Strategic Planning for Swine Disease Research

Transcription

Strategic Planning for Swine Disease
Research
Meeting Report
December 4, 2012 Washington, DC Strategic Planning for Swine Disease
Research
The National Pork Board and the Department of Homeland Security
(DHS) National Center for Foreign Animal and Zoonotic Disease
Defense (FAZD Center) convened a strategic planning meeting to
discuss swine disease research efforts supported by federal or
National Pork Board funds on December 4, 2012 in Washington, DC.
The overall goal of the meeting was to discuss current research
efforts focused on three key diseases of agricultural consequence
affecting swine, African Swine Fever (ASF), Classical Swine Fever
(CSF), and Foot and Mouth Disease (FMD), and to identify gaps in
research and funding.
Meeting participants addressed research and gaps in the following
areas:
•
Diagnostics
o Sampling procedures
o Utility of pen-side detection
o Reagent and technology development
•
Vaccines
o Virus-specific research and development
o Routes and methods of delivery
o Antigenic coverage and serotype protection
o Onset and duration of immunity
o Use of vaccines developed for/validated in cattle for use in
swine
o Domestic manufacture and formulation of vaccines
o Vaccine stockpiles and deployments
o Live attenuated and recombinant vaccines
o Adjuvant development
•
Biotherapeutics
o Interferon-enhanced protection against FMD
o Transmission studies
o Current models of experimental challenge versus natural
infection in swine
o Mechanisms of protection in swine
Page 1 of 32 Executive Summary
This report summarizes discussion points and research gaps
identified during a strategic planning for swine disease research
meeting convened by the National Pork Board and the FAZD Center
on December 4, 2012. Participants included 20 personnel
representing the FAZD Center, the National Pork Board, the DHS
Science and Technology Directorate (S&T), the United States
Department of Agriculture (USDA) Agriculture Research Service
(ARS), USDA Animal and Plant Health Inspection Service (USDA
APHIS), the Center of Excellence for Emerging and Zoonotic Animal
Diseases (CEEZAD), the American Association of Swine
Veterinarians (AASV), and Texas Veterinary Medical Diagnostic
Laboratory (TVMDL). A full list of attendees may be found in
Appendix A.
Meeting Objectives
The objectives of this meeting were to:
•
Review the current state of vaccine, diagnostics, and
biotherapeutics research involving three key foreign animal
diseases (FADs) of consequence to the swine industry: ASF, CSF,
and FMD
•
Identify gaps in funding and research involving these three key
diseases of consequence to the swine industry
•
Prevent duplication of efforts in order to best utilize available
research funding and resources
•
Identify other areas in need of attention from research,
government and industry representatives
•
Facilitate communication between academia, government, and
industry involved in research of diseases of consequence to the
swine industry
The meeting was structured as a round-table discussion organized by
disease. Each disease was discussed in context of current and past
Page 2 of 32 vaccine projects, as well as diagnostic and screening tools and
biotherapeutic projects. Following a summary of each project, gaps in
research and funding for each disease were identified. A list of current
and past research projects discussed may be found in Appendix B.
Page 3 of 32 Highlights of the Strategic Planning for
Swine Disease Research Meeting
Meeting Overview
Meeting discussions were structured by disease (ASF, CSF, FMD, or
multiple FADs), with each focus area further broken down to review
vaccine, diagnostics and screening tools, or biotherapeutics research
by pathogen of interest. Representatives or principal investigators
from each research entity provided an overview of current and past
research by project, followed by a discussion of gaps in research and
funding.
Discussion Summaries
Discussion Topic: African Swine Fever
ASF Vaccine Projects
Round table discussion of research involving ASF began with USDA
and DHS summarizing current and past research projects involving
vaccine development targeted against ASF (Table 1). This has been
identified as a high priority research area for the swine industry, as
there are currently no effective vaccines available to combat this
pathogen. The recent spread of outbreaks through the Caucuses and
near the Eastern European border makes the need for an efficacious
vaccine even more critical for protecting US industry. Several unique
and ongoing recent research efforts funded by DHS S&T are helping
address this need.
Table 1. ASF Vaccine Projects
Year Project Title Development of Multi-‐
2011 component Vaccines for African Swine Fever * Identification of African 2011 Swine Fever Candidates by Reverse Vaccinology Primary Investigator Institution Funding Agency Status Mwangi TAMU DHS S&T In progress Bounpheng TMVDL DHS S&T In progress Page 4 of 32 Development of a Proof of Concept Rationally 2011 Designed Live Attenuated African Swine Fever Virus Vaccines A Comprehensive Research Program in African Swine 2012 Fever Towards the Development of Novel Countermeasures Borca USDA ARS PIADC DHS S&T In progress Borca/Arzt USDA ARS PIADC DHS S&T In progress *Project includes monoclonal antibody development.
Gaps
The following gaps were identified in the areas of ASF vaccine
research and funding:
•
•
•
Significant, sustained funding commitment
o It is estimated that $2-3 million per year over the course of
several years is the minimum amount of necessary funding
to seamlessly support research
o Though DHS S&T has made significant investment in
vaccine research and development, sustained funding is
needed to take initially successful proof of concept studies
for lead candidate(s) through to final product
Immunogenicity/challenge studies
o Use attenuated and recombinant strains
o Focus on promising candidates
o Understanding host protective humoral and cellular immune
mechanism
o Understanding viral/host interactions at the cellular level
Increase workforce development
o Need to train the next generation of ASF experts
o PIADC is currently the only place in the U.S. that trains
people in ASF research and animal and lab capacity is
limited
o Dichotomy between post-doctoral starting salaries versus
student loan burden
o Lack of sustained training/development programs
ASF Diagnostics/Immunology Projects
Round table discussions continued with a review of ASF diagnostics
projects (Table 2). There was an active discussion about sampling for
Page 5 of 32 virus detection from swine oral fluids, which is currently done for
routine endemic disease testing as part of normal swine production
practices. Meat juice collected in slaughter plants is also used as a
routine sample for other pathogens, and could potentially be validated
for FADs as well, depending on whether detection of antibodies
versus isolation of virus is desired.
Table 2. ASF Diagnostics/Immunology Projects
Year Project Title Comparison of different sample source and sample 2011 pooling for the detection and surveillance of ASF Development of fluorescent recombinant antibodies to detect 2011 African swine fever virus in tissue samples and infected cells Development of 2012 Monoclonal Antibodies Specific to ASFV Proteins A Comprehensive Research Program in African Swine 2012 Fever Towards the Development of Novel Countermeasures Identification of genetic signatures for African 2012 swine fever virus serologic group specificity Primary Investigator Institution Funding Agency Status Dixon Pirbright DHS In progress Escribano Instituto Nacl. Investigacion National Pork In y Tecnologia Board progress Agraria y Alimentaria (INIA) Wu/Sayed USDA APHIS PIADC DHS S&T In progress Borca/Arzt USDA ARS PIADC DHS S&T In progress Rock University of National Pork In Illinois Board progress Gaps
The following gaps were identified in ASF diagnostics research and
funding:
Page 6 of 32 •
•
•
•
•
•
•
Understand the application of research results from strains with
differing virulence
o Studies with moderate strains are needed for comparison to
those with highly virulent strains (linear relationship?)
Develop US-based reagents
Develop and evaluate tools using moderate strains to improve
anti-mortem detection
o Matrices to best identify early infection
 Meat juice
 Oral fluids
 Tonsils
o Comparison of sample selection methods and time course
for window of detection
Standard diagnostic scoring tools
Active surveillance program in ASF endemic countries
o Identify circulating strains
Develop and use of a pen-side test (post-outbreak)
o Established policy on pen-side testing
o Outreach to producers
Additional deep sequencing to support targeted research efforts
(functional genomics)
Discussion Topic: Classical Swine Fever
CSF Vaccine Projects
Discussion of CSF vaccine projects included discussion of research
projects being performed by the FAZD Center and USDA ARS,
including several recent efforts involving novel development
technologies. The round table discussion emphasized the need for
vaccination against highly virulent strains of CSF. Table 3
summarizes CSF vaccine projects.
Page 7 of 32 Table 3. CSF Vaccine/Biotherapeutics Projects
Primary Investigator Institution Funding Agency Status Identification of host factors interacting with classical 2009 swine fever virus proteins: development of novel anti-‐
viral therapeutics. Borca USDA ARS NAA National Pork Board Completed FAD Countermeasure Development Roth/Riemser TABI, Inc. DHS S&T In progress Borca USDA ARS PIADC DHS S&T In progress Borca USDA ARS NAA National Pork Board In progress Holtz Caliber Biotherapeutics DHS S&T In progress Year 2009 Project Title Classical Swine Fever (CSF) Vaccine and Diagnostic 2010 Countermeasure Development Evaluation of Envelope Proteins for Rapid Induction 2011 of Protective Immune Responses Against Classical Swine Fever Plant-‐based expression vectors for rapid, high-‐
2012 throughput development of animal vaccines Gaps
The following gaps were identified in CSF vaccine research and
funding:
•
•
•
•
Information on the EU live, attenuated marker vaccine candidate
‘CP7_E2 alf’ (EU FP6 programme – CSF project SSPE-CT-2003501599)
o Onset of protection
o Plans for pursuit of U.S. license
Need for companion diagnostic assay to differentiate infected from
vaccinated animals (DIVA) for the ARS CSF marker vaccine
candidate
Deployment strategy/policy for the conventional, live attenuated Cstrain vaccine (non-DIVA) is not well defined
Evaluate onset of protection and duration of immunity of subunit
vaccines
o Protective dose(s)
Page 8 of 32 •
•
•
•
•
o Time-course of protection
o Combined efficacy with biotherapeutics
Potential to use early onset live attenuated vaccine at “ground
zero” and a subunit vaccine in surveillance areas?
Regulatory gaps
Better understanding of host/virus relationships in vaccine
production cells
Identification of virulence factors/mechanism of protection for antivirals
Evaluate use of amantadine to inhibit viral replication
CSF Diagnostics
Participants agreed that CSF endemic countries need diagnostic tools
and procedures to detect the virus. Virulence varies between
outbreaks and countries; however, establishing stronger international
collaborations will help in both capacity building and validation of tests
for domestic use. Table 4 summarizes CSF diagnostics projects.
