation
(CD38+, HLADR+, CD38+HLADR+), genital activation levels remained
high. Cytokine concentrations were significantly higher in CVL than blood.
Blood cytokine concentrations were generally lower after 1 month on
HAART, significantly so for IP-10 (p=0.05). In contrast, cytokine levels in
CVL remained unchanged, with the exception of IL-10 which was lower
after HAART (p=0.007). More than half of the HIV-infected women had
a bacterial STIs and 90% were infected with HPV before starting HAART.
Initiation of HAART did not change this prevalence. Whereas >85% of
women had detectable HIV in their genital secretions pre-HAART, 18%
still had detectable genital tract viral loads 1 month post-HAART.
Conclusions: On-going immune activation, inflammation and prevalent
STIs in the female genital tract of women initiating HAART may influence
risk for HIV transmission to uninfected partners, despite the beneficial
effect of HAART to the individual.
336
Kathleen E. Doherty1, Melis Anahtar2, Musie Ghebremichael2,
Christina Thogabekale3, Nikita Padavattan3, Krista Dong4, Bruce
D. Walker2, Thumbi Ndung’u3,4, Douglas S. Kwon2
Vanderbilt University, Nashville, TN, United States, 2Ragon Institute
of MGH, MIT and Harvard, Cambridge, MA, United States, 3HIV
Pathogenesis Programme, Durban, South Africa, 4KwaZulu-Natal
Research Institute for TB&HIV, Durban, South Africa
1
Background: Most HIV transmissions occur in women following
exposure to virus in the female genital tract (FGT). Because FGT
inflammation has been associated with increased risk of HIV acquisition,
it is critical to understand the biologic and behavioral factors that may
contribute to elevated FGT inflammation. The Kwazulu-Natal (KZN)
region in South Africa has one of the highest rates of intra-vaginal
drying agent (IDA) usage worldwide. However, little is known about the
immunologic effects of these agents, specifically as they relate to genital
inflammation and risk of HIV acquisition.
Methods: FGT samples and detailed behavioral data were collected from
a high-risk cohort of young, HIV-negative women in KZN. Cytobrush cells
were assessed by flow cytometry. FGT mononuclear cells and cervical
tissue explants were incubated with drying agents in vitro and assessed
for changes in immune activation.
Results: Overall, 15.2% of the cohort reported using IDAs. The most
commonly used agents were “ntsu” (powdered tobacco) and “china
fruit.” Associations were seen between IDA use and earlier sexual
debut (p=0.0305), older sexual partners (p=0.0073), frequency of
sex (p=0.0373) and, importantly, increased HIV acquisition rates
(p=0.0015). IDA use was also associated with altered frequencies and
activation states of immune populations at the cervicovaginal junction.
Several common IDAs were shown to have effects on immune activation
on FGT mucosal cells and cervical explants in vitro.
Conclusions: This study describes the behavioral and immunological
associations of IDA usage and their potential impact on HIV acquisition.
It reveals increased acquisition rates among those who use IDA that are
likely due to both behavioral and biological factors. This work lays the
foundation for future research on therapies and behavioral interventions
to modulate the immune landscape in the FGT to prevent HIV acquisition.
P40.09
P40.10
Acceptability of Using the Softcup for the
Collection of Genital Fluid for Mucosal Assays
in HIV Prevention Trials
Alterations in Genital Tract Soluble Immune
Mediators in HIV Positive Postmenopausal
Women: Implications for HIV Acquisition and
Transmission
Josie Delisle1, Ifeyinwa Benyeogor1, Mariel Jais1, Naji Younes1,
Mary Young2, Mimi Ghosh1
Centre for the AIDS Programme of Research in South Africa (CAPRISA),
1
Background: A simple, scalable and participant-friendly method for
genital specimen collection to measure mucosal immune responses
is needed for the HIV prevention field. The purpose of this study was
to assess the acceptability of using Softcup, a menstrual cup, for the
collection of genital fluid from women participating in an existing HIV
cohort study.
Methods: Forty HIV-infected antiretroviral-naïve women in a cohort
study were randomized to have their genital sampling either by
1) Softcup® (EuroFemPro, Netherlands) followed by cervico-vaginal
lavage (CVL), or
2) CVL alone, which is the current standard method for mucosal
sampling.
Softcups were clinician-inserted and were removed after 2 hours.
Questionnaires with 5-point Likert scales ranging from extremely
unacceptable/ uncomfortable to very acceptable/ comfortable were
distributed to all women after removal of the Softcup, and were
completed by the participants.
Results: Twenty women, median age 31 (IQR 28-32), had a Softcup
inserted. The average time from Softcup insertion to removal was
114 minutes (range 75-143). A total of 17/20 questionnaires were
returned (85% response rate). Sixteen (94%) of participants answered
that insertion and wearing the Softcup were very comfortable. All
participants stated that the removal procedure was very comfortable,
and that the Softcup procedure was very acceptable compared to their
previous experience of genital examinations with a speculum performed
by the clinician. All participants indicated their willingness to wear a
Softcup again, and all preferred to have the Softcup inserted instead of a
speculum. However, only 29% indicated that they would consider selfinserting the Softcup, if trained to do so.
Conclusions: Collecting genital secretions from women using Softcup
was highly acceptable and was preferred to speculum insertion used for
other mucosal sampling methods. The Softcup should be considered for
genital sampling in women in future HIV prevention trials.
Background: Heterosexual transmission accounts for the majority of
new HIV infections with women being more likely than men to be
infected during vaginal intercourse. Multiple soluble immune mediators
in the female reproductive tract (FRT) that are hormonally regulated are
protective against HIV. It is unclear as to whether loss of estradiol in
postmenopausal women results in a blunted innate immune response
in the FRT thereby making them more susceptible to acquiring and
transmitting HIV. Furthermore, it is unknown whether postmenopausal
women on hormone replacement therapy (HRT) might recover these
innate immune functions. In this study, we investigated changes in
soluble immune mediators, IL-6, TNF-a, IL-8, MIP3a, Elafin, SLPI and
Human beta defensin-2 (HBD-2) in cervical-vaginal lavage (CVL) of
HIV(+) and HIV(-) postmenopausal women compared to premenopausal
women and women on HRT.
Methods: CVL from HIV(+) and HIV(-) premenopausal, postmenopausal
and women on HRT were obtained from the Washington DC Women’s
Interagency HIV Study (WIHS) local repository and analyzed using
commercially available ELISA. None of the HIV(+) women were on
HAART. Statistical analyses were performed using the Kruskal-Wallis test
(Graphpad Prism).
Results: HIV(+) postmenopausal women had significantly higher
plasma viral load and lower CD4 counts compared to premenopausal
women. We also observed significantly lower levels of HBD2 and MIP3a
in postmenopausal women, from both HIV(+) and HIV(-) groups. In HIV(-)
postmenopausal women and women on HRT, significant decreases in
levels of TNF-a, IL-6, and anti-HIV protective factor SLPI was observed.
However, both TNF-a and IL-6 were increased in HIV(+) postmenopausal
women.
Conclusions: Our data indicate that both HIV status and menopausal
status can dysregulate levels of soluble immune mediators in the FRT.
A shift in the balance between proinflammatory factors and anti-HIV
protective factors can result in higher susceptibility to acquisition and
transmission of HIV in these women.
1
The George Washington University, Washington, DC, United States,
Women’s Interagency HIV Study, Washington, DC, United States
2
www.hivr4p.org
337
POSTERS
Nigel Garrett1, Lindiwe Mpanza1, Themba Cekwane1, Nivashnee
Naicker1, Lise Werner1, Nelisile Majola1, Adolphus Mntambo1,
Derseree Archary1
Posters
P40.11
P40.12
Altered Levels of Soluble Immune Mediators
in HIV-negative Postmenopausal Women:
Implications for HIV Acquisition in the Elderly
Bacterial Vaginosis and HIV: An Analysis of
the MDP301 Trial
Mariel Jais , Naji Younes , Susan Cu-Uvin , Mimi Ghosh
1
2
3
2
Milken Institute School for Public Health, The George Washington
University, Epidemiology and Biostatistics, Washington, DC, United
States, 2Milken Institute School fo Public Health, The George
Washington University, Epidemiology and Biostatistics, Washington,
DC, United States, 3Brown University, Miriam Hospital, Providence, RI,
United States
1
POSTERS
Background: The female reproductive tract (FRT) secretes immune
mediators protective against sexually transmitted infections, including
HIV. As multiple immune factors in FRT are hormone-responsive, the loss
of sex hormones with aging may undermine these defense mechanisms.
Women are disproportionately affected by the HIV/AIDS epidemic with
heterosexual contact being the major source of new infections. Reports
indicate older women are sexually active and often do not use protection
as pregnancy is a non-issue. Therefore, investigating the effects of sex
hormone-loss on FRT mucosal immune factors is an important target to
curtail HIV acquisition.
Methods: CVL samples were collected from 20 HIV-negative
premenopausal and postmenopausal women. Each postmenopausal
woman provided one sample; each premenopausal woman provided 3
samples collected during proliferative, ovulatory, and secretory stages of
menstrual cycle. Commercially available ELISA kits were used to assess
the levels of IL-6, IL-8, TNFa, Elafin, HBD-2, MIP3a/CCL20 and SLPI.
Samples were analyzed for their anti-HIV activity against HIV-1 IIIB and
BaL strains via the TZM-bl assay.
Results: We observed significantly lower levels of critical immune
mediators in CVL from postmenopausal women compared to those
from premenopausal women: TNFa (11.6 vs 51.5 pg/mL), MIP3a (1.0
vs 93.8 pg/mL), SLPI (39.6 vs 239.2 pg/mL) and HBD-2 (626 vs 6821
pg/mL). Levels of IL-6 and IL-8 displayed a trend toward lower levels in
postmenopausal samples whereas Elafin levels remained unchanged.
Inhibition of HIV-1 infection was observed for X4/IIIB and R5/BaL strains
in both pre and postmenopausal samples with inhibition of BaL stronger
in premenopausal samples (54.2 vs 37.6%).
Conclusions: Our findings indicate that levels of critical mucosal immune
factors and anti-HIV-1 activity in CVLs are affected by the hormonal
status of healthy HIV-negative women. This indicates the need for
specific therapeutic interventions to boost genital tract immunity against
HIV in older women.
338
Sarah Joseph1, Nathlee Abbai2, Sinead Delaney-Moretlwe3, Mitzy
Gafos1, Saidi Kapiga4, Maureen Chisembele5, Andrew Abaasa6,
Ute Jensch7, Suzanna Francis8, Sheena McCormack1, Richard
Hayes8, Angela Crook1
MRC Clinical Trials Unit at UCl, HIV Prevention, London, United
Kingdom, 2Medical Research Council, HIV Prevention Research Unit,
Durban, South Africa, 3Wits Reproductive Health & HIV Institute,
Johannesburg, South Africa, 4Mwanza Interventions Trials Unit,
Mwanza, Tanzania, United Republic of, 5University Teaching Hospital,
Obstetrics and Gynaecology, Lusaka, Zambia, 6MRC/UVRI, Uganda
Research Unit on AIDS, Entebbe, Uganda, 7Clinical Laboratory Services,
University of Witwatersrand, School of Pathology, Johannesburg, South
Africa, 8London School of Hygiene & Tropical Medicine, MRC Tropical
Epidemiology Group, London, United Kingdom
1
Background: Bacterial vaginosis (BV) has been shown to be associated
with increased susceptibility to HIV-1 infection. We have used data from
the MDP301 microbicide trial to estimate the effect of BV on the risk of
HIV acquisition.
Methods: 8491 HIV negative women, enrolled at 6 centres in 4 subSaharan African countries between 2005 and 2008, and with a BV
result at baseline, were included in the analysis. BV was assessed
using the Ison Hay method: 1 (negative, reference), 2 (intermediate) or
3 (positive) at weeks 0, 12, 24, 40 and 52. Cox proportional hazard
models were fitted to estimate the effect of BV on risk of HIV acquisition.
Data were censored at HIV sero-conversion or at the last visit attended.
The following baseline or time-updated covariates were considered
in a multivariate model: age, centre, gel group, condom use, sexual
frequency, abnormal vaginal discharge, HSV-2, other cervico-vaginal
infection (CT, NG, TV, syphilis) and reported contraception type.
Results: 384 women had HIV seroconverted by the end of follow-up
(8002 person-years). At baseline, 3150 (37%) women were positive
for BV, 2073 (24%) were intermediate and 3268 (38%) negative.
3066/8491 (36%) never tested positive for BV whereas 1821 (21%)
tested positive throughout. Adjusted hazard ratios (aHR) for the risk
of HIV were 1.45 (95% CI 1.23-1.88) for those testing positive for BV
and aHR 1.04 (95% CI 0.77-1.40) for those with an intermediate result.
Other covariates which remained independently associated with HIV
acquisition in the adjusted model were age, centre, abnormal vaginal
discharge, HSV-2 , other cervico-vaginal infection and use of injectable
DMPA contraception.
Conclusions: The results of this secondary analysis show positive BV
to be independently associated with increased risk of HIV acquisition,
further strengthening the case for more research into this potentially
neglected risk factor for HIV.
P40.13
P40.14
Dynamic and Divergent Bacterial Species
Similar to Bacterial Vaginosis Prior to SIV in
Pigtail Macaques
Impact of Persistent Human Papillomavirus
(HPV) Infections on Inflammatory Cytokine
Levels in the Female Genital Tract:
Implications for HIV Risk
Nichole R. Klatt1, Sujatha Srinivasan2, Laura Richert-Spuhler1,
Michael Koday1, David Fredricks2
University of Washington, Pharmaceutics, WaNPRC, Seattle, WA,
United States, 2Fred Hutchinson Cancer Research Center, Vaccine and
Infectious Disease Division, Seattle, WA, United States
1
Jean-Mari Kriek1, Shameem Z. Jaumdally1, Zizipho Mbulawa1,2,
Pamela P. Gumbi1, Lindi Masson1, Shaun L. Barnabas1,3, David
Coetzee2, Anna-Lise Williamson1,2, Francesca Little4, Jo-Ann S.
Passmore1,2
Institute of Infectious Disease and Molecular Medicine, University
of Cape Town, Cape Town, South Africa, 2National Health Laboratory
Service, Cape Town, South Africa, 3Desmond Tutu HIV Foundation,
IIDMM University of Cape Town, Cape Town, South Africa, 4University
of Cape Town, Department of Statistical Science, Cape Town, South
Africa
Background: A challenge in developing interventions to prevent
mucosal HIV transmission is incomplete understanding of correlates
of vaginal transmission, including mucosal inflammation. Vaginal
SIV/SHIV transmission in pigtail macaques is an excellent model for
HIV transmission, however little is known about how genital bacterial
species may influence inflammation and SIV transmission.
Methods: Broad-range 16S rRNA gene PCR and pyrosequencing was
performed on vaginal swabs for bacterial identification using a cohort of
pigtail macaques in a cross-sectional analysis (N=22), and a more limited
longitudinal analysis (N=3). Bacteria were identified to the species
or genus level using a custom designed reference set of sequences,
providing comprehensive characterization compared to previous studies
focused on genera and phylum.
Results: Pigtail macaques had diverse microbial communities at all
phases of the menstrual cycle, with a range of dominant bacteria in
different animals, including Prevotella, Porphyromonas, Dialister,
Clostridiales, Bacteroides, Streptococcus and Lactobacillus species. In
contrast to previous studies, we found three animals had a Lactobacillusdominant vaginal microbiota, all at peak sex skin swelling (indicating
ovulation). Several species were identified that are commonly found
in bacterial vaginosis (BV) in humans, including Atopobium vaginae,
Prevotella buccalis, BVAB2, Peptoniphilus lacrimalis, Prevotella timonensis
and Gardnerella vaginalis. Longitudinal sampling demonstrated that
vaginal bacterial communities were dynamic and possibly driven by
alterations in environment throughout the menstrual cycle.
Conclusions: The macaque vaginal microbiota is diverse, dynamic, and
can resemble the human vaginal microbiota, including BV-associated
bacteria as well as Lactobacillus spp. These data provide a foundation
for understanding how vaginal microbial communities may impact risk
of HIV/SIV transmission using these models.
Background: Women with persistent HPV-infections are at increased
risk for developing cervical cancer. Clearance of HPV-infections has been
associated with genital inflammation. HPV- infection has been shown as
a risk factor for HIV acquisition along with genital inflammation. The aim
of this study was to evaluate the impact of genital tract inflammation
and HPV persistence or clearance in HIV negative women.
Methods: Cervical samples were collected from 38 HIV-negative women
at two time points, six months apart. IL-8, IL-6, IL-10, IL-15, IL-12p40, IP10, MCP-1, MIP-1a, MIP-1b, IL-1a, IL-1b, eotaxin, fractalkine, and G-CSF
concentrations were measured by Luminex at enrollment. HPV types
were assessed at both time points using the Roche Linear Array HPV
Genotyping assay.
Results: There were 20/38 HPV-infected women at enrollment [9/38 had
high-risk (HR) and 11/38 had low-risk (LR) HPV types]. An additional 7
women acquired an HPV infection over the 6 months of follow-up. Of the
20 women initially infected with HPV, only 2/20 cleared their infections
while 18/20 had infections that persisted for 6 months. Women with
HR HPV at enrollment generally had higher cytokine concentrations in
their genital tracts than women with LR types, although none of the
cytokine evaluated were significantly different between groups. Women
acquiring an HPV infection over 6 months had lower overall genital
inflammation at enrolment compared to women who remained HPVnegative (p=0.005). In contrast, women with persistent HPV infections
generally had increased inflammation at enrolment compared to women
who remained negative (p=0.049).
Conclusions: Although HPV infection has been associated with
increased risk for HIV infection, we found that HPV infections generally
did not cause an inflammatory response in the female genital tract.
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339
POSTERS
1
Posters
P40.15
P40.16
Genital Tract Immunological Markers in SubSaharan African Women with Relevance to
HIV Risk and Prevention
Influence of Vaginal Microbiota on
the Diffusional Barrier Properties of
Cervicovaginal Mucus
Jordan K. Kyongo1, Tania Crucitti2, Joris Menten2, Liselotte
Hardy2,3, Piet Cools4, Johan Michiels1, Sinead Delany-Moretlwe5,
Mary Mwaura6, Gilles Ndayisaba7, Sarah Joseph8, Raina
Fichorova9, Janneke van de Wijgert10, Guido Vanham1,11,12, Kevin
K. Ariën1, Vicky Jespers3, Biomarkers Study Group
Kenetta Nunn1, Ying-Ying Wang2, Dimple Harit1, Richard Cone2,
Samuel Lai1
Institute of Tropical Medicine, Department of Biomedical Sciences,
Antwerp, Belgium, 2Institute of Tropical Medicine, Department of
Clinical Sciences, Antwerp, Belgium, 3Institute of Tropical Medicine,
Department of Public Health, Antwerp, Belgium, 4Ghent University,
Department of Microbiology, Immunology and Clinical Chemistry,
Ghent, Belgium, 5University of the Witwatersrand, Wits Reproductive
Health & HIV Institute, School of Clinical Medicine, Johannesburg,
South Africa, 6International Center for Reproductive Health, Mombasa,
Kenya, 7Project Ubuzima, Kigali, Rwanda, 8MRC Clinical Trials Unit at
University College London, London, United Kingdom, 9Brigham and
Women’s Hospital, Harvard Medical School, Department of Obstetrics,
Gynaecology and Reproductive Biology, Boston, MA, United States,
10
University of Liverpool, Institute of Infection and Global Health,
Liverpool, United Kingdom, 11University of Antwerp, Faculty of
Pharmaceutical, Veterinary and Biomedical Sciences, Antwerp, Belgium,
12
University of Brussels, Faculty of Medicine and Pharmacology,
Brussels, Belgium
University of North Carolina at Chapel Hill, Eshelman School of
Pharmacy, Chapel Hill, NC, United States, 2Johns Hopkins University,
Baltimore, MD, United States
1
1
POSTERS
Background: Data on immune mediators in the genital tract and the
behavioural and clinical factors that modulate them in Sub-Saharan
women are limited.
Methods: Cervicovaginal lavage (CVL) samples from 430 sexually active
women from Kenya, South Africa and Rwanda were analysed for twelve
soluble immune mediators. qPCR was used to quantify ten bacterial
species in vaginal swab samples. We also compared the anti-HIV activity
of CVL samples from bacterial vaginosis (BV)-positive women to those
from women with a Nugent score of 0.
Results: Pregnant women, adolescents, women engaging in traditional
vaginal practices and HIV-positive women differed in specific soluble
markers compared to reference groups of adult HIV-negative women.
An increase in cervical mucus, the presence of cervical ectopy, abnormal
vaginal discharge and having multiple sexual partners were each
associated with an increase in mediators of inflammation. Interleukin
(IL)-1α, IL-1β, IL-6, IL-12 and IL-8 were elevated and the IL-1RA/(IL-1(α+β)
ratio decreased in the CVLs of women with BV. Interferon gammainduced protein (IP)-10 was decreased in BV-positive compared to BVnegative women. Lactobacillus crispatus and Lactobacillus vaginalis were
associated with lower levels of pro-inflammatory cytokines and each
BV-associated species with increased pro-inflammatory cytokines. The
in vitro anti-HIV activity of CVLs from BV-positive women was stronger
than that of BV-negative women.
Conclusions: We found significant associations of factors that can
influence HIV susceptibility with the levels of soluble immune mediators
in the vaginal environment of sexually active women. These factors
need to be considered when establishing normative levels or pathogenic
cut-offs of biomarkers of inflammation and associated risks in African
women. IP-10 suppression may be one potential mechanism of immune
evasion by BV-associated bacteria. Lastly, cervicovaginal secretions of
women with BV may contain active anti-HIV substances that should be
examined more closely.
340
Background: To reach target cells underlying the vaginal epithelium,
HIV must penetrate cervicovaginal mucus (CVM) secretions. We
previously found that native CVM from women with healthy, lactobacillidominated vaginal flora (pH ~3.5-4) effectively trapped HIV, but the
same finding was not observed in a later follow-up study. We sought to
investigate whether the vaginal microbiota can differentially influence
the diffusional barrier properties of CVM against HIV and other sexually
transmitted viruses.
Methods: We evaluated the mobility of fluorescent HIV pseudoviruses
in fresh, undiluted CVM from different women subjects using high
resolution multiple particle tracking. We then correlated the observed
mobility to various biochemical and biophysical characterizations,
including pH, D- and
L- lactic acid levels and Nugent scores. We are also performing
microbiome analysis on a subset of the specimens.
Results: In a cohort of over 40 CVM specimens, the real-time mobility of
mCherry-Gag labeled HIV was not significantly correlated to native pH,
Nugent scores, total IgG/IgA or total lactic acid (LA) levels. Interestingly,
CVM samples that failed to trap HIV exhibited substantially lower D-LA
content, whereas CVM samples that effectively trapped HIV exhibited
both high D- and L- LA. The same phenomenon was observed with ΔEnv
virions, suggesting it may be universal among all enveloped viruses.
Addition of D-LA did not trap HIV, suggesting that low D- lactic acid
is likely a surrogate marker of the native CVM barrier. High L-LA/low
D-LA is consistent with vaginal microbiota that is populated with specific
strains of lactobacilli, which we are currently confirming using 16S rRNA
pyrosequencing.
Conclusions: The vaginal microbiota plays an underappreciated and yet
to be clarified role in modulating the innate diffusional barrier properties
of mucus against HIV. Our work may help identify women who are at
markedly enhanced susceptibility to acquiring HIV and other sexually
transmitted infections.
P40.17
P40.18
A Randomized Study Comparing Softcup and
Cervicovaginal Lavage Sampling to Measure
Genital Cytokine Concentrations in HIVinfected Women
Safety Studies of a Formulation Comprising
Recombinant Human Surfactant Protein D for
Prevention of HIV-1 Infection
Taruna Madan1, Hrishikesh Pandit1, Kavita Kale1, Uday Kishore2
Lenine J. Liebenberg1, Nigel Garrett1, Lise Werner1, Nelisile
Majola1, Nivashnee Naicker1, Natasha Samsunder1, Sarah
Dlamini1, Jo-Ann S. Passmore1,2,3, Salim S. Abdool Karim1,4,
Derseree Archary1
CAPRISA, Durban, South Africa, 2Infectious Disease and Molecular
Medicine, University of Cape Town, Cape Town, South Africa, 3National
Health Laboratory Services, Cape Town, South Africa, 4Department of
Epidemiology, Mailman School of Public Health, Columbia University,
National Institute for Research in Reproductive Health (ICMR), Innate
Immunity, Mumbai, India, 2Brunel University, Centre for Infection,
Immunity and Disease Mechanisms, London, United Kingdom
1
Background: Evaluating concentrations of genital cytokines are
key to understanding local immunity to HIV infections or infection
risk. Menstrual softcups may provide a more convenient method for
collection of genital specimens than cervicovaginal lavages (CVLs). We
compared softcup sampling to the CVL collection method for detection
of cytokines in genital fluid.
Methods: Forty ART-naïve HIV-infected women from a cohort study of
HIV infection were randomized to either have genital fluid collected by
softcup with subsequent CVL, or by CVL alone. Luminex was used to
measure the concentrations of 48 cytokines involved in hematopoiesis,
regulation, adaptive responses, inflammation and growth, in both CVL
and softcup specimens.
Results: In randomized softcup specimens, 42/48 cytokines were
consistently detected in all participants compared to 26/48 in CVL
(p=0.0003). Cytokines detected in softcups but not in CVL included those
important to HIV infection and pathogenesis (IL-8, MIP-1α, RANTES,
TNF-α). Differential concentrations were observed for 41/48 cytokines
in randomized softcup and CVL specimens, and 22 of these cytokines
remained significantly elevated in softcups after adjustment. Cytokine
concentrations in matched softcup secretions and CVL correlated
significantly (r=0.903; p< 0.0001), suggesting that proportions of
cytokines measured are retained in the two sampling methods. However,
even in paired specimens, 88% of cytokines were consistently detected
in softcup specimens from all participants compared to 69% in CVL
(p=0.0263).
Conclusions: This study demonstrates that the overall cytokine
composition in softcup secretions compares to CVL genital sampling.
However, softcup sampling allows for more reliable detection of
cytokines, particularly those present at the lowest concentrations.
Background: Owing to the lack of an effective vaccine for HIV-1,
efforts towards development of potent preventive strategies has gained
impetus. We have previously reported that human surfactant protein D,
an integral innate immune molecule has a potent anti-HIV-1 activity.
SP-D interferes with CD4 binding regions of gp120 and modulates
pro-inflammatory signaling cascade and cytokine production. Thus, in
view of its potential as a microbicidal agent, we determined in vitro
anti-HIV activity of a formulation comprising recombinant human SP-D
(rhSP-D) against several HIV-1 clinical isolates, with different tropism
and subtypes in various cell lines and primary cells. The formulation
did not show any adverse effects on the viability or lactic acid/peroxide
production of clinical strains of lactobacilli.
Methods: In vitro post-coital efficacy of rhSP-D in the presence of
vaginal lavage (VL) and seminal plasma (SP) was determined. Effect
on viability of VK2 cells was determined by MTT assay. In the rabbit
vaginal irritation model, 1ml gel containing rhSP-D formulation, placebo
and 0.1% SDS formulation as positive control was administered for 10
consecutive days. Vaginal tissues were examined by H&E staining for
infiltration of immune cells and inflammation and the serum cytokine
levels were determined.
Results: The rhSP-D inhibited HIV-1 infectivity in a dose-dependent
manner in the presence of VL and SP. We also observed that SP-D is
expressed by vaginal epithelial cells and is hormonally regulated. rhSP-D
had no adverse effect on the viability of VK2 cells with a therapeutic
index >10. The formulation is being evaluated using rabbit vaginal
irritation model and the results are being analyzed.
Conclusions: The study suggests that rhSP-D formulation is promising
with its efficacy and safety. Studies using human vaginal explants
models would be the next step towards its translational potential.
www.hivr4p.org
341
POSTERS
1
Posters
P40.19
P40.20
Mucosal Proteomic Profiles Associated with
Female Genital Tract Inflammation
Control of HIV-1 Infection in the Female
Reproductive Tract by Mucosal Innate
Immunity Determinants
Kelly Arnold1, Adam Burgener2, Kenzie Birse2, Laura Dunphy1,
Kamnoosh Shahabi3, Max Abou4, Jessie Kwatampora5, Billy
Nyanga5, Joshua Kimani2, Lenine Liebenberg6, Lindi Masson7,
Salim S. Abdool Karim6, Jo-Ann S. Passmore7, Douglas A.
Lauffenburger1, Rupert Kaul3, Lyle R. McKinnon6
MIT, Boston, MA, United States, 2University of Manitoba, Winnipeg,
MB, Canada, 3University of Toronto, Toronto, ON, Canada, 4Public
Health Agency of Canada, Winnipeg, MB, Canada, 5University of
Nairobi, Nairobi, Kenya, 6CAPRISA, Durban, South Africa, 7University of
Cape Town, Cape Town, South Africa
1
POSTERS
Background: Inflammatory cytokines in cervicovaginal lavage (CVL)
were associated with an increased risk of HIV infection in the CAPRISA004
trial. The mechanisms by which inflammation increases HIV risk are not
yet fully understood. We investigated mucosal biomarkers and pathways
associated with raised inflammatory cytokines in the genital tract.
Methods: We classified HIV-negative Kenyan women as ‘inflammation+’
(n=28) or ‘inflammation-’ (n=68) based on the elevation (upper quartile)
of at least three of 7 inflammatory cytokines MIP-3α, RANTES, IL-8, MIP1β, IL-1β, IL-1α, and GM-CSF in CVL. CD4+ T cells from endocervical
cytobrushes were enumerated in the same participants. CVL samples
were further analyzed by tandem mass spectrometry, and subjected
to several canonical and biofunctional pathway analyses. Partial
least squares discriminant analysis (PLSDA) and LASSO methods for
regression shrinkage/selection were used to model the minimum set of
biomarkers that best classified inflammation groups.
Results: Women with inflammation had a >2 fold increase in the
number of endocervical CD4+ T cells
(p< 0.001). A total of 716 proteins were quantified; 53 of these were
increased and 27 were decreased in inflammation+ women (p< 0.01). Of
these 80 biomarkers, 50 remained significantly different after applying
a 5% False Discovery Rate threshold. Hierarchical clustering suggested
significantly different proteomes associated with mucosal inflammation
(p< 0.01). Inflammation was associated with acute phase signaling,
complement activation, and remodeling of epithelial adherens junctions,
and the chemotaxis and adhesion of immune cells. Anti-inflammatory
peptidase inhibitors and keratinization factors were downregulated.
LASSO and PLSDA identified 13 biomarkers that distinguished the
inflammation+ group with 88% accuracy on calibration and 84%
accuracy on cross-validation.
Conclusions: These analyses highlight several potential underlying
molecular mechanisms that could link inflammation to HIV susceptibility.
342
Héloïse Quillay1,2, Hicham El Costa2, Claude Cannou2, Marion
Duriez2, Romain Marlin3, Claire de Truchis4, Anne Le Breton4,
Mona Rahmati5, Julien Ighil5, Olivier Schwartz2, Françoise BarréSinoussi2, Marie-Thérèse Nugeyre2, Elisabeth Menu2
Univ. Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Paris, France,
Institut Pasteur, Paris, France, 3Université Bordeaux, Bordeaux, France,
4
Antoine Béclère Hospital, Clamart, France, 5Pitié Salpétrière Hospital,
Paris, France
1
2
Background: Mucosas are the preferential portal for HIV-1 entry in the
body. It is thus crucial to identify the determinants necessary for an
efficient control of transmission in the mucosa.
The control of HIV-1 infection at the materno-fetal interface during the
first trimester of pregnancy is a good model to study natural protection
against transmission in the female reproductive tract. In the decidua
(uterine mucosa during pregnancy), macrophages (dMs) are the main
target cells of R5 tropic HIV-1. The aim of this study was to characterize
control mechanisms of infection in the decidua.
Methods: Deciduas were obtained from HIV-1 negative women
undergoing elective abortions (8-12 weeks of amenorrhea) with their
written informed consent. Purified dMs were infected with R5 HIV-1 or
HIV-1/VSV-G pseudotype. Natural killer (dNK) cells were added to dMs
(ratio 1:5) at different times. Cocultures were performed in the same
well or separately in a double chamber system. SAMHD1 expression
was determined by flow cytometry. In some experiments, VLP-Vpx was
added to dMs simultaneously with the virus.
Results: Cocultures of infected dMs with autologous dNK cells revealed
that dNK cells controlled dM HIV-1 infection in the early steps of
infection. Cell-to-cell contacts and soluble factors were necessary for
an efficient control. NKG2D and IFN-γ were involved in the control.
dMs highly expressed SAMHD1, a restriction factor blocking HIV-1
replication. Infection of dMs in presence of the viral accessory protein
Vpx, which degrades SAMHD1, increased dM infection.
Conclusions: These data demonstrate in vitro for the first time that 1)
dNK cells control HIV-1 infection by several mechanisms and 2) the
infection of dMs is restricted by SAMHD1. The determinants of protection
identified in the decidua are under investigation in the non pregnant
female reproductive tract. These studies give important information for
the development of future preventive strategies against HIV-1 mucosal
transmission.
P40.21
P40.22
Presence of Male Partner Semen Influences
the Inflammatory and Innate Cytokine
Environment in the Female Genital Tract
Acceptability of Multiple Mucosal Specimen
Collection in a Phase 1 HIV Vaccine Trial in
Rwanda
Sinaye Ngcapu1, Tracey Meiring2, Lindi Masson2, Lise Werner1,
Lenine Liebenberg1, Nigel Garrett1, Koleka Mlisana1,3, Carolyn
Williamson2, Quarraisha Abdool Karim1, Salim Abdool Karim1, JoAnn Passmore1,2,4
Julien M. Nyombayire1, Rosine Ingabire1, Jeannine
Mukamuyango1, Etienne Karita1, Dagna Laufer2, Harriet Park2,
Phillip Bergin3, Frances Priddy2, Susan Allen1,4
Background: In addition to spermatozoa, semen contains high
concentrations of anti-inflammatory cytokines (TGF-b, IL-10, PGE2),
inflammatory cytokines (IL-8, IL-1b, IL-6), and activated immune cells,
each potentially capable of influencing the immune environment of the
lower female genital tract. While several studies have quantified the
inflammatory cytokine composition of genital secretions from sexually
active women that may influence HIV risk, few have investigated
the influence of recent sexual intercourse on the cytokine milieu of
cervicovaginal secretions.
Methods: We investigated the semen exposure in cervicovaginal lavage
(CVL) samples collected from 72 high-risk HIV negative women by
investigating the presence of Y-chromosome by amplification of the
TSPY1 gene of the Y-chromosome by PCR, and by quantifying prostate
specific antigen (PSA) levels by ELISA. Levels of 42 cervicovaginal
cytokines, 5 MMPs and 4 TIMPs were measured by luminex and were
compared in women with or without semen exposure.
Results: Y-chromosomes were detected in 29% (21/72) of CVLs and
denoted sex within 7 days prior to sampling (Y-chromo+); and17%
(12/72) of participants were PSA+, denoting sex within 2 days prior
to sampling. Of the Y-chromo+ participants, 57% (12/21) were also
PSA+. Only 33% (7/21) of women who were Y-chromo+ and PSA+
had self-reported condom use at their last sex act. Y-chromo+ CVLs
had significantly higher levels of PDGF-AA, MMP-7 and MMP-10 levels,
after adjusting for multiple comparisons. In contrast, PSA+ CVLs had
significantly lower concentrations of IL-1Ra, IL-2Rα, GM-CSF and TNF-β.
Conclusions: These findings suggest that the presence of semen
influenced the cytokine and MMP profile in CVL and should be taken
into consideration when investigating biological markers in the female
genital tract.
Rwanda Zambia HIV Research Group-Projet San Francisco, Kigali,
Rwanda, 2International AIDS Vaccine Initiative, New York, NY, United
States, 3IAVI Human Immunology Laboratory, London, United
Kingdom, 4Emory University, School of Medicine, Department of
Pathology and Laboratory Medicine, Atlanta, GA, United States
1
Background: HIV infection is mostly acquired through mucosal surfaces
and an efficacious HIV vaccine would ideally elicit an immune response
at these surfaces.This study presents acceptability of the mucosal
sampling procedures among study volunteers enrolled in a phase 1 HIV
vaccine trial in Kigali, Rwanda.
Methods: Since January 2013, Project San Francisco (PSF) has been
participating in a multicenter phase 1 clinical trial to assess the safety
and immunogenicity of a Sendai HIV vaccine given intra-nasally and
an Ad35-GRIN HIV vaccine administered intra-muscularly in primeboost regimens.The study was discussed extensively with volunteers
during education and consenting sessions.Mucosal sampling included
saliva from parotid glands (Salimetrics Oral Swabs), oral fluids(Falcon
tubes),nasal secretions(FloQ swabs),cervico-vaginal secretions (Softcup
or Merocel sponge), and rectal secretions (Merocel sponge). Samples
were collected at baseline before vaccination and at 5 subsequent time
points during follow-up to date.The study is ongoing and will have 9
mucosal sampling time points in total.Consent for mucosal sampling
was reassessed at each collection visit.
Results: A total of 20 participants were enrolled (8 women and 12men),
and all 20 participants (100%) consented to each type of mucosal
secretion collection.As of April 2014, the 20 study volunteers have
completed the 6 mucosal collection visits each. All study participants
provided all protocol required specimens, both at baseline and at the 5
subsequent follow-up visits, resulting in a 100% acceptability rate.
Conclusions: This study confirms that multiple collection sites and
repeated mucosal specimen collection in HIV vaccine trials is highly
acceptable in Rwanda. The high acceptability of these procedures may
be a reflection of extensive counseling and mutual trust between study
participants and study staff.
www.hivr4p.org
343
POSTERS
CAPRISA, Durban, South Africa, 2Institute of Infectious Diseases and
Molecular Medicine, University of Cape Town, Cape Town, South Africa,
3
School of Laboratory Medicine and Medical Sciences, University of
KwaZulu Natal, Durban, South Africa, 4National Health Laboratory
Services, Cape Town, South Africa
1
Posters
P40.23
P40.24
Implementation of a Training Program to
Standardize Mucosal Sample Collection and
Processing at Multiple Laboratories in Eastern
Africa
Anti-HIV Activity of Vaginal Epithelial Cells
and Vaginal Secretions
Gloria Omosa-Manyonyi1, Robert Langat1, Elizabeth Mutisya1,
Harriet Park2, Philip Bergin3, Bashir Farah1, Hilda Ogutu1, Simon
Ogola1, Gina Ouattara1, Jackton Indangasi1, Rose Ndambuki1, Pat
Fast2, Dagna Laufer2, Fran Priddy2, Omu Anzala1
Kenya AIDS Vaccine Initiative Institute of Clinical Research, Nairobi,
Kenya, 2International AIDS Vaccine Initiative (IAVI), New York, NY,
United States, 3International AIDS Vaccine Initiative (IAVI)-HIL, Imperial
College, London, United Kingdom
Mickey V. Patel1, Mimi Ghosh2, Richard M. Rossoll1, John V.
Fahey1, Charles R. Wira1
Geisel School of Medicine at Dartmouth, Physiology and
Neurobiology, Lebanon, NH, United States, 2George Washington
University, Epidemiology and Biostatistics, Washington, DC, United
States
1
1
POSTERS
Background: HIV-1 transmission occurs predominantly across the
genital mucosa during sexual intercourse. Understanding host mucosal
responses to HIV-1 infection, and assessing protective mucosal responses
during microbicide and HIV vaccine clinical trials are important. For
comparable data across studies and research centers, mucosal samples
should be obtained in a standardized reproducible manner.
Methods: KAVI-ICR and IAVI developed standardized protocols for
collecting, processing and assay of cervico-vaginal, rectal and upper
respiratory tract mucosal samples. After proof of concept studies, KAVIICR developed a 3-day-hands-on training program on specific mucosal
sample collection and processing methods, and transferred these to
research staff at centers across Eastern Africa. Participant consenting,
community engagement, and access to supplies were discussed.
Results: Standardized mucosal sample collection and processing
procedures were applied to three Phase 1 studies at KAVI-ICR; mucosal
anti-HIV IgG/IgA responses were detectable. Forty-six research staff
from 5 centers had hands-on training on the standardized methods
(12 clinicians, 14 nurses, 20 lab staff). Projet San Francisco (PSF) Kigali
Rwanda, and MRC/UVRI Uganda Research Unit on AIDS had on-site
training; while staff from UVRI-IAVI HIV Program Uganda, KEMRI/
Walter Reed Project (KEMRI/WRP)-Kericho Kenya, and KEMRI-Centre
of Geographical Medicine Research Coast Kenya, visited KAVI-ICR
for training. PSF Rwanda and KEMRI/WRP-Kericho have applied the
methods learnt on mucosal studies, including mucosal sampling in HIV
vaccine trials. The mucosal studies at PSF Rwanda were multi-site and
the resulting mucosal data was comparable across all participating sites.
Conclusions: It is possible to standardize mucosal sampling across
research centers to ensure comparable samples and resulting data.
KAVI-ICR may become a regional mucosal training center for transfer of
these methods in South-South interactions.
344
Background: HIV is the leading cause of death for reproductiveage women, with the majority of transmission events occurring via
heterosexual intercourse. The first site of exposure is the vagina, and
understanding its immune environment, which changes with hormone
levels across the menstrual cycle, is crucial to developing effective
vaccines and prevention measures.
Methods: Using a menstrual cup, we developed a novel technique to
concurrently recover fresh undiluted vaginal secretions (VS) and vaginal
epithelial cells (VEC) at mid-proliferative, ovulatory and mid-secretory
stages of the menstrual cycle. VEC were isolated and treated in 96well plates with estradiol (E2: 5x10-8M), progesterone (P4: 1x10-7M), or
a combination of both for 48hrs after which conditioned media (CM)
were recovered. VEC CM and VS were analyzed for presence of anti-HIV
proteins by ELISA, and anti-HIV activity using a TZM-bl assay.
Results: CCL20, RANTES, elafin, HBD2, SDF-1α and IL-8 were present in
VS. VS demonstrated significant inhibition of X4 (IIIB) HIV and increased
infectivity of reporter cells by R5 (CH077.t) HIV. No inhibitory effect was
seen for BaL and CH058. No significant differences in either antiviral
protein concentration or anti-HIV activity with respect to menstrual cycle
stage were measured, but marked differences were observed in both
parameters over the course of the cycle between different women, and
in consecutive cycles from the same woman. For VEC, E2 significantly
decreased the secretion of HBD2 and elafin over 48hrs. CM from E2- or
P4-treated VEC had no anti-HIV activity. However, CM from E2/P4 cotreated cells significantly inhibited both R5 and X4 HIV.
Conclusions: There are multiple levels of protection in the vagina. The
sensitivity of VEC to E2 and P4 suggests that their contribution to immune
protection varies across the menstrual cycle. The variation in anti-HIV
activity in VS demonstrates that immune protection is not constant and
that factors in addition to hormones influence antiviral protection.
P40.25
P40.26
Antiviral Responses of Fibroblasts in the
Female Reproductive Tract
Soluble Toll-like Receptor 2 Is Significantly
Elevated in HIV-1 Infected Breast Milk and
Inhibits HIV-induced Cellular Activation and
Infection
Mickey V. Patel1, John V. Fahey1, Richard M. Rossoll1, Charles R.
Wira1
Geisel School of Medicine at Dartmouth, Physiology and
Neurobiology, Lebanon, NH, United States
1
Bethany M. Henrick1,2, Xiao-Dan Yao1,2, Anna G. Drannik1,2,
Alash’le Abimiku3, Kenneth L. Rosenthal1,2, INFANT Study Team
McMaster University, Department of Pathology & Molecular Medicine,
Hamilton, ON, Canada, 2McMaster University, McMaster Immunology
Research Centre, Hamilton, ON, Canada, 3Institute of Human Virology,
University of Maryland School of Medicine, Baltimore, MD, United
States
Background: The female reproductive tract (FRT) is the primary
location of heterosexual transmission of HIV. However, the intricacies of
immune protection at this site are not well understood, in particular the
contribution of stromal fibroblasts in preventing HIV infection. Fibroblasts
are the predominant cell type in the sub-epithelial layers of the FRT, and
the initial stages of HIV infection occur in their presence.
Methods: Benign FRT tissues from the endometrium (EM), endocervix
(Cx) and ectocervix (ECx), were recovered from HIV- women undergoing
hysterectomy. Following enzymatic digestion, cells were passed through
nylon filters to isolate purified populations of fibroblasts. These were
maintained in culture for 4-6 days prior to treatment with estradiol (E2:
5x10-8M) for up to 72hrs in the presence or absence of viral stimuli.
Conditioned media (CM) was recovered and analyzed for the presence
of anti-HIV proteins (ELISA) and anti-HIV activity (TZM-bl).
Results: The anti-HIV proteins RANTES, SDF-1α and CCL20 were
present in CM. While average RANTES levels were relatively constant
between the 3 tissue sites, SDF-1α was significantly higher in CM from
EM fibroblasts (2526 pg/ml) compared to the Cx (276 pg/ml) and ECx
(962 pg/ml). In contrast CCL20 was highest in CM from the Cx (500 pg/
ml) compared to the EM (40 pg/ml) and ECx (171 pg/ml). E2, via ERα,
significantly upregulated the secretion of SDF-1α by only EM fibroblasts,
with no effect on RANTES and CCL20. CM from all 3 sites showed potent
inhibition of BaL (R5 HIV). However, CM from E2-treated EM fibroblasts
demonstrated significantly greater inhibition of IIIB (X4 HIV) compared
to control CM. In contrast, control ECx CM significantly upregulated IIIB
infection of TZM-bl cells.
Conclusions: These results demonstrate that secretions from stromal
fibroblasts are capable of inhibiting HIV infection. Furthermore, their
antiviral activity is altered by E2 suggesting that it may vary across the
menstrual cycle and by anatomical location in the FRT.
Background: We previously demonstrated that immunodepletion
of soluble Toll-like receptor 2 (sTLR2) from breast milk significantly
increased HIV infection in vitro; therefore we next wanted to identify its
mechanism of action and characterize sTLR2 levels in HIV-infected and
uninfected breast milk.
Methods: Breast milk from HIV-infected and uninfected Nigerian and
Canadian women were blindly tested for levels of sTLR2, proinflammatory
cytokines, and viral antigenemia. In vitro experiments were conducted
using cell lines and primary cells to assess sTLR2 function on innate
responses and HIV infection.
Results: HIV-infected breast milk showed significantly increased levels
of sTLR2 compared to uninfected milk. Further, sTLR2 levels correlated
with HIV p24 and IL-15, thus suggesting a local innate compensatory
response in the HIV-infected breast. Given the increase in sTLR2 in
HIV-infected milk, we next demonstrated that mammary epithelial
cells (MECs) and macrophages, which are prevalent in milk, produced
significantly increased levels of sTLR2 following exposure to HIV-1
proteins p17, p24 and gp41 or the TLR2 ligand, Pam3CSK4. sTLR2 coimmunoprecipitated with p17, p24 and gp41 and inhibited HIV-induced
NFκB activation and inflammation. Importantly, binding of sTLR2 to HIV
proteins inhibited a TLR2-dependent increase in CCR5 expression, thus
resulting in significantly reduced HIV infection.
Conclusions: Our results demonstrate that sTLR2 inhibits HIV cellular
activation, inflammation, increased CCR5 expression and HIV infection
through direct interaction with the virus. Further, sTLR2 is significantly
elevated in HIV-infected breast milk and positively correlated with p24
and IL-15. Overall, our data suggests that sTLR2 may play a critical role
in inhibiting mother-to-child HIV transmission.
www.hivr4p.org
345
POSTERS
1
Posters
P40.27
P40.28
An Approach to Unravel Cellular Mechanisms
Responsible for Enhanced Neisseria gonorrhea
Induced HIV Acquisition and its Effect on
Microbicides
Unexpected Inflammatory Effects of
Intravaginal Gels (Universal Placebo Gel
and Nonoxynol-9) on the Upper Female
Reproductive Tract
Anwesha Sanyal1, Phalguni Gupta1, Ming Ding1, Deena Ratner1
Karen Smith-McCune1, Joseph Chen1, Ruth Greenblatt1, Barbara
Shacklett2, Uma Shanmugasundaram2, Joan Hilton1, Brittni
Johnson1,3, Fatima Barragan1,4, Juan Irwin1, Margaret Takeda1,
Jane Pannell1, Jean Perry1, Linda Giudice1
University of Pittsburgh Graduate School of Public Health, Infectious
Diseases and Microbiology, Pittsburgh, PA, United States
1
POSTERS
Background: Sexually Transmitted Infection, like Neisseria gonorrhea
(NG) in the female reproductive tract (FRT) increases sexual
transmission of HIV-1. Our goal is to identify NG induced cellular and
viral factors that are responsible for enhanced HIV acquisition.
Methods: Cervical tissues in an organ culture system were used to
measure inflammatory response to NG by real time RT-PCR and MSD
multiplex assay, respectively. HIV transcription in the presence of NG
was measured in a reporter gene assay in TZM bl cells. Microdisected
epithelial layer from cervical tissues following exposure to NG and HIV
were subjected to comprehensive transcriptome analysis using second
generation sequencing in an ion torrent platform. Effect of NG on
microbicides activity was evaluated in an organ culture system.
Results: Upon exposure to NG high levels of inflammatory cytokines
IL-1β and TNFα were detected in cervical tissues. Furthermore, the
supernatants from the organ cultures exposed to NG increased
transcription from the HIV- LTR in a TZM bl cell based assay and increased
transmission of HIV across cervical mucosa. From gene expression
analysis of the microdisected epithelial layer of the tissues exposed to
NG and the control, 30 genes have been identified to be differentially
upregulated with high statistical significance. CXCL10 and IL8 genes
were found to be common between tissue exposed to HIV and NG
group. In addition, the efficacy of microbicide (RC101 and CSIC), which
decreased HIV transmission in organ culture, was not compromised in
the presence of NG in the organ culture system.
Conclusions: The results suggest that NG infection induces higher
levels of CXCL10 and IL8 proteins that either by themselves or interact
with cellular factors responsible for HIV transmission and enhance
HIV acquisition. Furthermore, the presence of NG did not have any
effect on the antiviral activity of the microbicides indicating that these
microbicides would be effective for decreasing HIV transmission in the
presence of existing NG.
346
1
UCSF, San Francisco, CA, United States, 2UC Davis, Davis, CA, United
States, 3UCLA, Los Angeles, CA, United States, 4Michigan State
University, Grand Rapids, MI, United States
Background: Intravaginal microbicides could provide women with a selfcontrolled means for HIV prevention, but results from clinical trials have
been largely disappointing. We postulated that unrecognized effects
of intravaginal gels on the upper female reproductive tract (FRT) might
contribute to the lower-than-expected efficacy of HIV microbicides.
Methods: In this observational crossover study, 28 healthy female
volunteers used no product (control cycle) or nightly application of either
intravaginal nonoxynol-9 gel [N9] (an agent with known harmful effects
on the lower FRT) or the universal placebo gel [UPG] (an agent used as
a placebo control in many microbicide trials) from the end of menses
to the mid-luteal phase (intervention cycles). They then underwent
sample collection for measurements of T-cell phenotypes, transcriptional
profiling, and protein concentrations from 3 anatomic sites above the
vagina: the cervical transformation zone (TZ), the endocervix and the
endometrium. We used hierarchical statistical models to estimate mean
(95% CI) intervention:control fold-changes in relevant phenotype levels.
Results: Exposure to N9 and UPG generated a common “harm signal”
that included transcriptional up-regulation of inflammatory genes CCL20
and IL8 in the cervical TZ, decreased protein concentrations of secretory
leukocyte protease inhibitor in endocervical fluid, increased percentages
of terminally differentiated CD4+ effector T-cells in the endocervix,
and transcriptional up-regulation of inflammatory mediators KIR3DS1,
glycodelin-A, and osteopontin in the endometrium.
Conclusions: These results underscore the need to consider the effects
of gel vehicles as well as microbicide agents on the upper FRT in studies
of vaginal microbicides. Given the pro-inflammatory effects of UPG on
the upper FRT, it may not be a suitable placebo for microbicide trials.
Supported by NIH grants #AI083050 (PI: Warner Greene),
#U54HD055764 (to L.C.G.), and #1F32HD074423-02 (to J.C.C.)
P40.29
P40.30
Lactic Acid, a Vaginal Microbiota Metabolite,
Elicits an Anti-inflammatory Response from
Vaginal and Cervical Epithelial Cells
Gene Expression Analysis of Human Vaginal
Mucosal Response to Pro-inflammatory
Stimuli - Identification of Biomarkers of
Vaginal Inflammation
Burnet Institute, Centre for Biomedical Research, Melbourne, Australia,
Monash University, Melbourne, Australia, 3Boston University, Boston,
MA, United States, 4Johns Hopkins University, Baltimore, MD, United
States, 5The University of Melbourne, Department of Microbiology and
Immunology, Melbourne, Australia
1
2
Background: Lactobacilli sp. dominate the vaginal microbiota in about
1/3 of reproductive age women and HIV susceptibility increases with
a shift from lactobacilli to bacterial vaginosis associated bacteria.
Lactobacilli acidify the vagina to pH< 4.0 by producing ~1% lactic acid
(LA) in a nearly racemic mixture of D and L isomers. We have shown that
≥0.3% L-LA has potent HIV virucidal activity. Here we investigate if LA
elicits an anti-inflammatory response on epithelial cells from the lower
female reproductive tract (FRT) that might play a role in decreasing HIV
susceptibility.
Methods: The effect of LA in the apical medium was assessed on
vaginal (VK2), endocervical (End) and ectocervical (Ect) epithelial cells
grown in transwells. Toxicity effects were determined by viability
staining and diffusion of fluorescently labelled dextrans through the cell
layer. Cytokine profile from epithelial cells was determined following
stimulation with toll-like receptor (TLR) agonists ± LA.
Results: L-LA (0.3%; pH 3.9) had little impact on VK2, End and Ect
monolayer toxicity. Stimulation of all epithelial cell types with poly(I:C)
(TLR3) induced high-levels of pro-inflammatory cytokines IL-6 and IL8, and their variable induction with TLR agonists Pam(3)CSK(4)(TLR1/2)
and lipopolysaccharide (TLR4). In contrast, the presence of 0.3% L-LA
or D-LA either significantly abrogated or reduced TLR-induced IL-6 and
IL-8 secretion by epithelial cells. Irrespective of the TLR agonists, L-LA
and D-LA elicited high-levels of the anti-inflammatory cytokine IL-1RA
(~30,000 pg/ml) from all cell types. Neither 0.3% L-LA at neutral pH nor
acidity alone (HCl) increased IL-1RA or decreased TLR induced IL-6 and
IL-8 production.
Conclusions: LA at pH 3.9 found in lactobacillus-dominated vaginal
microbiota elicits an anti-inflammatory effect on lower FRT epithelial
cells and inhibits inflammation induced by bacterial and viral TLR
agonists suggesting a role in mitigating inflammation-induced HIV
susceptibility at the genital mucosa.
Irina A. Zalenskaya1, Andrea R. Thurman1, Neelima Chandra1,
Nazita Yousefieh1, Suzanne S. Jackson1, Sharon Anderson1,
Gustavo F. Doncel1
CONRAD, Eastern Virginia Medical School, OB/GYN, Norfolk, VA,
United States
1
Background: Understanding the mechanisms underlying mucosal
susceptibility to HIV infection and defining new biomarkers of vaginal
inflammation and immune activation are essential to developing
safe and effective HIV prevention products. To this end we analyzed
transcriptomes of human vaginal tissues after in vivo treatment with proinflammatory (PIC) and non-inflammatory (NIC) compounds.
Methods: Nineteen women (mean age= 32) applied 2-4 doses of PICs,
including imiquimod and nonoxynol-9 (N9), and NIC, hydroxyethyl
cellulose (HEC)-based placebo gel, intravaginally, in a crossover design.
Vaginal biopsies were taken at baseline and after treatment and preserved
in RNAlater or formalin. Gene expression (n = 93 tissue samples) was
analyzed by using Affymetrix U133 Plus 2 arrays. Data processing
was done using BRB-array tools software. Genes showing statistically
different expression (p< 0.001) between treatment and control groups
and fold change differences ≥2 were considered differentially expressed.
Functional analysis was done using Ingenuity Pathway Analysis and
published data. Immunohistochemical analysis (IHC) was performed on
paraffin embedded vaginal tissues.
Results: Transcriptomic analysis indicated that HEC did not alter
mucosal gene expression significantly. Conversely, imiquimod caused
dysregulation of 879 genes, and N9 of 89 genes. There were 66
genes common to both PIC treatments. Functional analysis indicated
27 genes involved in inflammatory and immune responses, 8 of which
were chemoattractants for immune cells. Upstream regulation analysis
revealed strong interferon signature. IHC showed PIC-induced influx of
immune cells, including CD3+ T cells, into the vaginal mucosa.
Conclusions: Intravaginal application of PICs induced a strong mucosal
immuno-inflammatory response and transcriptomic change with a
common profile of dysregulated genes which can serve as biomarkers
of vaginal inflammation and immune activation in the safety evaluation
of HIV prevention interventions.
www.hivr4p.org
347
POSTERS
Anna Hearps1,2, Raffi Gugasyan1, Daniela Srbinovski1,2, David
Tyssen1, Muriel Aldunate1,2, Deborah J. Anderson3, Richard Cone4,
Gilda Tachedjian1,2,5
Posters
P40.31
Cervical Epithelial Cells from HIV-1-Exposed
Seronegative Sex Workers Express a Distinct
Cytokine/Chemokine Profile upon Toll-like
Receptor Activation
Nyla Dil1, Joshua Kimani2, Makobu Kimani2, Frank Plummer3,
Blake T. Ball1
University of Manitoba/Public Health Agency of Canada, Medical
Microbiology and Infectious Diseases/National Lab for HIV
Immunology, Winnipeg, MB, Canada, 2University of Nairobi, Medical
Microbiology, Nairobi, Kenya, 3University of Manitoba/National
Microbiology Lab, Winnipeg, MB, Canada
1
POSTERS
Background: Majority of human HIV-1 infections are acquired through
heterosexual transmission across female genital mucosa. Epithelial cells
that line the genital mucosa express toll-like receptors (TLR) and act as first
line of defense against mucosal infections. Immune locale orchestrated
by TLR mediated activation of female genital epithelial cells can be a
critical determinant of HIV-1 resistance or susceptibility. In this study we
investigated the role of TLR signaling in female genital epithelial cells in
determining the local mucosal cytokine /chemokines milieu.
Methods: Endoervical cytobrsuh samples were obtained from Pumwani
CSWs cohort in Nairobi, Kenya. HESN (n=22), HIV- (n=24) and HIV+
(n=23).Cervical epithelial cells (CECs) were purified through a series of
nylon membrane filtrations. Purity and viability of CECs was assessed by
Ber-EP4 expression and MTS assay, respectively. CECs were cultured with
or without TLR3 ligand (poly:IC) or TLR1-9 ligand combined for 24 h.
Cytokine/chemokines levels in the supernatants were determined using
the Milliplex MAP multiplex kit (Human Cytokine/Chemokine I and III)
and analyzed on the BioPlex-200 according to manufacturer’s protocol.
Results: We tested 28 cytokines and chemokines (GM-CSF, IFNa2,
IFNgamma,IL-1a, IL-1b, IL-2, IL-6, IL-7, IL-8, IL-10, IL-12 (p40), IL-12(p70),
IL-15, IL-17, IP-10, MCP-1, MCP-3, MDC, MIP-1a, MIP-1b, RANTES,
Fractalkine, GRO, TNFa, I-TAC, MIG, MIP-3 a, and MIP-3 b). CECs from
HESN individuals expressed significantly lower levels of interferon-γinduced protein 10 (IP-10), IL-8, IL-1 a, MIG, and MIP-3 a upon TLR3 or
combined TLR1-9 stimulation compared with CECs from HIV- and HIV+
women. These cytokines and chemokines have important functions in
inflammatory and immune cell recruitment.
Conclusions: These results highlight the role of TLR signaling in CECs
and support the immune quiescence model of HIV-1 protection, whereby
lower target and inflammatory cell recruitment at the genital mucosa
reduces HIV-1 target cell numbers in HESN.
348
Posters 41: Novel Vaccine and Prevention Concepts
P41.01
P41.02
Immunogenicity of Recombinant Semliki
Forest Virus-based RNA Vaccines Expressing
HIV-1 Indian Subtype C Antigens in Mice
Antiviral Activity of 5-Hydroxytyrosol,
a Microbicidal Candidate against HIV-1
Transmission
Seema Ajbani1, Shilpa Velhal1, Ravindra Kadam1, Vainav Patel1,
Atmaram Bandivdekar1
Jose Alcami1, Luis-Miguel Bedoya1, Patricia Obregón1, Manuela
Beltran1, Eduardo Gomez-Acebo2, David Auñón2, Laura Capa1
1
National Institute for Research in Reproductive Health (ICMR),
Department of Biochemistry and Virology, Mumbai, India
1
Background: Genetic vaccinations have generally involved
immunization with antigen-encoding nucleic acid, usually DNA. RNA
vaccination based on engineered RNA replicons derived from several
RNA viruses is gaining increased attention and several vaccines are
currently being investigated for infectious diseases, cancer and allergies.
Present study investigates the potential of recombinant Semliki Forest
virus replicon RNA expressing HIV-1 antigens from Indian subtype C
isolates to generate cellular and humoral immune responses in mice.
Methods: Balb/C mice were immunized with recombinant Semliki Forest
virus (SFV2gen) RNA encoding HIV-1C gag or env antigen from Indian
primary isolates. Three intramuscular injections of rSFV2gen/gag RNA
or rSFV2gen/env RNA were given at 2 week intervals in the hind legs.
Splenocytes were harvested at necropsy and analyzed for antigenspecific T cell responses by IFN-γ ELISPOT assay and Intracellular
cytokine staining (ICCS) assay using antigen-specific peptide pools
representing HIV-1C Gag/Env consensus sequences. Sera of mice
injected with rSFV2gen/env RNA were evaluated for HIV-1C Env-specific
binding antibodies by gp120 ELISA.
Results: Gag-specific IFN-γ T cell responses were detected in all animals
vaccinated with rSFV2gen/gag RNA following three immunizations.
Surprisingly although Env-specific IFN-γ positive T cells were below the
detection limit of ELISPOT assay, an assessment of vaccine-induced IL-2
secretion by intracellular staining followed by flow cytometry revealed
Env-specific IL-2 production which was mediated by both CD4+ and
CD8+ T cells. Serological analysis revealed HIV-1C Env-specific binding
antibody responses in mice receiving rSFV2gen/env RNA.
Conclusions: These findings demonstrate that SFV replicon-based
recombinant RNA vaccines were capable of eliciting HIV-1C antigenspecific cellular and humoral immune responses in mice. These
constructs are currently being investigated for their immunogenicity as
a component of a multi-gene vaccine.
Background: We are currently developing an effective and low-cost
microbicide based on 5-Hydroxytyrosol (5-HT), a molecule not used as
microbicide previously that displays both anti-inflammatory and antiviral
properties. The objective of this work is to analyze the activity against
HIV-1 of 5-HT, alone or in combination with other antiviral molecules in
different cellular systems and its toxicty “in vitro” and “in vivo”.
Methods: The antiviral activity of 5-HT was assessed against SIV and
different HIV-1 strains (B and C clades) using different experimental
models: cell lines, lymphocytes and monocytes from peripheral blood
and autologous co-culture of DC-SIGN expressing cells and lymphocytes.
Anti-HIV activity of 5-HT was assessed alone or in combination with
Tenofovir. Synergism was analyzed according to T-C Chou and P. Talalay
method (Trends Pharmacol. Sci. 4, 450-454). Local tolerability at
vaginal mucosa of 5-HT was assessed in rabbits (n=6) at two different
concentrations (90 and 397mg/L) during 7 consecutive days by topical
route.
Results: 5-HT was active against SIV and different HIV-1 clades with
IC50 ranging between 20-30µM. 5-HT was also active in the context
of the immune synapse with IC50 in the 5µM range. “In vitro” toxicity
was not observed at doses of 1.000 µM. Topical administration of 5-HT
did not cause irritative responses or morphological alteration in the
vaginal mucosa of rabbit. A strong synergistic effect was displayed by
combination of 5-HT and Tenofovir (ED50 Combination index = 0.237).
Conclusions:
1. 5-HT was active against SIV and different HIV-1 strains “in vitro” in
2. 5-HT inhibited HIV infection of lymphocytes, monocytes and was
active in the context of the immune synapse,
3. Strong synergistic activity with Tenofovir was found,
4. No toxicity was observed “in vitro” and “in vivo” at the doses tested.
In summary 5-hydroxytyrosol is a new class of microbicide combining
both anti-inflammatory and anti-HIV properties and represents a
potential candidate for future clinical trials.
www.hivr4p.org
349
POSTERS
Instituto de Salud Carlos III, AIDS Immunopathogenesis Unit,
Majadahonda, Spain, 2Seprox Biotech SL, Madrid, Spain
Posters
P41.03
P41.04
Co-delivery of Antigen p24 and NOD-ligands
by PLA Nanoparticles to Human Dendritic
Cells Promote Highly Functional HIV-1Specific T-cell Responses
Vaccine Antigen Design to Maximize anti-HIV
CD4+ T-cell Responses: From Mice to Nonhuman Primates
Núria Climent1, Vincent Pavot2, Felipe García1, Thierry Lioux3, Eric
Perouzel3, Charlotte Primard2, Stéphane Paul4, Jose María Gatell1,
Montserrat Plana1, Bernard Verrier2, Teresa Gallart1
1
CELLEX-IDIBAPS-HIVACAT, Barcelona, Spain, 2Institut de Biologie
et Chimie des Protéines, UMR 5305 CNRS/UCBL, Cedex 07, Lyon,
France, 3CAYLA, InvivoGen, Toulouse, France, 4Groupe Immunité des
Muqueuses et Agents Pathogènes, INSERM CIE 3 Vaccinologie, Faculté
de Médecine, Saint-Etienne, France
POSTERS
Background: A recently explored approach for HIV vaccination is
the administration of a variety of antigens and adjuvants included in
nanoparticles targeted to dendritic cells (DC) to potentiate antigenspecific T-cell responses and a protective immunity without having
detrimental effects associated to viral vector. The use of TLR ligands
as mucosal adjuvant for vaccine administration has already been
largely studied, but the use of NOD-like receptor (NLR) ligands needs
to be further investigated. This study tested polylactic acid (PLA)
colloidal biodegradable nanoparticles, coated with HIV Gag (p24) and
encapsulating Nod ligands 1 or 2 (NodL1 and NodL2) as adjuvant.
Methods: We performed different ex vivo assays with monocyte derived
DC (MDDC) including capture of the different nanoparticle formulations
and induction of maturation. Co-cultures between MDDC and autologous
lymphocytes were performed in order to measure proliferation by CFSE
(FACS) and cytokine secretion (25-plex luminex) from asymptomatic
HIV+-infected patients (n=11).
Results: PLA-p24-NodL1 and PLA-p24-NodL2 nanoparticles were highly
captured by MDDC from HIV-1 individuals and induced a high degree
of MDDC maturation compared with p24 or PLA-p24 (p< 0.05). In
accordance, PLA-NodL1 and PLA-p24-NodL2 nanoparticles increased
HIV-specific CD4+ and CD8+ T-cell proliferation being PLA-p24-NodL2
more than 6-folds higher compared with p24 or PLA-p24 (p< 0.01).
These compounds induced highly functional cytokine secretion (IFN-γ,
IL-2R, IL-6 and MIP-1β/CCL4) in the autologous co-culture compared with
p24 or PLA-p24 (p< 0.05). In all the cases, the highest immunoresponse
was induced by PLA-p24-NodL2 (p< 0.01).
Conclusions: Co-delivery of antigen p24 and NOD ligands by PLA
nanoparticles to dendritic cells enhances antigen capture, DC-maturation
and improves highly functional HIV-specific cellular immune responses
from HIV+ patients. These compounds would be useful as novel
therapeutic and preventive approaches in the context of HIV-1 vaccine.
350
Edecio Cunha-Neto1, Susan Ribeiro1, Daniela Santoro-Rosa2,
Rafael Almeida1, Vinicius Santana1, Jorge Kalil1
1
University of São Paulo, São Paulo, Brazil, 2Federal University of São
Paulo, São Paulo, Brazil
Background: Purposely eliciting CD4+ T cell responses has been
essentially unexplored in the HIV vaccine field, despite increasing
evidence for the importance of the CD4+ T cell response in protection
against HIV.
Methods: We used rational vaccine design to develop a DNA vaccine
encoding HIV-1 B subtype or M-type conserved, multiple HLA-DRbinding CD4+ T cell epitopes, found to be recognized by multiple HIV1-infected patients.
Results: Vaccines elicited broad, polyfunctional, and long-lived CD4+
T cell responses in BALB/c and several HLA class II transgenic mice,
eliciting extensive cross-clade immunity. Immunization of BALB/c mice
increased CD8+ T cell responses against subsequent whole HIV protein
immunization, and reduced vital titers after challenge with a recombinant
vaccinia virus encoding HIV proteins. Immunization prior to recombinant
gp140 HIV envelope protein drastically increased the IgG2a/IgG1 ratio
of elicited anti-gp140 antibodies. Immunization of Rhesus macaques
under electroporation induced broad IFN-γ ELISPOT responses 10-fold
higher than those found in mice; we found a predominantly CD4+ T cell
intracellular cytokine response basically consisting of IFNγ, TNFα IL-2,
Granzyme B.
Conclusions: By virtue of inducing broad responses against multiple
conserved CD4+ T cell epitopes that can be recognized across widely
diverse, common HLA class II alleles, this vaccine concept may induce
T cell responses against multiple peptides in large proportion of the
genetically heterogeneous population. By increasing the chance of
matching the responses with multiple epitopes in the infecting HIV
isolate, the vaccine concept may also cope with HIV genetic variability.
The vaccine concept may be a candidate for standalone use or in
association with conventional immunogens, to increase the amplitude,
coverage and effectiveness of the induced response. Pending new results
with immunization of Rhesus macaques with different viral vectors and
prime-boosting with gp140, a Phase I clinical trial will be launched.
P41.05
P41.06
Vaccinia Virus with Selective Deletions
Enhances T Cell Response to HIV Antigens by
Specific Neutrophil Recruitment
Endosymbiont Trichomonas Vaginalis Virus as
a Target for HIV Prevention
Mauro Di Pilato , Ernesto Mejías-Pérez , Manuela Zonca , Beatriz
Perdiguero1, Carmen Elena Gómez1, Marianna Trakala3, Jacobo
Nieto1, José Luis Nájera1, Carlos Oscar S. Sorzano4, Lourdes
Planelles2, Mariano Esteban1
1
1
2
Spanish National Centre for Biotechnology (CNB), Department of
Molecular and Cellular Biology, Madrid, Spain, 2Spanish National
Centre for Biotechnology (CNB), Department of Immunology and
Oncology, Madrid, Spain, 3Spanish National Cancer Research Centre
(CNIO), Cell Division and Cancer Group, Madrid, Spain, 4Spanish
National Centre for Biotechnology (CNB), Biocomputing Unit, Madrid,
Spain
Titilayo Fashemi1, Hidemi Yamamoto1, Nibert Max2, Athe Tsibris1,2,
Raina Fichorova1,2
Brigham and Women’s Hospital, Boston, MA, United States, 2Harvard
Medical School, Boston, MA, United States
1
Background: In the context of vaccinia virus (VACV) infection, it was
previously known that neutrophil infiltration is tissue-protective and that
neutrophils generate virus-specific memory CD8 T cells, transporting
antigens from the dermis to the bone marrow. However, it was not
known how neutrophil recruitment is modulated by VACV, factors
involved and significance in triggering immune responses.
Methods: Here we describe the generation of an attenuated VACV strain
(NYVAC) that expresses HIV-1 clade C antigens but lacks three specific
viral genes (A52R, K7R and B15R). We analyze the capacity of NYVAC-C
ΔA52RΔB15RΔK7R to activate NFκB signaling pathway, to induce specific
innate immune response, and to generate adaptive immune response to
HIV-1 antigens compared to NYVAC-C.
Results: A52R, K7R and B15R cooperate to inhibit the NFκB signaling
pathway, and the triple ablation in modified virus restores NFκB function
in macrophages. After NYVAC-C ΔA52RΔB15RΔK7R virus infection,
NFκB pathway activation leads to expression of several cytokines and
chemokines that recruit specific neutrophil populations (Nα and Nβ) to
the infection site. Neutrophils display features of antigen-presenting
cells, and their trafficking to the infection site and to various lymphoid
organs is essential to enhance the T-cell response to HIV Gag and Pol
antigens.
Conclusions: Based on these inherent properties, the NYVAC-C
ΔA52RΔB15RΔK7R represents an effective vector vaccine candidate for
prophylactic and therapeutic settings, and provides a basis for the design
of new vaccines.
Background: Trichomoniasis caused by the genitourinary protozoan
parasite Trichomonas vaginalis (TV) is one of the leading risk factors
of genital HIV shedding, sexual and perinatal HIV transmission. The
majority of TV clinical isolates carry endosymbiont dsRNA viruses
(TVV). We have recently shown that TVV causes a profound TLR-3dependent inflammatory reaction by human cervical and vaginal
epithelial cells; however, the effects of TVV on HIV host cells and HIV
replication have not been investigated to date. We hypothesized that the
immunoinflammatory responses to TVV are at least partially responsible
for trichomoniasis-attributable HIV risk.
Methods: Peripheral blood mononuclear cells were stimulated with
clinical TV isolates and purified TVV, followed by infection with primary
isolates of CXCR4- and CCR5-tropic HIV-1. Supernatants were collected
at multiple time points over a period of 15 days to measure HIV-1 p24
by ELISA and immune mediators by multiplex immunoassays. ANOVA
was applied, and P< 0.05 was considered significant.
Results: TVV-positive TV induced significantly higher proinflammatory
responses as compared to TVV negative TV isolates. TVV-positive TV
up-regulated proinflammatory and Th1/Th2 cytokines and enhanced
those responses when applied together with other immune stimulants
e.g. IL-2 and PHA. While levels of proinflammatory responses (IL-1-beta,
TNF-alpha and IL-8) showed a tendency to subside after 72h, the levels
of immunoregulatory cytokines e.g. IFN-gamma, IL-4, IL-5, IL-13 and
IL-12p70 continued to rise. Purified TVV virions induced comparable
immune activation. TVV+TV pre-exposure caused a 50-fold increase in
p24 levels and the enhancement lasted for at least 15 days (p< 0.001).
Conclusions: Our data support the hypothesis that TVV virus causes
inflammation and immune cell activation capable of enhancing
and propagating HIV replication and thus should be further
investigated as a plausible target for HIV prophylaxis. (NIAID HU CFAR
5P30AI0600354-08, R56AI091889 and R01AI079085).
www.hivr4p.org
351
POSTERS
1
Posters
P41.07
P41.08
Deletion of Immunomodulatory A44L,
A46R and C12L Viral Genes from Modified
Vaccinia Ankara (MVA) Genome: Effect on its
Immunogenicity
Vaccine-induced Intestinal and Salivary IgA
Correlates with Reduced Viremia in Orallychallenged Neonatal Macaques
Maria Pia Holgado1, Cynthia Maeto1, Juliana Falivene1, Yanina
Ghiglione1, Maria Paula del Medico Zajac2, Gabriela Calamante2,
Maria Magdalena Gherardi1
Instituto de Investigaciones Biomedicas en Retrovirus y SIDA, Buenos
Aires, Argentina, 2Instituto de Biotecnologia, CICVyA - INTA, Castelar,
Argentina
1
POSTERS
Background: MVA still retains genes involved in host immune response
evasion. We have previously reported the optimization of its vaccine
potential after removing the C12L gene, coding an IL18 binding protein.
Here we analyze the immunogenicity of MVA vectors harboring the
simultaneous deletion of two viral genes: A44L, implicated in synthesis
of steroid hormones and A46R, which inhibits transduction signals
from TLR (MVAΔA44L-A46R: MVAd); or including C12L deletion also
(MVAΔC12L/ΔA44L-A46R: MVAt)
Methods: C57Bl/6 mice were immunized with MVAwt or deleted
MVAs (ΔMVAs). We evaluated the adaptive T-cell response to VV
(Vaccinia Virus) epitopes at acute (7 dpi) and memory phases (45 dpi) in
spleen and draining lymph nodes (DLNs). The amount of IFNγ and IL2
producing cells was measured by ELISPOT. We studied the percentage
of specific cytotoxic CD8 T-cells by flow cytometry, and the response of
memory T-cells among specific proliferating CD8 T-cells. To study the
innate response, we immunized mice with MVAwt or MVAt and pattern
of cytokines produced were evaluated between 0-24 hpi
Results: At 7 dpi both ΔMVAs induced significant increases in IFNγ
anti-VV CD8 and CD4-T cells vs MVAwt (1.5 to two-fold,p< 0.01;2.5
to five-fold, p< 0.01 respectively), and IL2 anti-VV CD8 T- cells (up to
five-fold higher;p< 0.01). Notably, ΔMVAs still elicited a higher response
than MVAwt at 45 dpi (p< 0.05). Proliferating anti-VV CD8 T-cells were
augmented from 1% (MVAwt) to 3% (MVAt). Moreover, this vector
elicited a higher proportion of specific TCM compared to MVAwt (45%
vs 20%). The percentage of specific cytotoxic CD8 T-cells secreting IFNγ
was also improved by MVAt. The innate response analysis showed that
mice who received MVAt produced higher levels of IFNγ, IL6 and IL12 in
DLNs compared to MVAwt
Conclusions: The deletion of interrelated immunomodulatory genes
from MVA genome is a proper approach to improve its vaccine potential
and it is a helpful tool that could contribute in the development of an
HIV vaccine
352
Kara Jensen1, Robert Wilson2, Michael Piatak3, Jeff Lifson3,
Uma Devi Ranganathan4, William Jacobs, Jr.4, Glenn Fennelly4,
Michelle Larsen4, Koen Van Rompay5, Pamela Kozlowski2, Kristina
Abel1
University of North Carolina at Chapel Hill and Center for AIDS
Research, Chapel Hill, NC, United States, 2Louisiana State University
Health Sciences Center, New Orleans, LA, United States, 3SAIC
Frederick, Inc., Frederick, MD, United States, 4Albert Einstein College of
Medicine, Bronx, NY, United States, 5CNPRC and UC Davis, Davis, CA,
United States
1
Background: HIV acquisition from breast milk is a significant route of
pediatric HIV infections. Neonates produce less IgA than adults and are
thus more susceptible to oral pathogens, including HIV. While serum
antibodies are critical for controlling HIV post-exposure, the importance
of mucosal IgA for the prevention of HIV transmission is not known. In
addition, it is unclear how best to induce the development of specific
mucosal antibodies through vaccination.
Methods: We tested whether oral pediatric vaccination could induce
SIV envelope (Env)-specific salivary and intestinal IgA, and protect
against oral SIV acquisition. Neonatal rhesus macaques were mucosally
immunized at birth with a live attenuated Mycobacterium tuberculosis
strain engineered to express SIV Env and Gag, and twice boosted with
MVAgpe. Animals were orally challenged beginning at week 9 using a
weekly regimen of low-dose SIVmac251 (5000 TCID50) to mimic HIV
exposure from infected breast milk.
Results: Vaccination induced SIV-specific plasma IgG and IgA antibodies
in all animals by week 9, plus salivary and intestinal SIV-specific
antibodies were detected in 8 of 8 and 3 of 8 animals, respectively.
Although vaccination did not prevent infection, infants producing
intestinal and salivary IgA (n=3, 37.5%) had significantly lower peak
and set-point viremias compared to other vaccinated animals (p=0.0019
and p=0.034, respectively). In addition, higher Env-specific mucosal IgA
activities and plasma IgG avidities inversely correlated with peak viremia
and viral set-point.
Conclusions: Therefore, mucosal and systemic antibodies produced
in response to neonatal oral vaccination contributed to controlled
virus replication. Vaccine strategies that promote mucosal antibody
development could help prevent oral HIV acquisition.
P41.09
P41.10
Construction and Evaluation of BCG and
Modified Vaccinia Ankara Vaccines Expressing
HIV-1 Subtype C Mosaic Gag
ADCC Measurements in Rabbits Immunized
with HIV-1 Vaccine Candidates
Tsungai Ivai Jongwe , Ros Chapman , Shivan Chetty , Niki
Douglass1, Anna-Lise Williamson1,2
Sanne S. Jensen1, Marie Borggren1, Gabriella Scarlatti2, Leo
Heyndrickx3, Anders Fomsgaard1, Ingrid Karlsson1, NGIN
consortium
1
University of Cape Town, Cape Town, South Africa, 2National
Laboratory Services, Cape Town, South Africa
1
Statens Serum Institut, Copenhagen, Denmark, 2San Raffaele Scientific
Institute, Milan, Italy, 3Institute of Tropical Medicine, Antwerp, Belgium
Background: Over 90% of people with HIV-1 in sub-Saharan Africa
are infected with the clade C virus. High mutagenesis rates and
diversity, even within clades, make it difficult to develop effective
vaccines. Mosaic immunogens have been computationally designed to
maximize inclusion of common T-cell epitopes. Compared to consensus
immunogens, polyvalent mosaic immunogens of HIV-1 group M have
increased breadth and depth of antigen-specific T-cell responses. Here,
we determined the immunogenicity of vaccines expressing HIV-1
subtype C Gag mosaic in mice.
Methods: BCG (BCG-GagM) and MVA (MVA-GagM) vaccines expressing
the HIV subtype C mosaic gag gene were constructed. p24 ELISA
and electron microscopy (EM) of MVA-GagM -infected cells was used
to determine the ability of the mosaic Gag to bud and form virus-like
particles (VLPs). Shuttle vector integrity in BCG-GagM was determined by
PCR. Mice were primed intraperitoneally with 2x107-cfu BCG-GagM and
boosted intramuscularly with 104-pfu MVA-GagM at week 10. Mice were
sacrificed 12 days later and T-cell responses analysed by IFN-γ ELISPOT,
CBA, and ICS assays.
Results: Gag VLP production was confirmed by EM and p24 ELISA in
the media of infected cells. A Th1 response was induced by the BCGGagM/MVA-GagM vaccination regimen, and both CD4+ and CD8+ IFN-γ
responses were detected. A potent effector memory phenotype was
detected from both CD4+ and CD8+ cells. Overall, the BCG-GagM prime
MVA-GagM boost induced robust HIV-specific CD4+ and CD8+ T cell
responses that were 3 fold higher than responses induced by a control
BCG prime MVA-GagM boost. Genetic integrity of the BCG-GagM vaccine
was confirmed 10 weeks post vaccination in mice.
Conclusions: These vaccines are immunogenic in Balb/c mice in a
prime-boost vaccination regimen. Either vaccine alone had a dominant
CD4 response while the combination of vaccines induced a greater CD8
response. The vaccines induce strong effector memory functions and
will be further evaluated in non-human primates.
Background: Induction of both neutralizing antibodies and nonneutralizing antibodies with effector functions e.g. antibody-dependent
cellular cytotoxicity (ADCC) are desired in the search for effective
vaccines against HIV-1. In the search for novel immunogens capable of
inducing an efficient antibody response, rabbits were immunized with
selected antigens using different prime-boost strategies within the NGIN
consortium.
Methods: We selected 30 different groups of rabbits immunized with
antigens from HIV-1 subtype A and B, including immunization with DNA
alone, protein alone and DNA prime with protein boost. Rabbit sera were
screened for ADCC activity using the GranToxiLux assay with human
PBMC as effector cells and CEM.NKRCCR5 coated with rgp120 as target
cells. This assay measures the proteolytic activity of Granzyme B after its
delivery into target cells, initiated by antibody recognition of rgp120 on
the target cell membrane.
Results: The groups of rabbits with the highest serum ADCC activity
were immunized with protein, monomeric gp120 or trimeric gp140,
in CAF adjuvant. Interestingly, the ADCC activity did not correlate with
neutralizing activity of purified IgG measured against SF162 and Bx08
with the pseudovirus-TZMbl assay, nor did it correlate with IgG gp120
ELISA binding titre.
Conclusions: The antigens and/or immunization strategies capable
of inducing antibodies with good ADCC activity do not necessarily
induce good neutralizing activity and vice versa. When searching for
an effective vaccine candidate it is important to evaluate the antibody
response using an assay measuring the desired function.
1
1
1
www.hivr4p.org
353
POSTERS
Posters
P41.11
P41.12
Eliciting Cytotoxic T-lymphocyte Responses
to HIV by Human Dendrocyte in vitro T-cell
Activation with Synthetic Peptide-containing
Microspheres
The Viral Vector Vaccine VSV-GP Boosts
the Immune Response upon Repeated
Applications
Sarah S. Killingbeck1,2, C V. Herst3, Craig Rouskey3, R M.
Rubsamen3
University of California, Division of Infectious Disease and
Vaccinology, School of Public Health, Berkeley, CA, United States,
2
University of California, Immunology, Davis, CA, United States,
3
Immunity Project, Palo Alto, CA, United States
1
POSTERS
Background: We describe a novel HIV vaccine consisting of synthetic
d,l poly(L-lactic-co-glycolic) acid (PLGA) microspheres encapsulating
HLA-specific cytotoxic T lymphocyte (CTL) HIV-1 Gag epitope KF11 in
combination with CpG oligodeoxynucleotide (ODN) as adjuvant. We
then demonstrate productive in vitro uptake and presentation of this
peptide by healthy donor PBMC-derived dendrocytes (DCs), as well as
the induction of specific CTL responses targeting KF11 when immunized
DCs are co-cultured with autologous PBMCs.
Methods: CTL responses to PLGA microsphere-encapsulated peptide
seven days post incubation with autologous DCs presented with the
microsphere-based vaccine in vitro were measured by interferon gamma
(IFN-γ) ELISPOT. CTL killing of HIV-infected CD4+ T cells was quantified
by p24 antigen ELISA specific for the p24 epitope within the HIV-1 Gag
protein.
Results: Antigen/peptide-experienced T cells are activated and
secrete IFN-γ in response to peptide KF11. In addition, CTLs elicited
by microsphere-encapsulated peptides were capable of significantly
restricting HIV replication in CD4+ T cells when compared to controls
treated with microspheres containing OVA peptide. We observed a
highly statistically significant (p< 0.0001) reduction in
CD4+ T cell viral load, approximately 40%, as assayed by p24
concentration in HIV-infected CD4+ T cells
(from 1026 pg/mL to 659 pg/mL) from one donor when co-cultured
in 5:1 effector:target ratio with autologous antigen/peptide-experienced
CD8+ T cells.
Conclusions: Taken together, our experiments show a versatile new
HIV vaccination method capable of eliciting robust HLA-specific
CTL responses that efficiently control viral replication in vitro. This
technique shows potential as a useful bioassay for a microsphere-based
T cell vaccine with applications in manufacturing quality control and
identification of a range of HLA types capable of presenting specific,
HLA-restricted epitopes.
354
Janine Kimpel1, Reinhard Tober1, Zoltan Banki1, Lisa Egerer1,
Alexander Muik2, Florian Kreppel3, Dorothee von Laer1
Innsbruck Medical University, Division of Virology, Innsbruck, Austria,
Georg-Speyer-Haus, Frankfurt, Germany, 3University of Ulm, Division
of Gene Therapy, Germany
1
2
Background: Vesicular stomatitis virus (VSV) is a potent candidate
vaccine vector for various diseases. However, VSV’s inherent neurotoxicity
has limited its clinical application. Additionally, VSV induces neutralizing
antibodies rapidly and is thus ineffective upon repeated applications.
Our group has recently shown that VSV pseudotyped with the
glycoprotein (GP) of the lymphocytic choriomeningitis virus, VSV-GP, is
not neurotoxic. Here, we evaluated the potential of VSV-GP as a vaccine
vector.
Methods: We used Ovalbumin (OVA) as a model antigen and analyzed
immunogenicity of GP-pseudotyped and wild-type VSV expressing OVA
(VSV-GP-OVA and VSV-OVA) in vitro and in vivo in mouse models.
Results: Mouse experiments revealed that both VSV-OVA and VSV-GPOVA induced functional OVA-specific CTLs and anti-OVA antibodies
upon single immunization. However, boosting with the same vector
was only possible for the GP-pseudotype but not for wild-type VSV. The
efficacy of repeated immunization with VSV-OVA was most likely limited
by the high levels of neutralizing antibodies, which we detected after
the first immunization. In contrast, no neutralizing antibodies against
VSV-GP were induced even after seven boost immunizations. CTL
responses induced by VSV-GP-OVA were as potent as those induced by
an adenoviral state-of-the-art vaccine vector. Additionally, immunization
with both vectors completely protected mice from infection with Listeria
monocytogenes expressing OVA.
Conclusions: Taken together, VSV-GP is non-neurotoxic, induces potent
immune responses, enables boosting and thus is a promising novel
vaccine vector.
P41.13
P41.14
Single and Combined Vaccination Modalities
Result in Distinct Immunological Profiles in
HIV-1 gp140-immunised Mice
Intracellular Antigen Trafficking Directs
Distinct Immune Responses Elicited by
Dendritic Cell-targeting HIV Vaccines
Deborah F.L. King1, Paul F. McKay1, Jamie F.S. Mann1, Bryn Jones1,
Robin J. Shattock1
Boon Kiat Lee1, Jingying Zhou1, Zhiwei Chen1
Background: Recent studies have indicated that co-immunisation
of protein antigens and DNA can induce both cellular and humoral
immune responses.
Methods: We investigated whether electroporated DNA and protein
vaccinations by intranasal (IN) and intramuscular (IM) routes alone or
as co-immunisations could be used to induce optimal HIV-1 gp140specific responses in mice. Serum and vaginal antigen-specific IgG and
IgA were measured by ELISA, and IFNγ+ T-cell responses were assessed
by ELIspot.
Results: IN Protein vaccination resulted in significantly higher antigenspecific IgG and IgA in the serum and IgA in the vagina, compared to
DNA vaccination. IM protein vaccination resulted in similar levels of
serum and vaginal IgG as IN vaccination. DNA vaccination induced
significantly higher frequencies of gp140-specific IFNγ+ T-cells than
IN or IM groups. Overall these results indicate that DNA vaccination
effectively induces cellular IFNγ responses, but only low-level humoral
responses, whereas IN vaccination optimally induces serum and vaginal
antibody responses.
DNA+IN co-immunisation resulted in significantly higher gp140specific serum IgG than either DNA or IM vaccination, but similar
levels to those observed with IN vaccination alone. The DNA+IN group
also had significantly higher serum IgG and IgA than DNA+IM and
DNA+IN+IM groups. The DNA+IN group exhibited significantly higher
vaginal IgA compared to DNA, IM or DNA+IM groups, but again was
not significantly higher than responses induced by IN vaccination alone.
IFNγ-specific responses were similar in all co-immunisation groups, but
significantly lower than DNA alone in DNA+IN, IN+IM and DNA+IN+IM
groups. DNA+IN vaccination induced similar levels of IFNγ+ T cells as
IN vaccination alone.
Conclusions: Cellular and humoral HIV-1 gp140-specific responses
were effectively induced by IN vaccination without the need for
additional DNA co-immunisation. Thus this single vaccination route may
be optimal for delivery of HIV vaccine antigens.
University of Hong Kong, AIDS Institute, Department of Microbiology,
Hong Kong, Hong Kong
1
Background: Improvement of HIV vaccine immunogenicity requires
deeper understanding of antigen processing in dendritic cells (DC). To date,
although antigen-targeting to DC has been explored as a novel strategy
of HIV vaccine design, few studies have investigated the intracellular
antigen processing and its influence on vaccine immunogenicity when
different DC surface receptors are engaged. In this study, we conduct
parallel experiments to determine the immunogenicity and possible
underlying mechanism of DC-targeting HIV vaccines via the native
ligands of PD-1 and CTLA-4, respectively.
Methods: A panel of DNA fusion vaccines was constructed including
soluble PD-1 (sPD1)-p24, soluble CTLA-4 (sCTLA-4)-p24, deletion mutant
sΔPD1-p24, sΔCTLA4-p24 and p24 alone. BALB/c mice were immunized
with each of these DNA vaccines at 100µg dose via intramuscular in
vivo electroporation following a prime/2-boost regimen with 3-week
intervals. Post vaccination, immunogenicity profiles were analyzed using
ELISA and ELIspot. Confocal microscopy was used to investigate p24
antigen trafficking to MHC-I or -II compartments in DC based on several
endosomal markers.
Results: DC-targeting HIV vaccines were significantly more potent
than corresponding deletion mutant controls. sPD1-p24 is superior to
CTLA4-p24 with following distinct immune responses: (1) significantly
enhanced IgG2a (Th1) antibody responses, (2) ~3.5-fold greater p24specific CD8+ T cell responses besides a 2-fold increase in CD4+ T cells
by IFN-γ ELISpots. Mechanistically, while p24 delivered by sPD1-p24
and sCTLA4-p24 routed to Lamp-1 endosomes for MHC-II presentation
to CD4+ T cells, sPD1-p24 also routed to Rab14 endosomes for MHC-I
cross-presentation to CD8+ T cell.