Table 4. CSF Diagnostics Projects
Year Project Title Foreign Animal Disease (FAD) Diagnostic Assay 2009 Development -‐ Task 2: Diagnostic Technologies for Classical Swine Fever Development of classical swine fever virus diagnostic 2011 assays for porcine oral fluid samples Primary Investigator Institution Funding Agency Status Mayr/Batonick USDA APHIS PIADC DHS S&T In progress Thanawongnuwech Chulalongkorn National Pork In University Board progress Gaps
The following gaps were identified in CSF diagnostics research and
funding:
•
•
Establish collaborations overseas for diagnostic field testing
Develop US-produced reagents
o Reagents needed for virus isolation and tissue staining
Page 9 of 32 •
Commercial development and licensure of an improved CSF
serology test
Discussion Topic: Foot and Mouth Disease
FMD Vaccine Projects
Discussion of FMD vaccine projects began with an overview of the
projects currently underway at Plum Island Animal Disease Center.
On May 31st, 2012, the first US-produced licensed vaccine for FMD
was approved. The focus is primarily on bovine vaccination; however,
DHS’ commercial partner has indicated they hope to work towards
commercial development of the Adenovector-based FMD vaccine
platform for swine. A summary of other past and present FMD vaccine
research efforts conducted at PIADC and elsewhere may be found in
Table 5.
Table 5. FMD Vaccine Projects
Year 2000 2004 2010 2010 Project Title Development and Testing of a Subunit Vaccine for Foot-‐and-‐
Mouth Disease Development of an Antiviral and Vaccine Approach to Control Foot-‐and-‐Mouth Disease Production of FMD Virus Pseudovirion Vaccine Candidates Using a Plant Transient System Research and Development of Molecular FMD Vaccines (historically cattle focused) Primary Investigator Institution Funding Agency Status Grubman USDA ARS NAA National Pork Board Completed Grubman USDA ARS NAA National Pork Board Completed Hiatt Kentucky BioProcessing DHS S&T In progress Brough and Butman GenVec DHS S&T In progress Page 10 of 32 Improved Challenge Systems for FMD Vaccine and 2011 Biotherapeutics Testing in Cattle and Pigs Arzt USDA ARS PIADC DHS S&T In progress Development of VLP Vaccine as a 2011 Countermeasure for FMD Massare Novavax DHS S&T In progress Kern Inovio DHS S&T (CRADA/No Direct Funding) In progress de los Santos USDA ARS NAA National Pork Board Completed David Merial DHS S&T (CRADA/No Direct Funding) In progress Miller Benchmark Biolabs DHS S&T In progress Mittal Purdue University DHS S&T In progress Weinberger Bavarian Nordic DHS S&T In progress 2011 FMD DNA Vaccine 2011 2011 2012 2012 2012 Rational design of attenuated foot-‐and-‐
mouth disease virus strains for development of improved disease countermeasures Testing and Evaluation of Human Adenovirus Replication Deficient Vectored FMD Vaccines (swine focus) Acquisition of Master Seed Virus (MSV) and Working Seed Virus (WSV) Lots for FMD Molecular Vaccine Countermeasures Bovine Adenovirus Vector-‐Based Vaccine for FMD Construction and Evaluation of Recombinant MVA-‐BN FMDV Candidates Page 11 of 32 Exploiting the potential of leader proteinase coding sequence of foot-‐and-‐
mouth disease virus to 2012 derive attenuated strains suitable for pathogenesis studies and development of improved countermeasures. Replicon-‐based Rapid 2012 Response Vaccines for FMD de los Santos USDA ARS NAA Kamrud Harrisvaccines National Pork Board In progress DHS S&T In progress Gaps
The following gaps were identified in FMD vaccine research and
funding:
•
•
•
•
•
Test additional vaccine platforms in pigs that have shown promise
or success in cattle
o Host cross-species protection studies
Conduct immunogenicity studies in pigs
o Duration of immunity (key for eradication efforts)
o Broad antigenic coverage
o Cross-serotype protection (broadening immunity)
o Protective dose(s)
o Route of inoculation (e.g., subcutaneous or intradermal)
o Method of delivery
o Adjuvants
o Antigen stability in different environments
Define mechanisms of host protection
o Utilize newly developed veterinary immunity research
reagents
o Utilize vaccinated animals from endemic countries as a
resource to investigate vaccine efficacy and immunity
Develop policies supporting vaccination and/or “stamping out”
FMD globally via eradication efforts
o Policies/schedules for available vaccines during different
phases of an outbreak
Stockpile and/or procure vaccine for emergencies
Page 12 of 32 o Number of vaccine doses
o Number of serotypes/topotypes covered
FMD Diagnostics
The discussion of FMD diagnostics projects began with a discussion
of current research efforts funded by DHS S&T by performers outside
the Federal government, including TVMDL, TABI, and national
laboratories. This was followed by a review of past and ongoing
efforts being performed at Plum Island. PIADC has a new monoclonal
antibody development initiative that they hope will lead to U.S.
production of reagents so they do not have to be purchased from
abroad. This project feeds into R&D support for developing vaccines.
FMD diagnostic projects at TVMDL, PIADC, and elsewhere are
summarized in Table 6.
Table 6. FMD Diagnostics Projects
Year Project Title Diagnostic Evaluation of Multiplexed Reverse Transcription-‐
PCR Microsphere 2008 Array Assay for Detection of Foot-‐
and-‐Mouth and Look-‐
Alike Disease Viruses Foot-‐and-‐Mouth Disease Multiplexed 2008 Nucleic Acid Assay Enhancements and Validation 2010 FAD Countermeasure Development Primary Investigator Institution Funding Agency Status Hindson University of California Lawrence Livermore National Laboratory DHS S&T Completed Beckham (collaboration with LLNL) USDA APHIS PIADC DHS S&T Terminated Roth (collaboration with Prionics) TABI, Inc. DHS S&T In progress Page 13 of 32 Development of 3ABC ELISA for Detection of FMD Antibody in Infected Animals 2011 Regardless of Vaccination Status (Prototype kit and bench validation) 2011 2011 2011 2012 USDA ARS PIADC/FADDL/TVMDL Deployment of FMD Serology to the McIntosh National Animal (collaboration USDA APHIS PIADC Health Laboratory with Network (swine and NAHLN/TVMDL) cattle together) Investigating potential existence of chronic, persistent foot-‐and-‐
mouth disease virus Arzt USDA ARS NAA infection in domestic pigs; implications for disease control strategies Negative Cohort Study for Validation of the Tomlinson/ USDA/FADDL/ NAHLN SVANODIP® FMDV-‐Ag McIntosh test Development and Characterization of Monoclonal TVMDL/ LLNL/ Antibodies to Foot-‐
Bounpheng BIOO Sci and-‐Mouth Disease Virus Structural Proteins Establishment of a FMD Antisera Bank Molecular Epidemiology and Biosurveillance of 2012 FMD In Endemic Regions of Middle East, Southeast Asia and Africa 2012 Rieder/Sayed/ Bounpheng DHS S&T In progress DHS S&T In progress National Pork Board In progress DHS S&T Completed DHS S&T In progress Wu USDA APHIS DHS S&T In progress Rodriguez USDA ARS PIADC DHS S&T In progress Page 14 of 32 Pen-‐side detection of Foot-‐and-‐Mouth 2012 Disease virus by a portable microfluidics PCR system Bounpheng/ McIntosh USDA/FADDL/ TVMDL DHS S&T In progress Gaps
The following gaps in FMD diagnostics research and funding were
identified:
•
•
•
•
•
Validate 3ABC ELISA in swine vaccinated with Ad5 vectored
vaccine
Develop US-based reagents
o Most reagents developed with bovine model
o Need sera to test diagnostics and detection methods
o Issues with continuity of supply/cost of international
reagents
Validate different sample types across multiple serotypes
Explore other modalities for FMD detection (aerosol, thermal
imaging)
Explore value of penside testing in outbreak situations with
industry
FMD Biotherapeutics
Discussion of FMD biotherapeutics began with a review of research
that had originally been done in cattle and is now being translated to
the swine model. USDA presented research highlighting the use of
interferon lambda as an adjuvant to FMD vaccines to generate a
protective B-cell response. In addition, USDA ARS has promising
research involving adenovirus vectors that can offer protection as an
adjuvant to the current Ad5 vaccine within one week. Challenges
involved in modeling natural infection were also discussed. Table 7
summarizes past and present FMD biotherapeutics studies conducted
by the USDA and others.
Page 15 of 32 Table 7. FMD Biotherapeutics Studies
Year 2009 2011 Project Title Primary Investigator Foreign Animal Disease (FAD) Countermeasure Development -‐ Identification Grubman of Biotherapeutic Candidates to Control FMDV Countermeasures for Foot-‐
and-‐Mouth Disease Utilizing Diagnostics, Biotherapeutics, a Novel Vaccine Platform, and de los Santos/ Improved Challenge Systems -‐ Grubman Identification of Biotherapeutic Candidates to Control FMDV Institution Funding Agency Status USDA ARS PIADC DHS S&T Completed USDA ARS PIADC DHS S&T In progress Gaps
The following gaps in research and funding were identified with
regards to FMD biotherapeutics in swine:
•
•
•
Establish biotherapeutic adjuvant utility with other vaccines
Evaluate site of replication/timecourse of infection in swine
o Oral challenge more closely models natural infection than
intradermal or foot
o Transmission studies need to be done in swine
o Identify tissues to target for biotherapeutics
Need to define persistence in swine versus cattle via carrier
studies
Discussion Topic: Multiple Foreign Animal Diseases (FADs)
Multiple FAD Diagnostic Projects
Several multiplex and cross-pathogen projects on FADs have been
conducted at Plum Island and elsewhere. All of the projects are
diagnostic in nature. The diagnostic projects conducted by PIADC,
TVMDL, and others are summarized in Table 8.
Page 16 of 32 Table 8. Multiple FAD Diagnostic Projects
Year Project Title Enhancements of High-‐
2008 Throughput Diagnosis for FADs 2009 2011 2011 2012 2011 & 2008 Primary Investigator Institution McIntosh Foreign Animal Disease Diagnostic Assay Development: Vesicular Jia Disease Reagent Production Development of a multiplex RT-‐qPCR assay for surveillance of FADs Bounpheng/McIntosh during routine testing of oral fluid samples The Matrix-‐Chaperone: Ambient Temperature Biospecimen Collection, Transport & Banking For Hogan Simplified Animal Disease Screening (swine and cattle). Assessment of Two Methods for Nucleic Acid Stability and Viral Mayr Inactivation of FAD Viruses Collected from Oral Fluid Rope Samples Development of Pan-‐
viral DNA Microarrays for the Detection of McIntosh Emerging and Foreign Animal Diseases Funding Agency Status USDA APHIS DHS S&T Completed USDA APHIS DHS S&T In progress TMVDL/ DHS S&T FADDL In progress IntegenX DHS S&T In progress USDA APHIS DHS S&T In progress USDA APHIS PIADC DHS S&T Completed Gaps
The following gaps were identified involving multiple FAD diagnostic
research projects and funding:
•
•
Develop universal protocols for sample collection and preparation
Establish working relationships and collaborations with
laboratories in countries where each disease is endemic
Page 17 of 32 •
•
•
•
•
Address questions surrounding transport of samples from outbreak
zones to laboratories for testing
Research usable/optimal sample matrices for multiplex assays
Develop capacity to work on FADs in other laboratories
o Use ABSL-2 containment where possible to make efficient
use of high containment space
Emphasize multi-institution collaborations in calls for proposals
Emphasize partnerships that facilitate transfer of technology earlier
in the process for projects that are leading to usable diagnostic
tools, vaccines or biotherapeutics
Outcomes and Next Steps
The meeting reported here was an unusual opportunity for scientists
from research entities involved in vaccine, diagnostics, and
biotherapeutics research for three diseases of consequence in swine,
African Swine Fever, Classical Swine Fever, and Foot-and-Mouth
Disease, to entertain open discussion with funding agencies and
industry representatives to identify gaps in research and funding.
Sharing information on technologies currently in development will help
prevent duplication of efforts and ensure responsible discharge of
research funding. Meeting participants agreed that the meeting was
fruitful in that it allowed participants to openly discuss research
projects conducted at multiple institutions and funded by multiple
agencies. The outcomes of the discussion will be used by the swine
industry to devise a national strategy for future ASF, CSF, and FMD
research and funding.
Page 18 of 32 Appendix A: Meeting Participants
Name
Mr. Bobby Acord
Organization
National Pork Board Council
Program Manager, Chemical and Biological Division, DHS
Dr. James Anthony
S&T
Director, FAZD Center and TVMDL
Dr. Tammy Beckham
Director, Swine Health Information and Research, National
Dr. Lisa Becton
Pork Board
Dr. Melissa Berquist
Associate Director, FAZD Center*
Dr. Manuel Borca
Head, Swine Diseases Research, PIADC
Dr. Mangkey Bounpheng Molecular Diagnostics Section Head, TVMDL
Scientific Consultant, Plum Island Animal Disease Center,
Dr. David Brake
DHS
Dr. Matthew Coats
Program Manager, Office of University Programs, DHS S&T
Agriculture Defense Branch Chief, Chemical and Biological
Dr. Michelle Colby
Division, DHS S&T
Director of Science, Plum Island Animal Disease Center,
Dr. Bruce Harper
DHS S&T
Dr. Mike King
Manager, Science Communications, National Pork Board
Microbiologist, Diagnostic Services Section, USDA APHIS
Dr. Greg Mayr
FADDL
Dr. Juergen Richt
Director, CEEZAD, Kansas State University
Research Leader, Plum Island Animal Disease Center,
Dr. Luis Rodriguez
USDA ARS
Dr. Abu Sayed
Microbiologist, USDA APHIS FADDL
Dr. Harry Snelson
Director of Communications, AASV
Dr. Paul Sundberg
Vice President, Science & Technology, National Pork Board
Dr. Sabrina Swenson
Virologist, USDA APHIS*
Dr. Patrick Webb
Director, Swine Health Programs, National Pork Board
* Participated via teleconference/webinar.
Page 19 of 32 Appendix B: Full List of Projects Sorted by Disease
All meeting participants received a full list of African Swine Fever, Classical Swine
Fever, and Foot-and-Mouth Disease projects, as well as multiple foreign animal disease
(FAD) projects, compiled by the FAZD Center, the National Pork Board, PIADC, and
USDA, sorted by disease. The collated list follows, beginning on the next page.
Page 20 of 32 Disease
ASF
ASF
ASF
Area
Diagnostics
Diagnostics
Diagnostics
Y ear
Project Title
2011
Comparison of different sample source
and sample pooling for the detection
and surveillance of ASF
2011
Development of fluorescent
recombinant antibodies to detect
African swine fever virus in tissue
samples and infected cells
2012
A Comprehensive Research Program in
African Swine Fever Towards the
Development of Novel
Countermeasures
Primary
Investigator
Institution
Funding Agency
DHS
Escribano
Instituto Nacl.
Investigacion y
Tecnologia
Agraria y
Alimentaria (INIA)
National Pork
Board
In progress
The present project pretends to develop new reagents to cover a
gap in the ASFV diagnosis.
DHS S&T Vx/Dx
In
progress.
2014 end
date.
Objective 1: Development of challenge model for ASF to
characterize pathogenesis and evaluate novel countermeasure
products. Objective 2: Determining immune mechanisms of
protection induced by attenuated strains. Objective 3: Functional
genomics.
In
progress.
2014 end
date.
The end product of this project is a battery of monoclonal
antibodies against ASF proteins p30, p72, p54, and 8-DR. These
will be produced and made available to ARS and DHS for their
research needs. They can also be potentially used in the
development or improvement of diagnostic assays in FADDL. It is
expected that the knowledge acquired from ARS and DHS
characterization will assist in this process.
Borca/Arzt
USDA ARS PIADC
2012
Development of Monoclonal Antibodies
Specific to ASFV Proteins
Wu/Sayed
USDA APHIS
PIADC
DHS S&T Vx/Dx
ASF
Diagnostics
2012
Identification of genetic signatures for
African swine fever virus serologic
group specificity
Rock
University of
Illinois
National Pork
Board
2011
Development of Multi-component
Vaccines for African Swine Fever
This project will compare and validate pooled sample matricies,
including oral fluids, nasal swabs, blood, and serum, for the
In progress molecular detection of ASFV. Project will include a methods
comparison between USDA NAHLN PCR, Pirbright PCR, and
serological assay for detection.
Pirbright
Diagnostics
Vaccine
Comments
Dixon
ASF
ASF
Status
Mwangi
Texas A&M
Research
Foundation
DHS S&T
Identify genetic signature (s) for ASFV serologic group specificity
In progress and determine serologic group specificity for currently untyped ASFV
isolates in VNIIVViM strain collection
In
progress.
2013 end
date.
The objective of this proposal is to clone and express 5 candidate
ASFv genes in eukaryotic (HEK) and adenovirus expression
platforms. The eukaryotic platform will be used to produce
recombinant proteins to serve as immunogens for MAb and rabbit
PcAb reagent production (2 ASFV targets). The adenovirus platform
will be used to produce (3) vaccine candidates for swine proof-ofconcept safety and immunogencity study at TAMU.
Disease
ASF
ASF
Area
Vaccine
Vaccine
Y ear
2011
2011
Project Title
Identification of African Swine Fever
Candidates by Reverse Vaccinology
Development of a Proof of Concept
Rationally Designed Live Attenuated
African Swine Fever Virus Vaccines
Primary
Investigator
Bounpheng
Borca
Institution
TMVDL
USDA ARS PIADC
Funding Agency
DHS S&T Vx/Dx
DHS S&T Vx/Dx
Status
Comments
In
progress.
2013 end
date.
The objective of this proposal is to evaluate a novel approach,
reverse vaccionology (RV), for the identification and development of
ASFV vaccine candidates. RV will be used to identify novel in silico
candidates and rank the DHS BAA candidates (i.e., p30, p54, p72,
CD2v). Two protein expression and delivery platforms will be
evaluated. The in silico and top ranked DHS candidates will be
expressed in mammalian HEK 293 cells and fused to an
immunogenic inducing tag. In addition these candidates will be
expressed in the highly safe poxvirus modified vaccinia Ankara
(MVA) vaccine vector to enhance cellular immunity. Recombinant
vaccines will be tested for immunogenicity and safety in pigs. The
PIs anticipate that these candidates (alone or synergistically
combined) will be highly immunogenic and induce strong humoral
and cellular responses. Protein vaccines are highly safe and can
induce strong humoral responses. In addition, MVA-based vaccines
are an attractive vaccine delivery technology because when
administered to the host they can induce in vivo expression of one
or more specific antigens. These newly expressed antigens are
processed and presented by professional antigen-presenting cells,
resulting in the induction of antibody responses with high avidity,
as well as major histocompatibility complex class I-restricted
cytotoxic T-lymphocyte (CTL) responses. This pattern of responses
is similar to that induced by live attenuated vaccines. In addition,
the induction of B and T cell immune responses eliminates the need
for adjuvants. These features are major advantages of viral vectors
when compared to other vaccine delivering platforms.
In
progress.
2014 end
date.
Objective: 1. Develop recombinant African Swine Fever Virus (ASFV)
strains by deletion of one or more viral genes already described as
responsible for inducting attenuation of highly virulent ASF strains.
2. Test attenuated ASFV strains for their ability to induce protection
against challenge with homologous, well-characterized, virulent
ASFV isolates. 3. Evaluate patterns of heterologous protection
among genetically heterogeneous ASFV strains. Approach: Develop
recombinant African Swine Fever Virus (ASFV) strains by deletion of
one or more viral genes already described as responsible for
inducting attenuation of highly virulent ASF strains. Critical ASFV
genes that are responsible for inducing attenuation of highly
virulent ASF strains will be deleted individually or as a group. This
deletion should confer virus replication but not disease production.
ASFV isolates. This will be done through: Testing strains for in vivo
attenuation, testing for efficacy against homologous challenge,
determination of minim protective dose response, evaluation of
protection profile for at least one attenuated ASFV vaccine
candidate, and the evaluation of lead vaccine candidate to induce
sterile immunity. 3. Evaluate and confirm cross-protection conferred
by lead vaccine candidate (obj. 2) by using genetically diverse ASFV
strains.