Conclusions: PD-1 and CTLA-4 lead to distinct immunogenicity
outcomes when used in our DC-targeting vaccines. Our findings provide
evidence that the intracellular antigen processing in DC influences
vaccine immunogenicity when different DC surface receptors are
engaged.
www.hivr4p.org
355
POSTERS
Imperial College London, Virology, London, United Kingdom
1
Posters
P41.15
P41.16
Vaccines for Therapeutic Cellular Immunity
that Target HIV Gag, Pol and Nef Epitopes
to CD40 and DCIR Elicit T-cell Responses in
Rhesus Macaques
Combined Approach of Vaccine and Antiviral
Drugs Can Achieve Better Protection in NHP
Model against Heterologous SHIV Challenges
Yves Lévy1,2,3, Sandra Zurawski1,4, Anne-Laure Flamar1,4, Christine
Lacabaratz1,2, Cécile Peltekian1,2, Andres Salazar5, Rodolphe
Thiébaut1,6, Gérard Zurawski1,4
Vaccine Research Institute (VRI), Créteil, France, 2Université ParisEst Créteil Val de Marne (UPEC), Inserm U 955, Creteil, France,
3
Assistance Publique-Hôpitaux de Paris, Groupe Henri Mondor-Albert
Chenevier, Service d’Immunologie Clinique, Créteil, France, 4Baylor
Institute for Immunology Research, Inserm U 955, Dallas, TX, United
States, 5Oncovir, Washington DC, WA, United States, 6INSERM U 897
Université Victor Segalen Bordeaux 2, Bordeaux, France
1
POSTERS
Background: We have developed αCD40.HIV5pep and αDCIR.HIV5pep,
two dendritic cell (DC)-targeting vaccines, containing 5 T cell epitope-rich
HIV regions of Gag, Pol, and Nef (G/P/N) directly fused to αCD40 (Flamar
et. al., AIDS, 2013) or αDCIR mAbs. In vitro, these vaccines expand multifunctional and polyepitopic specific CD4+ and CD8+ T cells.
Methods: NHP were primed twice at weeks (W) 0 and 8 with MVA
encoding G/P/N sequences encompassing the HIV5pep sequences and
boosted with either αCD40.HIV5pep (n=6) or αDCIR.HIV5pep (n=6)
(three ID injections of 250µg with 1 mg Poly-ICLC (Hiltonol) at W 12, 16
and 24). Two other groups of NHP received anti-DC vaccines at W 0, 4
and 12 (n=6 per group) followed by one MVA boost at W 22. Antibody
and HIV5pep-specific T cell responses were detected by ELISA and IFNγELISPOT performed two weeks after each immunization.
Results: IgG titers were weak (< 200) or undetectable following the two
MVA primes but were observed in all animals following the DC-targeting
vaccinations (~5,000 1/EC50). Two weeks after each of DC-targeting
vaccine boosts, HIV5pep-specific T cells were boosted to median 150,
503 and 388 SFC/106 PBMC for αDCIR.HIV5pep and 350, 1055 and
533 for αCD40.HIV5pep compared to after the second MVA vaccination
[145 and 98 for these two groups]. Globally, DC vaccines boosted
significantly T cell responses as compared to baseline (+648 SFC/106
PBMC, P< .0005). In non-MVA primed groups, median HIV5pep-specific
T cells responses following each DC-targeting vaccine were respectively
43, 135, 148 SFC/106 PBMC for αDCIR.HIV5pep and 118, 190, 198
SFC/106 PBMC for αCD40.HIV5pep. Those responses increased to 200
and 310 SFC/106 PBMC two weeks after MVA boost.
Conclusions: These DC-targeting vaccines elicited broad HIV peptidespecific T cell responses in both priming and boost settings. These
data support their potential for further development, in particular as a
component of a therapeutic vaccination strategy.
356
Ying Liu1, Zhou Zhang1, Yanling Hao1, Shuhui Wang1, Qiang
Wei2, Chuan Qin2, Chang Liu1, Hong Peng1, Yiming Shao1
1
National Center for AIDS/STD Control and Prevention, Chinese
Center for Disease Control and Prevention, Beijing, China, 2Institute
of Laboratory Animals Science, CAMS &PUMC, Beijing, China
Background: Vaccine is needed to stop the AIDS pandemic. Even
low efficacy vaccine can prevent millions HIV infections according to
mathematic modelling. Unlike the rapid advances in antivirals since
1990s and in biomedical interventions of recent years, HIV vaccine
research has been struggling due to the unprecedented challenges
it met. The best results of oral Prep of antivirals and RV144 vaccine
showed around 50% and 30% protective efficacy respectively in clinical
trials. The strategy of combining various prevention approaches, such as
vaccines and antivirals, should be tested to achieve higher prevention.
Methods: Rhesus macaques were immunized with DNA, recombinant
vaccinia (replicating Tiantan strain) and protein vaccines expressing HIV1 gag/pol/env and SIV gag. The immunized animals were given oral
TDF/FTC before challenged by multiple low dose SHIV SF162 P3 through
rectal mucosal. Immune responses and viral load were monitored for
evaluation.
Results: Among the 4 groups of monkeys (8 per group), high peak viral
loads (3.8 x 106) can be detected in all animal of the control group and
reduced peak viral load were detected in 4 (1.7x 105), 3 (2.9x 103) and
1 (1x 103) animals of the vaccine only, antiviral only and vaccine and
antiviral combined groups respectively. The reduced viral load in the
three treated groups also resulted in low viral set-point and faster felling
to undetectable levels, compared to the control group. Both strong
humoral (binding and V1V2 antibodies) and cellular (CD8 and CD4)
responses to HIV-1 antigens and SIV gag can be detected in immunized
animals. No viral load can be detected in the protected animal after
inoculation them with anti-CD8 antibody.
Conclusions: High protective efficacy (87.5%) can be achieved by the
combined use of vaccines and antivirals. Experiments are pending to
evaluate the mechanism of protection in protected animals, as well
to the impact of the treatment on the viral reservoirs in the infected
animals.
P41.17
P41.18
Longitudinal Analysis of SIVmac239
Mutations around the 12 Protease Cleavage
Sites and their Correlations with Viral Load
Reduction and CD4 counts
Sequences Surrounding the 12 Protease
Cleavage Sites are Good Targets for Both
Prophylactic and Therapeutic HIV Vaccines
National Microbiology Laboratory, National HIV and Retrovirology
Laboratory, Winnipeg, MB, Canada, 2University of Manitoba, Medical
Microbiology, Winnipeg, MB, Canada, 3National Microbiology
Laboratory, Winnipeg, MB, Canada, 4University of Santiago de
Compostela, Santiago de Compostela, Spain, 5University of NebraskaLincoln, Nebraska-Lincoln, NE, United States
1
Background: HIV-1 protease mediates the cleavage of Gag, Gag-Pol
and Nef precursor polyproteins in a highly specific and temporally
regulated manner. Because a total of 12 cleavage reactions are required
to generate a mature virion, generating focused immune response
targeting the sequences surrounding the protease cleavage sites (PCS)
could drive viral mutations to its disadvantage. We have conducted a
proof of concept study with Cynomolgus macaques and pathogenic
SIVmac239 as a model and used a modified recombinant vesicular
stomatitis vector and nanocarriers to deliver 12 20-amino acid antigens.
We showed that a vaccine targeting the sequences surrounding the 12
protease cleavage sites is promising at prevention of HIV-1 infection
and disease progression. In this study we systematically analyzed
breakthrough viruses of vaccinated and control macaques.
Methods: The sequences surrounding the 12 protease cleavage sites
were amplified from plasma RNA of all SIVmac239 positive samples.
The amplified PCR products were sequenced with 454 pyrosequencing
technology. The amino acid and frame shift mutations were analyzed
and correlated with viral load and CD4 counts. Regression analysis
was conducted to correlate the viral mutations with viral load and CD4
counts. WebLogo was used to plot the amino acid mutations.
Results: Extensive mutations were detected around PCS and both
conserved and non-conserved mutations are correlated with lower
viral load (p< 0.0001). The breakthrough viruses from the vaccinated
macaques carry significantly higher mutations than the controls.
Longitudinal analysis revealed that the high rate of non-conserved and
conserved amino acid mutations along the sequences surrounding the
PCS lead to the reduction and diminishing of viral load.
Conclusions: The pathogenic SIVmac239 is extremely vulnerable to any
amino acid alternations around PCS and focusing immune response to
sequences surrounding the PCS of HIV-1 can drive amino acid mutations
and lead to complete viral control.
National Microbiology Laboratory, National HIV and Retrovirology
Laboratory, Winnipeg, MB, Canada, 2University of Santiago de
Compostela, Santiago de Compostela, Spain, 3National Microbiology
Laboratory, Winnipeg, MB, Canada, 4University of Nebraska-Lincoln,
Nebraska-Lincoln, NE, United States, 5University of Manitoba, Medical
Microbiology, Winnipeg, MB, Canada
1
Background: HIV-1 protease mediates the cleavage of Gag, Gag-Pol
and Nef precursor polyproteins in a highly specific and temporally
regulated manner. Because a total of 12 cleavage reactions are required
to generate a mature virion, generating focused immune response
targeting the sequences surrounding the protease cleavage sites (PCS)
could drive viral mutations to its disadvantage. We have conducted a
pilot study to investigate the feasibility and effectiveness of a vaccine
targeting the sequences around the 12 PCS using Cynomolgus macaques
and pathogenic SIVmac239 as a model.
Methods: Twelve recombinant VSVpcs were used to immunize 12
Cynomolgus macaques and nanopackaged PCS peptides were used
as a boost. The immunized macaques and 6 controls were repeatedly
challenged intrarectally with an increased dosage of SIVmac239.
Antibody and T cell responses to the PCS peptides, CD4+ and CD8+ T
cell counts and challenge dosage were monitored. 454 Pyrosequencing
was conducted to analyze break-through viruses and the amino acid
mutations surrounding the PCS sites were correlated with viral load.
Results: Antibody and T cell responses to the 12 PCS protected
macaques against higher dosage of SIVmac239 intrarectal challenge
(p=0.005, R=0.42). The vaccine group maintained higher CD4+ counts
(p=0.0002) than the controls weeks after being infected. Analysis of
viral mutations around 12 PCS of 276 samples (14 to 20 sampling
points/monkey) detected extensive mutations. These mutations, both
conserved and non-conserved amino substitutions around PCS, are
correlated with lower viral load
(p< 0.0001).
Conclusions: Our study with nonhuman primates and pathogenic
SIVmac239 as a model showed that a vaccine targeting the sequences
surrounding the 12 protease cleavage sites is promising at prevention
of HIV-1 infection and disease progression. It demonstrated that the
pathogenic SIVmac239 is extremely vulnerable to any amino acid
alternations around PCS. Targeting PCS of HIV-1 could be an effective
vaccine approach.
www.hivr4p.org
357
POSTERS
Ma Luo1,2, David Tang1, David La1, Rupert Capina1, Xin-Yang
Yuan3, Jorge Correia-Pinto4, Cecilia Prego4, Maria Alonso4,
Christina Barry3, Richard Pilon1, Christina Daniuk1, Mikaela
Nykoluk1, Stephane Pillet3, Tomasz Bielawny1, Jeffrey Tuff1, Chris
Czarnecki1, Philip Lacap1, Gary Wong3, Shaun Tyler3, Ben Liang3,
Zhe Yuan5, Qingsheng Li5, Terry Blake Ball1, Paul Sandstrom1,
Gary Kobinger3, Francis Plummer2,3
David Tang1, David La1, Rupert Capina1, Xin-Yang Yuan1, Jorge
Correia-Pinto2, Cecilia Prego2, Maria Alonso2, Christina Barry3,
Richard Pilon1, Christina Daniuk1, Mikaela Nykoluk1, Stephane
Pillet3, Tomasz Bielawny1, Jeffrey Tuff1, Chris Czarnecki1, Philip
Lacap1, Gary Wong3, Shaun Tyler3, Ben Liang3, Zhe Yuan4,
Qingsheng Li4, Terry Blake Ball1, Paul Sandstrom1, Gary
Kobinger3, Francis Plummer3,5, Ma Luo1,5
Posters
P41.19
P41.20
Antibodies to CD52g, a Secreted Spermcoating Antigen, Agglutinate Seminal
Leukocytes and Prevent their Infiltration into
Vaginal Epithelium
Vaccine-induced CD107a+ CD4+ T-cells
Are Resistant to Killing Following
Immunodeficiency Virus Infection
Jai G. Marathe1, Joseph Politch2, Ayesha Islam2, Kevin Whaley3,
Thomas Moench4, Deborah J. Anderson5
Boston University, Medicine, Boston, MA, United States, Boston
University, Obstetrics and Gynecology, Boston, MA, United States,
3
Mapp Biopharmaceutical, Inc., San Diego, CA, United States, 4ReProtect
Inc., Baltimore, MD, United States, 5Boston University, Obstetrics and
Gynecology, Medicine, Microbiology, Boston, MA, United States
1
2
POSTERS
Background: Our program is studying the topical use of monoclonal
antibodies (mAbs) for contraception and HIV prevention. mAbs directed
against CD52g, an antigen secreted into the male genital tract and
inserted into sperm membranes, potently agglutinate sperm and are
being developed for contraceptive use. We are also seeking strategies
to prevent cell-associated HIV transmission mediated by HIV-infected
seminal white blood cells (sWBC). In this study, we investigated
whether CD52g also attaches to sWBC, and whether anti-CD52g mAbs
agglutinate these cells and/or inhibit their interaction with the vaginal
epithelium.
Methods: Dylight 633-conjugated MSH8, a mouse anti-CD52g mAb
(gift of J. Herr), was used in flow cytometry experiments to detect
CD52g on sWBC and passive insertion of seminal plasma CD52g into
the plasma membrane of monocyte-derived macrophages (MDMs).
HC4, a human mAb expressed in Nicotiana (Mapp Biopharmaceuticals
Inc.), was used in two functional assays: 1) agglutination of sWBC,
assessed by counting the percentage of CMFDA-labelled WBC
associated with sperm agglutinates in mAb-treated seminal fluid; and
2) cell attachment and infiltration assays, modeled with CD52g-coated
CMFDA-labeled WBCs and EpiVaginal™ tissue (MatTek Corp.), and
assessed by confocal microscopy.
Results: MSH8 bound to a majority of sWBCs and also to seminal
plasma-treated MDMs. HC4 trapped WBCs in sperm agglutinates. The
antibody also significantly inhibited the attachment and infiltration of
CD52g-coated WBCs into the vaginal epithelium. Control mAbs had no
effect in these assays.
Conclusions: Topical vaginal application of antibodies to CD52g could
serve a dual purpose use: prevention of pregnancy and inhibition of cellassociated HIV transmission.
358
Tetsuro Matano1,2, Kazutaka Terahara1, Hiroshi Ishii1, Takushi
Nomura1
National Institute of Infectious Diseases, AIDS Research Center, Tokyo,
Japan, 2University of Tokyo, Institute of Medical Science, Tokyo, Japan
1
Background: CD4+ T cell responses are crucial for effective antibody and
CD8+ T cell induction following virus infection. However, virus-specific
CD4+ T cells can be preferential targets for HIV infection. HIV-specific
CD4+ T cell induction by vaccination may thus result in enhancement of
virus replication following infection. We examined virus-specific CD4+
T cell responses before and after SIV infection in vaccinated macaques
and the sensitivity of vaccine-induced CD4+ T cells to killing following
infection.
Methods: We analyzed SIV-specific T-cell responses using frozen
PBMCs from vaccinated (n = 18) and unvaccinated (n = 21) Burmese
rhesus macaques. Vaccinated animals received a DNA prime and a
Gag-expressing Sendai virus vector boost, followed by an intravenous
SIVmac239 challenge (3 months post-boost). Eleven (61%) of
vaccinated animals controlled SIV replication. SIV-specific CD4+ T-cell
responses were examined at 1 or 2 months pre-challenge and 1 week
post-challenge by measurement of five markers, CD107a, MIP-1β, IFN-γ,
TNF-α, and IL-2 following SIV-specific stimulation.
Results: At week 1 post-challenge, both vaccinated and unvaccinated
animals had higher frequencies of SIV-specific CD4+ T-cell subsets
inducing CD107a than other markers. Comparison of virus-specific CD4+
T-cell responses in vaccinated animals pre- and 1-week post-challenge
showed a significant reduction in CD107a-negative but not in CD107a+
subsets after SIV challenge. The reduction in vaccinated non-controllers
was larger than controllers.
Conclusions: Vaccine-elicited CD4+ T cells with the potential to induce
CD107a are resistant to infection in a macaque AIDS model. Our results
indicate that vaccine-induced CD107a-negative
CD4+ T cells are efficiently killed following exposure, suggesting that
vaccine design for HIV protection should avoid CD107a-negative CD4+
T-cell induction.
P41.21
P41.22
Factors that Influence the Willingness of
Young Adults to Participate in Early Vaccine
Trials and Contraceptive Practices in Dar es
Salaam, Tanzania
Evaluation of Dendritic Cell Targeted
Consensus B and MOSAIC HIV Gag Protein
Vaccines in Vitro in PBMC of Treatment Naive
HIV-1 Infected People
Theodora Mbunda1, Muhammad Bakari1, Edith Tarimo1, Eric
Sandstrom2, Asli Kulane2
Godwin Nchinda1, Nadesh Nji2, Jules C Tchadji2, Georgia
Ambada2, Andres Salazar3, Ralph Steinman4, Michel
Nussensweig4, Tibor Keller5, Samuel Sosso2, Chae Gyu Park6,
Pierre Fouda7, Vitorro Colizzi8
Muhimbili University of Health and Allied Sciences, Dar es Salaam,
Tanzania, United Republic of, 2Karolinska Institutet, Stockholm, Sweden
1
Chantal Biya International Reference Center for HIV, Microbiology
and Immunology, Yaounde, Cameroon, 2Chantal Biya International
Reference Center for HIV, Yaounde, Cameroon, 3Oncovir, Jonesboro,
GA, GA, United States, 4Rockefeller University, New York, NY, United
States, 5Celldex Therapeutics, Hampton, NJ, United States, 6Yonsei
University College of Medicine, Seoul, Korea, Democratic People`s
Republic of, 7Central Hospital of Yaounde, Yaounde, Cameroon, 8Tor
Vergata, Roma, Italy
Background: HIV/AIDS destroys the lives of young people especially
in low-income countries. The inclusion of youths in HIV vaccine trials is
necessary in obtaining an effective vaccine that is acceptable to them.
Participation in HIV vaccine trials however necessitates participants not
to become pregnant or impregnate women. Therefore it is important
to study factors influencing willingness to participate and dynamics of
contraceptive practices among young adults
Methods: Four hundred and fifty young adults visited a youthfriendly Infectious Diseases Clinic (IDC) from February to September
2012 completed a self-administered questionnaire concerning sociodemographic information, contraceptive practices, knowledge and
perception of HIV vaccine studies, and the availability of social support.
Results: Of our participants, 50.6% expressed willingness to participate.
The willingness was positively correlated with having some knowledge
about HIV vaccine studies (AOR, 2.2; 95% CI: 1.4-3.4), a positive
perception toward such studies (AOR, 2.3; 95% CI: 1.5-3.6) having a
relationship with someone who could help them make a decision
(AOR, 2.5; 95% CI: 1.3-4.9), and age at the time of sexual debut (AOR,
2.6; 95% CI 1.0-6.7) for 15- to 19-year-olds and (AOR, 2.7; 95% CI
1.0-7.1) for older participants. 73% of those expressing WTP knew
contraception was for spacing children, 45% and 49% reported to ever
use contraceptives and to use them at time of last sexual intercourse
respectively. The reasons for not using contraceptives were not being
married; lack of knowledge on contraceptives; and having unplanned
sexual intercourse.
Conclusions: The participants exhibited a moderate willingness
to participate in HIV vaccine trials, less than half reported to use
contraceptives. More efforts should be made to inform the youths about
specific HIV vaccine trials, to engage significant others in the decisionmaking process, and to address impediments pertaining to contraceptive
use in the Tanzanian context.
Background: Our consortium has developed a first generation dendritic
cell (DC) targeted consensus B HIV gag protein vaccine for proof
of concept studies that selected delivery of proteins to DC will allow
proteins to be more immunogenic. This should provide a cheaper and
effective way to immunize people repeatedly with no negative impact
of pre-existing immunity. This vaccine is already in an ongoing phase 1
clinical trial in New York however it is not known whether this consensus
B based vaccine would work in sub Saharan African where unrelated
strians of HIV-1 are predominant.
Methods: To assess the consequense of targeting a consensus B and
MOSAIC gag protein vaccines to DC from people infected with unrelated
strains of HIV-1 in Africa we added the protein vaccines to their blood
cells in vitro and measured proliferation and IFNy production by bulk
PBMCs as well as a coculture between monocyte derived DC and T cells.
Results: Dendritic cell targeted Consensus B and MOSAIC HIV gag
Protein vaccines recalled pre-existing T cell responses in blood cells
of treatment naïve people infected with unrelated strains of HIV-1. DC
targeted MOSAIC gag vaccine was more efficient than the consensus B
vaccine and stimulated significant proliferation of HIV gag specific cells
(P< 0.001).
Conclusions: Thus dendritic cell targeted consensus B and MOSAIC
HIV gag protein vaccines could recall pre-exhisting CD4 and CD8 T
cell responses in vitro in PBMC of treatment naïve people infected
with unrelated strains of HIV-1 in AFrica. DC targeted MOSAIC gag
protein vaccine should be further evaluated for possible incorporation
in future vaccines.
www.hivr4p.org
359
POSTERS
1
Posters
P41.23
P41.24
Gp41 and p24 IgG Binding Antibody Titers Are
Associated with HIV-1 Viral Control during
Early HIV-1 Subtype C Infection
Host Restricted Poxviruses Produce Distinct
Host Responses in an in vivo Mouse Model
with Implications for Future Use as HIV-1
Vaccine Vectors
Bongiwe G. Ndlovu1, Amy Chung2, Jennifer Maroa3, Bruce D.
Walker1,2, Galit Alter2, Thumbi Ndung’u1,3
HIV Pathogenesis Programme, University of KwaZulu-Natal, Durban,
South Africa, 2Ragon Institute of MGH, MIT and Harvard, Cambridge,
MA, United States, 3KwaZulu-Natal Research Institute for Tuberculosis
and HIV, University of KwaZulu-Natal, Durban, South Africa
1
POSTERS
Background: The RV144 Thai HIV-1 vaccine trial showed 31%
protective efficacy and suggested a role for non-neutralizing antibodies
in protection against risk. However, the kinetics of development of
binding antibodies and their ability to mediate innate effector functions
in HIV-1 infection has not been fully characterized. The purpose of this
study was to characterize the evolution of anti-HIV binding antibodies
and to determine their antibody-dependant cellular phagocytic activity
(ADCP) in individuals with recent HIV-1 infection.
Methods: Thirty four individuals with acute HIV-1 infection (Fiebig I/
II) were identified and followed for one year in Durban, KwaZulu-Natal.
Viral loads were measured using the Roche Amplicor ver 1.5 assay.
Plasma HIV-1 specific gp120, gp41 and p24 total IgG and Ig subclasses
(IgG1-4) antibodies were measured using a customized Multiplex
Luminex assay at 1, 2, 3, 6 and 12 months post-infection. The capacity
of plasma antibodies to mediate ADCP in the presence of fluorescent
beads conjugated to gp120 protein was examined in a THP-1 cell based
in-vitro assay.
Results: Total gp120 and p24-specific IgG antibody titers significantly
increased over the follow up period (p< 0.0001, One-way ANOVA),
whereas gp41-specific IgG titers did not change over time compared
to titers at one month post-infection. Both gp41 and p24-specific total
IgG antibodies correlated negatively with contemporaneous viral load at
months 3 (p= 0.06, r=-0.38; p=0.0146, r=-0.457) and 12 (p= 0.0447,
r=-0.465; p=0.0524, r=-0.439) post-infection. Despite high IgG1
binding titers, neither anti-gp120, gp41 nor p24 IgG1 titers correlated
with viral load. Gp120-specific phagocytic scores correlated with higher
viral set point.
Conclusions: The kinetics of expansion of HIV-1 protein-specific IgG
titers were different. Gp41 and p24-specific IgG titers may have an antiHIV role in vivo which requires further investigation.
360
Kristy Offerman1, Olivia Carulei1, Armin Deffur2, Nicola Douglass1,
Anna-Lise Williamson1,3,4
University of Cape Town, Division Medical Virology, Department of
Clinical Laboratory Sciences, Cape Town, South Africa, 2University of
Cape Town, Clinical Infectious Diseases Research Initiative, Cape Town,
South Africa, 3Institute of Infectious Disease and Molecular Medicine,
Cape Town, South Africa, 4National Health Laboratory Service, Groote
Schuur Hospital, Cape Town, South Africa
1
Background: Host restricted poxviruses make promising candidates for
HIV vaccine vectors due to their safety profile and immunogenicity. The
comparison of host responses produced by different poxvirus vectors
would aid in the assessment and rational design of improved vaccines.
Methods: In order to investigate the ability of different poxviruses to
interact with the mammalian host, we compared host gene expression
profiles in the spleens of BALB/c mice in response to infection (105 pfu/
mouse) with four poxvirus species; Lumpy Skin Disease virus (LSDV),
Canarypox virus (CNPV), Fowlpox virus (DCEP 25 modified strain
(Merial)) (FWPV), modified Vaccinia Ankara (MVA).
Results: The findings presented here indicate that four, genetically
diverse host-restricted poxviruses, CNPV, FWPV, MVA and LSDV,
produce qualitatively and quantitatively distinct host responses in an
in vivo mouse model. Differential gene expression regulated by the
different poxviruses is discussed with particular emphasis on immunityrelated responses.
Of particular interest, CNPV and FWPV, and not MVA or LSDV induce
the upregulation of two immunoglobulin genes (Ighg and Ighg3 (IgG3))
with CNPV inducing a third, Ighm (IgM). Comparison of the immune
responses resulting from the partially effective clinical RV144 HIV1 trial and the ineffective VAX003 trial indicated that HIV-1-specific
IgG3 antibodies correlated with decreased risk of HIV-1 infection in the
RV144 trial. The upregulation of IgG3 by avipoxviruses in vivo, suggest
that the avipoxvirus-vector backbone may be involved in stimulation of
the clinically important IgG3 antibody subclass.
Conclusions: These findings suggest potentially important biological
differences in response to four clinically relevant poxviruses, especially
because small changes in gene expression may produce disproportionate
results in biological systems. These differences may affect the immune
response induced to vaccine antigen in vectors based on these viruses.
P41.25
P41.26
Generating an Anti-HIV Vaccine Using
Nucleoside-modified mRNA Encoding
Envelope
Comparative Neutralization Sensitivity of
Indian and South African HIV-1 Clade C
Viruses to Plasma Antibodies from Chronically
Infected Indian Donors
Norbert Pardi1, Katalin Kariko1, Michael Hogan1, Hiromi
Muramatsu1, James A. Hoxie1, Drew Weissman1
University of Pennsylvania, Department of Medicine, Philadelphia, PA,
United States
1
Shilpa Patil1, Sharda Gadhe2, Swapnil Sonawane2, Manish
Bansal1, Suprit Deshpande1, Tandile Hermanus3, Lynn Morris3,
K G Murugavel4, Suniti Solomon4, Seema Sahay2, Ramesh
Paranjape2, Bimal K. Chakrabarti1, Jayanta Bhattacharya1
HIV Vaccine Translational Research Laboratory, THSTI-IAVI HIV
Vaccine Design Program, Gurgaon, India, 2National AIDS Research
Institute, Pune, India, 3National Institute for Communicable Diseases,
Johannesburg, South Africa, 4Y.R. Gaitonde Centre for AIDS Research
and Education, Chennai, India
Background: There has been great progress in understanding the
detailed molecular mechanisms of HIV-1 infection but no effective
vaccine has been developed to date. mRNA has emerged as a very
promising new therapeutic agent in recent years and has already shown
progress as an effective method for generating potent vaccines.
Methods: To create a vaccine with maximal potency, in vitro transcribed
mRNAs were optimized for higher levels and extended translation by
incorporation of selected UTRs, modified nucleosides, 5’ cap, 3’ poly(A)tail, and other modifications and were HPLC-purified. To achieve both
strong T cell and B cell responses, heterologous mRNA prime - protein
boost vaccination regimens were used. For priming, naked mRNA
encoding HIV envelope gp160 was administered intradermally into
mice. Cell surface Env was used to increase exposure of neutralizing
epitopes. Four weeks after the second mRNA prime, Env protein was
injected intramuscularly as a boost. As nucleoside modified mRNA does
not activate RNA sensors, to increase the robustness of the immune
response, a series of adjuvant molecules and encoding mRNAs were
co-injected along with the antigen-encoding mRNA. Flow cytometry and
ELISA were used to evaluate T cell and B cell responses, respectively.
Results: Elevated levels of IFN-χ, TNF-α and IL-2 in antigen-specific CD4+
and CD8+ T cells and high gp120 antibody titers could be measured
following two rounds of mRNA prime - protein boost vaccination.
Conclusions: Our results demonstrate that antigen-encoding nucleoside
modified mRNA induces effective HIV-specific immune responses and
has great potential for vaccination against infectious diseases.
Background: HIV-1 clade C is the major subtype circulating in India
and South Africa. The present study was undertaken to examine the
extent of neutralization sensitivity of Indian and South African (SA) HIV-1
clade C viruses to plasma antibodies obtained from anti-retroviral naïve
chronically infected HIV-1 positive Indian patients.
Methods: Plasma samples from 150 anti retroviral naïve donors from
India chronically infected with HIV-1 were assessed for their degree of
neutralization of 9 Indian and 10 South African HIV-1 clade C envelopes
using a TZM-bl reporter cell assay. Reagents were shared between Indian
and SA laboratories for quality assessment. Antibody specificities were
determined by using TriMut core protein, mutant and chimeric viruses.
Results: 27/150 (18%) plasma samples were found to neutralize >50%
of the panel viruses at 1:100 dilution. Amongst these, seven were found
(4.66%) displayed maximum breadth and potency with median ID50s
ranging from 300-750. The BCN plasma antibodies were found to
neutralize Indian (57%) viruses slightly better than SA (43%) viruses.
None of the potent broadly cross neutralizing BCN plasmas was found
to show neutralizing antibodyspecificities targeting theCD4 binding site.
However, two of them showed N332 residuedependence in V3 loop
in both Indian and SA clade C Env backbones. While 3/7 BCN plasma
antibodies showed clade C MPER (HIV-2/HIV1 7312-C1C) dependence,
they did not show specificity to epitopes targeted by known MPER
directed monoclonal antibodies.
Conclusions: Our data suggest the presence of common epitopes in
Indian and South African HIV-1 clade C envelopesas most of the BCN
plasma antibodies cross-neutralized pseudotyped viruses expressing
clade C envelopes from both the countries. Identification of epitopes
common in both in Indian and SA clade C envelopes will help define
strategies to design vaccine that would be effective in both countries.
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361
POSTERS
1
Posters
P41.27
P41.28
Cell-surface Display and Panning of HIV-1
Derived Envelope Proteins
Development of Advanced Oligonucleotidebased Microbicides: Driving HIV into Suicide
Tim-Henrik Bruun1, Veronika Schmid1, Alexander Kliche1,
Ralf Wagner1
Maike Voges1, Joachim Hauber1, Karin Moelling1,2
University of Regensburg, Institute of Medical Microbiology and
Hygiene, Molecular Microbiology and Gene Therapy Unit, Regensburg,
Germany
1
POSTERS
Background: Available display systems allow the screening of millions
of candidate proteins and are the method of choice to identify optimized
antigen-antibody binding.
We established a mammalian cell-surface display to present HIV1 envelope derivatives in a natural, trimeric and membrane bound
environment. This allows us to generate affinity enhanced envelope
derivatives against broadly neutralizing antibodies (bNAbs) in order to
select potent Envelope (Env) based vaccine candidates.
Methods: An HIV-1 derived lentiviral vector was developed to infect
HEK293T cells at a low multiplicity of infection (MOI), in order to
correlate phenotype and genotype. The vector was designed and proven
to express both GFP and Env in a constant relationship, enabling indirect
normalization for Env expression by detecting GFP. After staining with
an appropriate bNAb, Env-displaying cells were selected for high affinity
binding via FACS-Sorting.
To adapt this system to the use of very large libraries, a refined vector
system was developed, allowing the stable genomic integration of both
ENV and GFP at a distinct integration site. This ensures that only one
envelope variant is expressed per cell, efficiently linking phenotype and
genotype. Due to the stringent linkage of ENV and GFP, GFP expression
can again be used as a means to normalize for Env expression in the
FACS-Sorting process.
Results: A small model library of Env variants with distinct binding
capacities towards the bNAb 447-52D was used to evaluate both
techniques.
The proof-of-concept experiment for the virus-based approach led to an
enrichment of up to tenfold for the Env variant with the highest affinity
toward 447-52D after two rounds of selection.
Using the stable cell line technology, the desired Env high affinity
variant could even be enriched up to 39-fold after only one round of
FACS-Sorting.
Conclusions: Hence, these techniques provide the possibility to screen
for membrane bound Env variants with high binding capacities towards
any bNAb applied.
362
1
Heinrich-Pette Institute, Antiviral Strategies, Hamburg, Germany, 2Max
Planck Institute for Molecular Genetics, Berlin, Germany
Background: HIV is transmitted primarily by sexual intercourse and
predominantly infects people in third world countries. An important
medical need is self-protection of women, particularly in societies
where condoms are not accepted. Therefore, we currently develop an
oligodeoxynucleotide (ODN)-based microbicide, which destroys HIV
prior to infection by activating the retroviral RNase H.
Methods: We performed RT/RNase H cleavage assays in vitro to
demonstrate RNase H-dependent cleavage of viral RNA in the highly
conserved polypurin tract after incubation with ODNs. Furthermore,
luciferase-based infection assays and p24 ELISA were used to analyze
de novo infection capacity by ODN-treated HIV. Finally, various ODN
stability assays were performed.
Results: It is shown that ODNs target cell-associated as well as cell-free
virions, leading to efficient degradation of the viral RNA genome and,
in turn, to strongly reduced infectivity. Moreover, pronounced antiviral
activity is demonstrated in vitro and in vivo (animal) models. Surprisingly,
ODNs are able to enter cell-free virions without any carrier and are also
stable over weeks at 37°C in different solutions (PBS, BSA, KY Jelly).
Conclusions: ODNs trigger highly effective destruction of the viral RNA
genome prior to infection without any carrier and are highly stable
for many weeks. Therefore, ODNs may be valuable components of
advanced microbicides to drive HIV into suicide.
P41.29
P41.30 LB
Induction of Broadly Neutralizing Antibodies
to HIV-1 via DNA Motif Immunization
IL-1R, WNT and γ-C Cytokine Receptor
Signaling Correlate with CD4 T Cell Effector
Function in Response to Anti-DEC205-HIV gag
p24/polyICLC Vaccination
Xilin Wu1, Zhiwei Wu2, Lin Cheng3, Zhiwei Chen3
The University of Hong Kong, Hong Kong, Hong Kong, 2Nanjing
University, NanJing, China, 3The University of Hong Kong, HongKong,
Hong Kong
1
Vaccine & Gene Therapy Institute of Florida, Port Saint Lucie, FL,
United States, 2Rockefeller University, New York, NY, United States,
3
Diseases, National Institutes of Health, Bethesda, MD, United States,
4
Diseases Vaccine Immune T-Cell and Antibody Laboratory, National
Institutes of Health, Bethesda, MD, United States, 5Case Western
Reserve University, Cleveland, OH, United States
1
Background: A randomized double-blinded, placebo-controlled phase
I study was conducted in healthy volunteers to evaluate the safety and
immunogenicity of the dendritic cell targeted mAb (3G9) fusion protein
vaccine, anti-DEC205-HIV gag p24 with polyICLC (DCVax). Subjects
were enrolled in three dose arms: Low(0.3mg), Mid(1mg) or High(3mg)
with 1.6mg of polyICLC subcutaneously. Control arms received either
polyICLC or Saline.
Methods: We performed microarray and bioinformatic analysis to
identify gene signatures and pathway activity differentially expressed in
whole blood from the different vaccine dose groups. Linear Regression
analysis was used to correlate gene expression with ICS data. Statistical
Inference was assessed using Wilcoxon rank sum test.
Results: DCVax induced greater IFN gene expression and IL-12
production in whole blood compared to polyICLC alone, and in a dosedependent fashion that correlated with PRDM1(BLIMP1) expression in
CD4 cells and total Gag-specific CD4 TH1 and CD8 IFNγ producing T cells
in the high dose vaccinees. Pathway analysis on top regression features
showed differential proinflammatory responses (e.g. IL1, RXR, LPS) at 1d
in subjects that mounted potent Gag-specific CD4 responses. High dose
subjects upregulated genes downstream of IL-1R signaling (e.g. IRAK3,
MAPK2K3) including NF-κB. Gag-specific CD4 TH1 responses correlated
with IL1R, WNT, and γ-C receptor cytokine signaling pathways that
might also predispose toward greater CD4 T cell memory development.
Conclusions: Synergy was seen with the high dose HIV Gag p24 dendritic
cell targeted vaccine and polyICLC for induction of both a quantitative
and qualitative greater Type I IFN response compared to lower vaccine
doses or polyICLC alone. This resulted in optimum CD4 TH1 and CD8
CTL responses. Of note, only the high dose and polyICLC combination
induced SOCS-1, a critical regulator of type I interferon signaling. These
results highlight that combinations of vaccine and adjuvant can trigger
quantitatively different responses.
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363
POSTERS
Background: HIV-1 is highly mutated, which poses a serious problem
to develop a prophylactic AIDS vaccine. To date, it remains elusive how
to induce broadly reactive neutralizing antibodies against genetically
divergent HIV-1 strains. We have previously invented peptide motif
immunization, which could induce broadly neutralizing antibodies
against highly conserved and functional domain of HIV-1 envelope
protein in mice, rabbits and rhesus monkeys. In this study, we aim to
invent a novel DNA motif immunization method.
Methods: The primers encoding 15 amino acids GPG motif were
synthesized, which were inserted in the plasmid by PCR as the gene of motif.
We constructed four groups of plasmid in the vector of Pvax, PD1-p24motif, PD1-ova-motif, CTLA4-p24 and PD1-p24 as control. Mice received
4 DNA immunizations by intramuscular injection plus electroporation.
Two weeks after the final immunization, ELISA, Neutralization, Elispot and
FACS were conducted to test the efficacy of such antigen in the responses
of humoral immunity and cellular immunity.
Results: The plasmid encoding the motif gene was characterized by
sequencing. It showed that the motif gene was constituted by nine
fixed nucleotides encoding three amino acid residues of GPG in the
center flanking with 36 random nucleotides. Plasmid expression was
confirmed by FACS. The plasmid containing the gene of motif all could
induce antibody specific for GPG with comparing to the control of
PD1-p24 group. In addition, the group of PD1-p24-motif could induce
GPG-specific antibodies in mice that neutralized 100% HIV-1 strains of
various subtypes tested with GPG motif in their V3 region. Moreover, it
could provide robust cellular immunity against gag protein of HIV.
Conclusions: Our findings firstly completed proof of concept of DNA
motif immunization. What´s more, the PD1-p24-motif plasmid could be
a potential candidate DNA vaccine against HIV infection with induction
of broadly neutralizing antibodies and robust cellular immunity.
Khader Ghneim1, Marina Caskey2, Christine Trumpfheller2, Gaelle
Breton2, Petra Stafova1, Stephanie Richards1, Richard Koup3, Bob
Bailor4, Sarah Schlesinger2, Jeff Ahlers1, Rafick P. Sekaly5, Ralph
Steinman2, Mark Cameron1
Posters
Posters 42: Participation in Trials: Willingness, Benefits and Challenges
P42.01
P42.02
Self-reported Benefits of Study Participation
in HVTN 503, “Phambili”
Impact of DSMB Outcomes on Participation
in HIV Prevention Trials: The VOICE Study
Experience in Kampala, Uganda
Mary A. Allen1, Barbara Metch2, Zoe Moodie2, Linda-Gail Bekker3,
Gavin Churchyard4, Koleka Mlisana5, Maphoshane Nchabeleng6,
Jim Kublin2, Glenda Gray7, HVTN 503 Study Team
NIH/NIAID, DAIDS, Bethesda, MD, United States, 2FHCRC, Seattle,
WA, United States, 3Desmond Tutu HIV Foundation, Cape Town,
South Africa, 4Aurum Institute, Klerksdorp, South Africa, 5University
of KwaZulu Natal, Durban, South Africa, 6Medunsa Clinical Research
Unit, Medunsa, South Africa, 7Chris Hani Baragwanath Hospital, PHRU,
Soweto, South Africa
1
POSTERS
Background: Participants (ppts) in HVTN 503 (“Phambili”), the only
HIV vaccine efficacy trial conducted in Africa to date, were asked about
possible benefits of study participation.
Methods: Social impact (SI) assessment was conducted at weeks 12,
78, 130, and 182, and included asking “In the last 6 months, has
participation in this study had a beneficial impact on your life?” Benefits
were analyzed by sex, age, study site and treatment group.
Results: Of the 801 ppts enrolled, 752 (94%) reported that study
participation had a beneficial impact on their lives, and only 48 (6%)
reported negative social impacts (NSI). Overall, 705 (88.0%) reported a
benefit and no NSIs; 48 (6.0%) reported neither; and 1 reported a NSI
and no benefit. Differences by site (n=5) were statistically significant (p<
0.001) with benefits reported by 89.1 to 98.1% of ppts. The percentage
of ppts reporting a benefit decreased slightly by age in years (yrs) at
enrollment (18-20yrs 229/238=96.2%, 21-25yrs 312/334=93.4%, 2630yrs 136/146=93.2% and 31-35yrs 75/83=90.4%) but the difference
was not statistically significant (p=0.23). Similarly high percentages of
both treatment groups, and both sexes reported a benefit at one or more
study visits (vaccine 374/400=93.5% vs. placebo 378/401=94.3%;
women 339/360=94.2% vs. men 413/441=93.5%). Frequently
reported benefits were receiving free HIV risk reduction counselling
and testing, and knowing one’s HIV status. Twenty male ppts identified
information about medical male circumcision was beneficial. Ppts also
reported pride in doing “something good for [their] community”, “helping
my nation”, and educating others about HIV/AIDS.
Conclusions: The majority of ppts, including those reporting negative SI
events, reported benefits of study participation on their lives. Differences
by site were statistically significant (p< 0.001). Additional examination
of reported benefits of study participation may be informative, and
potentially enhance future recruitment and retention efforts.
364
Juliane Etima1, Teopista Nakyanzi1, Samuel Kabwigu1, Flavia M.
Kiweewa1, Brenda G. Mirembe1, Carolyne A. Akello1, Lisa Rossi2,
Clemensia Nakabiito1
MU-JHU Research Collaboration, Kampala, Uganda, 2Microbicide Trials
Network, Pittsburgh, PA, United States
1
Background: Clinical trials are monitored and regulated by independent
bodies to ensure participants’ safety and ethical conduct. Depending on
study progress, these bodies may recommend a trial to continue as is,
be modified or stopped. Six such reviews were conducted during the
implementation of the VOICE study. We describe the impact of Data &
Safety Monitoring Board (DSMB) recommendations on continued study
participation in Kampala, Uganda, including retention, researchers’
motivation to continue the study, and community perceptions and
attitudes about the study.
Methods: Communication plans were used to disseminate DSMB
outcomes to 6 tiers of stakeholders including study staff, participants,
Institutional Review Board, Community Advisory Board, media, and other
key stakeholders, including civil society. A qualitative analysis of DSMB
outcome reports and review of participant retention was done. Discussion
about different scenarios was done in advance with stakeholders.
Results: Stakeholders across the 6 tiers expressed disappointment
with DSMB outcomes in VOICE. “We are ashamed because the doctors
treated many infections and we benefited. The tests were expensive and
the blood samples showed we did not use the products” (participant). “All
our effort has gone to waste” (staff). “HIV drugs disappoint researchers”
(media). “We told you that your things will not work, you want to
experiment on us” (community member). The number of missed visits
increased after dissemination of the Nov 2011 DSMB outcome when
a second study arm (tenofovir gel) was stopped; 24% (41/171) as
compared to 11% (18/171) in Sept 2011 when the first arm (tenofovir
tablet) was stopped (p< 0.001).
Conclusions: Communication about negative DSMB outcomes remains
a challenge, although communication plans make dissemination more
manageable for sites. Data from VOICE suggest that DSMB outcomes
may have had a significant impact on visit retention in Kampala.
P42.03
P42.04
Reporting of Adherence in the VOICE Trial:
Does Disclosure of Product Non-use Increase
at the Termination Visit?
Reporting of Challenges to Adherence in
VOICE: A Comparison of Quantitative and
Qualitative Self-reports among Women
during and after the Trial
1
Population Council, New York, NY, United States, 2RTI International,
San Francisco, CA, United States, 3SCHARP - FHCRC, Seattle, WA, United
States, 4University of Washington, Seattle STD/HIV Prevention Training
Center, Seattle, WA, United States, 5UZ-UCSF Collaborative Research
Programme, Harare, Zimbabwe, 6FHI 360, Durham, NC, United States,
7
National Institute of Allergy and Infectious Diseases, Bethesda, MD,
United States
Background: Results from VOICE indicated that ≥50% of women
assigned to active products (Truvada, oral or vaginal tenofovir) had
no drug detected in any plasma samples tested during the trial. Yet,
in response to questions on product use asked of participants during
the trial, ≥90% of doses were reportedly taken. To explore factors
associated with suboptimal adherence, a behavioral exit questionnaire
was developed after early closure of the oral tenofovir and vaginal
gel arms. The underlying rationale was that women might respond to
questions about adherence more candidly at termination than they
would during the course of the trial.
Methods: For the sub-sample of VOICE participants still enrolled in the
trial in December 2011, we compared self-reported adherence obtained
with the same/similar interviewer-administered questions at monthly/
quarterly visits and at the termination exit visit (TEV) using descriptive
statistics and regression models.
Results: A TEV questionnaire was administered for 2321 of the 5029
VOICE participants. Of the 1134 women who reported at the TEV that
they always used the product, 53.8% reported missing doses in one
of the monthly or quarterly follow-up interviews. Correspondingly, of
the 812 women who reported in a monthly/quarterly interview that
they always used the product, 34.6% reported in the TEV that they
sometimes missed doses. A comparison between a self-rating adherence
scale in the quarterly and TEV visits indicated a slightly lower reporting
of fair/poor/very poor adherence in the former (7% vs 12%) although
the difference was not statistically significant.