Page 22 of 32 Dis eas e
CSF
CSF
CSF
CSF
CSF
Area
Diagnostics
Diagnostics
Vaccine
Vaccine
Vaccine
Y ear
2009
2011
2009
2009
2010
Project Tit le
Diagnostic Technologies for Classical
Swine Fever
Primary
Inves t igat or
Mayr/Batonick
Ins t it ut ion
USDA APHIS
PIADC
Development of classical swine fever
Thanawongnuwe Chulalongkorn
virus diagnostic assays for porcine oral
ch
University
fluid samples
Identification of host factors
interacting with classical swine fever
virus proteins: development of novel
anti-viral therapeutics.
FAD Countermeasure Development
Classical Swine Fever (CSF) Vaccine
and Diagnostic Countermeasure
Development
Borca
Roth /Riemser
Borca
USDA ARS NAA
TABI, Inc.
ARS USDA
PIADC
Funding
Agency
DHS S&T
Vx/Dx
National Pork
Board
National Pork
Board
St at us
Comment s
In progress.
2013 end
date.
The current CSF antibody ELISA has limited specificity in excluding
antibody to ruminant pestiviruses. Identification and
characterization of the E2 glycoprotein epitopes that discriminate
between swine and ruminant pestivirus are essential in the design
of a next generation ELISA. For epitope mapping, a panel of
monoclonal antibodies was obtained in collaboration with the
Central Institute for Animal Diseases Control, Lelystad, the
Netherlands. The CSF target epitopes were defined, and the ELISA
was designed and is being optimized and validated. Such an ELISA
will have vital application in the current national surveillance
program and during recovery of an outbreak.
In progress
The primary objective of this project is to optimize and validate
technology capable of rapidly identifying premises infected with
classical swine fever virus (CSFV) following its introduction into
North America or other CSFV-free areas using oral fluid samples.
Achievement of this objective will also provide technology for
improved surveillance in CSFV endemic areas, thereby enhancing
elimination and control efforts. The specific objectives are focused
on demonstrating the feasibility of detecting CSFV in oral fluids by
modified PCR-based methods.
Completed
Results obtained enable the identification of several host proteins
interacting with CSFV structural protein Core. Core protein is the
major contributor to the virus capsid. Several of these interactions
have been studied in detail and the regions of the CSFV Core
protein interacting with the host proteins were identified. Mutant
CSFV viruses having altered these regions have been demonstrated
that have severely altered their ability to produce disease in swine.
Therefore, the manipulation of the identified host-virus interactions
allowed the development of attenuated strains of virus which may
constitute a tool for the further development of live attenuated
vaccine against classical swine fever. Additionally, this knowledge
may open the possibility of designing bio therapeutic compounds
that could alter those critical interactions that may limit the spread
of the disease.
DHS S&T
Vx/Dx
CVB Approval of Permittee License for Importation and Distribution
of CSF Modified Live Vaccine, Chinese strain (C strain). Riemser
Arzneitmittel AG (GDR) is currently producing a EU licensed vaccine.
In progress.
The vaccine (C strain) was recently excluded from the USDA APHIS
2013 end date
Select Agent List. Plans are in progress to import master seed virus
(modification
(MSV) and master seed cell bank (MSB) for safety testing in swine
to 2014 end
by FADDL, PIADC. If satisfactory, MSV and MSB will sent to USDA
date planned)
CVB for additional testing. Submission of various testing reports to
USDA CVB are in progress. GDR manufacturing facility has been
inspected by USDA CVB I&C.
DHS S&T
Vx/Dx
Objective 1: Develop a genetically stable version of FlagT4 virus
(FlagT4c). Objective 2: Viral-vectored vaccine (backup candidate):
In progress. POC safety and efficacy studies using baculovirus-vectored
2013 end date experimental vaccine. Objective 3: Obtain select agent exclusion
for FlagT4c vaccine candidate and transition to AH partner (CRADA
in place).
CSF
CSF
Area
Vaccine
Vaccine
Y ear
Project Title
2011
Evaluation of Envelope Proteins for
Rapid Induction of Protective Immune
Responses Against Classical Swine
Fever
2012
Plant-based expression vectors for
rapid, high-throughput development of
animal vaccines
Primary
Investigator
Borca
Holtz
Institution
USDA ARS NAA
Funding
Agency
National Pork
Board
Caliber
DHS S&T OUP
Biotherapeutics
Status
Comments
In progess
The main objective of the research project is to evaluate native and
modified forms of CSF envelope proteins for their capacity to induce
rapid protective immune response against the CSFV. CSFV envelope
proteins are expressed as fusion proteins along with different
immunological adjuvants. The improved efficacy of the novel
constructs will evaluated compared with the native version of the
proteins in terms of induced immune response and protection.
In progress
The main objective of this research project is to demonstrate proof
of principle for rapid expression, testing, and commercial-scale
production capacity using a novel plant-based approach. The CSF E2
protein is targeted for a potential recombinant subunit vaccine for
use in later stages of a disease outbreak as a possible compliment
to a live attenuated vaccination strategy.
FMD
FMD
Area
Biotherapeutics
Biotherapeutics
Primary
Investigator
Project Title
2009
Foreign Animal Disease (FAD)
Countermeasure Development Identification of Biotherapeutic Candidates
to Control FMDV
Grubman
USDA ARS
PIADC
DHS S&T
Completed
2011
Countermeasures for Foot-and-Mouth
Disease Utilizing Diagnostics,
Biotherapeutics, a Novel Vaccine Platform,
and Improved Challenge Systems Identification of Biotherapeutic Candidates
to Control FMDV
de los Santos/
Grubman
USDA ARS
PIADC
DHS S&T
In progress. Overarching Goal: Identification of one or more biotherapeutic candidates
2014 end
alone or with Ad-IFNs for transition to DHS S&T Targeted Advanced
date.
Development program for further evaluation and development.
Hindson
University of
California
Lawrence
Livermore
National
Laboratory
DHS S&T
Completed
A high-throughput multiplexed assay was developed for the differential laboratory
detection of foot-and-mouth disease virus (FMDV) from viruses that cause
clinically similar diseases of livestock. This assay simultaneously screens for five
RNA and two DNA viruses by using multiplexed reverse transcription-PCR (mRTPCR) amplification coupled with a microsphere hybridization array and flowcytometric detection. Two of the 17 primer-probe sets included in this multiplex
assay were adopted from previously characterized real-time RT-PCR (rRT-PCR)
assays for FMDV. The diagnostic accuracy of the mRT-PCR assay was evaluated
using 287 field samples, including 247 samples (213 true-positive samples and 35
true-negative samples) from suspected cases of foot-and-mouth disease collected
from 65 countries between 1965 and 2006 and 39 true-negative samples
collected from healthy animals. The mRT-PCR assay results were compared to
those of two singleplex rRT-PCR assays, using virus isolation with antigen enzymelinked immunosorbent assays as the reference method. The diagnostic sensitivity
of the mRT-PCR assay for FMDV was 93.9% (95% confidence interval [CI], 89.8
to 96.4%), and the sensitivity was 98.1% (95% CI, 95.3 to 99.3%) for the two
singleplex rRT-PCR assays used in combination. In addition, the assay could reliably
differentiate between FMDV and other vesicular viruses, such as swine vesicular
disease virus and vesicular exanthema of swine virus. Interestingly, the mRT-PCR
detected parapoxvirus (n = 2) and bovine viral diarrhea virus (n = 2) in clinical
samples, demonstrating the screening potential of this mRT-PCR assay to identify
viruses in FMDV-negative material not previously recognized by using focused
single-target rRT-PCR assays.
Roth (collaboration
with Prionics)
TABI, Inc.
DHS S&T
In progress.
CVB Approval of License for Importation and Distribution of Prionics
2013 end
PrioCHECK® FMD-NS 3ABC ELISA
date
FMD
Diagnostics
2008
Diagnostic Evaluation of Multiplexed
Reverse Transcription-PCR Microsphere
Array Assay for Detection of Foot-andMouth and Look-Alike Disease Viruses
FMD
Diagnostics
2010
Institution
Funding
Agency
Y ear
Status
Comments
Treatment of swine with poly IC alone or in combination with Ad5-pIFNα
confers early protection against FMD (Dias, C. C. A, Moraes, M. P., DiazSan Segundo, F., de los Santos, T., and Grubman, M. J. Porcine type I
interferon rapidly protects swine against challenge with multiple
serotypes of foot-and-mouth disease virus. J. Int. Cyt. Res. 31:227-236,
2011).
FMD
FMD
Area
Diagnostics
Diagnostics
Y ear
2011
2011
Project Title
Evaluation and Validation of 3ABC ELISA
for Detection of FMD Antibody in Infected
Animals Regardless of Vaccination Status
Development of 3ABC ELISA for Detection
of FMD Antibody in Infected Animals
Regardless of Vaccination Status
Primary
Investigator
Sayed (collaborative
with
Clavijo/Bounpheng
and Rieder projects
above and below)
Rieder (collaborative
with
Clavijo/Bounpheng
and Sayed projects
above)
FMD
Diagnostics
2011
Deployment of FMD Serology to the
National Animal Health Laboratory Network
(swine and cattle together)
McIntosh
(collaboration with
NAHLN/TVMDL)
FMD
Diagnostics
2011
Negative Cohort Study for Validation of the Tomlinson/McIntosh/
SVANODIP® FMDV-Ag test
Beckham
Institution
Funding
Agency
Status
Comments
DHS S&T
Project is directly linked to Diagnostic 2011 Bounpheng and Rieder
projects. The 3ABC ELISA kit (PrioCHECK® FMD-NS (3ABC ELISA) in used
in routine diagnosis. This test is costly, is imported from Europe, and
requires an overnight incubation to perform. Recent evaluations of this
commercial test done at FADDL on a conclusive panel of sera showed that
the sensitivity was 0.93 for pigs, 0.9 for cattle and sheep, and specificity
was 0.94 for pigs, 0.88 for cattle and 0.73 for sheep. In order to develop
an improved assay, USDA ARS at PIADC produced the recombinant 3ABC
protein. Test optimization was conducted at PIADC by ARS using the Mab
In progress.
from FAZD (Clavijo). The optimized assay along with standard reagents
2014 end
(3ABC clone and hybridomas), a detailed protocol describing optimization
date.
conditions and preparation of reagents, and evaluation data specifying
the analytical performance of the assays (specificity and sensitivity), were
transferred to FADDL for further analytical evaluation and full validation
for the test’s intended use. >100 positive and 500 negative samples
were evaluated in version 1.0 and showed improved performance over
PrioCHECK® FMD-NS (3ABC ELISA). Field evaluation is being planned for
version 2.0. The ideal test will be a differential ELISA to rapidly
demonstrate freedom from infection with greater performance
characteristics, less cost, and the capacity to be manufactured in the US.
DHS S&T
Completed
Project is directly linked to Diagnostic 2011 Sayed and Bounpheng
projects. Developed a cELISA that uses a FMDV 3ABC recombinant protein
and a monoclonal antibody (FAZD-Clavijo) specific for an immunodominant
B-cell epitope on the 3B protein that will be compatible with either next
generation FMD molecular vaccines (e.g., AdFMD platform) or current, high
quality inactivated vaccines in which NSPs have been removed. After
bench evaluation and assay optimization, the assay and associated
reagents were transitioned to APHIS FADDL and TVMDL for assay
validation (positive and negative samples).
USDA APHIS
PIADC
DHS S&T
This effort aims to deploy FMD DIVA Serology (Prionics PrioCHECK® FMDNS (3ABC ELISA) to a subset of the NAHLN within the first year of
funding with subsequent years to expand the capabilities to all
In progress.
appropriate NAHLN State Veterinary Diagnostic Laboratories. By
2013 end
deploying the 3ABC ELISA to the NAHLN, this effort will augment the
date.
national capacity to not only detect antibodies to FMDV during an
outbreak but to discriminate between vaccinated and unvaccinated
animals (DIVA) during recovery.
USDA/FADDL/
NAHLN/FAZD
DHS S&T
Completed
USDA APHIS
PIADC
USDA ARS
PIADC
The objective is to perform a negative cohort study for the SVANODIP®
FMDV-Ag FMD assay, utilizing samples from the US National herds.
Y ear
Project Title
Primary
Investigator
Institution
Funding
Agency
Status
Comments
FMD
Diagnostics
2011
Investigating potential existence of
chronic, persistent foot-and-mouth disease
virus infection in domestic pigs;
implications for disease control strategies
Arzt
USDA ARS NAA
National Pork
Board
In progress
(a) Determine optimal route of direct inoculation of donor pigs for contact
experiments; compare efficacy of intra-oral (IO) and heel bulb intradermal
(HBI) FMDV inoculation as administration route using FMDV, serotype O.
(b) Characterize FMDV acute pathogenesis parameters (shedding, viremia,
tissue-specific distribution) of infection in contact transmission studies
using FMDV, serotype O.(c) Characterize FMDV post-acute (i.e. suspect
persistent) pathogenesis parameters (shedding, tissue-specific
distribution) of infection in contact transmission studies using FMDV,
serotype O.(d) Characterize FMDV post-acute (i.e. suspect persistent)
pathogenesis parameters (shedding, tissue-specific distribution) of
infection in contact transmission studies using FMDV, serotypes A, and
Asia1.
FMD
Diagnostics
2012
Development of a Prototype and Bench
Validation of a 3BFMDV Competitive ELISA
Kit
Clavijo/ Bounpheng
(collaborative with
Sayed and Rieder
projects below)
TMVDL/Pirbright
DHS
In progress
2013 end
date.
The goal of this effort is to develop a competitive ELISA against the FMDV
3B non-structural protein for use in a diagnostic kit for early detection of
FMDV. Project is directly linked to Diagnostic 2011 Sayed and Rieder
projects.
FMD
FMD
Area
Diagnostics
Diagnostics
2012
Development of 3D ELISA for Detection of
FMDV Antibody in Infected Swine
Immunized with Ad5-FMD(-3D) Vaccine
2012
Pen-side detection of Foot-and-Mouth
Disease virus by a portable microfluidics
PCR system
FMD
Diagnostics
2012
Development and Characterization of
Monoclonal Antibodies to Foot-and-Mouth
Disease Virus Structural Proteins
FMD
Diagnostics
2008
Foot-and-Mouth Disease Multiplexed
Nucleic Acid Assay Enhancements and
Validation
Sayed
USDA APHIS
PIADC
Bounpheng/
McIntosh
USDA/FADDL/
TVMDL
DHS S&T
Develop an optimized and validated ELISA to discriminate Ad5-FMD
vaccinated from FMD infected swine with objectives: (A) Identify the
dominant antigenic region(s) of 3D protein in swine; (B) Develop Mab(s)
In progress.
against the identified antigenic fragment/peptide of 3D; (C) Develop an
2014 end
ELISA for detection of FMDV antibodies in infected swine; (D) Evaluate
date.
the suitability of the 3D ELISA as a DIVA test for Ad5-FMDV
vaccinated/challenged swine. It was discovered that different epitopes
are recognized in swine and in cattle.
DHS S&T
In progress
The overall objective of this effort is to perform a comprehensive
feasibility study of a novel pen-side PCR system (PanNAT™) for sensitive
detection of FMD virus in the field from oral swabs of infected cattle. This
proof of principle study will be performed using samples collected from
experimentally inoculated cattle and pigs.
Bounpheng
TVMDL/ LLNL/
BIOO Sci
DHS S&T
The objective of this project is to develop a set of Mabs to FMDV using
two protein expression systems and to evaluate a bioinformatics approach
to develop FMDV cross reactive Mabs to facilitate research and
development of FMDV vaccine and diagnostics. Cross reactive Mabs will be
developed using in silico ranked linear epitopes from VP1, VP2, and VP3
FMDV sequences. Diverse specificity Mabs will be developed using
recombinant proteins from the structural proteins VP1, VP2 and VP3 of
In progress.
selected serotypes of FMDV; these recombinant proteins will be produced
2013 end
in E. coli and baculovirus expression systems. The specificity of these
date.
Mabs will be evaluated and the epitope identified. These Mabs will have
the following specificities: 1. Linear epitopes of the structural proteins
VP1, VP2 and VP3. 2. Conformational epitopes from baculovirus
expressed structural proteins VP1 of FMDV serotypes O, A and Asia1. 3.
Cross reactive Mabs between all serotypes of FMDV. 4. Serotype (s) FMDV
specific. Mabs to linear and conformational epitopes of FMDV serotype A
and O will be produced.
Beckham
(collaboration with
LLNL)
USDA APHIS
PIADC
DHS S&T
Terminated
Bench validation was completed; no further work was authorized.
FMD
FMD
FMD
FMD
FMD
Area
Diagnostics
Diagnostics
Vacccine
Vaccine
Vaccine
Y ear
2012
Project Title
Establishment of a FMD Antisera Bank
Primary
Investigator
Wu
2012
Molecular Epidemiology and Biosurveillance
of FMD In Endemic Regions of Middle East,
Southeast Asia and Africa
2012
Development of novel foot and mouth
disease virus with multiple mutations for
de los Santos, Rieder
evaluation as live attenuated DIVA vaccine
candidates
2000
2004
Development and Testing of a Subunit
Vaccine for Foot-and-Mouth Disease
Development of an Antiviral and Vaccine
Approach to Control Foot-and-Mouth
Disease
Rodriguez
Grubman
Grubman
Institution
USDA APHIS
USDA ARS
PIADC
APHIS USDA
PIADC
USDA ARS NAA
USDA ARS NAA
Funding
Agency
DHS S&T
DHS S&T
Status
In progress
Comments
A steady supply of antisera is needed for at least two of DSS’s routine
diagnostic assays, ELISA’s, and virus neutralization tests. In addition to
the diagnostic applications, the availability of a wide range of well
characterized antisera has been identified by Reagent and Vaccine
Services Section (RVSS) as a gap in the development of new diagnostic
assays and the improvement of existing ones. This was recently
experienced by the RVSS in the development of the FMD liquid phase
blocking ELISA (LPBE), and it is expected to be even more critical during
the validation phase. The validation of serologic assays, a mission
critical element for Proficiency Validation Service Section (PVSS), requires
significant amounts and varieties of FMD antisera.
Goals: 1. Gain a better understanding of the epidemiology of FMDV
strains circulating in selected endemic regions of Central and Southeast
Asia and Africa. 2. Determine the phylogenetic relationships of FMDV
In progress.
strains in endemic regions. 3. Determine the antigenic relationship
2015 end date.
(vaccine matching) of FMDV strains circulating in selected regions with
currently available vaccine strains.
National Pork
Board
The objective of this project is to improve current strategies to generate
FMDV containing specific attenuating factors and antigenic markers and
In Progress, 1 evaluate their potential as vaccine candidates. This proposal builds upon
yr timeline
previous and ongoing research where mutations are being introduced into
the leader protein of FMDV in order to attenuate the virus and derive virus
strains that could be used as vaccine candidates
National Pork
Board
Completed
We have tested the potency and efficacy of replication-defective
recombinant human adenoviruses containing the capsid and 3C proteinase
coding regions of foot-and-mouth disease virus (FMDV) as vaccine
candidates in swine. This vaccine, which lacks the coding region of several
FMDV nonstructural proteins, has a number of advantages over the current
whole virus inactivated vaccine including the ability to readily distinguish
vaccinated from infected animals using current technology. Inoculation of
swine with this vaccine resulted in either complete protection from
disease after virulent homologous virus challenge or significant reduction
in clinical signs as compared to co-housed un-inoculated control animals.
The optimal vaccine regimen tested was a low dose initial inoculation
followed by a high dose boost. All animals inoculated by this regimen
were completely free of disease after challenge. Increased efficacy of the
two-dose regimen was associated with heightened levels of FMDV specific
IgG1 and IgG2 antibodies. Expression and processing of the FMDV capsid
precursor protein, requiring a biologically active 3C proteinase, was
essential for induction of FMDV-specific neutralizing antibodies and
protection from challenge. This vaccine is safe since there is no evidence
of sero-conversion of co-housed unvaccinated control animals. These
experimental results suggest that the Ad5-FMDV virus vector is a
promising vaccine candidate against FMD and additional efforts to improve
its potency and efficacy are underway.
Completed
In this study we have demonstrated that delivery of the combination of
our FMD subunit vaccine and interferon alpha can enhance the long-term
protection afforded swine with the vaccine alone. Furthermore, we are
developing additional antiviral reagents, i.e.., interferon beta, that may
allow us to supplement our current approach as well as develop a more
comprehensive understanding of the interactions between FMDV and its
host.