Conclusions: Participants in the VOICE trial were not more likely to
disclose non-adherence at the termination visit than during follow-up
visits. While a large proportion of women reporting perfect adherence
during the course of the trial reported missing doses at termination,
an even larger proportion of women reporting perfect adherence at
termination reported missing doses during the monthly/quarterly visits.
Barbara S. Mensch1, Ariane van der Straten2, Miriam
Hartmann2, Helen Cheng2, Barbara Miller1, Jeanna Piper3, Lisa
Levy4, Cynthia Grossman5, Elizabeth Montgomery2, on behalf of
the MTN 003D Team
Population Council, New York, NY, United States, 2RTI International,
San Francisco, CA, United States, 3National Institute of Allergy and
Infectious Diseases, Division of AIDS, Bethesda, MD, United States,
4
FHI 360, Washington, DC, United States, 5National Institute of Mental
Health, Center for Mental Health Research on AIDS, Bethesda, MD,
United States
1
Background: Results from VOICE revealed that drug was detected in
≤30% of plasma samples from women assigned to active arms. Since
participants indicated ≥90% of doses were taken, do self-reports on
challenges to product use provide useful information on adherence? If
so, when, during a trial, is it optimal to collect such information?
Methods: We investigate adherence challenges reported during and
after VOICE among 72 active arm participants enrolled in MTN-003D, a
qualitative study exploring aspects of the trial that affected product use.
Responses to questions on challenges to use and reasons for non-use
were compared:
1) at the first and last VOICE quarterly visits when participants reported
on product adherence (PA);
2) at a Termination Visit interview (TV) conducted about 8 weeks after
product use ended among 38 of the 72 participants still enrolled when
the questionnaire was designed following early closure of two arms; and
3) during the 003D Stage 2 in-depth interviews (IDI), conducted about
1.5 years after the TV, where participants were informed about their
drug detection pattern and classified into adherence groups.
Results: At the first PA 6 of 70 participants and, at the last PA, 4 of 68
indicated reasons they did not use product daily in the prior 4 weeks. At
TV, 15 of 38 participants responded that they were not always adherent;
on average 1.6 challenges were reported by these 15. During the IDIs
after drug results were revealed to participants, 65 of 72 reported
challenges; 38 listed 5+ challenges. Among the 43 in the low adherence
group, 4 reported no challenges; 26 reported 5+. Among the 20 in the
high adherence group, 3 reported no challenges; 9 reported 5+.
Conclusions: VOICE participants were reluctant to disclose reasons for
non-adherence when on product; after product use ended, willingness
to report challenges increased slightly. Only when presented with drug
detection patterns denoting whether product was taken, however, were
participants forthcoming about the challenges they faced.
www.hivr4p.org
365
POSTERS
Barbara S. Mensch1, Ariane van der Straten2, Elizabeth Brown3,
Karen Liu3, Jeanne Marrazzo4, Zvavahera Mike Chirenje5,
Kailazarid Gomez6, Jeanna Piper7, Karen Patterson3
Posters
P42.05
P42.06
Bone Mineral Density Changes among
Healthy African Pre-menopausal Women
Participating in a Tenofovir-based HIV PrEP
Study: The MTN-003B Study
Sharing of Investigational Drug among
Participants in the VOICE Trial
Brenda G. Mirembe1, Cliff Kelly2, Mgodi Nyaradzo3, Suzan
Greenspan4, James Dai2, Vivian Bragg5, Jeanna Piper6, Carolyne
A. Akello1, Flavia M. Kiweewa1, Magure Tsitsi3, Clemensia
Nakabiito1, Sharon Riddler4
1
Makerere University-Johns Hopkins University Research Collaboration,
Kampala, Uganda, 2SCHARP - FHCRC, Seattle, WA, United States, 3UZ
- UCSF, Harare, Zimbabwe, 4University of Pittsburgh, Pittsburgh, PA,
United States, 5FHI 360, Durham, NC, United States, 6DAIDS/NIAID/
NIH, Bethesda, MD, United States
Jeeva Moodley1, Leanne Vallabhjee1, Sarita Naidoo1, Jayajothi
Moodley1, Gita Ramjee1,2
South African Medical Research Council, HIV Prevention Research Unit,
Westville, South Africa, 2London School of Hygiene & Tropical Medicine,
Kingdom
1
POSTERS
Background: Limited data exists on the effect of tenofovir on bone
mineral density (BMD) in HIV negative women. This is a major concern
for long-term PrEP regimens. We evaluated effects of daily oral tenofovir
(TDF) and Truvada (FTC/TDF) compared to placebo on BMD among
women in a substudy of the VOICE trial.
Methods: HIV negative women in Uganda and Zimbabwe on oral
TDF, FTC/TDF, or placebo had BMD measurements of lumbar spine
and total hip via dual energy x-ray absorptiometry (DXA) at baseline
(BL) and every 24 weeks (w) until 48 w after study product completion.
Plasma tenofovir levels were assessed every 12 w for the first 48 w.
Student’s t-test and regression models were used to compare the mean
percentage (%) change from BL in hip and spine BMD between the
active and placebo arms.
Results: Of 518 women enrolled, 432 (155 Uganda, 277 Zimbabwe)
with DXA results at both BL and w 48 entered this analysis. Median
age and BMI at BL were 28 years and 24.6 kg/m2. History of depot
medroxyprogesterone acetate (DMPA) use was reported by 123 (28%)
and 94% had moderate to high BL physical activity levels. In the primary
analysis, there were no significant differences between arms in hip or
spine BMD % change, likely due to low product use in VOICE. Tenofovir
was detected in 75-100% of plasma samples in 43/118 (36%) TDF and
50/156 (32%) FTC/TDF recipients (median 4 samples tested). In these
women, mean % change from BL to w 48 in spine BMD was 1.0% to
1.3% lower than placebo and hip BMD was 0.5% to 0.8% lower after
adjusting for country, age, BMI, physical activity level, and history of
DMPA use (p< 0.05 for FTC/TDF and for oral active arms combined, but
not TDF). BMD increased after stopping product; 48 w change was 0.61.2% higher in TDF and FTC/TDF vs placebo recipients.
Conclusions: Small but significant reversible decreases in BMD were
observed among young African women with higher adherence on TDFbased oral PrEP. The observed differences were in the range seen in prior
studies of HIV-negative men and women.
366
Background: Randomised controlled trials rely on effective preservation
of the random allocation of interventions when tested against placebo for
a robust outcome. Product sharing among participants can compromise
the trial outcome. In resource poor settings, many participants attend
study visits together, travel together and often are neighbours or live
in the same household. In such circumstances product sharing poses a
real concern. We describe incidents of product sharing across 7 clinical
research sites in Durban conducting the VOICE trial and strategies
implemented to avert such occurrences.
Methods: The Durban sites enrolled 2750 women with 1647 and 1103
participants randomised to the oral and gel arms respectively. Product
use assessments were conducted at monthly visits. Product labels with
unique identifiers and product codes were checked to verify return of
correct products. Discrepancies prompted discussions with participants
and strategies were developed by pharmacists. Monthly product use
counselling with product sharing messaging was provided to participants.
Results: Thirty two incidents of product sharing were identified: 18
incidents involved participants returning products not assigned to them
and 14 incidents included product count discrepancies. Discussions with
participants suggested that the main source of product sharing involved
women residing in the same household with participants, 20/32 incidents
(62.5%). Four participants also reported that visitors had access to their
product. Topical gels were more commonly shared than oral tablets (72
% vs. 28 %). Investigations were done to rule out pharmacy dispensing
errors. Counselling, tailored messaging and unique identifiers were used
to address product use challenges.
Conclusions: In this trial, product sharing was common among women
residing in the same household. Unique labelling of products helped
avert repeat incidents of product sharing. For future trials, targeted
counselling and unique product identification is recommended.
P42.07
P42.08
An Exploration of Young Women´s
Perceptions and Experiences of Participating
in HV Vaccine Clinical Trials in Nyanga
Township, Cape Town
Racial Differences in Willingness to Participate
in HIV Prevention Clinical Trials amongst
University Students in KwaZulu-Natal, South
Africa
Norah Nandudu1, Johannes John-Langba1,2
Diantha Pillay1, Douglas R Wassenaar1
University of Cape Town, Department of Social Development, Cape
Town, South Africa, 2The Cairns Institute, Cairns, Australia
1
Background: HIV/AIDS has severely inflicted suffering on the global
population and reported to be the worst killer disease in sub-Saharan.
Because other preventive measures such as condom use among young
people is still low and less effective in preventing the spread of the
disease. On that basis, it was recommended by the United Nations
General Assembly Special Session on HIV/AIDS (2001) to accelerate the
development of HIV vaccine aimed at curbing the disease. This study
sought to explore young women’s experiences and perceptions about
HIV prevention vaccine clinical trials so as to inform the design and
implementation of vaccine trials in Africa.
Methods: The study employed purposive sampling to select participants
and qualitative research design was used to interview participants using
semi-structured interview schedule. A tape recorder was used to capture
data and coding procedures were used to analyze data.
Results: Findings drawn from participants’ responses and compared with
literature from previous studies on vaccine trials and social development
theories indicate that participants decide to join HIV prevention vaccine
clinical trials because they hope to be protected from HIV infection.
Most importantly participants hope to get access to medical care and
treatment, meanwhile some participants perceived HIV vaccines harmful
to humans hence they usually decline to participate. The study also
identified study participation challenges related to socio-cultural and
historical aspects.
Conclusions: Although vaccines have had some success stories in the
prevention and control of infectious diseases such as the eradication of
polio, smallpox and measles, prevailing challenges need to be addressed
if vaccine development is to be feasible. Providing more information,
reinforcement of community awareness and mobilization around
issues of HIV vaccine clinical trials at all levels of vaccine design and
implementation is required to ensure appropriateness and acceptability
of vaccine trial participation.
UKZN, Durban, South Africa
Background: Willingness to participate in clinical trials is a crucial
element in recruitment of suitable participants for intervention trials.
Measuring willingness to participate helps determine community
preparedness for clinical trials. Researchers in the USA developed a
Clinical Research Involvement Scale (CRIS) assessing willingness to
participate modelled on the Theory of Reasoned Action.
Methods: This study aimed to determine racial differences in willingness
to participate and explore potential factors associated with willingness
to participate in HIV prevention research using the CRIS. The CRIS was
administered online with a demographic questionnaire to university
students aged 18-45 at the University of KwaZulu-Natal, South Africa.
Associations between willingness to participate and age, gender,
relationship status, parity, religion, education status, student status,
employment status and access to private health care were examined.
Results: The study enrolled 636 participants, two thirds being female.
After data cleaning a sample of 509 was considered for analysis.
Results indicated that all students across all race groups were willing
to participate in HIV prevention research. However, there was a
statistically significant difference in factors affecting willingness of
participate. Based on the differences amongst these factors, Black
students expressed greater intention to participate compared to White
and Indian students. Racial differences in factors that affect willingness
to participate indicate differences in risk perception and seeking access
to better quality healthcare.
Conclusions: The CRIS is a reliable instrument in this population;
however in its current structure it does not show strong validity. Validity
improved if factors of motivation to comply and outcome evaluations
were removed. The CRIS should be used in other populations to assess
its validity.
www.hivr4p.org
367
POSTERS
1
Posters
P42.09
P42.10
Use of Visual Education Aids to Improve
Participant Understanding of HIV Prevention
Trials
Preparing for HIV Prevention Trials: From
Training to Service Delivery-lessons Learned
with Long Acting Reversible Contraception
(LARC) in Zambia
Nonzwakazi Mnqonywa1, Renee Street1
Medical Research Council, HIV Prevention Unit, Westville, South Africa
1
Background: The HIV Prevention Research Unit (HPRU) based in
KwaZulu-Natal (South Africa) has been involved in numerous HIV clinical
trials and has tested a range of products including pills, gels, diagrams
and rings. Over the years, visual aids have been created to improve
participant understanding of the study product, design and procedures
Methods: The use of educational materials such as flip charts and booklets
were developed to go hand in hand with visual aids to overcome potential
language barriers as well as to compensate for level of education when
explaining various aspects of the clinical trials. Visual aids are designed
by Family Health International in conjunction with input from clinical trial
research staff before they are sent for ethical approval.
Results: The study population typically consists of sexually active
females between the ages of 18-45. The socio-cultural, economic
& demographic determinants of the participants vary and level of
education ranges from illiterate to highly educated. The majority of
participants have a clearer understanding of the purpose of the study
after using visual aids. Through the years, visual aids have evolved from
pictures to real life models. These include sample applicators, vaginal
rings, dildos to demonstrate condom use, pelvic model and other
different types of family planning methods. Some studies use an audiocomputer-assisted-self interview system (ACASI) which includes visuals
to make understanding and operating more user friendly.
Conclusions: Illiteracy is no longer an excuse for HIV prevention
education. HIV clinical trial participants, their male partners and the
communities at large have demonstrated a clear understanding of study
products, designs and procedures with the use of visual aids.
POSTERS
368
Mubiana Inambao1, Naeemah Munir1, Kathleen Wu2, Danielle
Dang2, Nurilign Ahmed3, Bethelihem Getachew4, William
Kilembe2, Bellington Vwalika2, Amanda Tichacek5, Susan Allen5
Rwanda Zambia HIV Research Group, Zambia Emory HIV Research
Project, Ndola, Zambia, 2Rwanda Zambia HIV Research Group, Zambia
Emory HIV Research Project, Lusaka, Zambia, 3Rwanda Zambia HIV
Research Group, Projet San Francisco, Kigali, Rwanda, 4Rwanda Zambia
HIV Research Group, Emory University, School of Medicine, Atlanta, GA,
United States, 5Rwanda Zambia HIV Research Group, Emory University,
Pathology & Laboratory Medicine, School of Medicine, Atlanta, GA,
United States
1
Background: Zambia has a high burden of mother-to-child transmission
of HIV. The provision of long-acting reversible contraceptives (LARC),
specifically intra-uterine devices (IUDs) and Jadelle implants, is critical
to prevent unplanned pregnancy and perinatal transmission in HIV+
women and is also necessary to prevent pregnancy in HIV- prevention
trial participants. The UK Department for International Development has
funded a program to avert an estimated 13,900 new HIV infections
through integrated PMTCT with LARC and HIV couples-focused HIV
testing.
Methods: LARC training for providers began with a 3-day didactic
training followed by supervised IUD and Jadelle insertion and removal
practicums. Those showing proficiency in all 4 categories were
considered fully certified. Practicums were conducted at government
clinics in Copperbelt, Lusaka and Southern Province.
Results: From June 2013 to March 2014, ZEHRP trained 199 LARC
providers from 45clinics. 91% were certified in Jadelle insertions, 70%
in Jadelle removals, 55% in IUD insertions, 46% in IUD removals and
43% were fully certified. The main obstacles to completion of practicums
were not being assigned to the family planning department when one
of the few trainers was available, and coordinating these times with
availability of interested LARC clients. A Training-of-Trainers was held
for the fully certified nurses who were high performers, meetings were
held with clinic in-charges to coordinate trainee schedules, and satisfied
clients were recruited to give personal testimonials.
Conclusions: IUD and implants are rarely used in Africa, primarily
because lack of supply leads to lack of demand. This model illustrates an
effective way to harness successful LARC trainees as trainers and work
with clinic administrators and satisfied clients to coordinate practicum
training and demand creation. Repeating this cycle leads to a snowball
effect of progressive and simultaneous increase in supply and demand,
resulting in PMTCT and trial-ready clients with LARC.
P42.11
Implementation of an Adherence Counseling
Intervention in a Microbicide Trial: Challenges
in Changing Counselor Behavior
Iván Balán1, Alex Carballo-Diéguez1, Rebecca Giguere2, Javier
Lama3, Ross Cranston4
NYSPI/Columbia University, HIV Center for Clinical and Behavioral
Studies, New York, NY, United States, 2New York State Psychiatric
Institute, HIV Center for Clinical and Behavioral Studies, New York, NY,
United States, 3Asociacion Civil Impacta Salud y Educacion, Lima, Peru,
4
University of Pittsburgh Medical Center, Division of Infectious Diseases,
Pittsburgh, PA, United States
1
POSTERS
Background: Adherence counseling interventions have become
a critical component of biomedical HIV prevention trials. Yet, little is
known about how well counselors learn and deliver them. We present
interim findings about implementing the participant-centered adherence
counseling intervention being used in MTN-017.
Methods: Counselors at seven study sites completed two days of training
on delivering the standardized intervention. Next, they conducted two
practice sessions, at least one of which had to meet pre-determined
fidelity criteria for approval to see study participants. All sessions were
audio-recorded. For each counselor, the practice sessions, the first ten
study sessions, and one of every five subsequent sessions were rated
for fidelity. Ratings were used to guide monthly group coaching sessions
with the counselors.
Results: Twenty-six counselors completed the two-day training. For 15
counselors (58%), both practice sessions met fidelity criteria; for seven
counselors (27%), one session met criteria; and for four counselors
(15%), neither did. To date, 226 of 227 counseling sessions conducted
have been recorded. Of the eight counselors who have completed ten
study sessions, six met fidelity criteria on 80% or more of their sessions,
but two met criteria on 50% or less of the sessions. Fidelity ratings
varied over time. Fidelity ratings of the individual components of the
sessions suggest that lower ratings were mostly attributable to skipping
components of the session, not to conducting them incorrectly.
Conclusions: Recording adherence counseling sessions during
biomedical trials is feasible and acceptable to counselors and participants.
After 2 days of training, the majority of counselors, though not all, were
able to accurately deliver the intervention. Over time, counselors appear
vulnerable to lapses in fidelity. Findings highlight the need to assess
efficacy of training for such interventions and for continued monitoring
and coaching throughout the study to ensure fidelity.
www.hivr4p.org
369
Posters
Posters 43: Policy, Advocacy and Modeling
P43.01
P43.02
Should the United States Policy that Banned
MSM from Donating Blood be Repeal?
Streamlining Operational Processes by
Building a Centralized HIV Clinical Research
Centre
Olanrewaju Adedokun1
Southern Connecticut State University, Public Health, New Haven, CT,
United States
1
Jacqueline S. Burgess1, Krishnaveni Reddy1, Pranitha
Ramchuran1, Clare Dott1, Thesla Palanee1, Helen Rees1
Wits Reproductive Health & HIV Institute, School of Clinical Medicine,
University of the Witwatersrand, Johannesburg, South Africa
1
POSTERS
Background: Recently, the American Red Cross, which is the largest
single supplier of blood and blood products in the United States, issued
an emergency request for donors of all blood types because its blood
donations were down by about 10 percent across the country, with
about 50,000 fewer donations than expected. This led to the question
of whether the United States policy that banned men who have had sex
with other men (MSM), at any time since 1977 from donating blood
be repealed to allow gay and bisexual men with no HIV infection, who
meet the other conditions and are willing to donate blood be allowed to
do so. That permitting blood donation from the eligible MSM could help
reduce blood supply shortage in the United States. However, the initiator
of that policy, U.S. Food and Drug Administration maintained that the
life time ban will not be eased unless there is an evidence-based data
showing that a repeal of existing policy is necessary and will not put
the life of blood recipients at a significant risk of acquiring HIV infection
through blood transmission.
Methods: This study analyze the views of the interested stakeholders
of this issue by weighing the evidences presented by those who want
the FDA’s current policy to be repealed with the evidences of those who
does not.
Results: Having reviewed all the parties’ views in this case, three options
were identified and they are as follows: Not repeal the FDA’s current
policy (life time ban); Repeal the FDA’s current policy (life time ban) and
replace it with one- year deferral; and Repeal the FDA’s current policy
(life time ban) without deferral option.
Conclusions: Having carefully reviewed all the available evidences, the
best option would be to repeal the current policy (life time ban) and
replace it with one- year deferral. This option addresses the concerns
of every party involved. It safeguards the life of patients who received
donated blood from HIV infection and removed life time ban on the gay
community to one year deferral.
370
Background: Wits RHI is situated in Hillbrow, Johannesburg, a model
community to conduct HIV clinical studies, with its high prevalence
rates of HIV and developing infrastructure. Wits RHI has created a multifunctional Research Centre (RC) where 12 clinical and observational
research studies have been conducted over the past 4 years. Given the
financial constraints on investigator driven research and clinical trials,
the RC has formulated centralized operational services that support all
ongoing studies.
Methods: The centralized operations of the RC involves the sharing
of data, laboratory, pharmaceutical, regulatory, administration and
maintenance capacities across all studies. This includes the sharing of
fixed costs in terms of infrastructure, licensing, and equipment; offering of
voluntary counselling and testing; provision of concomitant medication;
shared data, administration, and laboratory centers; cross-trained
staff for back-up purposes; and support from centralized maintenance
and operational teams. The RC uses the MRC Biometric Co-Enrolment
Prevention System, which detects if participants are enrolled in other
studies, to eliminate the possibility of co-enrolments. The Division of
AIDS standards is used as a benchmark for quality across all studies.
Management decisions are aimed at balancing the needs of individual
studies and of the site.
Results: Centralizing services improves cost efficiencies for all studies,
while providing access to optimized laboratory, data, regulatory support,
clinical staff and equipment, pharmaceutical services, site infrastructure
and staff capacity. This approach has resulted in improved quality of
the site’s study implementation and overall outputs and has reduced
duplication of efforts in the organization.
Conclusions: With the current limited funding available for HIV clinical
research, developing centralized processes for research sites is a potential
mechanism for reducing costs and optimizing outputs and of benefit for
sites with financial limitations.
P43.03
P43.04
Policy Dialogue to Enhance the HIV Response
among MSM and Transwomen in Peru
through Combined Prevention Strategies: A
Baseline Stakeholder Assessment
Forging Consensus in the HIV-positive
Community on the Use of Treatment as
Prevention
Cayetano Heredia University, Unit of Health, Sexuality and Human
Development, Lima, Peru
1
Background: Despite significant changes in the HIV epidemic landscape
among MSM and transwomen (TW) to date, the national program
remains essentially as it was designed in the late 1990s, with limited
discussion of potential changes. Together with the Ministry of Health,
we are conducting, documenting and analyzing a process seeking to
enhance the program with combined prevention strategies, through
stakeholder involvement, health systems assessments and mathematical
modelling. Here we present the background stakeholder analysis.
Methods: We conducted in-depth interviews and focus groups
with community members (MSM/TW), in-depth interviews with
community leaders, health providers and decision makers, and two
policy discussion workshops (with civil society and government
representatives, respectively).
Results: Among community members and stakeholders alike, condom
use is still the epitome of prevention, and its limited use is explained
only as resulting from poor individual commitment and insufficient
prevention work. Knowledge of ETfP is almost absent, but there is some
knowledge of PrEP, together with resistance to its potential use. The
lack of information, misconceptions and confusion reflect limited public
discussion on new HIV prevention technologies. On an operational
level, participants consider the need of well-trained health providers as
part of a renovated program. People agreed with the idea of comibined
biomedical, behavioral and structural strategies based on appropriate
evidence. All recognized the need for continuing professional HIV
educaiton, and also for spaces to evaluate and improve current
programming. Renovated leadership will play a key role.
Conclusions: Community members and stakeholders in Peru agree on
the need to bring the HIV prevention response to date among MSM/TW
with combined strategies. Many feel that HIV prevention programming
is loosing momentum and needs new leadership and tools, and are
interested in an evidence-based process where the HIV response can be
reconceptualized.
NAM Publications, Editor, London, United Kingdom, 2European AIDS
Treatment Group, Brussels, Belgium, 3Waverley Care, Edinburgh, United
Kingdom, 4NAM Publications, London, United Kingdom, 5FHI 360,
Washington, DC, United States, 6Global Network of People Living with
HIV (GNP+), Amsterdam, Netherlands, 7AIDS Healthcare Foundation,
Amsterdam, Netherlands, 8Positive Voice, Athens, Greece
1
Background: HIV treatment policy has been moving closer towards
access to ART for all. This has not been universally welcomed among
HIV advocates.
Concerns have centred around whether TasP could lead to compulsory
and oppressive test-and-treat (T&T) policies and whether TasP is the
best use of resources when many needing ART for life still do not get
it. These exist alongside enthusiasm for TasP and determination that
disadvantaged populations should benefit from it.
Such tension in the community has led to the lack of a simple, strong
community-led policy on TasP.
Methods: EATG (www.eatg.org) and NAM (www.aidsmap.com) devised a
statement of broad principles for any TasP programme.
This sign-on statement is a guide to ensure that all ART programmes
centre on the interests of people with HIV.
Devising the statement was an exercise in forging community consensus.
The statement went through consultation with the EATG membership, a
public consultation on www.aidsmap.com, an international community
meeting and final consultation with key opinion leaders.
It was published in February 2014 on www.hivt4p.org.
Results: Feedback during consultation said that the statement was too
technical and its authorship and intended audience unclear; it needed
to be sited clearly within a human rights background; there should be
more mention of gender and power issues; and it needed to support
existing effective prevention methods.
Sections on the evidence for TasP and research needs became separate
appendices.
The statement now consists of the main sign-on statement of 30 clauses,
an 11-page fully-referenced statement including the appendices and a
Key Points summary.
Conclusions: The statement was presented at the 20 TasP Workshop
and in its own session at the International AIDS Conference and of an
article in Communicable Infectious Diseases. We aim to have the statement
incorporated into future guidelines and to work with partners to ensure
that TasP is firmly grounded in the rights of people living with HIV.
www.hivr4p.org
371
POSTERS
Carlos F. Caceres1, Ximena Salazar1, Alfonso Silva-Santisteban1,
Aron Nunez-Curto1
Gus Cairns1,2, Brian West2,3, Caspar Thomson4, A Cornelius Baker5,
Anna Zakowicz6,7, Nikos Dedes8
Posters
P43.05
P43.06
Do Civil Society Advocates’ Priorities Align
with HIV Prevention Research Landscape?
Making Treatment as Prevention Work
for People with Disabilities across the HIV
Prevention and Care Continuum in Zambia
Lisa Jacobs1, Manju Chatani1, Angelo Kaggwa-Katumba1
1
Clever Chilende1
Treatment Advocacy and Literacy Campaign (TALC), Programmes,
Lusaka, Zambia
1
POSTERS
Background: HIV prevention research-to-rollout processes require
effective community advocates who understand the evolving landscape.
The AVAC Advocacy Fellowship, launched in 2009, supports advocates
to engage around these issues in their countries. AVAC sought to
understand how applicants’ priorities relate to developments in the field.
Methods: Desk analysis was conducted on 284 eligible applications of
over 400 submitted in the five years of the program to determine which
interventions were of greatest interest each year and if any patterns
emerged related to events in the field.
Results: Voluntary medical male circumcision (VMMC) was identified
in 91 applications, 80 of them received from 2009-2011. Interest
waned as countries ramped up, and PEPFAR began funding, VMMC
programs. Microbicides were identified in 58 applications; however, 23
applications were received in 2009 and only 9 applications in 2010 after CAPRISA 004 results were announced and there was confusion
about what happens next. PrEP was identified in 57 applications with
a surge in 2012 after the FDA approved Truvada for PrEP. Treatment as
prevention was identified in 41 applications, all of which were received
in 2011-2012 after HPTN 052 results were announced. While vaccine
research has received relatively little interest, it was highest in 2009 just
after RV144 showed positive results.
Conclusions: Applicants’ interests seem to follow the events in the
field, demonstrating that advocates are aware of and interested to
localize global events. Since AVAC amends its application materials
every year and highlights priorities on its website, fluctuations in choice
of interventions may reflect a selection bias. However, the trends are
dramatic and reflect the evolving issues and interests of applicants to
the Fellows program. Since AVAC can only offer a few Fellowships each
year, more efforts are needed to support the large and informed pool
of HIV prevention research advocates who are interested in engaging
in the field.
372
Background: Since 2005, Zambia has developed a national universal
HIV prevention and testing program with free antiretroviral. Although
prior research in sub-Saharan Africa has shown that people with
disabilities are highly vulnerable to HIV, little research on this population
has been done in Zambia, and the national HIV program has only
recently included people with disabilities as a vulnerable population in
its national strategy.
Methods: In-depth interviews with adults, children and adolescents
with physical, sensory and psychosocial disabilities were conducted in
Zambia. Caregivers and parents of children with intellectual disabilities
were also interviewed, as were health workers, special education
teachers and representatives of disabled persons organizations and HIV
organizations. Legal and policy analysis was also conducted.
Results: Persons with disabilities experience pervasive stigma and
social marginalization compounded by economic, communication and
physical barriers to access to HIV services throughout the HIV care
continuum in Zambia. HIV prevention education and standard protocols
for assuring informed consent and confidentiality in the provision of
HIV testing and encouraging ART adherence need to be tailored to the
specific needs of individuals with disabilities. Existing HIV and health
policies do not identify specific strategies and interventions to address
people with disabilities. The equal right of people with disabilities to
sexual and reproductive health is not recognized and they remain
invisible to social protection services.
Conclusions: Efforts to achieve universal access to HIV prevention and
treatment in Zambia are undermined by the lack of programs addressing
the needs of individuals with disabilities, and current policies and
programs on HIV and disability lack adequate integration. Recognition
of people with disabilities as a highly vulnerable group within the
national HIV response has not been translated into implementation of
inclusive and targeted services.
P43.07
P43.08
Not without us: Mobilizing Communities to
Advocate for Women´s Involvement in PrEP
Implementation
Use of a Mathematical Model to Predict
Dissolution Profiles of Dapivirine Vaginal
Rings
Dazon D. Diallo1
Christopher Gilmour1, Stephen Ampofo1, Brid Devlin1
Background: As the US FDA considered the approval of the use of
Truvada as PrEP, women in the US organized to address the need
for greater community engagement in the implementation science
informing the introduction of the intervention in clinics, community
health centers and private care.Through an ad hoc coalition, US
women’s communities advocating for safe HIV prevention options for
women, including PrEP, convened a strategic working group dedicated
to providing input, community-based qualitative research , and
innovative strategies research to determine how to help women and
their healthcare providers make PrEP a viable option.
Methods: Community engagement in research includes activities beyond
the involvement of volunteers in trials, community advisory boards and
local partnerships with service organizations.Our methodology included
a diversity of initiatives including: informational webinars/community
symposia, key informant interviews, qualitative study with focus groups
and surveys; implementation advocacy; engaging federal partners.
Results: The work resulted in the following: convened 3 national/
international webinars for a total of 850 participants; captured qualitative
data on knowledge, attitudes and beliefs of 85 women at high risk for
transmission; published several articles/op-eds in newsprint; presented
papers, posters and workshops at national and international meetings; and
sustains a working group of nearly 80 interested parties/organizations.
Conclusions: The implementation of research, such as PrEP, requires
meaningful involvement and representation from the community most
affected by the research outcomes.In the US, and key countries where
PrEP is a priority, women’s education, understanding and acceptance of
PrEP is an important step to its use as a prevention option.The community
mobilization of women’s health/HIV advocates, clinical and behavioral/
social scientists & WLwH is a key component of engaging community in
research of a product before and after it has been approved.
International Partnership for Microbicides, Product Development, Silver
Spring, MD, United States
1
Background: The dapivirine vaginal ring is a sustained release
formulation containing dapivirine, a potent antiviral currently in clinical
evaluation for HIV prevention. Each ring contains 25 mg of dapivirine
in a platinum-cured silicone matrix. In vitro dissolution testing quantifies
the amount of dapivirine released from the ring over the 28-day use
period, but is labor intensive, time consuming and expensive. This paper
describes development of a mathematical model to predict average
cumulative drug release of a batch.
Methods: Empirical approaches and mechanistic considerations were
used to modify the Higuchi equation so as to assess the amount
of dapivirine released per unit area of the vaginal ring. The model
utilizes details of ring geometry, ring assay and a single attribute of
the silicone elastomer:
Q = (2A)1/2(Cp*Dp)1/2 X {polynomial (order 2) of t} Where
Q = amount of active ingredient released per unit area
A = concentration of active ingredient in the matrix
Cp = solubility of active ingredient in the matrix
Dp = diffusion coefficient of the active ingredient in the matrix
t = time, set at 281/2
The model was assessed for applicability to batch averages. Average 14day results were also used to estimate (Cp*Dp)1/2 for individual batches in
order to improve the model.
Results: The model resulted in errors typically within the range
+/- 4% for the predicted 28-day average cumulative release versus
actual average cumulative release. The accuracy of the model was
improved further by using the 14-day results to estimate (Cp*Dp)1/2 for
individual batches.
Conclusions: Data from mathematical models indicate that ring
attributes or results from earlier time points can predict 28-day
dissolution in terms of average cumulative amount of drug released per
batch or from an individual ring. These predictions provide a reliable and
efficient measure for product quality control.
www.hivr4p.org
373
POSTERS
SisterLove, Inc., Atlanta, GA, United States
1
Posters
P43.09
P43.10
Trends of Advocacy Journalism; a Case of the
HIV/AIDS Story in the Ugandan Press
Tenofovir Diphosphate Concentrations in
Human Vaginal Stroma after Different Dosage
Regimens with a Vaginal Gel: A Modeling
Approach
Kakaire A. Kirunda1,2, Angelo Kaggwa- Katumba3
Islamic University in Uganda, Mass Communication, Kampala, Uganda,
Makerere University College of Health Sciences, School of Public
Health, Health Policy Planning and Management, Kampala, Uganda,
3
AVAC: Global Advocacy for HIV Prevention, New York, NY, United
States
1
2
POSTERS
Background: The genre of advocacy journalism is one which journalists
can employ to advance HIV prevention efforts ranging from policy to
practice. Advocacy journalism deliberately but transparently adopts a
non-objective viewpoint, usually for some social purpose. Usually, in
advocacy journalism, practitioners have an opinion about the story they
are writing. We therefore sought to establish the extent to which the
strategically placed front page HIV/AIDS stories in Uganda’s leading
daily newspapers are a product of advocacy journalism.
Methods: A retrospective desk review of the Daily Monitor and New
Vision newspapers covering the period January 2013 through December
2013 was used. The two newspapers were purposively selected and
HIV/AIDS articles obtained from the cover pages were coded against
a set of variables. Two key informants were also interviewed. Data was
entered into SPSS (Statistical Package for the Social Sciences) software
for analysis while qualitative data from the articles was analysed using
thematic content analysis.
Results: The sample for both newspapers yielded a combined 2154
articles on the front pages. However, findings indicate that out of these,
only 28 articles (1.3%) were about HIV/AIDS with the Daily Monitor
having 16 while New Vision carried only 12. Overall, most of the articles
had an element of advocacy journalism. In the New Vision, 9 (75%)
front page articles were slanted towards advocacy, while in the Daily
Monitor this was exhibited in 14 (88%) articles. In both newspapers,
the journalists systematically employed a positive tone, the choice of
facts supported the cause the headlines were fronting and sourcing for
the articles was favourable. All these attributes are synonymous with
advocacy journalism.
Conclusions: With majority of HIV/AIDS articles that make it to the front
pages being products of advocacy journalism, prevention advocates
need seize the opportunity. Ways of working with the authors of these
articles should be devised to tap into advocacy opportunities.
374
Yajing Gao1, Andrew Yuan1, David F. Katz1,2
Duke University, Durham, NC, United States, 2Duke University Medical
Center, Obstetrics and Gynecology, Durham, NC, United States
1
Background: Different vaginal dosing (BAT24, daily) of the 1% Tenofovir
(TFV) gel gave differing pharmacokinetics and prophylactic efficacy.
The kinetics of stromal Tenofovir diphosphate (TFV-DP) production/loss
are governed by many factors, e.g.: mass transport kinetics of TFV to
stroma; concentration distribution of stromal host cells; TFV binding and
phosphorylation rates to/in host cells; and TFV-DP clearance rate in host
cells. Experimental PK studies give guidance on creation and persistence
of stromal TFV-DP levels, but optimization of dosing to achieve such
levels is limited. Modeling this process complements experimental
studies, providing insights on how the multiple factors govern TFV-DP
levels and suggesting optimal dosing strategies.
Methods: A mechanistic, mass transport-based model was created of
TFV delivery to human vaginal mucosa by a spreading gel, and coupled
TFV-DP production in stromal host cells. Parameters in the model were
obtained from in vitro measurements of gel rheology and TFV transport
properties, human vaginal morphometric and histological data, and
results of human PK studies for the 1% TFV gel. Single and multiple gel
application regimens were studied, including coitus.
Results: Results show, for example, that application of two doses 4 hours
apart (as found in BAT24) gives 40% higher maximum stromal TFV-DP
concentration than daily dosing. This elevated TFV-DP concentration is
sustained for 4 days vs daily dosing (due primarily to the long half life
of TFV-DP).
Conclusions: Modeling provides additional insights about interactive
effects of the multiple factors that govern optimal dosing by the TFV gel.
Results here illustrate pharmacokinetic benefits of multiple dosing within
a few hours vs sustained daily dosing. This approach can be used to help
optimize dosage regimen, accounting for factors such as gel volume and
drug loading.
P43.11
P43.12
Persistent HIV-related Stigma in Rural Uganda
during a Period of Increasing HIV Incidence
Using Advocacy, Guideline and Policy Change
to Increase Treatment Demand in Various
Parts of Africa and Asia
1
Massachusetts General Hospital, Boston, MA, United States, 2Brigham
and Women’s Hospital, Boston, MA, United States, 3UCSF, San
Francisco, CA, United States, 4Epicentre, Mbarara, Uganda, 5Fenway
Health, Boston, MA, United States
Background: Uganda is the only country in sub-Saharan Africa with
increasing HIV incidence during the period 2002-2013. Because HIVrelated stigma is associated with reduced uptake of HIV testing, increased
risk-taking behavior, decreased adherence to anti-retroviral therapy
(ART), and reduced HIV status disclosure, it is critical to understand how
changes in HIV incidence have correlated with HIV-related stigma during
this period.
Methods: We analyzed data from two sources:
1) the Uganda AIDS Rural Treatment Outcomes (UARTO) study during
2007-2012 and
2) the Uganda Demographic and Health Surveys (DHS) from 2006 and
2011 to estimate trends in internalized stigma among people living with
HIV (PLHIV) at ART initiation and trends in stigmatizing attitudes and
anticipated stigma among the general population. We fit regression
models adjusted for socio-demographic characteristics, with year of
cohort entry/DHS as the primary explanatory variable.
Results: Among people initiating ART in the UARTO study, we found
a statistically significant positive association between year of ART
initiation and internalized stigma score (adjusted b=0.17; 95% CI, 0.05
to 0.29), suggesting an 11% relative increase in the mean score in each
year of recruitment after the first year. From 1.4 (out of 4) at baseline,
the mean stigma score increased to a peak of 2.2 in 2011. Among the
general population comparing 2011 with 2006 DHS data, we found a
significantly higher odds of reporting anticipated stigma (adjusted OR
= 1.12; 95% CI, 1.08 to 1.17), despite a decreased odds of reporting
stigmatizing attitudes (adjusted OR = 0.90; 95% CI, 0.87 to 0.93).
Conclusions: Mean internalized stigma at ART initiation increased over
time among PLHIV in a rural Ugandan cohort in the setting of worsening
anticipated stigma among the general population. Further study is
needed to better understand the reasons for increasing HIV-related
stigma and its impact on HIV prevention efforts in Uganda.
Maureen Akumu Milanga1, Ntando Yola2, Oladayo Taiwo
Oyelakin3, Peter Michira4, Rumbidzai Mapfumo5, Josephine
Kamarebe6, Cai Lingping7
AIDS Law Project, Nairobi, Kenya, 2Networking HIV/AIDS Community
of South Africa, Cape Town, South Africa, 3Positive Action for Treatment
Access, Lagos, Nigeria, 4Partners in Prevention PrEP Thika Study Site,
Thika, Kenya, 5Centre for Sexual Health and HIV/AIDS Research,
Harare, Zimbabwe, 6Health Development Initiative, Kigali, Rwanda,
7
China HIV/AIDS Information Network, Beijing, China
1
Background: Through the AVAC fellowship, selected advocates from
Kenya, Rwanda, Nigeria, Zimbabwe, South Africa and China worked
with the aim of empowering the population with knowledge and to
advocate for treatment as prevention in their countries that targeted the
community and policy makers.
Methods: Advocacy took place in the span of a year where three
advocates from Kenya, Rwanda and China worked to increase the
demand and access of PrEP in their countries. They targeted key
populations consisting of sex workers, gay men, male sex workers
and sero-discordant couples, policy makers and civil society. In the
communities they created awareness for the program among the
recipients and other non-governmental organizations and advocated for
government to adopt the program for the country.
In Zimbabwe, South Africa, Nigeria and Kenya, four advocates increased
demand and awareness of treatment as prevention and good participatory
practice in clinical trials. The Kenya, Zimbabwe and Nigeria advocates
increased demand by advocating for early initiation of people living
with HIV, initiation of sero-discordant couples regardless of CD4 count
in Kenya, increase of post exposure prophylaxis access to sex workers
in Zimbabwe and in South Africa increase of effective engagement of
communities in trials through the development of treatment guidelines
and change in policy in Kenya, Zimbabwe and Nigeria and engaging
researchers and communities is south Africa.
Results: There is need for scale up of PrEP programs in countries with
key populations as consultations in the targeted countries showed
a demand for the services. There is need to create more community
engagement in programs to also increase access among populations.
Early initiation and lifelong treatment among mothers is timely and the
demand among communities is also high, strides should be taken to
fully roll out these programs.
Conclusions: Civil society groups in other countries should also seek to
create demand among communities on new prevention technologies.
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375
POSTERS
Brian T. Chan1,2, Sheri D. Weiser3, Yap Boum4, Mark J. Siedner1, A
R. Mocello3, Jessica E. Haberer1, Peter W. Hunt3, Jeffrey N. Martin3,
Kenneth H. Mayer5, David R. Bangsberg1, Alexander C. Tsai1
Posters
P43.13
P43.14
Ending HIV among Men who Have Sex
with Men (MSM) in Kenya: Community
Recommendations on Scaling up HIV
Prevention and Treatment
Using Advocacy for the V condom - A New
Female Condom Coming to South Africa Building Support for Female Condoms as an
MPT
Jeffrey W. Wambaya1, Leonard Mutisya2, Gaudensia Mutua3, Jack
Ndegwa4, George O. Owino5
Marion Lynn Stevens1,2, Penny Parenzee1, Jane Bennett2,
Kimberly Whipkey3
Ishtar MSM, Nairobi, Kenya, 2UHAI EASHRI, Nairobi, Kenya, 3KAVI,
Nairobi, Kenya, 4KANCO, Nairobi, Kenya, 5IAVI, Nairobi, Kenya
1
1
POSTERS
Background: MSM in Kenya contribute 15% of new HIV infections
yearly even when only 1% of men report same-sex sexual activity.
There has been no conclusive participatory strategy that has engaged
MSM nationally in developing HIV prevention responses for and by
MSM. Kenyan MSM community organisations purposed to design and
advocate for a combination package to advise planning, coordination
and evaluation of HIV prevention, treatment, research and advocacy and
development of the Kenya National AIDS Strategic Plan IV 2014-2019.
Methods: Between August and December 2013 with leadership of
NACC and NASCOP and support from Ishtar MSM, IAVI and LVCT Health,
participatory community discussions with 27 MSM-serving organizations
designed a package of approaches to ending HIV among MSM.
Results: There was consensus that a combination package for MSM be
defined and guidelines written. The package must prioritize strategies
to improve outreach and access to HIV testing, adopting a find-testtreat-retain model to immediately link HIV-infected MSM to treatment
regardless of CD4 count. Additionally, MSM be provided a broad range
of prevention options including scaling up access to condoms and
condom-safe lubricants, STI testing and treatment, post-rape care, PrEP
and PEP. Legal reforms and democracy are critical to fighting stigma and
discrimination in the public and among service providers. We must also
increasingly and meaningfully engage MSM in critical decision-making
in service delivery and research design and implementation, including
on PrEP, vaccines and microbicides.
Conclusions: Funding, research and advocacy strategies informed
by community insight should be prioritized to make HIV prevention
investments efficient and sustainable. MSM-led health organizations
should be supported to better sustain their work and to integrate and
scale up biomedical interventions and research in their existing package
of behavioural and structural interventions because of their community
familiarity and grassroots outreach strategies.
376
WISH Associates, Cape Town, South Africa, 2African Gender Institute,
University of Cape, Cape Town, South Africa, 3PATH, Washington, DC,
United States
Background: South Africa (SA) launched a new contraception policy in
2014 that highlights linkages between HIV prevention and sexual and
reproductive health (SRH). HIV-prevention campaigns in SA have focused
primarily on male condoms, while female condoms—the only available
woman-initiated multipurpose prevention technology (MPT)—have not
been strongly supported. Research suggests that knowledge about female
condoms is limited and myths and mistrust persist, which contributes
to limited access. The V condom (developed by PATH as the Woman’s
Condom) is being evaluated for early market introduction in SA.
Methods: WISH Associates, AGI/UCT, and PATH collaborated on
activities to strengthen the policy and advocacy environment for female
condoms overall, and for the V condom in particular. We conducted
a policy/advocacy analysis in SA, and identified opportunities for
strengthening female condom programming. From 2013-2014, we
implemented strategic activities engaging policymakers, health care
workers, journalists/media experts, researchers, SRH advocates, and
potential consumer groups. These activities raised awareness and
generated interest in female condoms overall, and the new V condom
specifically.
Results: A range of activities strengthened the environment for female
condom promotion and awareness, including: developing a cadre of
female condom ambassadors who promote female condoms among
diverse communities, organizing public awareness days, crafting
media guidelines to strengthen reporting on female condoms, and a
compilation of digital stories that describe why female condoms are
important to women and men in South Africa.
Conclusions: Advocacy partnerships and interest from decisionmakers for female condom programming suggest that support for
female condoms has gleaned media and public interest. Advocacy
helped pave the way for V condom introduction and offers new hope
in a country where women have limited options for protecting their
reproductive health.
P43.15 LB
Modeling the Impact of Targeting Treatment
and Prevention to the Migrant Population of
Male Miners in a Generalized Epidemic Setting
Daniel J. Klein1, Anna Bershteyn1, Kevin Oishi1, Philip Eckhoff1
Institute for Disease Modeling, Bellevue, WA, United States
1
POSTERS
Background: Migrant populations are thought to have played a
significant role in the initial spread of HIV in sub-Saharan Africa. Miners
in particular have been identified as a key driver of the HIV epidemic
in South Africa due to their circular migration patterns and elevated risk
behaviors. We used a computer model to quantify the potential impact
of targeting treatment and prevention to male miners.
Methods: We augmented an individual-based network model, EMODHIV v0.8, to include a migrant population representative of South
African miners. Simulated miners routinely migrate between their home
community and a mining community, where an external incidence
source replaces sexual transmission.
Results: Targeting miners with treatment, perfect prevention, or
combination treatment and prevention averted 2.7, 203.3, and 219.8
thousand infections over a 20-year period with 3% discounting,
respectively. These results come from baseline assumptions that 5%
of the male population engages in mining and that each miner visits
home monthly for one week. In comparison, universal test and treat
resulted in 3.8 million discounted infections averted over the same
time period. We increased the frequency of home visits, however the
impact changed only marginally. We then exaggerated the size of the
male mining population to 20%, resulting in 62.5, 890.6, and 938.2
thousand discounted infections averted from treatment, prevention,
and combination intervention. Finally, we changed the timing of the
prevention intervention from 2015 back to 1985 in 1-year increments.
To halve the incidence rate in 2020, prevention needed to start in
1989. Result may underestimate impact by not modeling transmission
at the mine.
Conclusions: While targeting miners can be cost effective, here we
see that these interventions will have a modest impact on the overall
incidence in a generalized epidemic. Nonetheless, targeting migrants
remains a promising tool for mitigating the future burden of HIV in lowlevel and concentrated epidemics.
www.hivr4p.org
377
Posters
Posters 44: Preclinical Evaluation of Biomedical Agents in Animal Models Tissues Explants
P44.01
P44.02
N-alkyl/aryl-4-(2-Substituted-3Phenylpropyl) Piperazine-1-Carbothioamide
as Vaginal Microbicide with RT Inhibition:
Synthesis, Docking and PK Studies
IND-directed Pharmacology and Toxicology
of IQP-0528, a Novel HIV-1 Topical
Microbicide
Veenu Bala1, Bhavana Kushwaha2, Yashpal S. Chhonker3,
Shagun Krishna4, Kavita Rawat5, Atul Krishna6, Praveen K.
Shukla6, Raj K. Tripathi5, Mohammad I. Siddiqui4, Rabi S. Bhatta3,
Gopal Gupta2, Vishnu L. Sharma1
CSIR-Central Drug Research Institute, Medicinal and Process Chemistry
Division, Lucknow, India, 2CSIR-Central Drug Research Institute,
Endocrinology Division, Lucknow, India, 3CSIR-Central Drug Research
Institute, Pharmacokinetics & Metabolism Div., Lucknow, India, 4CSIRCentral Drug Research Institute, Molecular & Structural Biology Division,
Lucknow, India, 5CSIR-Central Drug Research Institute, Toxicology
Division, Lucknow, India, 6CSIR-Central Drug Research Institute,
Microbiology Div., Lucknow, India
1
POSTERS
Background: The growing health and economic burden of STIs, HIV, and
population growth in resource-poor countries have led to integrate them
and vaginal microbicides have emerged as an option with additional
advantage of women control.
Methods:
Fifteen
N-alkyl/aryl-4-(3-substituted-3-phenylpropyl)
piperazine-1-carbothioamide (13-27) derivatives were synthesized
as topical vaginal microbicides and evaluated for anti-Trichomonas,
spermicidal, antifungal and reverse transcriptase (RT) inhibitory
activities. To assist the sulfhydryl (SH) binding molecular mechanism
of synthesized compounds, hexokinase and DTNB assays were carried
out because -SH group present over sperm and Trichomonas is vital for
their survival. All compounds were tested for safety through cytotoxic
assay against human cervical cell line (Hela) and compatibility with
vaginal flora, Lactobacillus. Docking study of most promising vaginal
microbicide (13) was carried out to find out its docking position and
orientation in comparison to known RT inhibitor Nevirapine. The
preliminary in vivo pharmacokinetics of compound 13 was performed
in female NZ-rabbits to evaluate systemic toxicity in comparison to
clinically used spermicide Nonoxynol-9.
Results: Compound 13 appeared as most favorable multiple active
scaffold, comprising of RT inhibition (72.30%), spermicidal (MEC 0.01%),
anti-Trichomonas (MIC 7.78 µg/mL) and antifungal (MIC 3.12-50 µg/mL)
actions. The compound’s low toxicity to HeLa cells and Lactobacillus
growth would eventually favor vaginal administration. Hexokinase and
DTNB assays proved the molecular mechanism involving SH binding.
Compound 13 docked on RT in a position and orientation similar to
the Nevirapine. In vivo pharmacokinetics of 13 suggested minimal side
effects in comparison to N-9.
Conclusions: The study resulted into novel compound (13) as women
controlled topical vaginal microbicidal spermicide possessing RT
inhibitory activity in order to empower women to deal independently
with their reproductive health and fertility.
378
Robert W. Buckheit Jr.1, Christian F. Freguia1, Anthony Ham1,
Karen W. Buckheit1
ImQuest BioSciences Inc., Frederick, MD, United States
1
Background: In the absence of a vaccine, topical microbicides are
considered one of the best strategies for HIV prevention. Here, we describe
the development of a novel microbicide, IQP-0528, formulated into a
vaginal gel for the pre-exposure prophylaxis of sexually transmitted HIV.
Methods: The efficacy and toxicity of IQP-0528 was evaluated using
in vitro cell-based assays. IQP-0528 vaginal gel was characterized by
rheological analysis. IND-enabling safety and toxicity studies were
performed as per FDA recommendations.
Results: IQP-0528 possesses good chemical and metabolic stability
and ease of synthesis. IQP-0528 potently inhibits the replication of all
clinical HIV subtypes (except O) with an EC50 of 0.2-20 nM and TC50
of approximately 400 uM and is active in the presence of seminal and
vaginal fluids with minimal toxicity to cell lines, explant tissues and
vaginal flora. Formulation into a gel for vaginal delivery showed favorable
physicochemical properties, long term stability, sub-nanomolar potency
and minimal toxicity. IQP-0528 did not cause acute toxicity in mice and
rats and exhibited no genotoxicity. IQP-0528 vaginal gel (up to 10%
w/v concentration) dosed daily for 28 days in rats and rabbits was well
tolerated with no adverse effects and minimal systemic absorption at a
clinical dose (1%). Vaginal tissue concentrations were in the ug range.
The gel did not elicit skin sensitization reactions in guinea pigs, no
teratogenic effects were observed in prenatal development studies, and
no cardiovascular effects were identified. The NOAEL of the IQP-0528
vaginal gel is a dose of 10%.
Conclusions: IQP-0528 is a novel and ideal clinical microbicide candidate
due to its high potency, ease of manufacturing, long-term stability, low
toxicity, poor systemic but favorable local drug absorption and low cost of
goods. Overall our data provide a rationale for continued advancement of
this molecule to human clinical trials.
P44.03
P44.04
Preventing Drug Resistant HIV Infection in
Colonic Tissue using Tenofovir and Maraviroc
Combination Topical Rectal Gels
Preclinical Evaluation of Multi-targeting
Antiretroviral Drug Based-combinations as
Candidate Microbicides
Charlene Dezzutti1,2, Julie Russo2, Lin Wang2, Brid Devlin3, Ian
McGowan1,2, Lisa C. Rohan1,2
Carolina Herrera1, Natalia Olejniczak1, Javier García Pérez2, José
Alcamí2, Charles Kelly3, Robin Shattock1
University of Pittsburgh, Pittsburgh, PA, United States, 2Magee-Womens
Research Institute, Pittsburgh, PA, United States, 3IPM, Silver Springs,
MD, United States
1
Background: The paradigm for topical microbicide development has
been a single agent product. Interest is now turning toward combination
products. A 1% tenofovir (TFV)/0.1% maraviroc (MVC) combination
rectal gel has been developed. We hypothesize the combination gel
should provide greater mucosal tissue protection against drug resistant
virus than single agent products.
Methods: Polarized colonic explants were treated apically with dilutions
(1:50 to 1:5000) of TFV only or TFV/MVC gel product along with 1×104
TCID50 HIV-1 wildtype or drug resistant (DR) viruses. Wildtype JR-CSF
and DR JR-CSF (K65R) and transmitter/founder viruses, wildtype THRO
and DR CH077 (K65R/Y181C), were tested. After an overnight culture,
the explants were washed and the basolateral medium was replenished.
Every 3 to 4 days medium was harvested, stored and replenished. HIV-1
replication was monitored in culture supernatant by p24 ELISA.
Results: For JR-CSF, TFV gel protected 5 of 6 explants at ≥328 µM
(1:100 dilution of the gel) while twice as much TFV was needed to
protect explants against JR-CSF (K65R); a 2-fold difference in activity.
The TFV/MVC gel was completely protective at ≥65.5 µM TFV/3.9 µM
MVC (1:500 dilution of the gel) and was more than 50% effective at
preventing HIV infection at 32.8 µM TFV/2.0 µM MVC (1:1000 dilution)
for both JR-CSF and JR-CSF (K65R). When tested against the transmitter/
founder viruses, ≥65.5 µM TFV/3.9 µM MVC was needed to protect
all explants against THRO. However, CH077 was more resistant to the
TFV/MVC protecting only 2 of 6 explants at 328 µM TFV/19.5 µM MVC
(1:100 dilution).
Conclusions: TFV/MVC gel was 5-fold more potent than TFV gel likely
reflecting the activity of MVC and it was equally potent against JR-CSF
and JR-CSF (K65R). While THRO showed similar activity to the TFV/MVC
gel as both JR-CSF viruses, CH077 showed marginal activity toward
the TFV/MVC combination which may reflect other compensatory
mutations. Additional testing is needed to confirm DR virus susceptibility
to TFV/MVC gel.
Background: Antiretroviral (ARV) drug combinations are highly effective
in HAART and may also be more effective as microbicides than single
drugs. This study aims to assess the activity of tenofovir, dapivirine
and darunavir combinations against wild-type and resistant HIV-1,
SIVmac32H and RT-SHIV isolates
Methods: Antiviral efficacy of dual and triple combinations of a
nucleotide reverse transcriptase inhibitor (NRTI), tenofovir, a nonNRTIs, dapivirine and a protease inhibitor, darunavir; was evaluated.
The combinations were assessed in cellular (TZM-bl cells, PM-1 CD4+T
cells, C8166 cells and activated PBMCs) and colorectal explant models.
Preincubation of cells or tissue with the drugs individually or in
combination, for one hour was followed by addition of virus. Wild type
isolates (BaL, YU.2, SIVmac32H and RT-SHIV) and NRTI-escape mutants
(point mutations K65R +/- M184V introduced in YU.2 and SIVmac32H)
were used. Infection was determined by measurement of luciferase
expression (in TZM-bl cells) or p24 viral antigen in culture supernatants
Results: All dual combinations tested in cellular and tissular models
against all isolates showed an increase of activity for at least one of the
drugs included in the combination compared to the drugs used alone.
The triple combination further augmented the inhibitory activity. All
combination tested inhibited NRTI-resistant isolates. The introduction
of point mutations affected the viral replication fitness in HIV and SIV.
Interestingly, dose-response curves were affected by the preclinical
model used, emphasizing the need to use multiple cellular and tissue
models during screening of candidate microbicides
Conclusions: The positive results obtained against HIV-1 and SIV/SHIV
isolates allow the design of in vivo studies in macaques before clinical
trials. The activity against HIV-1, SIV/SHIV indicate drug combinations
inhibiting reverse transcription and viral maturation have a great
potential as effective microbicides able to inhibit HIV-1 transmission
Instituto de Salud Carlos III, Madrid, Spain, 3King’s College London,
London, United Kingdom
2
www.hivr4p.org
379
POSTERS
1
Posters
P44.05
P44.06
Mini CD4-heparan Sulfate Mimetic Conjugates
Display Sub Nanomolar Anti-HIV-1 Activity
and Protect Macaques against a SHIV162P3
Vaginal Route Challenge
Engineering and Characterization of a
Bispecific Entry Inhibitor for Non-vaccine
Biomedical Prevention of HIV-1 Infection
Krystal Hamorsky1, Adam Husk1, Nobuyuki Matoba1
Françoise Baleux1, Kevin Arien2, Delphine Desjardins3, Jo
Michiels2, Yves-Marie Coic1, Bridgette J. Connell4, David Bonnaffé5,
Kawthar Bouchemal6, Roger Le Grand3, Guido Vanham2,
Nathalie Dereuddre-Bosquet3, Hugues Lortat-Jacob4
Institut Pasteur, unité de Chimie des Biomolécules, URA CNRS 2128,
Paris, France, 2Institute of Tropical Medicine, Virology Unit, Antwerp,
Belgium, 3CEA, Service d’Immunovirologie - IDMIT, Fontenay-auxRoses, France, 4Institut de Biologie Structurale, CNRS, CEA, Université
Grenoble-Alpes, UMR 5075, Grenoble, France, 5Université Paris-Sud,
Institut de Chimie Moléculaire et des Matériaux d’Orsay, Orsay, France,
6
Université Paris-Sud, Faculté de Pharmacie, Chatenay-Malabry, France
1
POSTERS
Background: The HIV-1 gp120 coreceptor binding site (CoRBS) can be
targeted by heparan sulfate (HS) or HS-like sulfopeptides, however its
cryptic nature limits its vulnerability. We hypothesized that molecules
comprising a CD4 mimetic covalently linked to HS-like compounds
would bind gp120 through the CD4 moiety, expose the CoRBS and
render it available to be blocked by HS (see Connell BJ. et al. Front.
Immunol. 2013).
Methods: CD4-mimetics (mCD4) were conjugated to synthetic HS or to
sulfopeptides (PS). A Surface Plasmon Resonance platform was developed
to analyze gp120 binding to HS, CD4, and detergent solubilized
CXCR4 and CCR5, functionally captured on biacore chips. Inhibition of
infectivity of primary HIV-1 by a series of mCD4-PS was measured in a
standardized assay with CD4 and CoR expressing TZMbl cells. Finally in
vivo activity of mCD4-PS, formulated in hydroxyethylcellulose-based gel,
was assessed with female cynomolgus macaque’s model of intravaginal
(IVAG) single high dose challenge with SHIV162P3.
Results: We demonstrated that mCD4-conjugates efficiently block the
HS, the CD4 and the CoR binding sites of gp120 and as such, mimic
several of the recently described broadly neutralizing antibodies. In cell
culture, mCD4-PS neutralize both R5- and X4- tropic HIV-1 of various
clades with pico to nano molar IC50 in the absence of cellular toxicity.
Moreover, cynomolgus macaques treated with mCD4.1-PS1 one hour
before IVAG challenge with SHIV162P3 were fully protected against
acquisition of infection whereas all control animals were infected.
Conclusions: We engineered a new class of compounds which have
the unique ability to target two critical and highly conserved regions of
gp120, i.e. the CD4 and the CoR binding sites, in addition to the V2, V3
loops. Chemically defined, these 5.5 kDa molecules which neutralize
both R5- and X4-tropic HIV-1 with very low IC50 are amenable to largescale production, and in view of their in vivo activity could be considered
for the development of a microbicide approach.
380
University of Louisville School of Medicine, Owensboro Cancer
Research Program, Owensboro, KY, United States
1
Background: Entry inhibitors could offer promising means for nonvaccine biomedical prevention against HIV infection, such as topical
microbicides. Here, we aim to create a novel entry inhibitor by
translationally fusing a broadly HIV-1 neutralizing monoclonal antibody
(mAb) and a carbohydrate-binding antiviral protein targeting the glycan
shield of viral envelope. We hypothesize that such a bispecific inhibitor
may effectively block and control HIV-1 infection.
Methods: We engineered a prototype bispecific fusion protein based on
the antigen-binding fragment (Fab) of the CD4 binding site-specific mAb
VRC01 and the oligomannose-specific, non-cytotoxic/inflammatory/
mitogenic antiviral lectin Avaren (VRC01-Av). VRC01-Av was produced in
Nicotiana benthamiana plants due to the prospective scalability and cost
effectiveness of plant-based expression systems. The fusion protein was
purified by a series of chromatography steps, and tested for the binding
properties and anti-HIV activity by surface plasmon resonance (SPR) and
Env-pseudotyped virus neutralization assays, respectively.
Results: The fusion protein was efficiently purified to >95% homogeneity
by a two-step chromatography process. SPR demonstrated that both
active sites of the VRC01-Av fusion retained affinity to a recombinant
HIV-1 Env protein gp120. VRC01-Av had significantly stronger HIV-1neutralizing activity than a 1:1 equimolar mixture of each component
(i.e., VRC01Fab and Avaren) and respective bivalent parental molecules
(i.e., VRC01 IgG and Avaren-Fc fusion) against multiple strains. We are
currently developing a dry powder formulation of VRC01-Av to enhance
stability and facilitate delivery system development.
Conclusions: These results highlight the advantages of VRC01-Av
bispecific fusion protein over a mixture of the two original entry inhibitors
in terms of efficacy, manufacturing and drug development. Our data
warrant further preclinical efficacy and toxicity studies of VRC01-Av.
P44.07
P44.08
Counteracting Semen-mediated
Enhancement of HIV Infection and Enveloped
Virus Infection by a Lysine-specific Molecular
Tweezer
Intravaginal Films Delivering
Aminoglycosides: Safety in a Macaque Model
Edina Lump1, Laura Castellano2, Thomas Schrader3, Gal Bitan4,
James Shorter2, Jan Münch1
1
Background: Semen contains amyloid fibrils that markedly enhance
HIV-1 infection by sequestering viral particles and increasing cell entry
rates. Thus, counteracting this proviral activity in semen presents a novel
strategy of reduce or block sexual HIV-1 transmission. It has previously
been shown that a lysine-specific molecular tweezer (CLR01) is capable of
preventing the formation of Aß and α-synuclein fibrils that are associated
with Alzheimer’s and Parkinson’s disease. Here, we explored whether
CLR01 also affects the formation and function of seminal amyloids.
Methods: The effect of CLR01 on seminal amyloid formation and its
degradation was studied by electron microscopy and spectrofluorometry;
interaction of CLR01-treated amyloid with viral particles was analyzed
by zeta potential measurements and confocal microscopy; the effect of
CLR01 on amyloid- and semen-mediated enhancement of HIV, HCV,
and HSV-2 infection was determined by infection assays;
Results: We demonstrate that CLR01 blocks the formation of seminal
amyloids and even slowly disaggregates pre-formed fibrils. Furthermore,
the tweezer neutralizes the positive surface charge of the fibrils
thereby preventing their interaction with virions and abrogating their
HIV enhancing activity. Most importantly, CLR01 also counteracts the
ability of semen to boost infection of HIV transmitted/founder viruses.
Unexpectedly, CLR01 also inhibits HIV and other enveloped viruses such
as HCV or HSV-2 by directly disrupting virion integrity.
Conclusions: CLR01 has previously been tested safe and therapeutically
active in mouse models of amyloid diseases. Thus, its low toxicity and
the combined anti-amyloid and anti-viral activities make CLR01 a
promising candidate for microbicide development to prevent HIV and
other sexually transmitted virus infection.
University of Washington, Obstetrics & Gynecology, Seattle, WA,
United States, 2Magee Women’s Research Institute, Pittsburgh, PA,
United States, 3University of Pittsburgh, School of Pharmacy, Pittsburgh,
PA, United States, 4University of Central Florida, Orlando, FL, United
States
Background: Aminoglycosides, which restore the natural production
of retrocyclins in the cervicovaginal mucosa, enhance innate antiviral
immunity. Tobramycin, an aminoglycoside antibiotic, can be delivered
by dissolvable vaginal films to heighten protection against sexually
transmitted HIV-1 infection. This study assessed the safety of
intravaginally delivered tobramycin in a macaque model for topical
microbicide use.
Methods: In test product vs placebo studies of n=6 per group, we
assessed the effects of a film formulation containing 5 mg tobramycin
during two weeks of daily applications. We performed baseline
colposcopy, vaginal microbiology, smear and pH measurements,
followed by intravaginal film placement in each animal. Thirty minutes
after film placement (test or placebo), repeat vaginal flora, smear and pH
measurements were made. These procedures were repeated daily for
five days in two successive weeks. On the following Monday (day 15), a
follow-up exam was conducted to document recovery.
Results: Colposcopic observations demonstrated that repeated
exposures to the 5mg tobramycin film did not lead to serious adverse
findings, i.e. cervicovaginal tissue abrasion and/or friability, in any of
the 6 animals. Microbiologic assessment revealed populations of H2O2
producing lactobacilli and viridans streptococci fluctuated over the
course of the experiment, but no clear trend of growth or suppression
was noted for either organism. The incidence of enteroccocus and E.
coli remained low, yet each was more prevalent at the end of the study
than at baseline. In both study arms, vaginal pH measures fluctuated
modestly throughout the study, and on average decreased from neutral
pH at baseline to pH 5.5-6.0 at the final visit. No product related
induction of neutrophil infiltration was noted by Gram stain.
Conclusions: Repeated vaginal exposures to the aminoglycoside film
were well tolerated in the macaque model. Studies assessing systemic
absorption and distribution of tobramycin in this experiment are underway.
www.hivr4p.org
381
POSTERS
Ulm University Medical Center, Institute of Molecular Virology,
Ulm, Germany, 2Perelman School of Medicine at the University of
Pennsylvania, Philadelphia, PA, United States, 3University of DuisburgEssen, Essen, Germany, 4University of California at Los Angeles, Los
Angeles, CA, United States
1
Dorothy L. Patton1, Yvonne Sweeney1, Tian Zhou2,3, Lisa C.
Rohan2,3, Alexander M. Cole4
Posters
P44.09
P44.10
RV306, an Evaluation of a 48 Week ALVACHIV AIDSVAX B/E Vaccination Regimen in
Thailand: Participation Rates for Optional
Specimen Collections
Candidate Microbicide 5-hydroxytyrosol (5HT) Inhibits Productive R5 HIV-1 Infection of
Human Cervical Tissue Explants (CTE)
Punnee Pitisuttithum1, Sorachai Nitayaphan2, Suwat
Chariyalertsak3, Nicos Karasavvas4, Jaranit Kaewkungwal5,
Viseth Ngauy4, Sandhya Vasan4, Merlin L. Robb6, Nelson L.
Michael6, J. Kendall Brown6, Charla Andrews6, Benjaluck Phonrat1,
Jittima Dhitavat1, Jean Louis Excler6, Jerome H. Kim6, Robert J.
O’Connell4, on Behalf of the RV306 Study Team
Vaccine Trial Centre, Faculty of Tropical Medicine, Mahidol University,
Bangkok, Thailand, 2Royal Thai Army Clinical Research Center, AFRIMS,
Bangkok, Thailand, 3Research Institute for Health Sciences (RIHES),
Chiang Mai University, Chiang Mai, Thailand, 4Armed Forces Research
Institute of Medical Sciences, Bangkok, Thailand, 5BIOPHICS - Center
of Excellence for Biomedical and Public Health Informatics, Bangkok,
Thailand, 6U.S. Military HIV Research Program, Walter Reed Army
Institute of Research, Bethesda, MD, United States
Elisa Saba1, Massimo Origoni1,2, Gianluca Taccagni1, Claudio
Doglioni1,2, David Auñón3, Eduardo Gomez-Acebo3, Josè Alcami4,
Guido Poli1,2, Elisa Vicenzi1
San Raffaele Scientific Institute, Milan, Italy, 2Vita-Salute San Raffaele
University, School of Medicine, Milan, Italy, 3Seprox Biotech SL, Madrid,
Spain, 4Instituto de Salud Carlos III University of Alcala, Madrid, Spain
1
1
POSTERS
Background: Vaccination with the RV144 regimen showed a waning
efficacy of 60% at one year and 31.2% three years post-last vaccination.
Risk correlated with plasma anti-Env binding antibody responses, but
specimen volumes and collection sites limited analysis. HIV-vaccine
naïve volunteers were vaccinated with the RV144 regimen plus a 12
month boost for immunologic assessment in multiple systemic and
mucosal compartments.
Methods: Volunteers from 3 sites in Thailand were randomly enrolled
into 4 placebo-controlled groups receiving the RV144 regimen plus
no boost, ALVAC-HIV+AIDSVAX B/E, AIDSVAX B/E, or ALVAC at 12
months. Immune responses are being assessed both systemically and
for additional site specimen collections in a subset of willing volunteers.
Results: Of 613 screened participants, 362 were enrolled (48% male
and 52% female) with a mean age of 28 years. Consent to provide rectal
secretions was provided by 69/172 (40%) of male volunteers, ranging
from 12-90% among the three sites, while 135/172 (78%) consented
to provide semen. Female participants consented for cervico-vaginal
secretion collection (VTC: 69/95 (73%); RTA: 39/76 (51%); RIHES: 9/19
(47%), all sites: 117/190 (62%). Volunteers in two sites also consented
for a single invasive procedure per participant as follows: Cervical
biopsy (VTC: 44/95 (46%); RTA: 15/76 (20%); all sites 59/171 (35%),
sigmoid biopsy in males [VTC: 29/57 (51%); RTA: 13/74 (18%); total
42/131 (32%)], leukapheresis: (VTC: 30/152 (20%); RTA 21/150 (14%);
total 51/302 (17%) and bone marrow aspiration [VTC: 9/152 (6%); RTA:
5/150 (3%); total: 14/302 (5%)].
Conclusions: Mucosal collections and invasive procedure participation
rates were high in this clinical trial conducted in Thailand, demonstrating
both favorable disposition to research participation and effective
counseling at the sites. Differences in participation in these procedures
among sites provide insights that may lead to strategies to improve
participation in future trials.
382
Background: Despite the encouraging results obtained in the CAPRISA
trial by using Tenofovir gel to prevent HIV-1 sexual transmission, the
development of an effective microbicide remains a major priority.
Among candidate new microbicides, we are investigating the phenolic
phytochemical compound 5-HT (previously shown to be possess antiHIV-1, anti-inflammatory and anti-oxidant activities in isolated cell
systems) in human cervical tissue explants (CTE).
Methods: CTE established from HIV-1 seronegative women undergoing
hysterectomy for benign tumors, were infected ex vivo with R5 HIV-1BaL
in the presence or absence of increasing concentrations of 5-HT or, as
control, of 3TC. Productive infection of HIV-1 was measured in terms
of p24Gag expression in the culture supernatants. In addition, total viral
DNA was quantified by RT-PCR in the different experimental conditions.
Results: A decrease of p24Gag release was observed in the culture
supernatant after incubation with 200 µM of 5-HT and this effect was
also associated with 50% decrease of total viral DNA, as determined on
tissue blocks after 12 days of culture.
No modulation of CD4 and CCR5 or of activation markers (CD69, CD25,
CD38) was observed in CD4+ lymphocytes incubated with up to 200
µM of 5-HT. Ongoing experiments are aimed at evaluating the potential
effect of 5-HT on the release of pro/anti-inflammatory cytokines and
chemokines in CTE culture supernatants.
Conclusions: These results confirm that 5-HT bears the potential to be
developed as an anti-HIV microbicide as it inhibits HIV-1 replication in
CTE, in addition to isolated cell systems, without inducing cytotoxicity or
promoting T cell activation.
P44.11
P44.12 LB
Mucosal Susceptibility to HIV-1 and Tenofovir
Protection in Explanted Cervicovaginal
Tissues from Postmenopausal Compared to
Premenopausal Women
Tissue Permeability and Drug-drug
Interactions of Darunavir in Intact Rabbit
Colo-rectal Tissue
Abhinav Kumar1, Magda Swedrowska1, Charles Kelly2, Ben Forbes1
Andrea R. Thurman1, Neelima Chandra1, Nazita Yousefieh1, Sharon
M. Anderson1, Susana Asin2,3, Christianne Rollenhagen2,3, Betsy C.
Herold4, Pedro Mesquita4, Ashley Huber4, Gustavo F. Doncel1
CONRAD, Eastern Virginia Medical School, Norfolk, VA, United States,
Department of Veteran’s Affairs, Department of Microbiology and
Immunology, White River Junction, VT, United States, 3Dartmouth
Medical School, Immunology and Microbiology, Hanover, NH, United
States, 4Albert Einstein College of Medicine, Infectious Diseases and
Pediatrics, Bronx, NY, United States
King’s College London, Institute of Pharmaceutical Science, London,
United Kingdom, 2King’s College London, Oral Immunology, London,
United Kingdom
1
1
Background: In order to obtain data on the mucosal effects of tenofovir
(TFV) on postmenopausal women, biologic endpoints related to HIV1 acquisition in cervicovaginal (CV) explants from pre (PreM) and
postmenopausal (PostM) women were compared.
Methods: CV explants from PreM and PostM women were obtained
within 1 hour of elective surgery and treated for 24 hours with growth
medium (GM) or 100 µg/mL of TFV. We assessed immune cell numbers
and phenotype in explants. Additional tissues were infected ex vivo with
HIV-1BaL and p24 antigen production was assayed over 21 days. Tissue
culture supernatants were assayed for inhibition of HIV-1 in TMZ-bl cells
in vitro.
Results: Based on preliminary data from ex vivo HIV-1 infection (mean
p24 endpoints and categorical analyses), PreM and PostM tissues
do not appear to show differences in their propensity to be infected
by HIV. TFV protected both tissues fully. Anti-HIV activity of PostM
explant supernatants demonstrated a trend (p = 0.06) toward reduced
HIV inhibition (PostM=38.5 ± 36.2% v. PreM=60.8 ± 19.1%). The
supernatants of explants from PreM and PostM women incubated with
TFV showed potent and equal activity against HIV (p = 0.61). Treated
with GM, both types of explants produced similar levels of cytokines and
chemokines. TFV increased the secretion of these immune mediators.
PostM explants had significantly greater number of HIV-1 target CD4+
cells (p = 0.02) and CD1a cells (p=0.03) and a trend towards higher
numbers of HLA-DR+ activated cells (p = 0.06).
Conclusions: PreM tissues have thicker epithelium, slightly fewer
epithelial immune cells (especially CD4 and CD1a), and similar basal
cytokine/chemokine production. They also produce secretions that
inhibited HIV infection in an in-vitro cell-based model to a greater
extent. Although these conditions would theoretically endow preM
mucosal tissues with higher resistance to infection, thus far, we have
not seen increased susceptibility of postM explanted tissues to ex vivo
HIV-1 infection.
Background: Cell culture models for studying drug disposition and drugdrug interactions are good but they cannot reflect the heterogeneous
cellular coexistence typical of colo-rectal mucosa. The aim of this study
was to establish an ex vivo excised tissue model which is closer to the
more complex in vivo situation and apply it for studying the transport
and interactions of darunavir (DRV) with other ARVs as reports suggest
that combination microbicides is more effective.
Methods: Colo-rectal tissue from a male New Zealand white rabbit was
excised and transferred to oxygenated ice cold Kreb’s buffer. A strip
of the colo-rectal segment was opened with a lateral cut thus making
a planar sheet, rinsed free of luminal contents with Kreb’s buffer and
stripped of the muscle layers before being mounted between twodiffusion half-cells containing oxygenated transport buffer solutions. The
permeability of darunavir was measured; interactions with transporter
inhibitor elacridar and effect of co-administration with tenofovir were
evaluated in 3 independent experiments.
Results: The respective absorptive and secretory permeability of
DRV across rabbit colo-rectal mucosa was 1.8±2.2×10-6 cm/s and
4.6±3.1×10-6 cm/s (ER-efflux ratio 3). Modulation of the transport of DRV
(10 µM) by elacidar (1 µM) showed that absorptive transport increased
by 4.4 fold and secretory transport decreased by 0.3 fold (ER 0.2). Coadministration of TFV (100 µM), with DRV resulted in 1.6 fold increase
in absorptive direction and 0.5 fold decrease in secretory direction as
compared to the transport of singlet agent (ER 0.8).
Conclusions: Transport of DRV was vectorial and affected by transporter
inhibitors, suggesting that DRV is a substrate of P-glycoprotein. DRV
transport was affected by the presence of tenofovir. The intact colorectal tissue model is a valuable technique and this could be extended
to include excised tissue samples from non-human primates as well as
applied for assessing cervico-vaginal drug transport.
www.hivr4p.org
383
POSTERS
2
Posters
Posters 45: Pregnancy and Prevention of Mother to Child Transmission
P45.01
P45.02
Role and Trend of Maternal Antiretroviral
Therapy in Preventing Mother-to-Childtransmission of HIV in Sub-Saharan Africa
Prevention of Mother-to-Child-Transmission:
Antiretroviral Coverage and Access to
Virological Testing during Breastfeeding
Olatunji O. Adetokunboh1, Mojisola Oluwasanu2
Olatunji O. Adetokunboh1, Tolulope Balogun1
Stellenbosch University, Division of Community Health, Cape Town,
South Africa, 2University of Ibadan, Faculty of Public Health, Ibadan,
Nigeria
1
1
POSTERS
Background: Pediatric HIV infections are largely due to mother-to-child
transmission (MTCT) in utero, during delivery, or via breastfeeding. The
highest burden of global pediatric HIV infections are in sub-Saharan
Africa (SSA) where many of the countries still have high MTCT rates
and new pediatric HIV infections. This study seeks to determine the role
and trend of antiretroviral therapy in HIV positive women with respect
to reduction in MTCT rate in the high burden countries of SSA countries
since 2009 when call for virtual elimination of pediatric HIV was made.
Methods: Data were obtained from the Joint United Nations Programme
on HIV/AIDS (UNAIDS) 2013 progress report on the Global Plan. The data
were that of 21 SSA priority countries from 2009 to 2012. Our analysis
focused on final MTCT rates, percent of women receiving antiretroviral
agents (ARVs) to prevent MTCT, percent of women or infants receiving
ARVs during breastfeeding to prevent MTCT and percent of HIV positive
pregnant women receiving ART for their own health.
Results: The final MTCT rate reduced from 27% to 19%, p=0.0001.
Percentage of women receiving ARVs to prevent MTCT was 63% in
2012, mean difference MD 30%, p=0.0001 and the percentage of
women or infants receiving ARVs during breastfeeding was 43%, MD
33%, p=0.0001. The percentage of HIV positive pregnant women
receiving ART was 54% in 2012, MD 34%, p=0.0001. The final MTCT
rate was strongly negatively correlated with the percentage of women
receiving ARVs to prevent MTCT (r=-0.9266; p< 0.0001).
Conclusions: There has been a significant increase in ART coverage
among HIV infected women in SSA in the last few years. Countries
that have higher ART coverage tend to have lower MTCT rates thereby
preventing many new infections. However there is still lot of work to be
done to achieve total elimination.
384
Stellenbosch University, Division of Community Health, Cape Town,
South Africa
Background: Sub-Saharan Africa accounts for about 90% of new HIVinfections in children with most of them contracting the infection via
mother-to-child-transmission (MTCT). Transmission during breastfeeding
accounts for about 40% of new infections. Access to antiretroviral
(ARV) regimens by the HIV positive mothers and their children during
breastfeeding will improve the chances of these children to be free of
HIV infection. Ensuring early infant virological diagnosis of HIV infection
will help in identifying those who are HIV-exposed but uninfected and
making sure they are free of the disease during breastfeeding period.
We will like to evaluate the coverage level of women or infants receiving
ARVs during breastfeeding to prevent MTCT and access to virological
testing in sub-Saharan countries.
Methods: Data were obtained from United Nations Children’s Fund
(UNICEF) 2013 Children and AIDS Sixth Stocktaking Report. We
analysed data from 21 SSA priority countries. Our focus was on the
percentages of mother or child on ARVs to prevent MTCT during
breastfeeding (2009-2012) and early virological diagnosis of HIV in
HIV exposed infants (2012).
Results: In 2012, 446471 infants had early virological testing for HIV
infection with South Africa recording 51.6% of the total number. The
average percentage of infants tested was 32.2% with 15 of the countries
recording < 50% coverage. Percentage of women or infants receiving
ARVs during breastfeeding increased from 10.2% to 42.5%. The mean
difference was 32.2%, 95%CI: 19.9-44.6, p = 0.0000. Ghana made a
remarkable progress with a difference of 95% while 2 countries were at
< 10% level in 2012.
Conclusions: The African priority countries made significant progress
in the provision of ARVs for mothers and infants during breastfeeding
period although some countries are still lagging behind. The level of
virological testing is still very low, additional effort is needed to improve
the early infant diagnosis in countries with poor coverage.
P45.03
P45.04
Ethics of HIV Prevention Research with
Pregnant Women
Barriers to Uptake of Polymerase Chain
Reaction Testing for HIV Exposed Infants at
Six Weeks among PMTCT Mothers at the
State Hospital Osogbo, Nigeria
Liza Dawson1
NIH/NIAID, Division of AIDS, Bethesda, MD, United States
1
Elizabeth Edoni1, PMTCT Mothers
Niger Delta University, Bayelsa, Nigeria
1
Background: Expand access to early infant diagnosis and earlier and
improved pediatric treatment are essential in order to improve survival
rates and health outcomes for children. Non-adherence in the first few
weeks may lead to the development of the premature development of
drug-resistant virus. This study therefore designed to document barriers
to Uptake of Polymerase Chain Reaction Testing (PCR) and initiation
of Cotrimoxazole (CTX) for HIV-exposed Infants at Six Weeks among
PMTCT Mothers at the State Hospital, Osogbo, Nigeria
Methods: A Case file of 30 PMTCT mothers who delivered two
months prior to this study were randomly selected and reviewed. The
HIV prevalence of pregnant women in this facility was 16.0%. Client
information was reviewed and telephone interviews were conducted
with each client. Counseling was provided to mothers to encourage a
clinic visit for infant HIV PCR testing and CTX prophylaxis.
Results: Few (17.8%) of the mothers were not aware that they need to
bring their infant for HIV PCR testing and these women had brought
their babies for immunizations but the infants were not identified as
HIV-exposed. Some (36.9%) of the infants had received HIV PCR testing,
but CTX was not dispensed. Reported barriers among respondents were
distance to the clinic (78.9%), spouse disapproval (43.7%), and stigma
and discrimination (39.0%).
Conclusions: In order to improve early diagnosis and treatment of
children, linking of children from PMTCT to care has to be strengthened.
This calls for more recruitment and training of Staff. Delivery facilities
should also have clear traceable records of the number of HIV-positive
women who deliver.
POSTERS
Background: Pregnant women need safe and effective HIV prevention
methods. In many countries, pregnant women are at high risk of HIV
acquisition; countries with high rates of HIV incidence and generalized
epidemics are also largely countries with high fertility rates. Also, social
and structural barriers to women’s autonomous decision-making may
hinder pregnant women from protecting themselves.
While the HIV prevention field is moving rapidly, testing prevention
interventions in trials with pregnant women remains daunting. Ethical
and regulatory barriers are significant, and community, provider, and
individual women’s perspectives are complex.
The consequences of failing to conduct research with pregnant
women are serious. Absent robust data, pregnant women must either
eventually use these products without knowing if they are safe and
effective, or avoid the products and continue to be exposed to high
risks of HIV acquisition.
Methods: Significant barriers and opportunities must be identified.
Human research regulations place stringent restrictions on research
with pregnant women. Although the heaviest burden of HIV lies in
sub-Saharan African countries, each with its own research regulations,
US regulations have a significant impact. The US is a major funder of
biomedical HIV prevention research, and any study funded by the US
government must adhere to US regulations. Researchers may have
difficulty meeting criteria for approval under this rule.
Results: To overcome these challenges, a more robust ethical approach
to research with pregnant women is needed. This paper explores the key
ethical parameters that must be considered in finding an appropriate
balance for inclusion of pregnant women in research, and discusses how
this might fit under current regulatory standards.
Conclusions: Research with pregnant must advance to extend the
benefits of HIV prevention to this key population. A new and more
appropriate ethical framework will help accomplish this goal.
www.hivr4p.org
385
Posters
P45.05
P45.06
The MTN-016 Pregnancy Registry: Baseline
Characteristics of Enrollees from the VOICE
Study and Reasons for Non-enrollment of
Eligible Women
Impact of ARVs and Public Health Awareness
on the Trend of HIV Infections among Infants
Born to HIV Infected Mothers
Samuel Kabwigu1, Lisa Noguchi2, Jothi Moodley3, Thes Palanee4,
Kenneth Kintu1, Lulu Nair5, R Panchia6, Pearl Selepe7, Jennifer
E. Balkus8, Kristine Torjesen9, Jeanna Piper10, Rachel Scheckter9,
Rohan Hazra11, Richard Beigi12
MU-JHU Research Collaboration, Kampala, Uganda, 2MTN/Johns
Hopkins University, Baltimore MD USA, Baltimore, MD, United States,
3
MRC, Durban, South Africa, 4Wits Reproductive Health & HIV Institute,
Johannesburg, South Africa, 5CAPRISA, Durban, South Africa, 6PHRU,
Soweto, South Africa, 7Aurum Institute, Klerksdorp, South Africa,
8
FHCRC and University of Washington, Seattle WA, WA, United States,
9
FHI 360, Durham, NC, United States, 10US NIH/DAIDS, Bethesda,
MD, United States, 11US NIH/NICHD, Bethesda, MD, United States,
12
University of Pittsburgh/Magee-Womens Hospital, Pittsburgh, PA,
United States
1
POSTERS
Background: As pregnant women are at risk for HIV and women
at risk of HIV may become pregnant, it is important to assess the
safety of candidate HIV prevention products in both non-pregnant
and pregnant women.
Methods: The MTN-016 pregnancy registry is a prospective
observational cohort in which participants who became pregnant
during MTN effectiveness studies or those with planned exposures
in pregnancy safety studies are monitored for adverse obstetric
outcomes; infants from these pregnancies are followed through
the first year of life. Registry enrollment systematically excludes
termination of pregnancy and early pregnancy loss, including early
non-viable pregnancies with a transient positive test at a monthly visit,
as these data are captured in parent protocols. For VOICE participants
enrolled into the registry from Uganda, Zimbabwe and South Africa,
we describe baseline demographic characteristics and key reasons for
non-enrollment as reported by site investigators.
Results: Among 5029 VOICE participants with over 5425 person-years
(py) of follow-up, there were 424 pregnancies (7.8/100 py) and 201 live
births. Of these pregnancies, 261 (62%) were eligible for MTN-016.
The average age of women who became pregnant during VOICE was
24, with 24% of women married at baseline. Of these, 213/261 (82%)
women and 185/201 (92%) of their infants enrolled in MTN-016 (average
maternal age 25 years, 33% married). Reasons for non-enrollment of
eligible participants into MTN-016 included additional study visit burden
and employment. Cultural customs related to temporary relocation
to rural areas during the postnatal period and the limitation of public
access to newborns were also cited as barriers to enrollment.
Conclusions: In this pregnancy exposure registry for candidate HIV
prevention agents, the majority of eligible women from VOICE and their
infants were enrolled. Study visit burden and local cultural customs may
impact the enrollment of mothers and their infants into a pregnancy
registry protocol.
386
Olipher Makwaga1, Anne Muigai2, Freda Andayi1, Tom Mokaya1,
Matilu Mwau3
Kenya Medical Research Institute - CIPDCR, Busia, Kenya, 2Jomo
Kenyatta University of Agriculture and Technology, Nairobi, Kenya,
3
Kenya Medical Research Institute - CIPDCR/Jomo Kenyatta University
of Agriculture and Technology, Nairobi, Kenya
1
Background: Although ARVs and public awareness is being used to
eliminate new HIV infections among infants, limited information
indicating their impact on the trend of HIV prevalence among infants
born to HIV infected mothers exists. This study determined the trend
of HIV infection among infants born to HIV infected mothers in relation
to specific ARV administration. Also assessed the knowledge about HIV
transmission between mothers and infants.
Methods: In a cross-sectional study, dried blood spot samples
(2010,n=4210; 2011,n=4093; 2012, n=4686; 2013, n=3080) collected
from infants aged ≤18 months. These samples were analyzed at KEMRICIPDCR using PCR assay. Some mothers who received public health
education were pregnant. All mothers were put on anti-retroviral drugs
during and after delivery.
Results: Out of the total number of samples tested 9.7%; 8.5%; 7.9%;
7.2% were HIV positive in 2010; 2011; 2012; 2013 respectively. The
prevalence of HIV in infants whose mothers had been put on ARVS was
as follows: AZT+NVP+3TC was 10.4%, 9.1%, 6.3%, 6.0% in 2010; 2011;
2012, 2013 respectively. HAART was 7.1%; 6.1%; 4.7%; 3.9% in 2010;
2011; 2012, 2013 respectively. SdNVP was 10.3%; 7.4%; 6.2%; 6.0% in
2010; 2011; 2012; 2013 respectively. Those mothers who had not been
given any ARV had their infants with HIV prevalence reaching 11.2%;
12.6%; 12.7%; 12.9% in 2010; 2011; 2012; 2013 respectively. Those
mothers who were aware that ARVs reduce transmission of HIV from
mother to child were 30%vs70%; not breastfeeding were 37%vs81%
and Delivery at the hospital were 43%vs89% before and after public
health awareness respectively.
Conclusions: Direct relationship between specific ARV administration
and HIV infection among infants exists. This is evident by the fact that
HIV prevalence appeared to decrease with subsequent years of provision
of specific drugs. HAART seemed to provide greater impact in HIV
prevention compared to others. There is need for intensified awareness
among reproductive women to help in reduction of transmission.
P45.07
P45.08
Obstetric and Infant Outcomes Following
Planned Maternal Third Trimester Exposure
to Tenofovir 1% Vaginal Gel
A Case Control Study of Factors Associated
with HIV Infection Despite Overall Low
Transmission Rates in HIV Exposed Infants in
Rural Kenya
Johns Hopkins Bloomberg School of Public Health, Epidemiology,
Baltimore, MD, United States, 2Microbicide Trials Network, Pittsburgh,
PA, United States, 3University of Alabama at Birmingham, Birmingham,
AL, United States, 4University of Pittsburgh/Magee-Womens Hospital,
Pittsburgh, PA, United States, 5Statistical Center for HIV/AIDS Research
and Prevention, Seattle, WA, United States, 6FHI 360, Durham, NC,
United States, 7MU-JHU Research Collaboration, Kampala, Uganda,
8
CONRAD/EVMS, Arlington, VA, United States, 9San Francisco General
Hospital/UCSF School of Medicine, San Francisco, CA, United States,
10
Baylor College of Medicine, Houston, TX, United States, 11Office of the
Global AIDS Coordinator, U.S. Dept. of State, Washington, DC, United
States, 12US NIAID/Division of AIDS, Bethesda, MD, United States
1
Background: Thorough drug safety evaluation includes assessing
potential impact of use on obstetric (OB) and infant outcomes. The MTN016 study is the first pregnancy exposure registry for anti-HIV PrEP and
microbicide agents. We evaluated OB and infant outcomes for registrants
enrolled from third trimester TFV gel exposure studies.
Methods: Data were restricted to registrants enrolled from studies with
planned TFV vaginal gel exposure: MTN-002 (open label, single dose
prior to cesarean) and MTN-008 (2:1 placebo-controlled, 7-day use).
Registry study visits occurred before delivery when possible, and at <
1, 1, 6 and 12 months for infants. Infant malformation endpoints were
determined by geneticists via independent review of physical exam (PE)
and photo data.
Results: All 16 MTN-002 and 90% (88/98) of MTN-008 mothers were
registered, with 25% (n=4) of MTN-002 and 97% (n=86) of MTN-008
participants enrolling prior to known pregnancy outcome. Demographics
were similar for MTN-008 enrollees and non-enrollees in the registry.