National Pork
Board
Y ear
Project Title
Primary
Investigator
Institution
Funding
Agency
Status
Comments
Brough and Butman
GenVec
DHS S&T
Research milestones are associated with the construction, in vitro and in
vivo (cattle focus). Several AdFMD vaccine candidates have been made
and tested in cattle. Development milestones are associated with
In progress.
manufacturing process improvements and stability and potency
2013 end date.
assessment of AdFMD vaccine candidates producted under improved
manufacturing processes. Development milestones are aligned with DHS
S&T Vx and Dx CRADA with Merial.
Massare
Novavax
DHS S&T
Goals is construction and testing (cattle focus) of virus-like particle
In progress.
recombinant, subunit FMDV vaccine produced in insect cells. POC cattle
2014 end date.
study planned for Spring 2013.
Kern
Inovio
DHS S&T
(CRADA/No
Direct
Funding)
Goal is construction and testing (swine focus) of FMD DNA vaccines based
In progress.
on Inovio's SynCon technology and Cellectra delivery (electroporation)
End date TBD.
device. POC swine study planned for Spring 2013.
Merial
DHS S&T
(CRADA/No
Direct
Funding)
Goal is testing and evaluation of (3) AdFMD vaccine candidates in swine,
production of master seed vacine viruses, and improvement of vaccine
In progress.
manufacturing process. One POC efficacy study in swine has been
End date TBD.
completed and target efficacy level was not achieved. Efforts are ongoing
to improve vaccine potency.
FMD
Vaccine
2010
Research and Development of Molecular
FMD Vaccines (historically cattle focused)
FMD
Vaccine
2011
Development of VLP Vaccine as a
Countermeasure for FMD
FMD
Vaccine
2011
FMD DNA Vaccine
2011
Testing and Evaluation of Human
Adenovirus Replication Deficient Vectored
FMD Vaccines (swine focus)
Miller
Benchmark
Biolabs
DHS S&T
In progress.
2014 end date
(if all option
years awarded)
Mittal
Purdue
University
DHS S&T
Goal is construction and testing (cattle focus) of replication competent
In progress.
and replication deficient bovine adenovirus vectored FMD vaccine
2014 end date.
candidates.
FMD
Vaccine
FMD
Vaccine
2012
Acquisition of Master Seed Virus (MSV) and
Working Seed Virus (WSV) Lots for FMD
Molecular Vaccine Countermeasures
FMD
Vaccine
2012
Bovine Adenovirus Vector-Based Vaccine
for FMD
FMD
Vaccine
2012
Construction and Evaluation of
Recombinant MVA-BN FMDV Candidates
FMD
Vaccine
2012
Replicon-based Rapid Response Vaccines
for FMD
2010
Production of FMD Virus Pseudovirion
Vaccine Candidates Using a Plant Transient
System
2011
and Improved Challenge Systems Improved Challenge Systems for FMD
Vaccine and Biotherapeutics Testing in
Cattle and Pigs
FMD
FMD
Area
Vaccine
Vaccine
David
Goal is construction of MVA-BN-FMDV vaccine candidate for POC testing
(cattle focus). Major tasks include i) evaluation and identification of
optimal vector parameters, ii) construction and evaluaton of two
recombinatn MVA-BN-FMDV constructs expressing 3C in trans, and iii)
production and formulation of optimal MVA-BN FMDV candidate (research
grade).
Weinberger
Bavarian Nordic
DHS S&T
In progress. optimal vector parameters, ii) construction and evaluaton of two
2014 end date. recombinatn MVA-BN-FMDV constructs expressing 3C in trans, and iii)
grade).
Kamrud
Harrisvaccines
DHS S&T
In progress. Goals is rapid, large-scale (50-100K doses) of VEE replicon particle FMDV
2014 end date. vaccine candidate (cattle focus).
Hiatt
Kentucky
BioProcessing
DHS S&T
Vx/Dx
Arzt
USDA ARS PIADC
DHS S&T
In progress.
The key goal of the project is to produce FMDV pseudovirion structures in
plants and develop scaleable manners for purification and vaccine
production. To date, efforts to produce pseudovirion structures in tobacco
plants have failed. Modified SOW is in progress to use an alernative
strategy to improve capsid production and solubility.
The primary goal of this proposed research is the optimization of new,
simulated natural systems for challenging cattle and pigs with foot-andmouth disease virus (FMDV). These novel systems will be optimized in
In progress.
studies using naïve animals and subsequently validated in vaccinate2014 end date.
challenge experiments. Additionally, the novel systems will be used to
assess pigs’ and steers’ ability to transmit FMDV in the pre-clinical phase
of disease.
FMD
FMD
Area
Vaccine
Vaccine
Y ear
Project Title
Primary
Investigator
Institution
Funding
Agency
Status
Comments
2011
Rational design of attenuated foot-andmouth disease virus strains for
development of improved disease
countermeasures
de los Santos
USDA ARS NAA
National Pork
Board
Completed
Our goal is to develop alternative control strategies that could improve current
FMD countermeasure programs. An attenuated vaccine is expected to elicit more
rapid innate immunity and a long lived adaptive immunity to effectively control
disease. Moreover, induction of innate immunity could result in early protection
against multiple FMDV serotypes. Importantly, attenuated strains are excellent
new tools to study the interactions between FMDV and the host immune system
and ultimately could lead to the development of novel strategies to counteract
FMD. During the past year and with the support of NPB we have successfully
derived a mutant strain of FMDV that did not cause disease in swine (FMDV-SAP
mutant). Interestingly inoculation of swine with this mutant strain induced a strong
immune response that protected animals against infection with the parental (wild
type) virus, as early as two days post vaccination. Studies in animals and in
cultured swine cells demonstrated that, in contrast to the parental wild type virus,
the mutant variant was unable to block some inflammatory responses thus limiting
dissemination of the virus beyond the original site of inoculation. Furthermore we
have started studies to add more mutations to this virus aiming to increase the
stability of the original mutations therefore decreasing the probability of reversion
to virulence. Our results indicated that manipulation of the viral genome in the
region that encodes for the leader protein is a viable alternative to derive less
pathogenic FMDV strains that could be used as the basis for live attenuated
vaccines against FMD or as seeds to grow the virus for manufacturing safer
inactivated vaccines.
2012
Exploiting the potential of leader
proteinase coding sequence of foot-andmouth disease virus to derive attenuated
strains suitable for pathogenesis studies
and development of improved
countermeasures.
de los Santos
USDA ARS NAA
National Pork
Board
In progress
Goal is to develop alternative control strategies that could improve
current FMD countermeasure programs. An attenuated vaccine is expected
to elicit more rapid innate immunity and a long lived adaptive immunity to
effectively control disease.
Multiple
Agents
Multiple
Agents
Multiple
Agents
Multiple
Agents
Multiple
Agents
Area
Diagnostics
Diagnostics
Diagnostics
Diagnostics
Diagnostics
Y ear
2008
2011
2012
Project Title
Enhancements of High-Throughput
Diagnosis for FAD
Development of a multiplex RT-qPCR
assay for the simultaneous detection of
endemic and foreign animal diseases in
oral fluid samples.
Assessment of Two Methods for Nucleic
Acid Stability and Viral Inactivation of
FAD Viruses Collected from Oral Fluid
Rope Samples
2009
Vesicular Disease Diagnostic Reagent
Production
2011
The Matrix-Chaperone: Ambient
Temperature Biospecimen Collection,
Transport & Banking For Simplified
Animal Disease Screening (swine and
cattle).
Primary
Institution
Investigator
McIntosh
Beckham/
McIntosh/
Bounpheng
Mayr
Jia
Hogan
USDA
APHIS
PIADC
TMVDL/FAD
DL
USDA
APHIS
USDA
APHIS
PIADC
IntegenX
Funding
Agency
Status
Comments
DHS S&T
Vx/Dx
Construction and select agent certification of the highthroughput lab that will allow diagnostic work using
Completed
PCR and serology was completed. Validation of the lab
is underway.
DHS S&T
The objective of this effort is to develop an optimized
oral fluid nucleic acid purification method and a
multiplex reverse transcriptase quantitative PCR assay
(mpRT-qPCR) that can be used to simultaneously
detect several economically important swine endemic
In progress and foreign animal diseases from the same sample.
APHIS will conduct bench validation of a multiplex RTqPCR assay for the simultaneous detection of endemic
and foreign animal diseases in oral fluid samples. The
multiplex is currently under development by FAZD under
a separate agreement
DHS S&T
Two (or more) methods for the preservation of nucleic
acid while neutralizing virus infectivity will be
examined. Whitman FTA paper and PrimeStore MTM
media will be examined to determine if they are
capable of stabilizing viral nucleic acids, RNA and DNA,
In progress from rope oral fluid samples while at the same time
inactivating/neutralizing any viable virus present in the
oral fluid so as to make transport of the samples to
laboratories outside of the outbreak zone risk free.
The recent approval to fill this position will result in the
job posting within the month.
DHS S&T
Vx/Dx
DHS S&T
USDA APHIS FADDL established an in-house reagent
production capacity to produce large quantities of
highly specific rabbit and guinea pig antisera against
homologous FMDV, SVDV, and VSV to be used in the
In Progress. vesicular disease Ag ELISA, as well as to fulfill the
2013 end diagnostic surge capacity required in the event of a
vesicular disease outbreak. A result of this effort is
date.
that this lab was recognized as a FMD Reference lab by
FAO and the World Organization for Animal Health,
resulting in significantly less reliance on foreign labs.
This project will develop and validate a new technology
for the collection, transport and banking of
In progress biospecimens for animal disease screening that is
based on ambient-temperature, dry-state sample
preservation
Area
Y ear
Multiple
Agents
Diagnostics
2011 &
2008
Multiple
Agents
Post-Harvest
2012
Project Title
Development of Pan-viral DNA
Microarrays for the Detection of
Emerging and Foreign Animal Diseases
Disinfection of foreign animal disease
viruses on surfaces relevant to the Pork
Packing Industry
Primary
Institution
Investigator
McIntosh
Rodriguez and
Krug
USDA
APHIS
PIADC
USDA
APHIS
PIADC
Funding
Agency
Status
Comments
DHS S&T
Vx/Dx
An enhanced version of the microarray platform is being
Completed validated that will detect more viruses isolated from a
variety of organisms.