Infant retention at 12 months was 88% (MTN-002) and 80% (MTN-008).
One defect (ear canal) was noted in MTN-002, a rate (6%) comparable
to the 3% US background rate for malformations (p=0.51); no defects
were noted in infants from MTN-008. Compared to placebo (n=30), TFV
gel (n=58) was not associated with preterm delivery (1/58 (2%) vs. 2/30
(7%), p=0.27), postpartum hemorrhage (11/58 (19%) vs. 3/30 (10%),
p=0.36), non-reassuring fetal status (3/58 (5%) vs. 1/30 (3%), p=1.0),
chorioamnionitis (1/58 (2%) vs. 2/30 (7%), p=0.27), gestational diabetes
(0/58 (0%) vs. 1/30 (3%), p=0.34), or abnormal infant PE findings in the
first year of life (14/58 (24%) vs. 8 (27%), p=1.0).
Conclusions: This first report from a novel pregnancy registry suggests
third trimester TFV gel exposure is not associated with infant malformation
or adverse OB or infant outcomes. Future HIV chemoprevention studies
should include safety evaluation, including registry participation, for
pregnant mothers and their infants.
Nicollate Awuor Okoko1, Kevin Owuor1, Jayne Lewis Kulzer1,2,
George Owino1, Irene Ogolla1, Ronald Wandera3, Elizabeth Anne
Bukusi1,2, Craig R. Cohen1,2, Lisa Abuogi1,4
Family AIDS Care and Education Services (FACES), Research Care and
Training Program (RCTP), Centre for Microbiology Research (CMR),
Kenya Medical Research Institute (KEMRI), Kisumu, Kenya, 2University
of California San Francisco (UCSF), Departments of Obstetrics,
Gynaecology and Reproductive Sciences; Medicine, San Francisco,
CA, United States, 3Rongo District Hospital, Ministry of Health, Rongo,
Kenya, 4University of Colorado, Department of Paediatrics, Denver,
Denver, CO, United States
1
Background: Despite the availability of Prevention of Mother-to-Child
HIV Transmission (PMTCT) interventions and donor and government
investments in developing country implementation, vertical HIV
transmission persists. In Kenya, an estimated 37,000 to 42,000 infants
are infected with HIV annually due to mother-to-child transmission
(MTCT). This study explored the reasons for MTCT persistence in areas
with overall low transmission rates and PMTCT service provision.
Methods:
A case-control study of HIV-exposed infants (HEI) enrolled in follow-up
care between January and June 2012 was conducted at 20 rural health
facilities supported by Family AIDS Care and Education Services (FACES)
in Nyanza Province, Kenya. Cases were HEI who turned HIV positive,
controls were HEI who remained negative at last test; an equal number
of controls were randomly selected after matching based on birth month
and gender. Data were abstracted from routine PMTCT registers, HEI
cards, and infant forms. Logistic regression analysis was conducted to
determine factors associated with HIV infection.
Results: Forty-five cases and 45 controls were compared. Maternal,
clinical and infant factors associated with HIV-infected infants included
poor PMTCT service uptake including late enrolment of infant to follow
up, (OR = 0.14, 95%CI: 0.06 - 0.38), poor infant prophylaxis adherence
(OR=8.32, 95%CI 3.24 -21.38), and fewer antenatal (ANC) visits (OR
= 0.62, 95% CI: 0.41 - 0.96). Mothers of cases were significantly less
likely to report having received clinic level HIV education and counseling
compared to controls (OR 0.13, 95%CI 0.04 - 0.54 and OR 0.12, 95% CI
0.03 -0.46). Maternal disclosure, gestation at first ANC visit, and infant
feeding type were not significantly associated.
Conclusions: Poor PMTCT service uptake and a reported absence of
clinic level HIV education and counseling were associated with MTCT.
More emphasis on PMTCT service provision including counseling and
education are needed to minimize HIV transmission to infants.
www.hivr4p.org
387
POSTERS
Lisa M. Noguchi1,2, Joseph Biggio3, Katherine Bunge2,4, James
Dai5, Karen Isaacs6, Kristine Torjesen6, Samuel Kabwigu7, Jill
Schwartz8, Juan Vargas9, Fernando Scaglia10, Cindy Jacobson2,
D H. Watts11, Jeanna M. Piper12, Richard H. Beigi2,4, for the MTN002, MTN-008 & MTN-016 (EMBRACE) Study Teams
Posters
Posters 46: PrEP Trials: Preparing for Demos, Participant Experiences
P46.01
P46.02
Preparing for Risk-reduction Counseling on
Pre-exposure Prophylaxis for Women in
Kenya and South Africa
Guidelines on Informed-Choice Counseling
for Women Using Pre-exposure Prophylaxis
(PrEP)
Amy Corneli1, Emily Namey1, Khatija Ahmed2, Kawango Agot3,
Jennifer Headley1, Kevin Mckenna1, Joseph Skhosana2, Jacob
Odhiambo3
Amy Corneli1, Irina Yacobson1, Emily Namey1, Kawango Agot2,
Khatija Ahmed3, Joseph Skhosana3, Jacob Odhiambo2
FHI 360, Durham, NC, United States, 2Setshaba Research Centre,
Soshanguve, South Africa, 3Impact Research and Development
Organization, Kisumu, Kenya
1
POSTERS
Background: Our previous survey research on pre-exposure prophylaxis
(PrEP) and risk compensation with women at HIV risk in Bondo, Kenya,
and Soshanguve, South Africa, showed that a considerable minority
of women may engage in riskier sexual behavior if taking PrEP. We
conducted a small qualitative study to explore perceptions of an
informed-choice approach to PrEP counseling. This approach promotes
condoms but empowers women who are unable or unwilling to use
condoms to make informed decisions regarding their sexual health
when taking PrEP.
Methods: We conducted eight focus groups with women in Bondo and
Soshanguve (four per site). We introduced the counseling approach
and presented participants with multiple messages on PrEP and on risk
compensation when using PrEP, with some concepts framed as gain
versus loss. Responses were coded on aspects of the messages that
were perceived to be effective and ineffective and on perceptions of the
counseling. Themes were identified and summarized.
Results: Preliminary analysis suggests that participants appreciated
the counseling’s autonomy framework and believed messages on risk
compensation would be understood by women in their communities.
Participants generally liked the positive tone of the gain-frame messages
and found them polite and encouraging. The loss-frame messages were
viewed as harsh and threatening, but powerful. Many believed these
messages’ cautionary tone and description of consequences would more
effectively influence women’s behavior, primarily in Soshanguve. In
Bondo, many participants preferred the softer gain-frame messages and
described clarity and comprehension as critical aspects of messaging.
Conclusions: Both gain- and loss-frame messages were generally
understood, although variation existed in their acceptability and
perceived influence. Research among actual PrEP users is needed to
understand the effectiveness of message framing and of the informedchoice counseling approach, particularly in African contexts.
388
1
FHI 360, Durham, NC, United States, 2Impact Research and
Development Organization, Kisumu, Kenya, 3Setshaba Research Centre,
Soshanguve, South Africa
Background: Our previous survey research with women at risk of
HIV infection in Kenya and South Africa showed that a considerable
minority of women may engage in riskier sexual behavior if they
take PrEP. Enhanced prevention efforts are needed to minimize risk
compensation and promote informed decision-making on sexual
health when using PrEP.
Methods: We systematically reviewed best practices in informed choice
counseling and considered how they could be applied to strengthen
the decision-making process within HIV risk-reduction counseling. We
also integrated essential knowledge and key counseling messages to
support sexual health decisions about PrEP based on findings from a
study on PrEP and risk compensation.
Results: Counseling guidelines were developed for African women who
are considering initiating, continuing, or stopping PrEP. The guidelines
are intended to be used in conjunction with evidence-based behavioral
change strategies and are framed using a decision tree approach. They
take a woman’s individual needs and circumstances into consideration
and guide her through the decision-making process by asking essential
questions and communicating key sexual health messages. The guidance
encourages condom use while on PrEP, but also empowers women who
are unable or unwilling to use condoms consistently to make betterinformed decisions by considering contraception, considering regular
screening and treatment for STIs, and placing additional emphasis on
PrEP adherence. Guidance for maintaining sexual health is also provided
for women who transition off of PrEP.
Conclusions: Risk-reduction counseling for women considering,
continuing, or stopping PrEP must acknowledge that consistent condom
use may not always be a realistic option. These women need specific
knowledge and individualized messages to inform their sexual health
decisions. The counseling guidelines will be disseminated to PrEP
demonstration projects and country governments considering the
approval of PrEP.
P46.03
P46.04
Perceptions and Practice of Heterosexual Anal
Sex amongst VOICE Participants in Uganda,
Zimbabwe and South Africa
Preparing for PrEP & Immediate Treatment:
Focus Group Discussions in Advance of a
Demonstration Project in South Africa
Zoe Duby1,2, Miriam Hartmann3, Elizabeth T. Montgomery3,
Christopher J. Colvin1, Barbara Mensch4, Ariane van der Straten3
Robyn Eakle1,2, Goitse Manthata1, Jonathan Stadler1, Judie
Mbogua1, Maria Sibanyoni1, W.D. Francois Venter1, Helen Rees1
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, 4Population Council,
1
Background: In VOICE, a phase IIB study testing vaginal gel and oral
tablets for HIV prevention, 5029 female participants in 3 sub-Saharan
African countries used Audio Computer-Assisted Self-Interviewing
(ACASI) to report adherence to study products and sexual behaviour. At
baseline 17% of participants reported anal intercourse (AI) in the past
3 months. MTN-003D, a qualitative ancillary study, explored reasons,
motivations and context of engaging in AI and rectal gel use.
Methods: In-depth interviews (IDIs) were conducted in participants’
preferred language (Luganda, Shona, Zulu or English) with 88
systematically selected exited VOICE participants (26 Zimbabwean, 22
Ugandan and 40 South African).
Results: Perceptions of heterosexual AI practice varied between countries.
While AI was generally perceived to be male initiated, perceptions of
women’s ability to refuse AI, and narratives of pleasure and pain during
AI differed between countries. Participants listed motivating factors for AI
including male pleasure, relationship security, self-perceptions of having
an over-stretched or over-lubricated vagina, coercion and financial
incentives. Findings suggest low levels of condom use for AI and poor
knowledge of condom compatible lubricant. One participant disclosed
using study gel rectally; and one participant cited the belief that vaginal
use of gel would provide protection for AI. Reactions to AI questions
elicited varied responses amongst participants, ranging from shock,
disgust, embarrassment to amusement.
Conclusions: Findings provide insight into the under-researched
taboo topic of heterosexual AI in sub-Saharan Africa, shedding light
on relationship contexts and gendered power dynamics in which AI
takes place. Describing attitudes towards and practices surrounding
heterosexual AI such as condom and lubricant use, the findings inform
current HIV prevention priorities aimed at women in these countries, as
well as future prevention efforts to address HIV transmission through AI,
such as rectal microbicides.
Wits Reproductive Health & HIV Institute, Johannesburg, South Africa,
London School of Hygiene & Tropical Medicine, Infectious Disease
Epidemiology, London, United Kingdom
2
Background: The WHO and others have called for demonstration
projects to understand the deliverability of ARV-based prevention
methods. One project will be in South Africa through The Sex Worker
Project, offering oral PrEP and immediate treatment to female sex
workers (FSWs). In advance of the demonstration project, information
will be collected to gain insight into how these interventions can be
effectively delivered to FSWs.
Methods: Focus Group Discussions (FGDs) are currently being
conducted amongst FSWs at 2 demonstration project clinics. Participants
are being recruited from the clinic and surrounding communities. FGDs
are exploring FSWs’ perceptions of: optimal prevention and treatment
delivery, attitudes towards & perceptions of the use of oral PrEP and
immediate treatment, how a comprehensive HIV prevention & treatment
programme could function as an incentive to access services, & the
acceptability & feasibility of regular HIV testing every three months.
Results: Preliminary data from FGDs and local stakeholder consultations
point to several themes to be considered for the demonstration project,
and future implementation. Risk disinhibition is a concern, as some
FSWs could stop using condoms. This may affect the market rates of sex
for money, as those not using condoms could charge higher prices. HIVpositive FSWs feared that clients could favour those on PrEP in instances
where no early treatment was available. Possible resistance, in the case
of inconsistent adherence, was also a concern. Women felt positive about
the proposed service delivery mechanism, including the frequency of
testing and were willing to participate even without financial incentives.
Conclusions: The FGDs raised important issues for consideration. It will
be crucial to develop careful messaging, especially on the importance of
adherence and consistent condom use as combined protection. Overall,
FSWs have been positive about the deliverability of PrEP and immediate
treatment, with the right messaging.
www.hivr4p.org
389
POSTERS
1
Posters
P46.05
P46.06
Characteristics of HIV Discordant Couples who
Separated while Enrolled in a HIV Prevention
Clinical Trial in Kenya
Following the Doctor’s Advice: Experiences of
HIV Serodiscordant Couples Enrolled in a PrEP
Demonstration Project in Kenya
Lawrence Mwaniki Mwihaki1, Elizabeth Mugoiri Irungu2, John
Njoroge Mwathi3, Snaidah Ongachi Ayub3, Kenneth Kairu
Ngure4, Nelly Rwamba Mugo4
Kenneth Ngure1,2, Kathyrn Curran3, Sophie Vusha4, Maria Ngutu4,
Nelly Mugo2,5, Connie Celum2,6,7, Bettina Shell-Duncan8, Renee
Heffron2, Jared M. Baeten2,6,7
1
Parters in Prevention Thika- University of Washington, Data, Nairobi,
Kenya, 2Parters in Prevention Thika- University of Washington,
Clinic, Nairobi, Kenya, 3Parters in Prevention Thika- University of
Washington, Counseling, Nairobi, Kenya, 4Parters in Prevention ThikaUniversity of Washington, Managerial, Nairobi, Kenya
1
POSTERS
Background: Relationship dissolution among HIV serodiscordant
couples enrolled in clinical trials bears a negative impact on adherence to
study product and retention to clinic visits. We evaluated characteristics
of couples that separated while enrolled in HIV prevention clinical trial
in Kenya
Methods: We conducted univariate analysis to determine factors among
HIV discordant couples associated with ever separating while enrolled
at the Thika site in the Partners PrEP Study, a randomized controlled
trial conducted in Kenya and Uganda. Participants reported relationship
status at each visit. Data on relationship stability was extracted from the
counselors’ chart notes.
Results: Of the 496 HIV discordant couples enrolled, 98 (20%) reported
ever separating. In relationships reporting separation, HIV uninfected
persons were 34 [IQR 28, 39] years, 77% were male and duration of
relationship was 3 [IQR1, 7] years. Compared to couples with children
together, those without were likely to separate (30.1% vs 15.0%, OR=2.4,
95%CI 1.6-3.8, p< 0.0001). Couples who did not consider themselves
to be married (36.4% vs 18.6%, OR=2.5, 95%CI 1.2-5.2, p=0.016) and
couples who had known their discordant status for less than a year at
baseline (21.7% vs 13.1%, OR=1.8, 95%CI 1.0-3.4, p=0.05), were more
likely to separate. Compared to monogamous relationships, people in
polygamous unions were twice as likely to separate (OR=1.9, 95%CI
1.3-3.02, p=0.001). Couples who had stayed together longer were less
likely to separate (P=0.001).
Conclusions: A significant proportion of HIV discordant couples
enrolled in HIV prevention trials are likely to separate. The impact of this
separation on retention to care should be explored.
390
Jomo Kenyatta University of Agriculture and Technology, Institute of
Tropical Medicine and Infectious Diseases, Nairobi, Kenya, 2University
of Washington, Global Health, Seattle, WA, United States, 3World Health
Organization (WHO), HIV/AIDS, Geneva, Switzerland, 4Partners in Health
Research and Development, Thika, Kenya, 5Kenya Medical Research
Institute, Center for Clinical Research, Nairobi, Kenya, 6University of
Washington, Epidemiology, Seattle, WA, United States, 7University
of Washington, Medicine, Seattle, WA, United States, 8University of
Washington, Anthropology, Seattle, WA, United States
Background: Pre-exposure prophylaxis (PrEP) for HIV-uninfected
persons is highly efficacious for HIV prevention. Evidence-based social
science research is urgently needed to inform PrEP rollout. We used
qualitative methods to gather insights into couples’ early experiences of
using PrEP within a PrEP demonstration study.
Methods: From June-December 2013, we conducted 16 in-depth dyadic
interviews with heterosexual HIV discordant couples participating at
the Thika, Kenya site of the Partners Demonstration Project, a project
evaluating uptake of and adherence to PrEP and ART. PrEP is offered
when couples enroll in the study until the HIV-infected partner initiates
ART and achieves viral suppression [i.e. as a “bridge”]. We developed
and applied deductive and inductive codes and identified key themes
related to early experiences of initiation and use of time-limited PrEP.
Results: Of the couples interviewed, all HIV-uninfected partners had
initiated PrEP and 7 of the 16 HIV-infected partners interviewed had
initiated ART at time of interview. Prior to joining the study, no couples
had heard of PrEP. Interviewees often described their decision to take
PrEP as “following the doctor’s advice,” and believed that “doctor knows
best” due to the trust that had been established between the health
providers and couples. The friendly environment at the clinic reportedly
enabled a majority of the couples to make shared decision to initiate
PrEP. Many couples reported that PrEP could reduce the risk of HIV
transmission, meet their aspirations for fertility and cope with HIV
discordance (i.e. a solution to their discordance challenges). Remaining
HIV negative in the follow-up visits reinforced couples’ decisions and
motivations to continue adhering to PrEP.
Conclusions: Among these early adopters of PrEP for HIV prevention,
confidence in provider’s advice and patient-friendly services were critical
to decisions to initiate PrEP. PrEP responded to couples’ values and
preferences for reducing their HIV risk.
Posters 47: Resource Tracking and Economics
P47.01
P47.02
Tracking Investments and Expenditures in
HIV Prevention Research and Development:
Sustaining Funding in a Shifting International
Development Landscape
Local Investment in HIV Prevention Research:
The Contributions of African Countries that
Host Biomedical HIV Prevention Clinical Trials
Emily D. Donaldson1, Kevin Fisher1, Reuben Granich2, Thomas
Harmon3, Polly Harrison1, Arne Post Uiterweer3, Mitchell Warren1,
HIV Vaccines & Microbicides Resource Tracking Working Group
AVAC, New York, NY, United States, 2UNAIDS, the Joint United Nations
Programme on HIV/AIDS, Geneva, Switzerland, 3International AIDS
Vaccine Initiative, New York, NY, United States
Emily D. Donaldson1, Kevin Fisher1, Mitchell Warren1, Julien
Burns1
1
Background: Since 2004, the HIV Vaccines and Microbicides Resource
Tracking Working Group has employed a comprehensive methodology
to track trends in research and development (R&D) investments and
expenditures for biomedical HIV prevention, including HIV vaccines,
microbicides, PrEP, treatment as prevention and medical male circumcision.
Methods: R&D data were collected on annual disbursements by public,
private and philanthropic funders for product development, clinical trials
and trial preparation, community education and policy advocacy efforts
to estimate annual investment in HIV prevention R&D. Investment trends
were assessed and compared by year, prevention type, research phase,
funder category and geographic location.
Results: In 2013, investment in HIV prevention research reflected
decreased public sector budgets, revised investment strategies by
philanthropic donors and a continued retreat from prioritizing HIV
prevention by industry. The Working Group collated and analyzed 2013
data for all areas of HIV prevention R&D. International development
agency funding trends were analyzed and disaggregated by research
type showing a move towards funding large-scale clinical trials and
implementation research, following the research pipeline and trends
towards results-based financing.
Conclusions: Shifts taking place in 2013 in the international development
landscape in both policy and strategy may have profound effects on
HIV prevention research funding. The reorganization of Canada’s and
Australia’s development agencies into their respective departments
of foreign affairs and trade and the trend towards country-ownership
models for the HIV/AIDS response could have profound effects on these
department’s priorities. It is increasingly important to ensure continued
prioritization of HIV prevention research on the global development
agenda by understanding and evaluating research in the context of
public, private and philanthropic funding.
Background: The HIV Vaccines & Microbicides Resource Tracking
Working Group collects annual data on trends in research and
development (R&D) investments and expenditures for biomedical HIV
prevention options, including HIV vaccines, microbicides, PrEP, treatment
as prevention and medical male circumcision from public, private and
philanthropic sources predominantly in donor countries. There is a need
to track all investments in research, including those from countries and
communities in which trials take place.
Methods: Investment and expenditure data were collected from
countries in sub-Saharan Africa which undertake HIV prevention R&D.
Data were collected on annual disbursements by public, private and
philanthropic funders for product development, clinical trials and trial
preparation, community education and policy advocacy. Data were also
collected on non-monetary investments—in-kind and trial volunteer
commitment—to report on the full amount invested in HIV prevention
R&D. Policy and environmental considerations were analyzed by country.
Results: With numerous clinical trials and demonstration projects taking
place in countries across sub-Saharan Africa and nearly 70 percent of
all HIV prevention trial participants located in these countries, the region
has invested considerably in HIV prevention R&D. Promising results
in pre-exposure prophylaxis, treatment as prevention and adult male
circumcision have pushed these countries to support studies that will aid
in the rollout and uptake of new prevention modalities.
Conclusions: HIV prevention R&D could not advance without the
contributions of the countries in which trials take place, including
financial, collaborative and especially, the commitment of trial
participants and communities. As the state of donor funding is
uncertain, engagement and investment of funders and end-users from
the global South in the outcomes of R&D taking place in their countries
is imperative to advance the science needed to discover, develop and
deliver new HIV prevention options.
www.hivr4p.org
391
POSTERS
1
Posters
P47.03
P47.04
Resource Tracking to Ensure Accountability
and Sustainability: HIV Prevention, Treatment,
Diagnostics and Cure Research Investment
Analysis
Cost Benefit Analysis of the Centralisation of
Pharmacy Services in a Multi-study Clinical
Research Site in Hillbrow, Johannesburg
Clare Dott1, Krishnaveni Reddy1, Thesla Palanee1, Helen Rees1
Emily D. Donaldson1, Kevin Fisher1, Mitchell Warren1
1
POSTERS
Background: Since 2004, AVAC has tracked resource flows in biomedical
HIV prevention research and development (R&D) as the secretariat of
the HIV Vaccines & Microbicides Resource Tracking Working Group. As
the field evolves, AVAC employs a comprehensive methodology to track
investments and expenditures across the HIV prevention field, from
basic science to implementation research. By forming partnerships with
HIV advocacy, research and funding organizations to ensure accuracy
in reporting and grant allocation, AVAC expanded its resource tracking
efforts to including investments towards an HIV cure, treatment as
prevention, therapeutics and diagnostics.
Methods: Annual investment data are collected from public, private and
philanthropic funders, as well as principle investigators and scientific
experts, for product development, clinical trials and trial preparation,
implementation research, community education and policy advocacy
efforts. Investment trends are compared by year, prevention technology
type, research phase, funder category and geographic location.
Partnerships with public and private agencies and organizations enable
close review and analysis.
Results: By employing a continuous and consistent methodology,
forming partnerships and expanding its data collection areas AVAC
has overcome the challenges of collection of funding information
and maintained over a decade of investment data. Methods stayed
consistent since 2004 allowing for accurate and consistent year-to-year
comparisons of grants and overall investment figures.
Conclusions: Resource tracking data is an important tool for advocates
and policymakers to ensure accountability with public, philanthropic and
industry funders. As donor funding flatlines and industry involvement in
HIV research declines, it is increasingly important to ensure continued
prioritization of HIV science on the global development and research
agenda by understanding and evaluating research in the context of
public, private and philanthropic funding.
392
1
Wits Reproductive Health and HIV Institute, School of Clinical
Medicine, University of the Witwatersrand, Johannesburg, South
Africa
Background: In financially constrained research settings, it is critical to
explore and implement methods for optimising resource allocation. The
Wits RHI Clinical Research Site was renovated with the explicit intention
of accommodating multiple concurrent studies under one roof, reducing
duplicative effort and maximising resource use while providing quality
care to the population it serves. Key to the design efficiency has been
centralisation of pharmacy services across several studies. While clinical
services are often study-specific, pharmacy services provide broad
coverage and can be shared.
Methods: The Wits RHI Research Centre Pharmacy services several
ongoing studies including MTN 020, MTN 015, MTN 016, HARP and HIMSA. Operational expenses are fixed regardless of the number of studies
serviced by the pharmacy. While only MTN 020 is an Investigational New
Drug (IND) study, participants enrolled in all five studies are provided
with non-IND medications by the pharmacy. The fixed operational
expenses of this multi-study pharmacy were investigated for 2013 by
reviewing expenses incurred for staffing, pharmacy licence, electricity,
maintenance, calibrations and temperature monitoring.
Results: The fixed operational costs for the pharmacy were ±R800 000
(±$80 000). If an additional pharmacy was established, operational
costs would be approximately doubled, as most of these costs would be
duplicated. By sharing pharmacy services, the consequent economies of
scale are significant, where cost per unit of work output decreases with
an increase in scale as fixed costs are spread out over several studies
and operational efficiency is increased.
Conclusions: Centralisation of Pharmacy Services significantly reduces
costs when the service is shared across multiple budgets. It also allows for
the rolling over of studies, where new studies can start before older studies
have ended and staff can be retained. In addition, it reduces study staffing
requirements, as pharmacy staff can be cross-trained on all studies.
P47.05
P47.06
Getting Multi-purpose Prevention to Women
Now! A Business Case for Female Condoms
Is a HIV Vaccine a Viable Option in South
Africa and at What Cost?
Anna Forbes1,2, Sarah Thurston1
Nishila Moodley1,2, Glenda Gray1, Alex Welte3, Melanie Bertram4
Global Health Visions, Brooklyn, NY, United States, 2Independent
Consultant, Kensington, MD, United States
Perinatal HIV Research Unit, Vaccine Research Centre, Johannesburg,
South Africa, 2School of Public Health, Faculty of Health Sciences,
University of the Witwatersrand, Johannesburg, South Africa,
3
The South African Department of Science and Technology/
National Research Foundation (DST/NRF) Centre of Excellence in
Epidemiological Modelling and Analysis (SACEMA), University of
Stellenbosch, Stellenbosch, South Africa, 4Health Systems Governance
and Finance, World Health Organization, Geneva, Switzerland
Background: In 2012, the UN Commission on Life-saving Commodities
for Women and Children cited female condoms (FCs) as a high-impact
tool that, more widely used, could save millions of lives. Pilot projects
show that creating sustained demand for FCs is feasible. With strategic
investment, they could become as familiar as anti-malarials, oral
rehydration salts, male condoms, and other products widely used in the
developing world. This business case, commissioned by the Universal
Access to Female Condoms Joint Programme, shows the economic
impact that such investment could have in reducing health care costs
and lost economic productivity associated with unprotected sex.
Methods: The business case uses peer-reviewed, open-source modelling
tools to examine the return on investment (ROI) achievable by funding
FC programmes and the cost effectiveness of this investment. With
key data from four countries, it assesses the impact that providing one
million FCs (and the requisite marketing) could have on population
health status and real economy variables. Desk research and interviews
also shed light on the likely economic implications across society of
such investment.
Results: Excellent ROI was shown for Cameroon, Nigeria and Kenya. In
Myanmar, the ROI fell short of total costs due to lower HIV prevalence.
Using WHO-CHOICE recommended cost effectiveness thresholds, FCs
offered a highly competitive cost per DALY in all four countries, compared
to other investments. FCs provide wide-reaching economic and social
benefits by improving method choice and coverage, supporting women’s
empowerment and reducing health disparities.
Conclusions: Broad macroeconomic benefits are associated with higher
rates of protected sex, lower fertility, and reduced HIV prevalence, all
achievable through greater access to a broad basket of contraceptive
choice, including FCs. Investment in female condoms now can unleash a
virtuous cycle of increased utilization, sustained demand, better health
outcomes, and stronger economies.
1
Background: South Africa is the epicentre of the global HIV epidemic.
Improved access to anti-retroviral therapy has done little to counter
balance the 2.5 million people infected with HIV annually. Competing
disease burdens, such as trauma, place considerable demands on
limited South African health budgets. While vaccination may be the most
effective way to prevent infectious disease, it is not without challenges or
an opportunity cost. The study aims to define the maximum HIV vaccine
price that would prove cost-effective in South African public health
sector. The sensitivity of cost-effectiveness to vaccine characteristics and
risk-behaviour changes among vaccine recipients will be assessed.
Methods: Markov models will estimate the costs and health outcomes of
existing South African HIV treatment and prevention strategies with and
without HIV vaccination. Estimates will be based on a lifetime horizon
period and employ a governmental perspective. Cost utility analysis
will be conducted with uncertainty analysis of assumed parameters.
Uncertainty will be assessed by one-way sensitivity and probabilistic
sensitivity analysis. Expected value of perfect information (EVPI) analysis
will determine the importance on individual parameters.
Results: The analysis will indicate the price at which the HIV vaccine will
become most cost-effective in the South African setting and will explore
the factors that contribute the most to these costs.
Conclusions: The methodology will allow for the evaluation of health
related interventions not available in healthcare settings. Apart from HIV
vaccine research value, the study will advise decision-makers on the
economic impact of HIV vaccine adoption into policy.
www.hivr4p.org
393
POSTERS
1
Posters
P47.07
Exploring the Impact of and Requirements
for Adding a Vaccine to the Updated UNAIDS
Investment Framework to End AIDS
Arne Näveke1, Emily Donaldson2, Chaitra Gopalappa3, Kevin
Fisher2, Thomas Harmon1, Margaret McGlynn1, John Stover3,
Mitchell Warren2
International AIDS Vaccine Initiative, New York, NY, United States,
AVAC, New York, NY, United States, 3Futures Institute, Glastonbury, CT,
United States
1
2
POSTERS
Background: An updated 2011 UNAIDS Investment Framework
reflecting 2013 WHO treatment guidelines modeled how scaling up
available treatment and prevention interventions, and adding new
prevention tools, including a vaccine, could reduce new HIV infections
and AIDS-related deaths until 2050. This study explores the specific
impact of an AIDS vaccine depending on product and program
characteristics. It also explores cost-effectiveness as a major criterion for
policy makers to consider a new tool for public health programs.
Methods: We used the Goals model to project the impact of vaccines of
various characteristics in 24 low- and middle-income countries (LMICs)
for various scenarios of scale-up of existing interventions. Vaccine
characteristics explored include efficacy, duration of protection, uptake
and cost.
Results: Vaccines under all scenarios strongly reduce HIV incidence
and AIDS-related deaths, with more efficacious vaccines of longer
protection providing greatest benefit. Added to an optimized scale-up of
existing interventions, a 60% efficacious and well implemented vaccine
could avert up to 8.9 million infections by 2050, reducing annual new
infections to 184,000 in LMICs. Added to a 50% scale-up of existing
tools, the vaccine could avert up to 16 million new infections, reducing
annual new infections to 367,000. Cost-effectiveness is highly sensitive
to efficacy, cost, and duration of protection, but less sensitive to uptake.
Added to an optimized scale-up of existing interventions the vaccine
would be cost-effective at costs per regimen of less than $40 in lowincome countries. A vaccine added to 50% scale-up would be costeffective at costs per regimen below $80.
Conclusions: With existing and other emerging treatment and
prevention options, AIDS vaccines could be fundamental to achieving
and sustaining the end of AIDS. These data provide strong evidence for
sustained research and development toward efficacious AIDS vaccines
at acceptable cost for incorporation into comprehensive programs.
394
Posters 48: Risk Perception
P48.01
P48.02
Significant Prevalence of Heterosexual Anal
Sex among FSWs in India Highlights the Need
for Specifically Tailored Interventions for HIV
Prevention
Knowledge and Perception on HIV
Discordance, Transmission and Prevention
among Couples in Durban, South Africa
National AIDS Research Institute, Clinical Trial Unit, Pune, India,
National AIDS Research Institute, Pune, India, 3National AIDS Research
Institute, Director, Pune, India
1
2
Background: Receptive anal sex increases HIV transmission risk by
20-fold per act as compared to receptive vaginal intercourse. Reports
on Heterosexual Anal Sex (HAS) in Female Sex Workers (FSW), an
important core population for HIV transmission are limited. HAS and
its association with awareness, vulnerability, social empowerment and
effect of focused intervention was studied in a large cross sectional
survey among FSWs in India.
Methods: Data from 8109 FSWs from 19 districts of three high prevalence
States of India from Round 2 Integrated Behavioural and Biological
Assessment (IBBA) are presented. IBBA was aimed at studying the impact
of “Avahan”, one of the largest HIV prevention programmes in India.
Results: HAS was reported by 15.6%. FSWs who were literate (AOR 1.28
(1.09-1.49)) with good knowledge of STI symptoms (AOR 1.85(1.532.25)) reported HAS. No other source of income (AOR 1.32(1.10-1.57))
and high volume of sex work in a week in terms of number of days (AOR
1.43(1.16-1.77)) and clients (AOR 1.25(1.02-1.54)) were associated with
HAS. Accessing STI treatment from Avahan clinic (AOR 2.01(1.71-2.36))
and greater sense of empowerment (4.06(3.52-4.67)) were positively
correlated. Membership to Avahan Self Help Group (AOR 0.70(0.59-0.82))
and STI testing at Avahan clinic (AOR 0.83(0.71-0.98)) were protective.
Conclusions: Ignorance of FSWs of HIV transmission potential through
anal sex and hence need for specific interventions is highlighted. Higher
report of anal sex by FSWs who accessed STI treatment emphasises need
for doctors and nurses to openly discuss anal sex and treat anal STIs. Lifeskill training sessions aimed at empowering FSWs could be designed as
a platform for safe anal sex messages as indicated by higher reporting
of HAS by those with greater sense of empowerment. Focused service
delivery and structural intervention through trained NGOs working with
the core group will help in addressing HIV transmission through HAS.
Findings also stress the need for rectal microbicide for women too.
HIV Pathogenesis Programme, Durban, South Africa, 2Rwanda Zambia
HIV Research Group, Lusaka, Zambia, 3Medical Research Council,
Durban, South Africa, 4Emory University School of Medicine, Atlanta,
GA, United States, 5Simon Fraser University, Burnaby, BC, Canada
1
Background: Most incident HIV infection in sub-Saharan Africa occurs in
cohabiting discordant heterosexual couples. Though Couples Voluntary
Counseling and Testing (CVCT) is an effective, well studied intervention
in Africa, < 10% of couples have been jointly tested.
Methods: As part of a 2013 pilot project to expand CVCT in Durban,
South Africa, a survey was conducted pre and post-CVCT intervention
to assess change in knowledge on HIV discordance, HIV transmission
and prevention. Partners of a couple were interviewed separately using
true/false responses to statements. The McNemar Chi -square test using
STATA 11 was applied to assess change in knowledge from baseline to
post-CVCT for each statement.
Results: This study included 318 couples (636 individuals), all black
South African; mostly Zulu ethnicity (86%), Christian (84%) and at least
secondary level educated (76%). The mean age and range for men and
women was 31 (17 - 59) and 29 (16 - 59) respectively. There was low
level knowledge of possibility of HIV discordant results for a couple; only
195 individuals (31%) thought this was possible at pre‐CVCT compared
to 578 (91%) at post-test. The survey also assessed knowledge on the
benefit of CVCT; 208 (33%) thought there was at least one benefit at
pre‐CVCT and this increased to 572 (90%) at post‐CVCT. Overall, there
were statistically significant positive changes in knowledge on HIV
transmission and prevention. In comparison between pre and post CVCT
responses, most people thought that all HIV positive mothers give birth
to babies with AIDS (63% and 58%; p=0.002). Most people also thought
that Male circumcision does not protect HIV negative men against HIV
(70% and 66%; p=0.05).
Conclusions: Though the respondents had a positive attitude towards
CVCT, the majority were initially unaware of the possibility of HIV
discordance and there were misconceptions about HIV prevention and
transmission. Future messages should target gaps in knowledge and
provide information about CVCT services.
www.hivr4p.org
395
POSTERS
Mallika Alexander1, Shweta Chidrawar1, Sucheta Deshpande2,
Ramesh Paranjape3
Mammekwa Mokgoro1, William Kilembe2, Hildah Shumba2,
Miriam Kamusoko1, Tarylee Reddy3, Elisabeth Dissen2, Jonathan
Davitte4, Annie Mwaanga2, Mark Brockman5, Thumbi Ndung’u1,
Susan Allen4
Posters
P48.03
P48.04
The Resonance Project: How Canadian Gay
Men Understand, View and Incorporate
Biomedical Knowledge of HIV into their
Sexual Practices
Mobility Patterns and Concurrent Sexual
Relationships among Fisher Folk along Lake
Victoria, Kenya
Marc-André LeBlanc1, Ed Jackson2, Barry Adam3, San Patten4,
Len Tooley2, James Wilton2, Shayna Buhler5, Greg Penney6, Kim
Thomas7, Wayne Robert8, Jody Jollimore8, Joshun Dulai8, Owen
McEwen9, Daniel Pugh9, Robert Rousseau10, Gabriel Girard10,
Alexandre Dumont-Blais10
Resonance Project, Gatineau, QC, Canada, 2CATIE, Toronto, ON,
Canada, 3University of Windsor, Windsor, ON, Canada, 4San Patten
& Associates, Halifax, NS, Canada, 5Interagency Coalition on AIDS
and Development, Ottawa, ON, Canada, 6Canadian Public Health
Association, Ottawa, ON, Canada, 7Canadian AIDS Society, Ottawa, ON,
Canada, 8Health Initiative for Men, Vancouver, BC, Canada, 9Gay Men’s
Sexual Health Alliance, Toronto, ON, Canada, 10RÉZO, Montreal, QC,
Canada
1
POSTERS
Background: In the last 10 years, our biomedical knowledge of HIV
prevention has grown tremendously and several new prevention tools
are now at our disposal. Historically, gay men have been early adopters
of risk reduction strategies, such as condoms. The Resonance Project is
a 3-year community-based research project, funded by the Canadian
Institutes of Health Research, seeking to understand how biomedical
HIV knowledge is entering the discourses, prevention strategies and folk
wisdom of gay men and their service providers.
Methods: Using ‘vignettes’ of typical online profiles and dating
scenarios, we conducted 12 semi-structured focus groups with 86 gay
men in Montreal, Toronto and Vancouver. The focus groups included 4
categories:
1) gay men connected to the HIV sector (other than employment),
2) HIV + and - gay men in serodiscordant relationships,
3) HIV+ men who are sexually active, and
4) HIV- men who are sexually active and at high risk.
We then conducted additional in-depth individual interviews with 10
of the gay men. Topics we explored included: seroadaptive behaviours,
rapid and home-based testing, acute HIV infection, ARV-based prevention
options, vaccines, and cures. The audio recordings were transcribed,
coded, and analyzed using Interpretive Description (Thorne, 2008).
Results: Gay men who participated reflected diverse attitudes,
perspectives and backgrounds, based on ethnocultural origins, age,
serostatus, relationship status, and city location. Depending on the
extent to which the new biomedical knowledge had resonated with
them, they demonstrated different levels of uptake of the information
and resulting personal calculations of risk.
Conclusions: Effective HIV prevention interventions need to enable
gay men to develop their own risk management strategies in ways
that resonate with their sexual lives. They must also take into account
the ways in which gay men’s biomedical knowledge of HIV can be
at once sophisticated and incomplete, innovative and inaccurate,
complementary and contradictory.
396
Stella Njuguna1, Zachary Kwena2, Everlyn Ombati1, Margaret
Mburu2, Elizabeth Bukusi1, Raphael Ondondo, Betty Njoroge
1
Kenya Medical Research Institute, Nairobi, Kenya, 2Kenya Medical
Research Institute, Kisumu, Kenya
Background: HIV prevalence in Kenya dropped from 7.4% in 2007
to 5.6% in 2012. Despite this decrease, the HIV prevalence in Nyanza
region remains the same, 15.1%. This high prevalence may be driven
by the highly migratory “high risk” fishing communities residing along
the shores of Lake Victoria in Nyanza, with HIV prevalence at 25.6%.
We examined the association between mobility patterns and perceived
sexual practices of the fisher folk.
Methods: Beach management units’ registers were used to identify
and determine the number of fisher folk in every fish landing beach
on the shores of Lake Victoria in Kenya. Probability proportionate to
size sampling was used to draw a random sample of 2638 across
308 beaches for participation in a cross-sectional survey. Data was
collected on sociodemographics, mobility patterns and reported sexual
concurrency. Descriptive statistics and logistic regressions were used for
data analysis.
Results: In 2013 mobility was mainly to Homabay and Siaya counties
which are high HIV prevalent areas, ~36% and 31% respectively.
Mobility was highest between April and December, 167 (33.3%) and
119 (23.7%) in Homa Bay and Siaya respectively. Approximately 36%
suspected their partners had concurrent partners and 566(46.1%)
of the respondents reported that they were currently in a concurrent
relationships. Mobility was significantly associated with reported sexual
concurrency (OR 1.423 95% CI 1.09-1.85 p = 0.009). After controlling
for age, gender, education level, income, reported condom use, age of
last born child, marital status, alcohol use and duration of relationship
with the most recent sexual partner, mobility was significantly associated
with concurrency (AOR 1.74 95% CI 1.01-2.98 p=0.046).
Conclusions: Mobility and concurrent sexual practices among fisher folk
may contribute the sustained high HIV prevalence in Nyanza and calls
for targeted HIV prevention and care services.
P48.05
P48.06
How Canadian Service Providers Understand,
and Incorporate, Biomedical Knowledge of
HIV into their Prevention Work and their Own
Sexual Practices
Risk Disinhibition: The Effect of Microbicide
Communication Materials on Future
Prevention Behavior
San Patten and Associates, Inc, Halifax, NS, Canada, 2Resonance
Project, Halifax, NS, Canada, 3CATIE, Toronto, ON, Canada, 4University
of Windsor, Windsor, ON, Canada, 5Resonance Project, Gatineau, QC,
Canada, 6Interagency Coalition on AIDS and Development, Ottawa,
ON, Canada, 7Canadian AIDS Society, Ottawa, ON, Canada, 8Canadian
Public Health Association, Ottawa, ON, Canada, 9Health Initiative for
Men, Vancouver, BC, Canada, 10Gay Men’s Sexual Health Alliance,
Toronto, ON, Canada, 11REZO, Montreal, QC, Canada
1
Background: In the last 10 years, our biomedical knowledge of HIV
prevention has grown tremendously and several new prevention tools
are now at our disposal. Historically, gay men have been early adopters
of risk reduction strategies, such as condoms. The Resonance Project is
a 3-year community-based research project, funded by the Canadian
Institutes of Health Research, seeking to understand how biomedical
HIV knowledge is entering the discourses, prevention strategies and folk
wisdom of gay men and their service providers.
Methods: We conducted 3 semi-structured focus groups with 22 service
providers who work with gay men in Montreal, Toronto and Vancouver.
This was followed by 20 one-on-one interviews with service providers,
10 of which were with service providers who identify as gay men.
Topics we explored included: the challenges of conveying biomedical
information to gay men; the organizational supports and constraints
faced by service providers; and, the complex dual professional/personal
position of service providers who are gay men. The audio recordings
were transcribed, coded, and analyzed using Interpretive Description
(Thorne, 2008).
Results: Service providers reflected a wide variety of attitudes, level
of understanding, degree of comfort and willingness to incorporate
biomedical aspects of HIV prevention into:
1) their HIV prevention messages and interventions; and
2) in the case of service providers who are gay men, their own
sexual practices.
Some of the latter service providers experienced challenges in reconciling
their own sexual practices with the advice they give to gay men in their
communities, vis-à-vis their emerging biomedical knowledge of HIV risk
and prevention.
Conclusions: Service providers play a critical role in implementing
HIV prevention interventions that are reflective of gay men’s lives and
new biomedical understandings of HIV. Special attention is required to
recognize and mobilize the dual position of service providers who are
gay men in shaping prevention responses.
FHI 360, Social & Behavioral Health Sciences, Durham, NC, United
States, 2FHI 360, Quantitative Sciences, Durham, NC, United States,
3
FHI 360, Social Marketing and Communication, Washington, DC,
United States, 4FHI 360, Global Health, Population & Nutrition,
Washington, DC, United States, 5FHI 360, Country Office, Nairobi,
Kenya, 6National AIDS & STI Control Programme, Nairobi, Kenya
1
Background: Vaginal gel use may provide a new HIV prevention
tool for women who cannot negotiate condom use. Yet, current
microbicide formulations are less effective than consistent condom
use. Before introducing such options, it is essential to develop and test
communication materials that encourage gel use in the absence of
condom use, while promoting condoms-only or gel+condoms among
those already using condoms to avoid reduced protection. In Kenya, we
developed and assessed the effect of a minimum package of materials
on preferences for future HIV prevention and possible risk disinhibition.
Methods: We compared participants’ reports of “ever condom use
with a current/recent partner” to future HIV prevention preferences (no
product, gel-only, condoms-only, or gel+condom) among 746 sexuallyactive women and men in two Kenyan cities, who received informationonly, or viewed either HIV-framed or non-HIV framed awarenessraising materials (posters, TV and radio spots). We also assessed future
prevention preferences among 99 young women and female sex
workers (FSWs) pre- and post- participation in educational sessions using
tailored flipcharts.
Results: Risk disinhibition was low regardless of framing or material.
After viewing awareness-raising materials, 80% of participants would
maintain condom use, or increase their protection by adding gel to
condom use or moving from no protection to gel, condoms or both.
Married monogamous participants were significantly less likely than
others to increase protection; 21% reporting past condom use preferred
using no protection or gel-only. Preference for less protection after
participating in educational sessions remained low (2% pre and post-test
for FSWs) or decreased (from 7.5% to 2.5% in young women).
Conclusions: Microbicide introduction must position use as possible,
even desirable, by stable couples, while reinforcing the idea of gel as
added protection for condom users or casual couples. Accomplishing
these goals requires careful message development and testing.
www.hivr4p.org
397
POSTERS
San Patten1,2, Ed Jackson3, Barry Adam4, Marc-Andre Leblanc5,
Len Tooley3, James Wilton3, Shayna Buhler6, Kim Thomas7, Greg
Penney8, Wayne Robert9, Jody Jollimore9, Joshun Dulai9, Owen
Mcewen10, Daniel Pugh10, Robert Rousseau11, Gabriel Girard11,
Alexandre Dumont-Blais11
Elizabeth E. Tolley1, Allison P. Pack1, Sam Field2, Elizabeth Ryan3,
Bockh Emily4, Caroline Mackenzie5, Alice Olawo5, George Githuka6
Posters
P48.07
Conceptualizations of Uncertainty and
Risk and Implications for Biomedical HIV
Prevention Technologies in Sub-Saharan
Africa: A Systematic Review
Emily Warren1, Pauline Paterson1, Shelley Lees2, Robyn Eakle3,
Jonathan Stadler3, Heidi Larson1
London School of Hygiene and Tropical Medicine, Epidemiology
and Population Health, London, United Kingdom, 2London School of
Hygiene and Tropical Medicine, Public Health and Policy, London,
United Kingdom, 3Wits Reproductive Health & HIV Institute, Hillbrow,
South Africa
1
POSTERS
Background: Two HIV pre-exposure prophylaxis (PrEP) trials, Voice
and Fem PrEP, have recently been stopped due to lack of efficacy,
often attributed to poor adherence. While oral PrEP was shown to be
effective in other settings, its lack of efficacy, particularly for African
women has prompted an urgent re-examination of assumption
about HIV risk perception and the quotidian context in which sexual
transmission of HIV occurs.
Our aim is to understand how healthcare providers and individuals
considered in the “general” or “high-risk” populations define and classify
risk and navigate uncertainty in relation to sexual transmission of HIV,
and identify how those conceptualizations are associated with the use
biomedical prevention interventions.
Methods: We are conducting a systematic review and meta-ethnography
of qualitative findings, which allow us to keep the explanatory context
intact. Six databases, Medline, Embase, PsychInfo, Africa Wide Info,
CINAHL, and Global Health, were searched. To be included, papers
employ qualitative methods to explore people’s perception of risk,
uncertainty, hope, trust, fear and/or decision making in relation to
biomedical HIV prevention interventions. Because of feasibility concerns,
we limited our included articles to only those that examined sexual
transmission of HIV.
Results: After duplicates were removed from our search results, two
authors screened 8,826 articles. Screening is ongoing and more detailed
results will be available before October.
Conclusions: To the best of our knowledge, this is the first systematic
review to explore different conceptions of risk and uncertainty in
relation to the use of HIV biomedical technologies. This is especially
important given the increased number of prevention interventions
currently being researched.
398
Posters 49: Sexually Transmitted Infections
P49.01
P49.02
Socio-demographic and Behavioural
Characteristics Associated with HSV-2 Seroprevalence in High Risk Women in KwaZuluNatal, South Africa
Burden and Predictors of HIV /Hepatitis B Coinfection in Rural Uganda
South African Medical Research Council, HIV Prevention Research
Unit, Durban, South Africa, 2The Kirby Institute, University of New
South Wales, Sydney, Australia, 3London School of Hygiene & Tropical
Medicine, Department of Epidemiology and Population Health, London,
United Kingdom
1
Background: The World Health Organization estimated that 536
million people aged 15-49 are infected with Herpes simplex virus type
2 (HSV-2), the causative agent of genital herpes. The aim of this study
was to investigate the role of behavioral and demographic factors
that contribute to the high HSV-2 sero-prevalence among women
participating in a HIV prevention trial.
Methods: The Methods for Improving Reproductive Health in Africa
(MIRA) study assessed the effectiveness the latex diaphragm and
lubricant gel on HIV prevention among women in South Africa and
Zimbabwe. At screening an interviewer administered questionnaire
on demographics and sexual behaviour was obtained. HSV-2 serum
antibodies was detected using HerpeSelect TM2 ELISA IgG. Statistical
analysis was performed using STATA release 12.0. Univariate and
multivariate analyses were performed using logistic regression for both.
Results: In this study, the prevalence of HSV-2 was estimated at 73%
with 41% of the women also co-infected with HIV. In the multivariate
analyses, older women (35 years and older) were more likely to be
HSV-2 sero-positive as compared to younger women (OR: 3.49 95% CI:
2.71, 4.49 and OR: 1.82 95% CI: 1.49, 2.22) respectively. Women with
a lower level of education were also considered to be more likely to be
HSV2 positive (OR: 1.26 95% CI: 1.03, 1.53). Additionally, having >1
life-time sexual partners (OR: 2.48, 95% 1.92, 3.20) parity >1 (OR: 1.95
95% CI: 1.92, 3.20) and being HIV positive (OR: 6.31, 95% CI 5.06,
7.88) were all significantly associated with HSV-2 infection.
Conclusions: The high sero-prevalence of HSV-2 in the studied
population is of great public health importance since this high risk
population could act as a reservoir for future infections particularly HIV
transmission. The identification of risk factors for HSV-2 infection could
aid in the development and implementation of personalized prevention
messages for this infection.
MRC/UVRI, Uganda Research Unit on AIDS, Kampala, Uganda,
Wellcome Trust Sanger Institute, Hinxton, United Kingdom, 3University
of Cambridge, Public Health and Primary Care, Cambridge, United
Kingdom
1
2
Background: Hepatitis B (HBV) and HIV have similar transmission
patterns and thus HBV/HIV co-infection is common and this increases
morbidity, mortality and risk of transmission. We estimated the burden
of HBV/HIV co-infection and determined the associated factors in rural
Uganda to inform the design of targeted preventive interventions.
Methods: : In 2011 a population based survey was conducted among
residents of an established rural general population cohort (aged ≥13
years) in SW Uganda. Data on demographic characteristics, risky sexual
behaviour (multiple and casual sex partners, condom use) were collected
and all participants were tested for HIV (ELISA+ Western Blot), HBV
(HBsAg) and liver function. Participants with confirmed infections were
referred for care where viral loads, CD4 count (HIV+ only) were done.
We determined the proportion of HBV/HIV co-infection and explored
associated factors by logistic regression.
Results: Of 8078 participants tested, 7% had HIV infection only; majority
were female (66%) and the mean age was 38 years (±11.5SD), 36%
were on ART with a mean CD4 count of 367.6cell/mm3 (±1.8SD). The
HBV prevalence was 4.6% (27/584) among those infected with HIV and
3.0% among HIV uninfected participants. Compared to those infected
with HIV only, HBV/HIV co-infected participants were more likely to
have had sex with a casual partner in the last 12 months (31.8%vs
14.4% p=0.04) and to have elevated liver transaminases (p=0.04). Sex,
number of sexual partners, CD4 count, HIV RNA viral load and use of
ART were not associated with co-infections.
Conclusions: In this population we found high risk sexual behaviour as a
predictor of HBV/HIV co-infection. The presence elevated liver enzymes
may be a surrogate marker for acute hepatitis reflecting a higher risk of
liver damage among co-infected participants. Preventive interventions
that target both HIV and HBV are needed among high risk groups.
www.hivr4p.org
399
POSTERS
Nathlee S. Abbai1, Handan Wand2, Ramjee Gita1,3
Clara Wekesa1, Gershim Asiki1, Ivan Kasamba1, Rebecca N.
Nsubuga1, Robert Newton1, Liz H. Young2,3, Manjinder Sandhu2,3,
Alex Karabarinde1, Anatoli Kamali1
Posters
P49.03
P49.04
Factors Associated with Sexually Transmitted
Infections and HIV Risk in the Carraguard
Phase 3 Trial
Patterns of Drug Use among People who
Inject Drugs (PWID) and their Implications for
Sexually Transmitted Infections in Northern
Nigeria
Barbara A. Friedland1, Doug Taylor2, Khatija Ahmed3, Thesla
Palanee4, Marlena Plagianos5, Lauren Katzen5
Population Council, HIV and AIDS Program, New York, NY, United
States, 2FHI 360, Durham, NC, United States, 3Setshaba Research
Centre, Soshanguve, South Africa, 4Wits Reproductive Health & HIV
Institute, Johannesburg, South Africa, 5Population Council, New York,
NY, United States
1
POSTERS
Background: To describe the burden of sexually transmitted infections
(STIs), correlates of prevalent infection, and associations between
prevalent infection and HIV risk among South African women regularly
tested and treated for curable STIs in an HIV prevention trial.
Methods: We conducted a secondary analysis of data from 6004
women who participated in the Carraguard Phase 3 trial between 2004
and 2007 to further assess the burden of STIs and related risk factors.
Women were treated for STIs based on positive test results or if clinically
indicated, and were given referral letters for partners to access testing and
treatment. We classified prevalent gonorrhea, chlamydia, trichomoniasis
and syphilis in 6-month follow-up periods (up to 24 months). We used
adjusted logistic regression and survival analysis to assess relationships
between time in study, types of partnerships, previous STI status and
other time-varying factors on prevalent and incident STIs.
Results: STI prevalence fell significantly from baseline (25%) to Months
1-6 (21%) and Months 7-12 of follow-up (12%), after which prevalent
infection with one or more STIs remained steady through Month 24.
Younger age was associated with higher prevalence of gonorrhea
and chlamydia (p< 0.01); the opposite relationship was observed for
syphilis. Previous STI (except syphilis) and reports of new partners were
positively associated with prevalence of STIs. Prevalent gonorrhea,
chlamydia and trichomoniasis were also positively associated with
risk of HIV seroconversion in adjusted models (HR=3.0, 2.7 and 2.4,
respectively; P< 0.01).
Conclusions: We observed a significant reduction in prevalence of curable
STI over time in a trial with routine testing, treatment and partner referral.
The overall STI burden remained substantial, however, emphasizing
the ubiquitous nature of STIs in South Africa. Further reductions in STI
prevalence would require including tests of cure and actively ensuring
partner treatment, which may not be feasible in trial settings.
400
Desmond A. Iriaye1, Ibrahim Suleiman1, Otibho Obianwu1,
Sylvia Adebajo1, George Eluwa1, Jean Njab1, Ayodeji Oginni1,
Babatunde Ahonsi1
Population Council, HIV/AIDS, Abuja, Nigeria
1
Background: Drug use impairs judgment and leads to sexual risk
behaviours. The prevalence of HIV among PWID in Kano (4.8%) and
Kaduna (5.8%) respectively are higher than the national HIV prevalence
among PWID (4.2%). However the national prevalence of sexually
transmitted infections (STI) among PWID is about 6%. In this study we
sought to determine drug use profile, prevalence and correlates of STIs
among PWID in Northern Nigeria.
Methods: Data of drug use, sexual and injecting risk behaviors were
collected from structured risk assessment forms between May and
October 2013 and evaluated in a cross sectional study. Chi-square test
was used to compare differences between categorical variables and
logistic regression analysis was conducted to identify factors associated
with patterns of drug use and sexual risk behaviour.
Results: A total of 385 drug users (9% Females and 91% Males) with
mean age of 30 years were interviewed. More than 94% injected
drugs in the last 12 months. Mean age of drug use debut was15.4
years, while mean duration of drugs use was 15 years. Types of drugs
injected included Heroine (30%), Cocaine (24%), Amphetamine (8%),
Crack (10.4%) and Pentazocine (39.2%). Over 80% of heroin users and
72.3% of cocaine users, had unprotected sex with casual partner in
the last 6 months. About 28% of drug users experienced one form of
sexually transmitted infection in the last 6 months. Logistic regression
analysis showed that amphetamine users were 11 times more likely to
have STI (Adjusted OR= 11.52, 95%CI=4.27-31.10) followed by Heroin
users (Adjusted OR= 4.99, 95%CI=2.46-10.13) and Pentazocine users
(Adjusted OR=2.86, 95%CI=1.41-5.81).
Conclusions: This study strongly suggested that drug use significantly
predisposes PWID to sexual risk behaviours and STIs. In addition,
exposure to STIs seems to significantly vary with the type of drugs used.
Therefore, there is a need for appropriate HIV/STI prevention programing
for PWID in Northern Nigeria
P49.05
P49.06
Prevalent and Incident Gonorrhea
and Chlamydia Infections among
Oral Contraceptive and Depot
Medroxyprogesterone Acetate Users in MTN003 (VOICE)
Uptake of HBV Vaccination and Incident HBV
Infection in Women of Reproductive Age and
at Risk of HIV-1 Infection in the VOICE (MTN
003) Study
2
3
MU-JHU Research Collaboration, Kampala, Uganda, 2Johns Hopkins
University, Baltimore, MD, United States, 3SCHARP-FHCRC, Seattle, WA,
United States, 4MRC, Durban, South Africa, 5Wits RHI, Johannesburg,
South Africa, 6CAPRISA, Durban, South Africa, 7Aurum Institute,
Klerksdorp, South Africa, 8PHRU, Soweto, South Africa, 9University of
Washington, Seattle, WA, United States, 10Magee-Womens Research
Institute, Pittsburgh, PA, United States, 11RTI International, San Francisco,
CA, United States, 12FHI 360, Durham, NC, United States, 13DAIDS/
NIAID/NIH, Bethesda, MD, United States, 14UZ-UCSF, Harare, Zimbabwe
1
Background: Oral contraceptive (OC) and depot medroxyprogesterone
acetate (DMPA) use are common in areas with high sexually transmitted
infection (STI) incidence. Recent STI increases risk of HIV acquisition and
evidence suggests Chlamydia trachomatis (CT) risk may be higher among
OC and DMPA users compared to users of no hormonal contraception.
Data are limited regarding whether STI incidence differs between OC
and DMPA users. We compared CT and Neisseria gonorrhoeae (NG)
rates directly between OC and DMPA users.
Methods: We analyzed prospective data from VOICE, a randomized
trial of oral and topical HIV chemoprophylaxis in Uganda, Zimbabwe
and South Africa. Analysis was restricted to 2,548 women who used
OC (756) or DMPA (1,792) exclusively during follow-up (f/u). Screening
and treatment for CT and NG occurred at baseline (BL), annually, when
clinically indicated, and at exit. We used site-stratified Andersen-Gill
proportional hazards models to quantify CT and NG risk, excluding
periods of no injectable or oral contraception and censoring at pregnancy
and HIV seroconversion.
Results: Compared to DMPA users, OC users were older (26.2 vs.
25.4 years, p< 0.001) and had lower BL prevalence of CT, although
NG prevalence was similar (CT: 9.9% vs. 14.0%, p=0.01) (NG: 2.5% vs.
3.5%, p=0.23). Over 2,810 person-years (py) of f/u, 405 cases of CT
(14.4/100 py) and 93 of NG (3.3/100 py) were detected. While incidence
for both CT and NG was lower among OC compared to DMPA users (CT:
9.4 vs. 16.5/100 py; hazard ratio [HR] 0.59, 95% CI 0.46-0.76) (NG: 1.5
vs. 4.1/100 py, HR: 0.37, 95% CI 0.20-0.68), no significant differences
were noted for CT or NG after adjusting for age, recent partner change,
and other factors (CT adjusted HR [aHR]: 0.71, 95% CI 0.47-1.08; NG
aHR: 0.54, 95% CI 0.20-1.46).
Conclusions: Differences in CT and NG risk between OC and DMPA
users may be related to demographic and behavioral factors. Providers
should emphasize condoms and other STI prevention strategies for all
women and their partners.
Nyaradzo M. Mgodi1, Cliff W. Kelly2, Brenda Kakayi3, James Y. Dai2,
Gonasagrie Nair4, Thesla Palanee5, Kailazarid Gomez-Feliciano6,
Jeanne Marrazzo7, Jeanna Piper8, Zvavahera Mike Chirenje1
University of Zimbabwe - University of California San Francisco
Collaborative Research Programme, Harare, Zimbabwe, 2SCHARP FHCRC, Seattle, WA, United States, 3Makerere University-Johns Hopkins
University Research Collaboration, Kampala, Uganda, 4University
of KwaZulu Natal, CAPRISA, Durban, South Africa, 5University of
the Witwatersrand, Wits Reproductive Health and HIV Institute,
Johannesburg, South Africa, 6FHI 360, Durham, NC, United States,
7
University of Washington, Seattle, WA, United States, 8DAIDS/NIAID/
NIH, Bethesda, MD, United States
1
Background: In 1991, the Global Advisory Group of the Expanded
Programme on Immunization recommended that in areas of high HBV
endemicity like Sub Saharan Africa (SSA), HBV vaccine be integrated
into national immunization programs. Most SSA countries found this
roll-out challenging given financial constraints and competing public
health priorities. VOICE, a multi-site HIV PrEP trial, was conducted in SSA
where high HBV and HIV prevalence rates exist. We aimed to determine
the uptake of HBV vaccine and HBV incidence in VOICE.
Methods: A total of 5029 women enrolled into VOICE. Prior HBV exposure
was determined at screening (HbsAg, HBsAb). Screening exclusion criteria
included HIV infection and HBV infection. Eligible HBsAg negative women
were randomized equally to daily oral tenofovir disoproxil fumarate (TDF),
TDF/emtricitabine, oral placebo, vaginal tenofovir or placebo gel; all were
offered HBV vaccine if not immune (HBsAb negative) or counselling only
if immune (HBsAb positive). We summarize HBV testing, vaccination
uptake/completion and HBV incidence.
Results: Three thousand six hundred and sixty seven (72.9%) women
were not immune at enrolment. Of these, 3648 initiated the vaccination
schedule at baseline or sometime during follow-up, yielding a total
rate of HBV vaccination uptake of 99.5%. Two thousand nine hundred
and eighty eight (81.5%) completed the 3-dose vaccination course as
scheduled. A total of 3310 (90.3%) completed > 3 doses, including
cases where the vaccination course was interrupted and later restarted.
Among the 3667 who were susceptible at baseline, 5(0.14%) acquired
HBV during follow-up, with each HBV infection occurring after receipt of
> 2 doses of vaccine. Of the 5, 1(20%) had HBV and HIV co-infection.
Conclusions: Among VOICE participants, most were susceptible to HBV
at enrolment, and uptake of vaccination was high. Despite HIV incidence
of 5.7% during the study, the incidence of HBV in women at risk for HIV
was low, presumably due to high uptake of effective vaccination.
www.hivr4p.org
401
POSTERS
Flavia Matovu Kiweewa , Lisa Noguchi , Barbra Richardson ,
Jen Balkus3, Betty Kamira1, Brenda Mirembe Gati1, Clemensia
Nakabiito1, Gita Ramjee4, Thesla Palanee5, Gonasagrie Nair6,
Pearl Selepe7, Ravindre Panchia8, Patrick Ndase9, Edward Livant10,
Ariane Van der Straten11, Kailazarid Gomez12, Devika Singh9,
Mary Glenn Fowler1,2, Jeanna Piper13, Zvavahera Mike Chirenje14,
Jeanne Marrazzo9
1
Posters
P49.07
P49.08
Analysis of Griffithsin (GRFT) Binding by
Proteins in Cervical Vaginal Lavage and the
Influence of Vaginal Microflora
Prevalence of HIV, Sexual Transmitted
Infections and High Risk Sexual Behaviors
among Female Sex Workers in Kigali, Rwanda
Bernard J. Moncla1,2, Lara K. Mahal3, Brian M. Debo2, Catherine
Chappell4, Katherine E. Bunge4, Leslie Meyn4, Jordan J. Szpara2,
Lisa C. Rohan2,5, Sharon L. Hillier2,4
Jeannine Mukamuyango1, Rosine Ingabire1, Etienne Karita1, Julien
Nyombayire1, Robertine Sinabamenye1, Nuri Ahmed1, Jean Nepo
Nduwamungu1, Sukaina Radjabari1, Marydale Oppert2, Matt Price3,
Frances Priddy3, Pat Fast3, Amanda Tichacek1,2, Susan Allen1,2
University of Pittsburgh, Ob./Gyn and Reproductive Sciences,
Pittsburgh, PA, United States, 2Magee-Womens Research Institute,
Microbiology, Pittsburgh, PA, United States, 3New York University,
Chemistry, New York, NY, United States, 4University of Pittsburgh,
Ob/Gyn and Reproductive Sciences, Pittsburgh, PA, United States,
5
University of Pittsburgh, Pharmacy, Pittsburgh, PA, United States
1
POSTERS
Background: Griffithsin (GRFT), an anti-HIV protein isolated from the
red alga Griffithsia sp., is being developed as a microbicide. GRFT
binds N-linked glycans on gp120 to prevent HIV entry. Earlier studies
evaluating GRFT binding in cervicovaginal lavage (CVL) samples
using lectin microarrays led us to investigate how GRFT interacts with
glycoproteins in the vaginal microenvironment
Methods: CVL was collected in PBS from 135 healthy asymptomatic
reproductive aged women and stored at -70̊ C until used. Vaginal smears
were evaluated using the Nugent criteria for bacterial vaginosis (BV).
The samples were evaluated for GRFT binding in an ELISA using antiGRFT primary antibody and a horseradish peroxidase labeled secondary
antibody. The ELISA data were analyzed using one-way analysis of
variance. GRT binding proteins in the CVLs were identified by SDS-PAGE
and Western blotting.
Results: GRFT bound at higher levels in the CVLs obtained from
women having a Lactobacillus-predominant flora (Nugent score 0-3) or
intermediate flora (Nugent 4-6) compared to BV flora (Nugent 7-10) in
pg/ng protein: 10.74 + 5.82, 10.56 + 8.09, 4.03 + 3.22 pg/ng protein,
respectively, P value = < 0.001(Normal vs BV P< 0.001; Intermediate vs
BV P=0.001). Western blots of normal CVL revealed five GRFT binding
proteins at apparent molecular weights (in kDa) of 185, 142, 120, 81
and 71, while 2/4 BV CVL demonstrated no GRFT binding proteins and
two had binding to proteins with apparent MWs of 54 to>250. The CVL
from women with BV had reduced levels of these GRFT binding proteins.
Conclusions: GRFT binds to at least 5 proteins found in CVL, with
different protein patterns in women with normal flora vs those with BV.
The lower binding of GRFT in the CVL of women with BV suggests a
protective function for GRFT binding proteins. It also suggests that a
GRFT based microbicide may be bound to endogenous vaginal proteins
reducing availability to compete with the HIV gp120 protein in women
with normal flora.
402
Rwanda Zambia HIV Research Group-Projet San Francisco, Kigali,
Rwanda, 2Emory University, School of Medicine, Department of
Pathology and Laboratory Medicine, Atlanta, GA, United States, 3IAVI,
1
Background: Female sex workers (FSW) are at high risk of transmitting/
acquiring HIV and other Sexually Transmitted Infections (STIs).
Despite widespread education, counselling, and condom distribution
interventions among FSW, unprotected sexual intercourse persists. This
study aimed to assess the level of HIV prevalence among FSWs who
consulted VCT services at Projet San Francisco in Kigali, Rwanda.
Methods: From September 2012-March 2014, Projet San Francisco
provided HIV voluntary counseling and testing, STI screening and
treatment as well as Long Acting and Reversible Contraceptive (LARC)
methods to FSW invited from known hot spots of commercial sex
activity in Kigali. HIV negative women are offered enrollment into a
prospective study on HIV incidence, while HIV positive women are
referred to government health centers for care and treatment.
Results: During the study, 605 FSW received services at PSF, of whom
263 (43%) tested HIV negative. The mean age was 31 years with a range
of 16-51. HIV prevalence rose with age from 31% in < 24 year olds to
56% in those aged 25-29, 58% in 30-34 year olds and 70% among
those >35. At baseline, 264 (44%) FSW were diagnosed with at least
one STI. The most commonly diagnosed STI was Syphilis (151 cases;
25%), followed by Trichomoniasis (84 cases; 14%), Pelvic Inflammatory
diseases (33 cases; 5%), and Gonorrhea (5 cases; 1%). 12 women were
treated for more than one STI. The average number of sex clients per
month was 44 (range: 0-500), the median was 20 clients per month and
the proportion of FSW who reported consistent condom use for all sex
acts in the last month was 75%.
Conclusions: FSW in Kigali have a very high prevalence of HIV and
other STI. Increasing HIV testing and counseling is especially needed at
an early age to decrease risk of new HIV infections. Specific interventions
for this high-risk population are needed to limit the spread of the HIV/
AIDS epidemic.
P49.09
P49.10
Sexually Transmitted Infections: Co-infections
and Link to Variations in HIV Prevalence
and Disease Management Outcomes across
Regions in Kenya
Prevalence and Determinants of Hepatitis
B Virus (HBV) Infection in a Cohort of HIV1 Discordant Couples in Western Kenya:
Implications for HIV Care
Joseph Mwangi1,2, Joyceline Kinyua1, Nancy Lagat1, Raphael
Lihana1, HIV Coinfection and Molecular Study Group
Dismas C.O. Oketch1, Eunice Kaguiri1, Nereo Murgor2, Cosmas
Apaka3, Paul Ayuo4, Edwin Were5, Kenneth Fife6
1
Kenya Medical Research Institute, Nairobi, Kenya, 2Insititute for
Tropical Medicine and Infectious Disease, Jomo Kenyatta University of
Science and Technology, Nairobi, Kenya
Moi University School of Medicine, Partners in Prevention, Eldoret,
Kenya, 2Moi University School of Nursing, Eldoret, Kenya, 3Moi
University College of Health Sciences, Center of Excellence for
Cardiovascular & Respiratory Diseases, Eldoret, Kenya, 4Moi University
School of Medicine, Department of Internal Medicine, Eldoret, Kenya,
5
Moi University School of Medicine, Department of Reproductive
Health, Eldoret, Kenya, 6Indiana University School of Medicine,
Department of Medicine & Infectious Diseases, Indianapolis, IN, United
States
Background: Hepatitis B is a major global health problem with over
350 million people suffering from chronic HBV infection. Infection with
HBV negatively impacts the outcome of HIV infection and yet there are
no national clinical guidelines for diagnosis, treatment and care for HBVHIV co-infection. This study sought to determine the prevalence and
determinants of HBV infection in a cohort of HIV-1 discordant couples
in Western Kenya.
Methods: A cross sectional study of healthy heterosexual HIV-1
discordant couples from Western Kenya referred for possible recruitment
into the Partners PrEP Study was conducted between September 2008
and October 2010. All participants were screened for hepatitis B surface
antigen (HBsAg) and socio-demographic data were collected. Descriptive
statistics were used to determine frequencies while the association
between HBsAg and the independent variables were evaluated using
logistic regression.
Results: Data on 1834 adults aged 18-64 are presented. HBsAg was
positive in 77 [4.2%] individuals. Men were 40% less likely to be
infected compared to women [OR 0.61; 95% CI 0.379-0.972; p value
0.0380]. Those from rural areas were almost 2 times more likely to be
infected than those from urban areas [OR 1.92; 95% CI 1.190-3.099;
p value 0.008]. Hepatitis B prevalence did not differ by HIV status [HIV
positive 3.9% vs negative 4.5% [OR=1.15; 95% CI of 0.725-1.810 p
value = 0.561]. There was also no significant correlation between
hepatitis B and participants´ age, alcohol intake, CD4 counts, income or
level of education. Immunity to hepatitis B (as measured by the presence
of antibody to HBsAg) was identified in 25.6% [469] with the majority
[73%] of the participants being in their 3rd and 4th decades.
Conclusions: Whereas HBV vaccination should be scaled up for both
adult men and women irrespective of their HIV status, more effort should
be directed towards adult rural women of child bearing age especially
those below 25 years.
www.hivr4p.org
403
POSTERS
Background: Whereas many STIs have gained significance over the
years and especially with regard to HIV and AIDS, Herpes Simples
Virus 2 and Syphilis are some of the most important. Herpes simplex
virus 2 (HSV-2) and syphilis are important etiologic agents of sexually
transmitted infections. In this era of HIV and AIDS, the two infections
have become a major public health concern due to their association
with HIV acquisition and pathogenesis by driving viral replication and
facilitating transmission. In this study testing for the two infections was
carried out to determine levels of infections and possible associations
including CD4 counts since differences in HIV infections have been
indicated across regions in Kenya.
Methods: Three sites across 3 regions in Kenya were included in the
study where 711 HIV positive participants were included. CD4 count,
HSV and Syphilis status were tested using FacsCalibur and serological
tests respectively.
Results: Among the study participants, 66.9% were female and 33.1%
Male. Test results indicated 67.5% of the participants were HSV Positive
while 32.5% were Negative with females having a higher prevalence.
Syphilis sero-status was at 12.4% and 87.6% positive and negative
respectively.CD4 count ranged from 1 to 1700 counts /ul of blood
with differences noted between genders, test results status as well as
between sites/regions.
Conclusions: Higher prevalence of STIs, including HSV and syphilis
has potential to contribute to increased HIV transmission and could
reverse the gains being made with treatment for preventions especially
in HIV discordant relations and increased incident noted among certain
population groups and regions in Kenya.
1
Posters
P49.11
P49.12
Burden of Sexually Transmitted Infections
Among High Risk Women Screened for a
Phase III Microbicide Trial in Southwestern
Uganda
Number Needed to Screen for Chlamydial and
Gonococcal Infection among Asymptomatic
Men Who Have Sex with Men in Bangkok,
Thailand, 2006-2010
Martin Onyango1, Andrew Abaasa1, Anita Kabarambi1, Faith
Naddunga1, Sylvia Kusemererwa1, Neliette Van Neikerk2,
Annalene Nel2, Anatoli Kamali1
Sarika Pattanasin1, Punneeporn Wasinrapee1, Jaray Tongtoyai1,
Wannee Chonwattana1, Anuwat Sriporn1, Pikunchai Luechai1,
Wichuda Sukwicha1, Anupong Chitwarakorn2, Timothy H.
Holtz1,3, Marcel Curlin1,3
Medical Research Council/ Uganda Virus Research Institute, Uganda
Research Unit on AIDS, Entebbe, Uganda, 2International Partnership for
Microbicides, Paarl, South Africa
1
POSTERS
Background: Sexually Transmitted Infections (STIs) facilitate
transmission and acquisition of HIV infection. Screening for STIs is
recommended in HIV microbicide trials and untreated STIs delay
enrolment. We determined the burden of STIs among high risk women
screened for a Phase III microbicide trial in Southwestern Uganda.
Methods: The Medical Research Council/Uganda Virus Research
Institute (MRC/UVRI) in collaboration with International Partnership for
Microbicides (IPM) is evaluating safety and efficacy of a dapivirine vaginal
ring among healthy HIV-negative high risk women in a multicentre Phase
III trial. These women were identified from bars, lodges, restaurants,
and hair salons in townships and fishing communities within 50 km
from the research centre. High risk was defined by presence of STIs,
self-reported unprotected sex with multiple sex partners, and frequent
use of recreational drugs (marijuana, alcohol) in the past 3 months.
All consenting female volunteers were screened for STIs regardless of
clinical symptoms. Cervico-vaginal samples were taken for T. vaginalis
(OSOM Rapid test), C. trachomatis/N. gonorrhoeae (Roche PCR test), and
blood samples taken for syphilis (RPR/TPHA). Treatment was as per
the Uganda National guidelines for syndromic management and CDC
STI treatment guidelines. The proportion of STIs among the women
screened was determined.
Results: Of the 236 women screened between August 2013 and March
2014, the mean age was 28 years (range 18 to 42 years). About half (111,
47%) had T. vaginalis, 23 (10%) C. Trachomatis, 20 (8%) N.gonorrhoeae
and 12 (5%) syphilis (RPR and TPHA positive). A total of 43 (18.2%) had
at least 2 or more STIs.
Conclusions: There is a high burden of STIs among high risk women.
Though such populations are considered suitable for vaginal microbicide
efficacy trials, the high STI burden impacts greatly on trial enrolment.
There is a need for regular STI screening, partner identification and
treatment, and condom promotion in this population.
404
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
Background: Asymptomatic infection caused by Chlamydia trachomatis
(CT) and Neisseria gonorrhoeae (NG) are common among men who have
sex with men (MSM). The U.S. Centers for Disease Control and Prevention
(CDC) recommends screening sexually active MSM at least annually for
CT and NG infection. We calculated the number of asymptomatic MSM
needed to screen (NNS) to detect one MSM with CT and/or NG, stratified
by sexual role.
Methods: Between 2006 and2010, we enrolled Thai MSM, age ≥18
years, from the Bangkok metropolitan area. Participants completed sexual
behavior questions using audio computer-assisted self-interviews, and
underwent physical examination, and rectal, urethral, and pharyngeal
screening for CT and NG using nucleic acid amplification testing (NAAT).
We calculated NNS as the reciprocal of the proportion of asymptomatic
MSM with unrecognized infection detected by NAAT.
Results: Of 1,744 participants enrolled, 1593 (91%) had no symptoms
or signs of CT/NG at baseline. We detected CT infection in 216 (14%),
NG infection in 99 (6%), and CT/NG co-infection in 40 (2.5%). The
overall NNS to detect CT and/or NG at any sites was 5 (95% Confidence
Interval [CI] 5-6). Among insertive-only MSM (n=285), the chance of
detecting urethral CT infection (NNS 11, 95% CI 8-19) was higher than
detecting urethral NG infection (NNS 71, 95% CI 36-2646, p< 0.05).
Among versatile MSM (n=1284), the NNS for infection at any site was
7 (95% CI 6-8) for CT, and 14 (95% CI 12-18) for NG. Among receptive
only MSM (n=287), the NNS for infection at rectal was 6 (95% CI 5-9).
Conclusions: Asymptomatic CT and NG infection in this population
is high. Given that the NNS in asymptomatic sexually active MSM is
low, annual screening using NAAT for all sites can diminish progression
of diseases and disrupt transmission. In settings where screening all
specimen sites is not available, the rectal swab test should be a priority.
P49.13
P49.14
ABSTRACT WITHDRAWN
HIV-1+/HSV-2+ Co-infection Is Associated with
Persistence of CD14+ and DC-SIGN+Antigen
Presenting Cells at the Mucosa Independent
of HSV Recurrences
Gloria C. Preza1, Marla J. Keller2, Maria T. Ochoa1, Betsy Herold3
University of Southern California, School of Medicine, Los Angeles,
CA, United States, 2Albert Einstein College of Medicine, Pediatric
Infectious Diseases, Bronx, NY, United States, 3Albert Einstein College
of Medicine, Pedriatic Infectious Diseases, Bronx, NY, United States
Background: Prior studies demonstrate that HSV-2 seropositivity is
associated with higher HIV plasma viral loads & increased genital HIV
shedding. We hypothesize that in HIV+/HSV-2+ women, genital HSV
outbreaks will be associated with an increase in antigen presenting cells
(APCs), which may facilitate recruitment of HIV target cells & contribute
to replication & transmission in HIV+/HSV-2+ women.
Methods: A 12-week study was conducted among HIV+/HSV-2+, HIV+/
HSV-2-, HIV-/HSV-2+& HIV-/HSV-2- women to examine the impact of HSV2 reactivation on mucosal immunity. Vaginal biopsies were collected at
baseline &from a lesion & contralateral uninvolved site during a clinical
recurrence. Sections from 4 HIV+/HSV-2+& 4 HIV-/HSV-2+ women were
evaluated for APCs & results quantified by in situ IHC imaging analysis.
Results: At baseline, co-infected women had significantly more CD209+
(DC-SIGN) & CD14+ (monocytes/macrophages) cells compared to HIV+/
HSV-2-&HIV-/HSV-2+ women. After an outbreak, there was an increase in
these APCs in HIV- women at the lesional but not the contralateral site
(p< .02). However, the response in co-infected women was blunted. For
example, CD209+ expression increased from 1002.6 ± 78.6 cells/mm2
at baseline to only 1132.3 ± 99.4 cells/mm2 in lesions, but increased
in lesions from HIV- women from 251.3 ± 92 cells/mm2 at baseline to
795.8 ± 21 cells/mm2 (p< .004). Similarly, there was little or no increase
in CD14+ cells in lesions in co-infected women whereas CD14 increased
in HIV- women from 514.5± 118.9 to 1291.6 ± 119.7 cells/mm2 (p< .04).
Conclusions: Co-infection is associated with an increase in CD209+ &
CD14+ APCs in biopsies independent of HSV disease. During clinical
HSV recurrences, there is an increase in APCs at the lesional but not the
contralateral site, which is more pronounced in HIV- women. The largest
APCs observed were CD14+ monocytes & CD209+ macrophages. These
findings provide a potential mechanism for the increased risk of HIV
transmission in the setting of HSV-2 co-infection.
www.hivr4p.org
405
POSTERS
1
Posters
Posters 50: Tenofovir Gel: Acceptability and Adherence
P50.01
P50.02
FACTS 001: Characteristics of Participants
Enrolled in a Phase III Randomised Controlled
Trial of Tenofovir Gel for Prevention of HIV-1
and HSV-2
Meta-analysis of the Effectiveness of
Tenofovir Gel in Preventing HIV Infections
Anneke Grobler1
CAPRISA, Congella Durban, South Africa
1
Sinead Delany-Moretlwe1, Emilee Smith2, Landon Myer2, Khatija
Ahmed3, Busi Nkala4, Rebone Maboa5, Cynthia Gama6, Sydney
Sibiya7, Modulakgotla Sebe8, William Brumskine9, Linda-Gail
Bekker10, Maposhane Nchabeleng11, Carl Lombard12, Glenda
Gray4,12, Helen Rees1, FACTS 001 Study Group
University of the Witwatersrand, Wits Reproductive Health & HIV
Institute, Johannesburg, South Africa, 2University of Cape Town,
Centre for Infectious Diseases Epidemiology & Research, Cape Town,
South Africa, 3Setshaba Research Centre, Soshanguve, South Africa,
4
University of the Witwatersrand, Perinatal HIV Research Unit, Soweto,
South Africa, 5University of the Witwatersrand, Wits Reproductive
Health and HIV Institute, Johannesburg, South Africa, 6University of
the Witwatersrand, MATCH, Durban, South Africa, 7Qhakaza Mbokodo
Research Centre, Ladysmith, South Africa, 8Aurum Institute, Tembisa,
South Africa, 9Aurum Institute, Rustenburg, South Africa, 10University
of Cape Town, Desmond Tutu HIV Centre, Cape Town, South Africa,
11
Medunsa, Medunsa Clinical Research Unit (MeCRU), Ga-Rankuwa,
South Africa, 12Medical Research Council of South Africa, Cape Town,
South Africa
1
POSTERS
Background: FACTS 001 is a phase III licensure trial to assess the safety
and effectiveness of tenofovir 1% gel applied intravaginally before and
after sex in preventing HIV-1 and HSV-2 in women in South Africa. We
report on the baseline characteristics of the trial population.
Methods: Participants were screened for eligibility for the trial at nine
sites across South Africa from October 2011 until March 2014. Women
aged 18-30 years, who were HIV-1 negative, sexually active, using a
reliable modern contraceptive, with no travel plans, no recent history of
enrolment in another HIV prevention trial, and without evidence of renal
or pelvic disease, raised liver enzymes, or pathological bone fractures,
who consented to all study procedures, were considered eligible for the
trial. Participants were enrolled irrespective of HSV-2 status at enrolment.
Results: 3846 participants were screened and 2059 were enrolled in
the trial. The main reasons for exclusion were HIV-1 positive, not willing
or able to consent to all procedures, and evidence of proteinuria or
glycosuria. At the time of enrolment, the mean age of participants was
23 years, and the majority of participants were unmarried (88%), lived
with parents (62%), had completed school (43%), were unemployed
and earned < R1000 in the past three months (73%). Most participants
reported having only one regular partner (89%) and a median of 9 (IQR
5-16) sex acts in the past 28 days. The overall prevalence of HSV-2 at
enrolment was 42%.
Conclusions: The baseline characteristics of FACTS 001 enrolled trial
population reflect those most at risk for HIV infection in South Africa,
and in need of new HIV prevention technologies.
406
Background: Three clinical trials have been conducted to test whether
1% Tenofovir gel prevents heterosexual transmission of HIV. The
CAPRISA 004 study found a reduction of 39% and the VOICE study
found a reduction of 14.7% in HIV incidence comparing the Tenofovir
gel arm to the placebo arm. The FACTS study is still under way and
results are expected early next year. The objective is to estimate the
combined efficacy of tenofovir gel using all three studies.
Methods: A meta-analysis was done of the CAPRISA and VOICE results.
Additional meta-analyses also included hypothetical results of the
FACTS study. The Cochran-Mantel-Haenszel common relative risk was
calculated for each meta-analysis. The CAPRISA and FACTS studies had
coitally dependent dosing (the so-called BAT24 strategy), while the
VOICE study had daily dosing.
Results: A meta-analysis of the gel arms in VOICE and CAPRISA
004, estimates that Tenofovir gel provides 24% protection (95%CI:
2-41%,p-value=0.03). If the results of the FACTS study was similar to
the VOICE study, the meta-analysis estimates 21% protection (95%CI:
2-36%,p-value= 0.03) and if it was similar to CAPRISA 004 the metaanalysis estimates 28% protection (95%CI:11-42%,p-value=0.003). If
the FACTS study showed a null-result (the same number of infections in
the Tenofovir gel and placebo arms) then the meta-analysis estimates
that Tenofovir gel provides 18% protection (-1-34%,p-value=0.068).
In all instances the test for heterogeneity did not detect heterogeneity
between the studies.
Conclusions: If the results of the FACTS study lie between the results of
the CAPRISA and the VOICE studies, i.e. provides between 14.7% and
39% protection from HIV infection, one expects the combined efficacy
estimate of the three studies to lie between 21% and 28%, with a p-value
smaller than 0.03. This would indicate that Tenofovir gel is efficacious in
preventing HV infection, when all available efficacy trials are combined.
P50.03
P50.04
Tenofovir Gel as a Component of the
HIV Prevention Mix: Perspectives from
Participants, Partners and Community Men in
KwaZulu-Natal, South Africa
Tenofovir Gel Acceptability and Adherence
among Pregnant Women in the United States
1
FHI 360, Durham, NC, United States, 2CAPRISA/University of Kwa Zulu
Natal, Durban, South Africa
Background: Many questions exist with regard to how 1% tenofovir
gel could be integrated with existing HIV prevention methods, if it is
approved. We explored perceptions of women and men about the gel
relative to other prevention methods in the context of CAPRISA 008,
an ongoing open-label follow-on trial of tenofovir gel that enrolled
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 women in the CAPRISA 008 trial. Perspectives on HIV prevention
options including tenofovir gel were assessed using an applied thematic
analysis approach.
Results: Male condoms and male circumcision were perceived to be
the most effective means of HIV prevention by both female and male
respondents; however, the difficulty of consistent condom use often
negated perceived efficacy while male circumcision faced cultural
aversion and was viewed as mainly protecting men. Few respondents
reported use of female condoms; most were averse to the thought of
using them. Tenofovir gel was generally viewed as an effective means
of HIV prevention by both women and men, though there was concern
that it is still not licensed by the government and that it is “not 100%
effective.” Use of tenofovir gel was particularly considered beneficial for
women if their partners refuse to wear condoms and was described as
easy to use relative to other HIV prevention methods. Few participants
reported cases of condom migration. Some men noted that their partner’s
use of tenofovir gel provided risk reduction benefits to them as well.
Conclusions: Both men and women in KwaZulu-Natal, South Africa
perceive tenofovir gel as offering the potential to address limitations in
the current HIV prevention method mix.
RTI International, Womens Global Health Imperative, Los Angeles,
CA, United States, 2University of Pittsburgh Medical Center and
Magee-Women Research Institute, Pittsburgh, PA, United States,
3
SCHARP - FHCRC, Seattle, WA, United States, 4University of Alabama
at Birmingham, Birmingham, AL, United States, 5FHI 360, Durham, NC,
United States, 6NIH/NICHD, Bethesda, MD, United States, 7CONRAD/
EVMS, Arlington, VA, United States, 8NIH/NIAID, Bethesda, MD, United
States
1
Background: The MTN-008 trial, the first multi-dose study of a
microbicide gel (2:1 randomized to tenofovir 1% or HEC placebo gel)
during pregnancy, included daily pregnant participant gel insertion at
home for 5 days. Because pregnancy may pose unique challenges to
consistent gel use and acceptability these factors were assessed.
Methods: Participants completed a web-based computer-assisted selfinterview (CASI) at Days 0 and 6 about gel attitudes and behaviors. At
Day 6 trained research staff conducted a short interviewer-administered
questionnaire with both structured and open-ended questions.
Frequencies of quantitative data were tabulated in SAS; qualitative data
were manually summarized.
Results: CASI data include 96 participants in both active and placebo
gel arms. Self-reported adherence was high with 88% reporting daily
gel use and the majority reporting no difficulty with daily use. The
most common reason for nonuse was forgetting. Participants reported
generally neutral perceptions of gel characteristics. Two-thirds thought
the gel was runny, many complained of bothersome gel leakage and
several cited this reason for not inserting a full dose. A small number
of women (6-8%) reported pain, other physical discomfort, or mental
discomfort associated with the process of applicator insertion, and
fewer than 5% reported the same in regards to the gel itself. Although
the majority were not worried the gel would cause problems for their
pregnancy or babies, one-quarter to one-third felt somewhat worried
about these issues. Ten percent of women said they would not use the
gel in the future when not pregnant, while 38% would prefer to use gel
only before sex and 32% had no preference between daily and precoital gel use.
Conclusions: Self-reported adherence to gel for 5 days at home was
high. Gel was generally acceptable, but many complained of a runny
consistency and leakage. There were no frequent or strong concerns
about the effects of the gel on the pregnancy/ fetus reported.
www.hivr4p.org
407
POSTERS
Kathleen M. MacQueen1, Sarah Dlamini2, Brian Perry1, Eunice
Okumu1, Steve Sortijas1, Chitra Singh2, Diantha Pillay1, Sharon
Watson1
Elizabeth T. Montgomery1, Lisa Noguchi2, James Dai3, Jason Pan3,
Joseph Biggio4, Karen Isaacs5, Heather Watts6, Jill Schwartz7,
Jeanna Piper8, Richard Beigi2
Posters
P50.05
P50.06 LB
Group Adherence Support (GAS) - Experiences
from CAPRISA 008
A Phase 1 Evaluation of the Rectal Safety,
Acceptability, Pharmacokinetics, and
Pharmacodynamics of Three Formulations of
Tenofovir 1% Gel
Chitra Singh1, Nelisiwe Ngcobo1, Sarah Dlamini1, Carl Montague1,
Fanele Ntombela1, Nabeela Warrasally1, Kershia Sunjeevan1,
Quarraisha Abdool Karim1, Leila Mansoor1
Centre for AIDS Programme of Research in South Africa (CAPRISA),
1
POSTERS
Background: Adherence is key to ensuring effectiveness of ARV based
microbicides. Adherence patterns may differ in a placebo controlled trial
where efficacy is uncertain compared to an open-label trial of a product
of known efficacy and also with increasing duration of follow-up.
Participant experiences and challenges in adhering to using 1% tenofovir
gel in CAPRISA 008 was assessed as part of ongoing adherence support.
Methods: Five group adherence support (GAS) sessions were conducted
between January and February 2014 by a trained staff member.
Participants who were enrolled in the study for >12 months volunteered
to take part in these GAS sessions that comprised groups of 6-10
participants and lasted 60-90 minutes. Detailed notes transcribed
during these sessions were reviewed to identify common themes and
differences within and across sessions.
Results: A total of forty six women aged between 23-45 years
participated in these GAS sessions. Common issues emerged regarding
factors affecting adherence to gel use over time. Initial confidence about
product use started to be eroded with time as they learnt of participants
becoming infected with HIV. In four out of five sessions it was reported
that partner disclosure of gel use enhanced adherence. In three out of five
sessions participants reported that unplanned coitus, partner complaints
of vaginal wetness and leakage of gel, influenced adherence over time.