National Pork
Board
The main objective of this proposal is to determine the
efficacy of chemical disinfectants against foreign
animal disease (FAD) viruses dried on surfaces relevant
to the Pork Packing industry. A second objective of this
Completed project is to ensure that necessary surface-specific
changes are made to existing protocols to ensure valid
results are generated. A third objective of this proposal
is to validate the use of non-FAD viruses as surrogates
for FAD viruses in disinfection assays.
ASF
ASF
ASF
Diagnostics
Diagnostics
Diagnostics
Y ear
Project Title
2011
Comparison of different sample source
and sample pooling for the detection
and surveillance of ASF
2011
Development of fluorescent
recombinant antibodies to detect
African swine fever virus in tissue
samples and infected cells
2012
A Comprehensive Research Program in
African Swine Fever Towards the
Development of Novel
Countermeasures
Primary
Investigator
Institution
Funding Agency
Escribano
Instituto Nacl.
Investigacion y
Tecnologia
Agraria y
Alimentaria (INIA)
National Pork
Board
In progress
The present project pretends to develop new reagents to cover a
gap in the ASFV diagnosis.
DHS S&T Vx/Dx
In
progress.
2014 end
date.
Objective 1: Development of challenge model for ASF to
characterize pathogenesis and evaluate novel countermeasure
products. Objective 2: Determining immune mechanisms of
protection induced by attenuated strains. Objective 3: Functional
genomics.
In
progress.
2014 end
date.
The end product of this project is a battery of monoclonal
antibodies against ASF proteins p30, p72, p54, and 8-DR. These
will be produced and made available to ARS and DHS for their
research needs. They can also be potentially used in the
development or improvement of diagnostic assays in FADDL. It is
expected that the knowledge acquired from ARS and DHS
characterization will assist in this process.
Borca/Arzt
USDA ARS PIADC
Development of Monoclonal Antibodies
Specific to ASFV Proteins
Wu/Sayed
USDA APHIS
PIADC
DHS S&T Vx/Dx
ASF
Diagnostics
2012
Identification of genetic signatures for
African swine fever virus serologic
group specificity
Rock
University of
Illinois
National Pork
Board
Development of Multi-component
Vaccines for African Swine Fever
This project will compare and validate pooled sample matricies,
including oral fluids, nasal swabs, blood, and serum, for the
In progress molecular detection of ASFV. Project will include a methods
comparison between USDA NAHLN PCR, Pirbright PCR, and
serological assay for detection.
DHS
2012
2011
Comments
Pirbright
Diagnostics
Vaccine
Status
Dixon
ASF
ASF
Area
Mwangi
Texas A&M
Research
Foundation
DHS S&T
Identify genetic signature (s) for ASFV serologic group specificity
In progress and determine serologic group specificity for currently untyped ASFV
isolates in VNIIVViM strain collection
In
progress.
2013 end
date.
The objective of this proposal is to clone and express 5 candidate
ASFv genes in eukaryotic (HEK) and adenovirus expression
platforms. The eukaryotic platform will be used to produce
recombinant proteins to serve as immunogens for MAb and rabbit
PcAb reagent production (2 ASFV targets). The adenovirus platform
will be used to produce (3) vaccine candidates for swine proof-ofconcept safety and immunogencity study at TAMU.
ASF
ASF
Area
Vaccine
Vaccine
Y ear
2011
2011
Project Title
Identification of African Swine Fever
Candidates by Reverse Vaccinology
Development of a Proof of Concept
Rationally Designed Live Attenuated
African Swine Fever Virus Vaccines
Primary
Investigator
Bounpheng
Borca
Institution
TMVDL
USDA ARS PIADC
Funding Agency
DHS S&T Vx/Dx
DHS S&T Vx/Dx
Status
Comments
In
progress.
2013 end
date.
The objective of this proposal is to evaluate a novel approach,
reverse vaccionology (RV), for the identification and development of
ASFV vaccine candidates. RV will be used to identify novel in silico
candidates and rank the DHS BAA candidates (i.e., p30, p54, p72,
CD2v). Two protein expression and delivery platforms will be
evaluated. The in silico and top ranked DHS candidates will be
expressed in mammalian HEK 293 cells and fused to an
immunogenic inducing tag. In addition these candidates will be
expressed in the highly safe poxvirus modified vaccinia Ankara
(MVA) vaccine vector to enhance cellular immunity. Recombinant
vaccines will be tested for immunogenicity and safety in pigs. The
PIs anticipate that these candidates (alone or synergistically
combined) will be highly immunogenic and induce strong humoral
and cellular responses. Protein vaccines are highly safe and can
induce strong humoral responses. In addition, MVA-based vaccines
are an attractive vaccine delivery technology because when
administered to the host they can induce in vivo expression of one
or more specific antigens. These newly expressed antigens are
processed and presented by professional antigen-presenting cells,
resulting in the induction of antibody responses with high avidity,
as well as major histocompatibility complex class I-restricted
cytotoxic T-lymphocyte (CTL) responses. This pattern of responses
is similar to that induced by live attenuated vaccines. In addition,
the induction of B and T cell immune responses eliminates the need
for adjuvants. These features are major advantages of viral vectors
when compared to other vaccine delivering platforms.
In
progress.
2014 end
date.
Objective: 1. Develop recombinant African Swine Fever Virus (ASFV)
strains by deletion of one or more viral genes already described as
responsible for inducting attenuation of highly virulent ASF strains.
ASFV isolates. 3. Evaluate patterns of heterologous protection
among genetically heterogeneous ASFV strains. Approach: Develop
recombinant African Swine Fever Virus (ASFV) strains by deletion of
one or more viral genes already described as responsible for
inducting attenuation of highly virulent ASF strains. Critical ASFV
genes that are responsible for inducing attenuation of highly
virulent ASF strains will be deleted individually or as a group. This
deletion should confer virus replication but not disease production.
ASFV isolates. This will be done through: Testing strains for in vivo
attenuation, testing for efficacy against homologous challenge,
determination of minim protective dose response, evaluation of
protection profile for at least one attenuated ASFV vaccine
candidate, and the evaluation of lead vaccine candidate to induce
sterile immunity. 3. Evaluate and confirm cross-protection conferred
by lead vaccine candidate (obj. 2) by using genetically diverse ASFV
strains.
Page 23 of 32 Dis eas e
CSF
CSF
CSF
CSF
CSF
Area
Diagnostics
Diagnostics
Vaccine
Vaccine
Vaccine
Y ear
2009
2011
2009
2009
2010
Project Tit le
Diagnostic Technologies for Classical
Swine Fever
Primary
Inves t igat or
Mayr/Batonick
Ins t it ut ion
USDA APHIS
PIADC
Development of classical swine fever
Thanawongnuwe Chulalongkorn
virus diagnostic assays for porcine oral
ch
University
fluid samples
Identification of host factors
interacting with classical swine fever
virus proteins: development of novel
anti-viral therapeutics.
Classical Swine Fever (CSF) Vaccine
and Diagnostic Countermeasure
Development
Borca
Roth /Riemser
Borca
USDA ARS NAA
TABI, Inc.
ARS USDA
PIADC
Funding
Agency
DHS S&T
Vx/Dx
National Pork
Board
National Pork
Board
St at us
Comment s
In progress.
2013 end
date.
The current CSF antibody ELISA has limited specificity in excluding
antibody to ruminant pestiviruses. Identification and
characterization of the E2 glycoprotein epitopes that discriminate
between swine and ruminant pestivirus are essential in the design
of a next generation ELISA. For epitope mapping, a panel of
monoclonal antibodies was obtained in collaboration with the
Central Institute for Animal Diseases Control, Lelystad, the
Netherlands. The CSF target epitopes were defined, and the ELISA
was designed and is being optimized and validated. Such an ELISA
will have vital application in the current national surveillance
program and during recovery of an outbreak.
In progress
The primary objective of this project is to optimize and validate
technology capable of rapidly identifying premises infected with
classical swine fever virus (CSFV) following its introduction into
North America or other CSFV-free areas using oral fluid samples.
Achievement of this objective will also provide technology for
improved surveillance in CSFV endemic areas, thereby enhancing
elimination and control efforts. The specific objectives are focused
on demonstrating the feasibility of detecting CSFV in oral fluids by
modified PCR-based methods.
Completed
Results obtained enable the identification of several host proteins
interacting with CSFV structural protein Core. Core protein is the
major contributor to the virus capsid. Several of these interactions
have been studied in detail and the regions of the CSFV Core
protein interacting with the host proteins were identified. Mutant
CSFV viruses having altered these regions have been demonstrated
that have severely altered their ability to produce disease in swine.
Therefore, the manipulation of the identified host-virus interactions
allowed the development of attenuated strains of virus which may
constitute a tool for the further development of live attenuated
vaccine against classical swine fever. Additionally, this knowledge
may open the possibility of designing bio therapeutic compounds
that could alter those critical interactions that may limit the spread
of the disease.
DHS S&T
Vx/Dx
CVB Approval of Permittee License for Importation and Distribution
of CSF Modified Live Vaccine, Chinese strain (C strain). Riemser
Arzneitmittel AG (GDR) is currently producing a EU licensed vaccine.
In progress.
The vaccine (C strain) was recently excluded from the USDA APHIS
2013 end date
Select Agent List. Plans are in progress to import master seed virus
(modification
(MSV) and master seed cell bank (MSB) for safety testing in swine
to 2014 end
by FADDL, PIADC. If satisfactory, MSV and MSB will sent to USDA
date planned)
CVB for additional testing. Submission of various testing reports to
USDA CVB are in progress. GDR manufacturing facility has been
inspected by USDA CVB I&C.
DHS S&T
Vx/Dx
Objective 1: Develop a genetically stable version of FlagT4 virus
(FlagT4c). Objective 2: Viral-vectored vaccine (backup candidate):
In progress. POC safety and efficacy studies using baculovirus-vectored
2013 end date experimental vaccine. Objective 3: Obtain select agent exclusion
for FlagT4c vaccine candidate and transition to AH partner (CRADA
in place).