In order to address these barriers to adherence participants suggested
the following: inserting the gel earlier in the day to reduce wetness at
the time of coitus especially if they have not disclosed gel use to their
partner and storage of gel in convenient and multiple locations (i.e.
cosmetic bag/pencil case) for easy access particularly when unplanned
coitus takes place.
Conclusions: These pilot GAS sessions identified both barriers and
facilitators to product adherence. Participants welcomed suggestions
made by peers to enhance their product adherence levels.
408
Ian McGowan1, Kathy Duffill2, Alexiy Nikiforov2, Charlene
Dezzutti1, Nicola Richardson-Harman3, Kaleab Abebe1, Amy
Herrick4, Mark Marzinke5, Ross Cranston1, Lisa Rohan6, Craig
Hendrix5, Hirot Hiruy5, Julie Elliott7, Christine Mauck8, Peter
Anton7, and the CHARM Program Research Team
University of Pittsburgh School of Medicine, Pittsburgh, PA, United
States, 2Magee Womens Research Institute, Pittsburgh, PA, United
States, 3Alpha StatConsult LLC, Damascus, MD, United States,
4
University of Pittsburgh Graduate School of Public Health, Pittsburgh,
PA, United States, 5Johns Hopkins University School of Medicine,
Baltimore, MD, United States, 6University of Pittsburgh School of
Pharmacy, Pittsburgh, PA, United States, 7University of California at
Los Angeles, Los Angeles, CA, United States, 8CONRAD, Arlington, VA,
United States
1
Background: CHARM-01 was undertaken to characterize the rectal
safety, acceptability, pharmacokinetics (PK), and pharmacodynamics (PD)
of three formulations of tenofovir (TFV) 1% gel: a vaginal formulation
(VF), a reduced glycerin vaginal formulation (RGVF), and a rectal
formulation (RF) with osmolalities of 3,111, 846, and 479 mOsmol/kg
respectively. The VF gel was previously used in the CAPRISA 004 and
VOICE vaginal microbicide Phase 2B trial. The VF gel was used in the
RMP-02/MTN-006 Phase 1 rectal safety study. The RGVF gel was used
in the MTN-007 Phase 1 rectal microbicide trial and is currently being
used in the MTN-017 Phase 2 rectal microbicide trial.
Methods: Participants received 4 mL of study product in a blinded,
randomized, crossover design:
(i) 6 daily doses of a HEC placebo followed by a final single dose of
the VF gel,
(ii) 7 daily doses of the RGVF gel, and
(iii) 7 daily doses of the RF gel.
Safety, acceptability, compartmental PK, and explant PD were monitored
throughout the trial.
Results: Thirteen participants were enrolled into the CHARM-01
study. All three gels were found to be safe and acceptable. Median
rectal tissue homogenate TFV-diphosphate (DP) concentration was
significantly greater with the RF (10.3 ng/mg) versus the VF (below the
limit of quantification) gel. Median mucosal mononuclear cell TFV-DP
was significantly greater with the RF (1136 fmol/106 cells) versus VF (91
fmol/106 cells) gel (p ≤ 0.05). Use of each gel in vivo was associated
with significant inhibition of ex vivo colorectal explant HIV infection.
There was also a significant relationship between tissue levels of TFV/
TFV diphosphate (DP) and inhibition of explant infection.
Conclusions: All three formulations were safe and acceptable. There
was a trend towards higher tissue levels of TFV associated with exposure
to the RF gel but and all three gels were associated with significant
inhibition of explant infection. The RGVF or the RF gel could be advanced
into later stage development as a rectal microbicide.
P50.07 LB
Introducing Microbicides: Estimated Cost of
QI Approach and Tenofovir Gel Delivery with
Family Planning Services in the CAPRISA 008
Trial
Rick Homan1, Sarah Dlamini2, Leila Mansoor2, Imraan Valodia3,
Steve Sortijas1, Sharon Watson1, Kristine Torjesen1
1
FHI 360, Durham, NC, United States, 2CAPRISA/University of
KwaZulu Natal, Durban, South Africa, 3University of Witwatersrand,
POSTERS
Background: The CAPRISA 008 implementation trial of 1% tenofovir
(TFV) gel randomizes participants to receive TFV gel with family planning
(FP) services in a primary health care (PHC) or research clinic setting in
South Africa. The CAPRISA 106 ancillary study estimated the cost of
using a quality improvement (QI) approach to strengthen FP services and
integrate TFV gel provision in the PHC clinic at Vulindlela between June
2012 and January 2014.
Methods: QI activity data were extracted from 86 documents and
estimates of eligible client volume, gel uptake and FP visit recall were
developed from PHC and CAPRISA service statistics. Unit costs were
applied to resources using KwaZulu-Natal Department of Health (DOH)
data and other sources. Financial labor costs (additional payment
required) were separated from opportunity labor costs (time spent during
routine work).
Results: The value of resources required at the district level per PHC clinic
for the first year of gel delivery is estimated at 911,946 ZAR ($89,500)
and for the QI approach is 190,194 ZAR ($18,660). Recurring annual
financial cost for gel delivery is estimated at 895,264 ZAR ($87,850) per
clinic. Major financial cost at the district level is for TFV gel procurement
with minor labor costs for staff training. The value of resources required
at the PHC level per clinic for the first year of gel delivery is 309,587
ZAR ($30,400) and for the QI approach is 126,961 ZAR ($12,460). For
gel delivery, recurring annual financial cost is 282,714 ZAR ($22,740)
per clinic for additional labor, HIV test kits and other supplies. For the
QI approach, recurring annual financial cost is 1,545 ZAR ($152) with
opportunity costs estimated at 1.25 nurse days/month and 2.25 data
clerk days/month.
Conclusions: The resources required for introducing TFV gel are
dominated by cost of the product, as well as additional labor to provide
services at PHC clinics. Compared to the costs of TFV gel, introducing a
QI approach to support gel delivery imposes minimal cost to the DOH.
www.hivr4p.org
409
Posters
Posters 51: Therapeutic Vaccine and Viral Latency
P51.01
P51.02
Partial Protection by Vaccination Does Not
Prevent Chronic Neuro-inflammation in an
SIV Model
In vitro Latent Virus Reactivation Potential
of a Novel Triterpenoid Compound and Two
Cytostatic Gold(III) Complexes
Debbie H. Ferguson1, Claire Ham1, Atze Das2, Ben Berkhout2,
Andrea Meiser3, Steve Patterson3, Neil Berry1, Neil Almond1
Pascaline Fonteh1, Petrina Kapewangolo2, Debra Meyer1
National Institute of Biological Standards and Control, Virology,
Potters Bar, United Kingdom, 2Academic Medical Centre, University
of Amsterdam, Amsterdam, Netherlands, 3Imperial College London,
London, United Kingdom
1
POSTERS
Background: With antiretroviral therapy (ART), HIV is now a chronic
viral infection. However, suppression of viral load does not prevent
chronic inflammation and the development of co-morbidities such as
HIV-associated neurocognitive impairment. Using a non-accelerated
ART-free SIV model that exhibits reproducibly controlled viral kinetics,
we have demonstrated early neuroinvasion and chronic progressive
neuropathology occurs even when peripheral viremia is well controlled.
Here we investigated the impact on chronic neuroinflammation of either
switching off a conditional live SIV or modifying early viremia.
Methods: Immunohistochemical analysis of macaque brain sections
was performed. Firstly, groups of macaques were infected with SIVrtTA,
a doxycycline (DOX) dependent, conditional-live SIV. We compared
groups on daily DOX treatment for 40 weeks with those where DOX
was removed at week 3. Secondly, we compared groups of macaques
immunised with an SIVgag based vaccine regimen. Vaccination did not
reduce set point viral loads, however, it did significantly blunt primary
viremia compared with naïve challenge controls.
Results: Study 1. Chronic neuroinflammatory responses were detected
in all SIVrtTA infected macaques even 37 weeks after DOX treatment
was removed. Indeed, there was evidence astrocytosis and microgliosis
were lower in macaques receiving the anti-inflammatory doxycycline
throughout the study.
Study 2. Suppression of primary viremia reduced neuroinflammation
compared with that seen in naïve challenge controls.
Conclusions: In conclusion, primary viremia alone is sufficient to
initiate chronic neuroinflammation. Blunting primary viremia modulates
subsequent neuropathology but does not prevent it. The neuroinflammatory legacy of acute infection even with subsequent potent
viral control implies patients may suffer significant co-morbidities despite
improved ART or even following eradication of HIV by functional cure.
410
University of Pretoria, Department of Biochemistry, Pretoria,
South Africa, 2University of Namibia, Department of Chemistry and
Biochemistry, Windhoek, Namibia
1
Background: Virus elimination by combining latency activators and
highly active anti-retroviral therapy (HAART) is one way of eradicating
HIV that is currently under investigation. Here, we present a comparative
study of the virus reactivation potential of a novel triterpenoid plant
product (1) derived from Ocimum labiatum to that of two synthetic
cytostatic bis(thiosemicarbazonate) gold(III) complexes (2 and 3).
Methods: Viral reactivation was quantified by measuring p24 antigen
production from a chronic infection and viral latency model of HIV (U1
cells). To exclude toxicity effects in the reactivation study, a tetrazolium
dye was used to determine cytotoxicity. The mechanism of viral
reactivation was determined by measuring the effect of 1, 2 and 3 on
protein kinase C (PKC) and histone deacetalyse enzyme activity in an
ELISA and a fluorometric assay respectively. A cytometric bead array kit
was used to quantify the endogenous production of TNF-α.
Results: The CC50 of 1, 2 and 3 on the U1 cells was found to be 8.2
±0.1µg/mL, 0.53±0.12 µM and 1.0±0.4 µM respectively. Non-toxic
concentrations (exhibiting >90% viability) of 1, 2, and 3 significantly
reactivated virus (p< 0.05) by 3.5, 2.7 and 2.3 fold from U1 cells.
Compound 1 and complexes 2 and 3 minimally activated PKC levels
but did not inhibit HDAC indicating that the reactivation mechanism
does not involve these enzymes. A notable observation was the increase
in TNF-α by up to 7 fold, possibly as a result of generalised immune
activation by these compounds.
Conclusions: The upregulation of TNF-α by 1, 2 and 3 is considered
an off target effect which is part of a non-specific mechanism of viral
reactivation. These findings suggest that the novel triterpene and the
cytostatic bis(thiosemicabazonate) gold(III) complexes could potentially
be incorporated in drug cocktails aimed at “activation/elimination”
of latent HIV but should be modified using techniques such as
nanotechnology to improve selectivity and exclude off target effects.
Posters 51: Therapeutic Vaccine and Viral Latency
P51.03
P51.04
Reactivation of Latent HIV-1 by Bryostatin-1
in the Presence of Antiretroviral Drugs
Bryostatin Activates HIV-1 Latent Expression
in Human Astrocytes through a PKC and NFkB-Dependent Mechanism
Research Center of Bioengineering, Madrid, Spain, 2Hospital
Universitario Ramón y Cajal, and IRYCIS, Madrid, Spain, 3Instituto
Maimónides de Investigación Biomédica de Córdoba, Universidad de
Córdoba, Cordoba, Spain
1
Background: Despite the fact that the antiretroviral therapy (ART)
successfully controls HIV-1 in most infected patients, virus latency
established early upon infection impedes HIV-1 eradication. Bryostatin-1
(BRYO) in vitro inhibits HIV-1-infection of CD4+T lymphocytes and, at
the same time, reactivates HIV-1 through PKC- NF-kB pathway. The
potential effects of BRYO in combination with antiretrovirals, such as
maraviroc (MVC) and Atripla® (ATP) is needed to be determined in vitro
prior to design clinical trials.
Methods: Jurkat-LTRGFP-R5 cell line and two latent and reactivatable
HIV-1-infected J89GFP lymphocytic or THP89GFP monocytic clonal cell
lines were used as latency models.
Results: BRYO reactivates HIV-1 reaching levels >80% in THP89GFP
cells, even in combination with MVC or ATP. Moreover, when MVC or
ATP pre-treated reporter TZM-bl cells were co-cultured with BRYO treated
with THP89GFP cells, new infections of reactivated X4/R5-HIV-189.6 were
inhibited 50% or 80%, respectively. Remarkably, when antiretrovirals
were combined with BRYO, the combinations maintained the antiviral
effect of the drug with the maximum rate of inhibition on its own. Also,
BRYO-mediated downregulation of surface CD4 and CXCR4 in primary
blood mononuclear cells (PBMC) was not affected when it was used
along with the other antirretrovirals and no hiperactivation or high
proliferation effects were observed in these PBMC. Significantly, BRYO
was also tested ex vivo for HIV-1 induction in CD4+ T lymphocytes
isolated from HIV-1 infected patients receiving ART and was found to
exhibit potent viral induction activity.
Conclusions: This work is the first to demonstrate that antiretrovirals
combinations with BRYO do not interfere in the BRYO activity neither the
BRY combination with antiretrovirals interferes in the antiviral activity of
the antiretrovirals. Thus, we propose BRYO to purge the viral reservoirs,
and this treatment should be combined with present ART.
Laura Diaz1, Marta Martinez-Bonet1, Javier Sanchez-Rodriguez1,
Alejandra Fernández-Pineda1, Esther Alonso1, Jos Luis Jiménez1,
Eduardo Muñoz2, Santiago Moreno3, Susana Alvarez1, Mª
Ángeles Muñoz-Fernández1
Research Center of Bioengineering, Madrid, Spain, 2Instituto
Maimónides de Investigación Biomédica de Córdoba, Universidad de
Córdoba, Cordoba, Spain, 3Hospital Universitario Ramón y Cajal, and
IRYCIS, Madrid, Spain
1
Background: Previous studies have suggested that the central nervous
system (CNS) is an important reservoir for HIV and that proviral DNA
and productive infections can be detected in this region. In particular,
the brain represents a sanctuary for HIV-1 latency because the provirus
can persist in this area due to the variable and poor penetration of
antiretrovirals (ARV). Despite an undetectable viral load in patients
treated with potent ARV, current therapies are unable to purge the virus
from these latent reservoirs. Combinatory strategies to eliminate HIV-1
reservoirs using selective activators of viral expression could lead to a
decline in HIV-1 levels in reservoirs that would be sufficient to efficiently
control the infection.
Methods: Human primary astrocytes (NHA) isolated from the cerebrums
of 5-month-old human foetuses and astrocytic cell line U-87 were used
in all experiments.
Results: To broaden the inhibitory range and effectiveness of current
ARV, the potential of bryostatin was investigated as a latent HIV-1
activator. Bryostatin reactivates latent viral infection in NHA and in U-87
via activation of protein kinase C (PKC)-alpha and -delta because the
PKC inhibitors rottlerin and GF109203X abrogated the bryostatin effect.
We did not find any alteration in cell proliferation. Moreover, bryostatin
strongly stimulated LTR transcription by activating the transcription
factor NF-κB. The effect of bryostatin could be especially important in
a cellular reservoir such as astrocytes because it may be a beneficial
adjunct to the treatment of HIV-infection.
Conclusions: Bryostatin can directly increase HIV-1 LTR activity in
human astrocytes in vitro via the PKC pathway and an NF-κB-dependent
mechanism. Thus, it is plausible that HIV-1-infected astrocyte cells
exposed to bryostatin may contribute to HIV-1 latency activation and
will provide a foundation for future novel HIV-1-purging strategies from
tissue reservoirs such as the CNS.
www.hivr4p.org
411
POSTERS
Marta Martínez-Bonet1, Maria Isabel Clemente1, Susana Alvarez1,
Laura Diaz1, Dolores García-Alonso1, Ana Lopez1, Santiago
Moreno2, Eduardo Muñoz3, Mariangeles Munoz-Fernandez1
Posters
Posters 52: Treatment as Prevention: Initiation, Adherence, and Monitoring on ARV
P52.01
P52.02
Couples’ Voluntary HIV Counseling and
Testing (CVCT) followed by Treatment as
Prevention (TasP) for Discordant Couples: The
Impact of Each Step
The Impact of Biological Monitoring by CD4
and Viral Load in Guiding the Decision to
Initiate ART Therapy in HIV-1 Infected People
Georgia Elna Ambada Ndzengué1
Susan Allen1, William Kilembe2, Mubiana Inambao3, Rachel
Parker4, Tyronza Sharkey2, Naeemah Munir3, Linda Kimaru3, Sam
Scherber2, Kristin Wall5,6, Amanda Tichacek4, Bellington Vwalika2,
Eric Hunter7, Joseph Mulenga8, Elwyn Chomba9
Rwanda Zambia HIV Research Group-Emory University, Pathology
& Laboratory Medicine, School of Medicine, Atlanta, GA, United
States, 2Rwanda Zambia HIV Research Group, Zambia Emory HIV
Research Project, Lusaka, Zambia, 3Rwanda Zambia HIV Research
Group, Zambia Emory HIV Research Project, Ndola, Zambia, 4Rwanda
Zambia HIV Research Group, Department of Pathology & Laboratory
Medicine, SOM, Emory University, Atlanta, GA, United States, 5Emory
University School of Public Health, Epidemiology, Atlanta, GA, United
States, 6Rwanda Zambia HIV Research Group, Lusaka, Zambia, 7Emory
University School of Medicine, Department of Pathology & Laboratory
Medicine, Atlanta, GA, United States, 8National Blood Transfusion
Services, Lusaka, Zambia, 9Ministry of Community Medicine, Maternal
& Child Health, Permanent Secretary, Lusaka, Zambia
1
POSTERS
Background: Most new adult HIV infections in Africa are acquired
from steady partners and WHO recommends that couples be tested
and counseled together as an HIV prevention strategy. ART reduces
transmission from the HIV+ partner in discordant couples.
Methods: CVCT programs established in 70 Zambian government
clinics include follow-up testing for discordant and concordant negative
(M-F-) couples. We assess seroincidence in discordant couples, confirm
linkage through sequencing, and compare rates before (antigen positive
at CVCT or seroconverting within 6 weeks of CVCT) and after CVCT and
on or off ART. We also compare the overall number of seroconversions
in discordant and M-F- couples, and compare the cost of averting one
HIV infection through CVCT vs. TasP.
Results: 148,839 couples were tested, of whom 17,619 were discordant
(3,229 or 18.3% on ART at the time of CVCT) and 109,677 were
concordant negative (M-F-). Of 112 seroconversions, 62 occurred in
discordant (5.4/100PY, 95% CI 4.1-6.9) and 50 in M-F- (0.98/100PY,
95% CI 0.73-1.30) couples. Among discordant couples, excluding
infections acquired from outside partners, the rate before CVCT was
8.9/100PY (6.5-11.8) compared with 2.3/100PY after CVCT (95% CI
1.2-3.8. The overall transmission rate in discordant couples not on ART
was 8.6/100PY (95% CI 5.6-12.4), compared with 3.7 (95% CI 2.2-5.9)
when the infected partner was on ART. Among M-F- couples, incidence
was 1.4/100PY (95%CI 1.0-1.9) before and 0.12/100PY (95% CI 0.010.44) after CVCT. Using program costs and rates, the cost of averting
one infection with CVCT was <$500, compared with an annual cost of
>$10,000 for TasP in discordant couples.
Conclusions: CVCT prompted a 74% reduction in new infections
among discordant couples and a 91% reduction in M-F- couples. Among
discordant couples TasP was associated with a 58% reduction. The
combination of CVCT + TasP in discordant couples is ideal, but CVCT
has significant impact even when TasP is not available, and at 5% of
the cost of TasP.
412
CIRCB, Yaoundé, Cameroon
1
Background: Helper CD4 T cell count is utilized in the CDC categorization
of HIV/AIDS disease progression and in guiding the decision to initiate
antiretroviral treatment (ART) in many low income countries. However,
in disease intense region exclusion of people from ART by CD4 values
might not reflect the reality.
Methods: From 2011 to 2013, HIV disease progression was monitored
in a cohort of treatment -naïve HIV-1 infected people by CD4 count
and plasmatic VL. Eligibility for treatment was assessed according to the
WHO 2010 guidelines.
Results: Amongst 249 participants analyzed 40 % were eligible to
treatment during the first visit when both helper CD4 count (257±170)
and HIV-1 VL (5.12±0.79) was taken into consideration. However, when
CD4 count alone was considered we recorded just 32% eligibility to
treatment implying that 7.63% of eligible participants are left out. During
the course of three years a further 15.3 % of the remaining participants
became eligible to therapy by VL (5.15±0.77) and CD4 count (275±71)
with 12 % being eligible by CD4 counts. With regards to the 2010
WHO recommendations well over 49 % of participants were eligible to
treatment during the course of three years.
Conclusions: When the recent WHO 2013 recommendations for the
initiation of HAART are considered well over 40% of the remaining
members of the cohort should be eligible to treatment. This suggests
that close to 90 % of treatment naïve HIV-1 infected people are eligible
within three years by this new recommendation. Thus a significant
number of people would be eligible to treatment within a short period
of time implying that the test and treat strategy should be envisaged
in Cameroon.
P52.03
P52.04
Treatment as Prevention (TasP) in MSM and
Transgender Women: Successful Detection of
Acute/Recent Infection and Linkage to Care in
Lima, Peru
Geographic Utilization of Gift Cards Used
for Financial Incentives to Encourage Viral
Suppression: Findings from HPTN 065
Fred Hutchinson Cancer Research Center, Seattle, WA, United States,
IMPACTA, Lima, Peru, 3Epicentro, Lima, Peru, 4Via Libre, Lima, Peru
1
2
Background: The overall objective of this study is to improve the efficacy
of Treatment as Prevention (TasP) strategies by increasing detection of
acute and recent HIV infection. This research is being conducted in men
who have sex with men (MSM) and transgender women (TW) in Lima,
Peru, where knowledge about the symptoms of acute HIV infection and
its high infectivity is low.
Methods: We will enroll 1800 HIV-uninfected MSM and TW at risk for
HIV infection (STI in the past 6 months, self-identify as a sex worker, no
condom use at last anal intercourse, anal intercourse with >5 partners in
past 6 months, sex with an HIV-infected partner) into an ‘acute infection’
cohort. Cohort participants are tested for HIV every month using a pointof-care third-generation EIA; negative specimens are pooled and tested
for HIV RNA twice daily by real-time PCR. Participants who test positive
for HIV RNA are contacted the next business day by peer counselors for
linkage to the study site.
Results: 2,027 MSM/TW were screened between July 1, 2013 and
April 30, 2014. HIV-infected MSM/TW (15%) were referred to care and
HIV-negative MSM/TW were enrolled into the cohort. In this period, we
detected 27 acute HIV infections (HIV seronegative, HIV RNA positive).
Thirty-eight participants had recent infections (seropositive: 7 had tested
HIV negative in the past 1 month and 31 within the past 3 months).
Nine HIV+ participants did not continue in the study: one elected to be
treated at a non-study clinic, two had psychiatric problems that precluded
enrollment, 5 refused to participate due to logistics and other reasons,
and we were unable to re-contact 1. Slightly over half of the HIV-infected
participants were linked to care within 7 days of HIV diagnosis.
Conclusions: Detection of acute and recent HIV infection and rapid
linkage to care is possible. Successful linkage of such individuals to HIV
care including ART can maximize TasP intervention and individual clinical
benefits. This approach may be important for future cure strategies.
1
FHI 360, Durham, NC, United States, 2Massachusetts Institute of
Technology, Cambridge, MA, United States, 3Whitman-Walker Health,
Washington, DC, United States, 4Morris Heights Health Center, Bronx,
NY, United States, 5Columbia University Mailman School of Public
Health, ICAP, New York, NY, United States, 6Harlem Hospital, New York,
NY, United States
Background: HPTN 065 (TLC-Plus) evaluated the feasibility and
effectiveness of providing quarterly $70 financial incentive gift cards
to HIV‑infected patients on antiretroviral therapy who maintained viral
suppression (HIV RNA< 400 copies/mL). A total of 39,359 FI gift cards
were dispensed over 2 years by 19 HIV care sites in the Bronx, NY and
Washington, DC.
Methods: Data for each gift card disbursed included dispensing site,
transaction amount, and location of transaction (zip code). Cards never
used and transactions without valid zip codes were excluded. ZIP Code
Tabulation Areas were used to map the location of transactions by
venue (Bronx/DC) and dispensing site. Python programing and Microsoft
Excel 2010 were used for all analysis and visualization. For transactions
that occurred outside of the local jurisdictions of DC and the Bronx, a
random sample of transactions was examined to identify transactions
made in person versus by internet/phone.
Results: Of 78,529 transactions for gift cards distributed in the Bronx,
3,611(4.6%) transactions occurred outside of NY, linked to 1,852 unique
gift cards of the 23,268 distributed. Of 62,022 transactions for gift
cards distributed in DC, 3,928 (6.3%) occurred outside of DC, Maryland
(MD) and Virginia (VA), linked to 1,987 unique gift cards of the 16,091
distributed. Transactions occurred in all 50 US states, Puerto Rico, the
Virgin Islands and international locations. Of gift cards distributed in the
Bronx but used outside of NY state, 62% were used in person and 74%
of the gift cards distributed in DC, but used outside of DC, MD and VA,
were used in person.
Conclusions: Gift cards distributed in this study were primarily used
locally; however, about 5% of transactions were outside the local
jurisdictions and mostly in person. These data suggest that HIV‑infected
individuals in the Bronx and DC travel throughout the US and beyond;
thus, research is needed to understand their migration/travel patterns
and the implications for interventions using ART for prevention.
www.hivr4p.org
413
POSTERS
Ann Duerr1, Javier Lama2, Hugo Sanchez3, Robinson Cabello4,
Jorge Sanchez2
Theresa Gamble1, Padraig Corcoran2, Jill Stanton1, Phaedrea
Watkins1, Elizabeth Greene1, Jennifer Farrior1, Richard Elion3,
Susan Amenichi-Enahoro4, Wafaa El-Sadr5,6, for the HPTN 065
Study Team
Posters
P52.05
P52.06
HIV Ascertainment through Repeat Homebased Testing in the Context of a Treatment as
Prevention Trial (ANRS 12249 TasP) in Rural
South Africa
Frequency of Achieving Viral Suppression
among HIV-infected Persons in
Serodiscordant Partnerships Initiating
Antiretroviral Therapy
Joseph Larmarange1,2, Éric Balestre3, Joanna Orne-Gliemann3,
Collins Iwuji2, Nonhlanhla Okesola2, Marie-Louise Newell4,
François Dabis3, France Lert5, TasP ANRS 12249 Group
Andrew Mujugira1, Connie Celum1, Allan Ronald2, Nelly Mugo1,3,
Jared Baeten1
IRD, CEPED (UMR 196 Paris Descartes Ined IRD), Paris, France,
University of KwaZulu Natal, Africa Centre for Health and Population
Studies, Mtubatuba, South Africa, 3Inserm, ISPED (Unité 897 Inserm
Université Bordeaux Segalen), Bordeaux, France, 4University of
Southampton, Faculty of Medicine, Faculty of Social and Human
Sciences, Southampton, United Kingdom, 5Inserm, CESP (Unité 1018),
Villejuif, France
1
2
POSTERS
Background: The ANRS 12249 TasP cluster-randomised trial evaluates
whether HIV testing of all members of a community, followed by
immediate antiretroviral treatment (ART) for infected people, will prevent
onward sexual transmission and reduce HIV incidence at population
level. Ascertaining the HIV status of a high proportion of the population
regularly and repeatedly is key to the success of any universal test and
treat strategy, as the first step of the HIV cascade.
Methods: Between March 2012 and March 2014, we implemented
three six-monthly rounds of home-based HIV counselling and testing
in ten local communities (clusters). At each home visit, individual
questionnaires were administered and a rapid HIV test offered to all
trial participants. We report early results on rates of HIV ascertainment,
defined as undergoing a rapid HIV test or HIV-positive self-report.
Results: Of 12,911 eligible individuals (resident in the trial area and ≥16
years), 10,007 were successfully contacted at least once. At first contact,
HIV status was ascertained for 7,628 (76.2% [95% CI: 75.4-77.1])
individuals. At second contact, among the 5,885 individuals contacted
a second time, HIV status was ascertained for 2,829 (85.0% [95% CI:
83.7-86.2]) of the 3,328 tested negative at first contact and for 543
(45.7% [95% CI: 42.9-48.6]) of the 1,188 who refused a rapid test at first
contact. Overall, HIV ascertainment rate was 89.0% (5,239/5,885 [95%
CI: 88.2-89.8]) among trial participants contacted twice.
Conclusions: Repeat home-based HIV testing is acceptable and feasible
in this rural area. Socio-demographic characteristics, behaviours,
attitudes, household characteristics and experience of HIV infection and
ART in the household will be explored for their association with HIV
ascertainment uptake. This will inform whether this intervention reaches
the individuals at higher risk in a rural South African region.
414
1
University of Washington, Seattle, WA, United States, 2University of
Manitoba, Winnipeg, MB, Canada, 3Kenya Medical Research Institute,
Nairobi, Kenya
Background: Antiretroviral therapy (ART) markedly reduces the risk of
HIV transmission in serodiscordant partnerships. The concentration of
HIV RNA in plasma is the principal measure of ART adherence and is
also the prime determinant of HIV transmission risk.
Methods: We used data from a prospective study of 928 HIV-infected
persons from Kenya and Uganda who had a known heterosexual HIVuninfected partner, to assess viral suppression after ART initiation.
Plasma was archived 6-monthly and later tested for HIV RNA quantity.
Kaplan-Meier methods, and Cox regression models, were used to
identify independent predictors of viral suppression.
Results: The median age was 35 years (interquartile range [IQR] 29,
41), and 502 (54%) were women. The median CD4 count and HIV
RNA plasma concentration prior to ART start were 270 cells/µL (IQR
213, 370) and 4.34 log10 copies/ml (IQR 3.65, 4.85), respectively. The
cumulative probabilities of achieving viral suppression at 6 and 12
months were 82.8% and 90.8% for HIV RNA concentration < 400
copies/ml. Female gender (adjusted hazard ratio [AHR] 1.19; 95% CI:
1.02, 1.39; p=0.03) predicted viral suppression. Older age (AHR 1.04
per-5 year increase; 95% CI: 0.99, 1.08; p=0.03), higher education
(AHR 1.14; 95% CI: 0.99, 1.31; p=0.07 for >7 years), and higher CD4
count (AHR 1.16; 95% CI: 0.97, 1.38 for >200 compared with < 200
cells/µL) were of borderline significance.
Conclusions: Three-quarters of HIV-infected persons with known HIVuninfected partners achieved viral suppression within 6 months of ART
initiation. These results support World Health Organization guidance
recommending ART initiation for persons with known HIV-uninfected
partners, which could be optimized by viral load monitoring to identify
those needing adherence support.
P52.07
P52.08
The Cascade of HIV Care: The Tool for
Assessing Treatment as Prevention in Russian
Federation
Patient Retention in Antiretroviral Therapy
Programs up to Three Years on Treatment in
Uganda, 2006-2009: A Retrospective Review
Anastasia V. Pokrovskaya1, Vadim V. Pokrovsky1, Natalia N.
Ladnaya1, Oleg G. Yurin1, Karl Emerole2
Livingstone Ssali1, Charles Ngobi2, Bernard Michael Etukoit2,
Aggrey Egessa3, Celestine Bakanda4, Jonathan Wangisi5, Stephen
Okoboi5
Background: Success of treatment as prevention (TasP) depends on
effective engagement of HIV-positive persons in care. The objective of
this study was to estimate the cascade of HIV care and to identify gaps
that impede realization of TasP in Russia.
Methods: We defined 7 steps in the cascade of care framework: HIV
infected (estimation data), HIV diagnosed, Linked to HIV care, Retained
in HIV care, Need ART, On ART, Viral suppressed (VL < 1,000 copies/ml
during 12 month ART). Information was extracted from the Federal AIDS
Centre database (period 1989 year - 31st of December 2012) and also
from the National monitoring forms of Rospotrebnadzor.
Results: 611,858 HIV diagnosed Russian residents were alive by the end
of 2012, which consisted 47% of the estimated 1,300,631 people living
with HIV. Among the HIV diagnosed patients, 470,892 (77%) were linked
to care while 428,279 (70%) were retained. Of 129,817 (21% of HIV
diagnosed) patients who were eligible for ART, 125,623 (97%) were on
treatment and 102,383 (81%) of them had viral suppression. However,
only 17% of HIV diagnosed patients acheived viral suppression which
is necessary to prevent viral transmission. The most significant leakages
of patients were on steps: “HIV infected → HIV diagnosed” (loss - 53%),
“HIV diagnosed → Linked to care” (-23%) and “Retained in care → Need
ART” (-70%).
Conclusions: The stages of HIV diagnosis and estimation of ART
eligibility were the most vulnerable to leakage. Additional efforts are
needed to encourage HIV testing to reduce the number of undiagnosed
cases. Also earlier initiation and enlarging the recommendations to
commence ART among patients retained to care is one of the key aims
to optimize treatment as HIV prevention initiative in Russian Federation.
The AIDS Support Organisation,TASO, Monitoring and Evaluation,
Kampala, Uganda, 2The AIDS Support Organisation,TASO, Programme
Management, Kampala, Uganda, 3The AIDS Support Organisatio,TASO,
Programme Management, Kampala, Uganda, 4The AIDS Support
Organisation,TASO, Planning and Strategic Information, Kampala,
Uganda, 5The AIDS Support Organisation,TASO, Research, Kampala,
Uganda
1
Background: Uganda is considered a success story in HIV and AIDS
programming, but the extent to which adolescents have been retained in
care still not well understood. Retention in care is important for positive
clinical outcomes. The major of objective of the study was to assess the
socio-demographic factors associated with retention in care, treatment
and support programs among adolescents.
Methods: We identified cohort of patients within TASO Management
Information System, who were initiated on Antiretroviral Therapy in
2006. We conducted a retrospective review of data of adolescents aged
10-19 years who received ART services at 11 TASO service centers from
2006 to 2009 Using Cox proportional hazard model, Hazard ratios (HR),
and Adjusted Hazard ratios were computed. Confidence Intervals and
appropriate p-values were computed to control for confounding. We
estimated the overall retention level at 36 months after ART initiation
including patients who died, lost to follow-up, transferred patients.
Results: Female 390(63%), Male 227 (37%), age disaggregation 1014 years 344(56%), 15-19 years 273(44%). In multivariate analyses,
male clients had reduced probabilities to attrition adjusted hazard ratio
(AHR) 0.79 (95% CI: 0.64 to 0.98), (p=0.040) compared to female.
Adolescents who received ART at the health facility had increased risk
to attrition, adjusted hazard ratio (AHR) 1.31 (95%CI: 1.05 to 1.61), (p
= 0.013) compared to those who received services at the Community
drug distribution points (CDDPs). Adolescents with higher CD4 at ART
initiation (cells/ml) CD4 100-259 cells/ml AHR 1.36 (95%CI: 1.05 to
1.77)) and >=250 cells/ml, AHR 1.33 (95%CI: 1.02 to1.75), (p=0.036)
had a high risk to attrition compared to adolescents with lower CD4
Conclusions: The study indicated that using Community Drug
Distribution Points (CDDP) is an effective way to improve patient
retention amongst adolescents receiving ART in a low-income setting.
www.hivr4p.org
415
POSTERS
Federal Budget Institution of Science “Central Scientific Research
Institute of Epidemiology” of Rospotrebnadzor, Russian Federal AIDS
Centre, Moscow, Russian Federation, 2Peoples’ Friendship University of
Russia, Department of Infectious Diseases, Moscow, Russian Federation
1
Posters
Posters 53: Vaginal Rings: Baseline Characteristics, Impact on Condoms and Flora
P53.01
P53.02
Male Condom Functionality in the Presence of
a Vaginal Ring
Baseline Characteristics of HIV-negative
Women Enrolled into the Ring Study - A
Clinical Trial of the Dapivirine Vaginal Ring for
HIV-1 Prevention
Mildie Leuvennink1, Neliette Van Niekerk1, Terri Walsh2, Ron
Frezieres2, Annalene Nel1
International Partnership for Microbicides, Clinical Affairs, Paarl, South
Africa, 2California Family Health Council, Los Angeles, CA, United States
1
POSTERS
Background: IPM is evaluating the safety and efficacy of a dapivirine
vaginal ring for the prevention of male to female HIV-1 transmission.
Once the safety and efficacy of the ring have been proven and it is
available for general use, women will be counselled to use male
condoms together with the ring for maximum protection against HIV-1
infection. The compatibility of male condoms with the vaginal ring was
evaluated.
Methods: The total clinical failure rate of male condoms in the
presence and absence of a placebo ring was assessed in an openlabel, randomized, crossover non-inferiority trial. Seventy healthy,
monogamous, heterosexual couples, 18-45 years (females) or 18-55
years (males) were enrolled, and used 4 condoms when the female was
wearing the ring and 4 condoms when the female was not wearing the
ring. The total clinical failure rate was defined as the number of condoms
that slipped off the penis or broke during intercourse, divided by the
number of condoms used. The safety, tolerability and acceptability of
male condoms in the presence of the ring were assessed.
Results: 275 condoms were used with and without the ring. The total
clinical failure rate was 2.2% with the ring and 4.0% without the ring.
The difference “with ring-without ring” was -1.9% (95% CI: -5.3%; 1.5%).
The upper bound of the CI was less than the pre-defined non-inferiority
margin of 3%. One male urogenital discomfort (severe burning) occurred
during condom use without the ring, due to the condom being too tight.
One participant reported a product-related adverse event (AE) of mild
penile pain during the period of vaginal ring use. No serious AEs were
reported; no AE led to trial discontinuation. More male than female
participants felt the vaginal ring during intercourse, but the majority
reported not being bothered by the ring.
Conclusions: Male condom use was safe and well tolerated with vaginal
ring use. The presence of the ring did not negatively affect the total
clinical failure rate of male condoms or condom use experience.
416
Annalene Nel1, Neliette Van Niekerk1, Allison Carter1, Hannelie
Carstens1, Michelle Isaacs1, Val Kidd1, John Steytler1, Linda-Gail
Bekker2, Cynthia Gama3, Katherine Gill2, Anatoli Kamali4, Philip
Kotze5, Cheryl Louw6, Nokuthula Miti7, Hugo Tempelman8,
Saidi Kapiga9,10
International Partnership for Microbicides, Clinical Affairs, Paarl, South
Africa, 2Desmond Tutu HIV Foundation, Masiphumelele Clinic, Cape
Town, South Africa, 3Maternal, Adolescent and Child Health, Edendale,
South Africa, 4MRC/UVRI Uganda Research Unit on AIDS, Masaka,
Uganda, 5Qhakaza Mbokodo Research Clinic, Ladysmith, South Africa,
6
Madibeng Centre for Research, Brits, South Africa, 7Prevention for HIV/
AIDS Project, Pinetown, South Africa, 8Ndlovu Care Group, Elandsdoorn,
South Africa, 9London School of Hygiene and Tropical Medicine,
London, United Kingdom, 10Mwanza Intervention Trials Unit, Mwanza,
Tanzania, United Republic of
1
Background: Multiple HIV prevention trials are being conducted in
sub-Saharan Africa as women remain vulnerable to HIV-1 infection.
The dapivirine vaginal ring, currently being evaluated for safety and
efficacy, shows potential as a topical microbicide to prevent HIV-1
infection in women.
Methods: The Ring Study (IPM 027) is a Phase III, multi-centre,
randomized, double-blind, placebo-controlled safety and efficacy trial of
a dapivirine vaginal ring in healthy women. Initiated in March 2012, HIV1 sero-negative women, aged 18-45 years, are recruited at 7 research
centres in South Africa and Uganda. Eligible participants must be sexually
active, have normal serum chemistry and haematology profiles and a
normal gynaecological examination. Women must not be pregnant, be
on stable contraception, and not planning to become pregnant for the
trial duration. Post-enrolment, participants attend monthly visits for HIV1 counselling and testing, ring adherence counselling and are issued
with an HIV risk-reduction care package and a new vaginal ring. STI tests
are done 12-weekly and infections treated appropriately.
Results: As of March 2014, enrolment is ongoing with 1483 women
enrolled. The screen:enrolment ratio is 2:1, the most common reason
for screen failure being HIV-infection (up to 55%) and other sexually
transmitted infections (up to 4%). The median age is 26 years with 57%
< 25 years and 9% >35 years. At enrolment 64% reported secondary
school education, and 6% tertiary education. 91% of the participants are
unmarried, and 100% reported a primary partner in the last 3 months;
38% reported no condom use with the last sex act. The most commonly
used contraceptive at baseline is long-acting injectable progestins (83%)
followed by oral contraceptive pills (9%).
Conclusions: It is imperative that HIV prevention methods be available
to women to end the HIV epidemic. It is possible to recruit, screen and
enrol eligible HIV-negative participants at risk for acquiring HIV into a
Phase III microbicide vaginal ring trial.
Posters 53: Vaginal Rings: Baseline Characteristics, Impact on Condoms and Flora
P53.03
P53.04
Baseline Characteristics of HIV-1 Negative
Women Enrolled into a Clinical Trial of
Dapivirine Vaginal Ring for HIV-1 Prevention
Effects of a Vaginal Ring Containing Maraviroc
and or Dapivirine Worn for 28 Days on the
Vaginal Microflora
Thesla Palanee1, Katie Schwartz2, Elizabeth Brown3, Vaneshree
Govender4, Nyaradzo Mgodi5, Flavia Matovu Kiweewa6,
Gonasagrie Nair7, Felix Mhlanga5, Samantha Siva4, Linda-Gail
Bekker8, Zakir Gaffoor4, Nitesha Jeenarain4, Sarita Naidoo4,
Francis Martinson9, Jessica Phillip4, Arendevi Pather4, Bonus
Makhanini10, Lydia Soto-Torres11, Jared Baeten12, on behalf of the
ASPIRE Study Team
Lorna Rabe1, Leslie Meyn1, Beatrice A. Chen2, Lori Panther3, Craig
Hoesley4, Sharon L. Hillier1,2
Wits Reproductive Health and HIV Institute, Johannesburg, South
Africa, 2FHI 360, North Carolina, NC, United States, 3SCHARP - FHCRC,
Seattle, WA, United States, 4Medical Research Council, Durban, South
Africa, 5University of Zimbabwe-University of California San Francisco
Collaborative Research Programme, Harare, Zimbabwe, 6Makerere
University-Johns Hopkins University Research Collaboration, Kampala,
Uganda, 7CAPRISA/University of Kwa Zulu Natal, Durban, South Africa,
8
Desmond Tutu HIV Foundation, Cape Town, South Africa, 9UNC,
Lilongwe, Malawi, 10College of Medicine-John Hopkins University
Research Project, Blantyre, Malawi, 11DAIDS/NIAID/NIH, Washington, DC,
United States, 12University of Washington, Seattle, WA, United States
Magee-Womens Research Institute, Pittsburgh, PA, United States,
University of Pittsburgh, Obstetrics, Gynecology, and Reproductive
Sciences, Pittsburgh, PA, United States, 3The Fenway Institute/Fenway
Community Health, Boston, MA, United States, 4University of Alabama at
Birmingham, Birmingham, AL, United States
1
2
Background: Women’s vulnerability to HIV-1 remains high in subSaharan Africa where transmission occurs mainly through heterosexual
sex.Antiretroviral prophylaxis, is a promising biomedical HIV-1
prevention strategy.
Methods: ASPIRE - A Study to Prevent Infection with a Ring for Extended
Use - is a phase III, randomized, double-blind, placebo-controlled trial
testing the safety and effectiveness of the dapivirine vaginal ring for
prevention of HIV-1 infection. Initiated in August 2012, HIV-1 seronegative women between 18-45 years are being recruited from 15 trial
sites in Malawi (MW), South Africa(SA), Uganda(UG), and Zimbabwe(ZIM).
Eligible participants must be sexually active, have normal serum
chemistry and hematology profiles, gynecologic examinations, not be
pregnant nor planning to fall pregnant for the duration of participation.
Post enrolment, participants attend monthly visits for HIV-1 counseling
and testing, provision of a HIV-1 risk-reduction care package, adherence
counseling, and provision of a new vaginal ring, to be worn continuously
for the following month.
Results: As of March 2014, enrolment is ongoing with 2303 HIV-1
negative women enrolled in a screen:enrol ratio of 2.1:1. The median
age is 26 years with 39%< 25 years and 14%>35 years. Overall, 100%
reported having a primary partner in the 3 months prior to enrolment and
18% reported ≥1 other partner. Eight percent of SA participants report
being married, with 84% in ZIM, 82% in MW and 66% in UG. Forty
percent report no condom use with the last sex act prior to enrolment.
Chlamydia and gonorrhoeae prevalence is 14% and 4% respectively.
Baseline contraceptive use reflects a wide method mix: 41% injectable
depot medroxyprogesterone acetate, 19% contraceptive implants, 14%
injectable Norethisterone enanthate, 13% intrauterine devices, and 11%
oral contraceptive pills.
Conclusions: African seronegative women at risk of HIV -1 infection
can be successfully enrolled into a trial of dapvirine vaginal ring for
HIV-1 prevention.
Background: Vaginal rings with the capacity to release drugs over
time may help with adherence to a microbicide for prevention of HIV.
The objective of this study was to assess the impact of vaginal rings
containing a microbicide on the microflora of the vagina and to assess
the quantity of biofilm adherent to the rings.
Methods: 48 Non-pregnant, HIV negative, sexually abstinent women
aged 18-40 were enrolled in a double-blinded, 4 armed (12 per arm)
randomized controlled trial. Women were recruited in Pittsburgh, PA;
Boston, MA; and Birmingham, AL. Vaginal rings containing 25 mg
dapivirine, 100 mg maraviroc, a combination of the two drugs, or
placebo were worn for 28 days. The rings were removed and a subset
of 16 was assessed for biofilm formation with electron microscopy.
Vaginal swabs and smears were collected at baseline, 7, 28, and 52
days and cultured for aerobic and anaerobic bacteria and assessed for
Nugent score. Modified Poisson regression and generalized estimating
equations were used to assess the effect of ring use on the prevalence
and concentration of vaginal microflora.
Results: 47 women had cultures for at least 3 visits. The prevalence
of pigmented anaerobic Gram negative rods increased significantly
during the use of the placebo ring (RR 2.28, CI 1.15-4.49, P= 0.02)
and dapivirine ring (RR 1.67, CI 1.02-2.73, P=0.04). Overall there was a
decrease in prevalence of E. coli during ring use (RR 0.50, CI 0.26-0.96,
P=0.04). However, no arm was statistically significant. There was no
significant change of the Nugent score over the 52 days. The amount
of biofilm ranged from scant to confluent but there was no significant
difference in the quantity of biofilm stratified by the type of ring.
Conclusions: Although there was an increase in prevalence of anaerobic
pigmented Gram negative rods in the placebo and dapivirine users,
there was no overall change in biofilm development or in Lactobacillus
or microflora when assessed by Nugent score.
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