CSF
CSF
Area
Vaccine
Vaccine
Y ear
Project Title
2011
Evaluation of Envelope Proteins for
Rapid Induction of Protective Immune
Responses Against Classical Swine
Fever
2012
Plant-based expression vectors for
rapid, high-throughput development of
animal vaccines
Primary
Investigator
Borca
Holtz
Institution
USDA ARS NAA
Funding
Agency
National Pork
Board
Caliber
DHS S&T OUP
Biotherapeutics
Status
Comments
In progess
The main objective of the research project is to evaluate native and
modified forms of CSF envelope proteins for their capacity to induce
rapid protective immune response against the CSFV. CSFV envelope
proteins are expressed as fusion proteins along with different
immunological adjuvants. The improved efficacy of the novel
constructs will evaluated compared with the native version of the
proteins in terms of induced immune response and protection.
In progress
The main objective of this research project is to demonstrate proof
of principle for rapid expression, testing, and commercial-scale
production capacity using a novel plant-based approach. The CSF E2
protein is targeted for a potential recombinant subunit vaccine for
use in later stages of a disease outbreak as a possible compliment
to a live attenuated vaccination strategy.
FMD
FMD
Area
Biotherapeutics
Biotherapeutics
Primary
Investigator
Project Title
2009
Foreign Animal Disease (FAD)
Countermeasure Development Identification of Biotherapeutic Candidates
to Control FMDV
Grubman
USDA ARS
PIADC
DHS S&T
Completed
2011
and Improved Challenge Systems Identification of Biotherapeutic Candidates
to Control FMDV
de los Santos/
Grubman
USDA ARS
PIADC
DHS S&T
In progress. Overarching Goal: Identification of one or more biotherapeutic candidates
2014 end
alone or with Ad-IFNs for transition to DHS S&T Targeted Advanced
date.
Development program for further evaluation and development.
Hindson
University of
California
Lawrence
Livermore
National
Laboratory
DHS S&T
Completed
A high-throughput multiplexed assay was developed for the differential laboratory
detection of foot-and-mouth disease virus (FMDV) from viruses that cause
clinically similar diseases of livestock. This assay simultaneously screens for five
RNA and two DNA viruses by using multiplexed reverse transcription-PCR (mRTPCR) amplification coupled with a microsphere hybridization array and flowcytometric detection. Two of the 17 primer-probe sets included in this multiplex
assay were adopted from previously characterized real-time RT-PCR (rRT-PCR)
assays for FMDV. The diagnostic accuracy of the mRT-PCR assay was evaluated
using 287 field samples, including 247 samples (213 true-positive samples and 35
true-negative samples) from suspected cases of foot-and-mouth disease collected
from 65 countries between 1965 and 2006 and 39 true-negative samples
collected from healthy animals. The mRT-PCR assay results were compared to
those of two singleplex rRT-PCR assays, using virus isolation with antigen enzymelinked immunosorbent assays as the reference method. The diagnostic sensitivity
of the mRT-PCR assay for FMDV was 93.9% (95% confidence interval [CI], 89.8
to 96.4%), and the sensitivity was 98.1% (95% CI, 95.3 to 99.3%) for the two
singleplex rRT-PCR assays used in combination. In addition, the assay could reliably
differentiate between FMDV and other vesicular viruses, such as swine vesicular
disease virus and vesicular exanthema of swine virus. Interestingly, the mRT-PCR
detected parapoxvirus (n = 2) and bovine viral diarrhea virus (n = 2) in clinical
samples, demonstrating the screening potential of this mRT-PCR assay to identify
viruses in FMDV-negative material not previously recognized by using focused
single-target rRT-PCR assays.
Roth (collaboration
with Prionics)
TABI, Inc.
DHS S&T
In progress.
CVB Approval of License for Importation and Distribution of Prionics
2013 end
PrioCHECK® FMD-NS 3ABC ELISA
date
FMD
Diagnostics
2008
Diagnostic Evaluation of Multiplexed
Reverse Transcription-PCR Microsphere
Array Assay for Detection of Foot-andMouth and Look-Alike Disease Viruses
FMD
Diagnostics
2010
Institution
Funding
Agency
Y ear
Status
Comments
Treatment of swine with poly IC alone or in combination with Ad5-pIFNα
confers early protection against FMD (Dias, C. C. A, Moraes, M. P., DiazSan Segundo, F., de los Santos, T., and Grubman, M. J. Porcine type I
interferon rapidly protects swine against challenge with multiple
serotypes of foot-and-mouth disease virus. J. Int. Cyt. Res. 31:227-236,
2011).
FMD
FMD
Area
Diagnostics
Diagnostics
Y ear
2011
2011
Project Title
Evaluation and Validation of 3ABC ELISA
for Detection of FMD Antibody in Infected
Animals Regardless of Vaccination Status
Development of 3ABC ELISA for Detection
of FMD Antibody in Infected Animals
Regardless of Vaccination Status
Primary
Investigator
Sayed (collaborative
with
Clavijo/Bounpheng
and Rieder projects
above and below)
Rieder (collaborative
with
Clavijo/Bounpheng
and Sayed projects
above)
FMD
Diagnostics
2011
Deployment of FMD Serology to the
National Animal Health Laboratory Network
(swine and cattle together)
McIntosh
(collaboration with
NAHLN/TVMDL)
FMD
Diagnostics
2011
Negative Cohort Study for Validation of the Tomlinson/McIntosh/
SVANODIP® FMDV-Ag test
Beckham
Institution
Funding
Agency
Status
Comments
DHS S&T
Project is directly linked to Diagnostic 2011 Bounpheng and Rieder
projects. The 3ABC ELISA kit (PrioCHECK® FMD-NS (3ABC ELISA) in used
in routine diagnosis. This test is costly, is imported from Europe, and
requires an overnight incubation to perform. Recent evaluations of this
commercial test done at FADDL on a conclusive panel of sera showed that
the sensitivity was 0.93 for pigs, 0.9 for cattle and sheep, and specificity
was 0.94 for pigs, 0.88 for cattle and 0.73 for sheep. In order to develop
an improved assay, USDA ARS at PIADC produced the recombinant 3ABC
protein. Test optimization was conducted at PIADC by ARS using the Mab
In progress.
from FAZD (Clavijo). The optimized assay along with standard reagents
2014 end
(3ABC clone and hybridomas), a detailed protocol describing optimization
date.
conditions and preparation of reagents, and evaluation data specifying
the analytical performance of the assays (specificity and sensitivity), were
transferred to FADDL for further analytical evaluation and full validation
for the test’s intended use. >100 positive and 500 negative samples
were evaluated in version 1.0 and showed improved performance over
PrioCHECK® FMD-NS (3ABC ELISA). Field evaluation is being planned for
version 2.0. The ideal test will be a differential ELISA to rapidly
demonstrate freedom from infection with greater performance
characteristics, less cost, and the capacity to be manufactured in the US.
DHS S&T
Completed
Project is directly linked to Diagnostic 2011 Sayed and Bounpheng
projects. Developed a cELISA that uses a FMDV 3ABC recombinant protein
and a monoclonal antibody (FAZD-Clavijo) specific for an immunodominant
B-cell epitope on the 3B protein that will be compatible with either next
generation FMD molecular vaccines (e.g., AdFMD platform) or current, high
quality inactivated vaccines in which NSPs have been removed. After
bench evaluation and assay optimization, the assay and associated
reagents were transitioned to APHIS FADDL and TVMDL for assay
validation (positive and negative samples).
USDA APHIS
PIADC
DHS S&T
This effort aims to deploy FMD DIVA Serology (Prionics PrioCHECK® FMDNS (3ABC ELISA) to a subset of the NAHLN within the first year of
funding with subsequent years to expand the capabilities to all
In progress.
appropriate NAHLN State Veterinary Diagnostic Laboratories. By
2013 end
deploying the 3ABC ELISA to the NAHLN, this effort will augment the
date.
national capacity to not only detect antibodies to FMDV during an
outbreak but to discriminate between vaccinated and unvaccinated
animals (DIVA) during recovery.
USDA/FADDL/
NAHLN/FAZD
DHS S&T
Completed
USDA APHIS
PIADC
USDA ARS
PIADC
The objective is to perform a negative cohort study for the SVANODIP®
FMDV-Ag FMD assay, utilizing samples from the US National herds.
Y ear
Project Title
Primary
Investigator
Institution
Funding
Agency
Status
Comments
FMD
Diagnostics
2011
Investigating potential existence of
chronic, persistent foot-and-mouth disease
virus infection in domestic pigs;
implications for disease control strategies
Arzt
USDA ARS NAA
National Pork
Board
In progress
(a) Determine optimal route of direct inoculation of donor pigs for contact
experiments; compare efficacy of intra-oral (IO) and heel bulb intradermal
(HBI) FMDV inoculation as administration route using FMDV, serotype O.
(b) Characterize FMDV acute pathogenesis parameters (shedding, viremia,
tissue-specific distribution) of infection in contact transmission studies
using FMDV, serotype O.(c) Characterize FMDV post-acute (i.e. suspect
persistent) pathogenesis parameters (shedding, tissue-specific
distribution) of infection in contact transmission studies using FMDV,
serotype O.(d) Characterize FMDV post-acute (i.e. suspect persistent)
pathogenesis parameters (shedding, tissue-specific distribution) of
infection in contact transmission studies using FMDV, serotypes A, and
Asia1.
FMD
Diagnostics
2012
Development of a Prototype and Bench
Validation of a 3BFMDV Competitive ELISA
Kit
Clavijo/ Bounpheng
(collaborative with
Sayed and Rieder
projects below)
TMVDL/Pirbright
DHS
In progress
2013 end
date.
The goal of this effort is to develop a competitive ELISA against the FMDV
3B non-structural protein for use in a diagnostic kit for early detection of
FMDV. Project is directly linked to Diagnostic 2011 Sayed and Rieder
projects.
FMD
FMD
Area
Diagnostics
Diagnostics
2012
Development of 3D ELISA for Detection of
FMDV Antibody in Infected Swine
Immunized with Ad5-FMD(-3D) Vaccine
2012
Pen-side detection of Foot-and-Mouth
Disease virus by a portable microfluidics
PCR system
FMD
Diagnostics
2012
Development and Characterization of
Monoclonal Antibodies to Foot-and-Mouth
Disease Virus Structural Proteins
FMD
Diagnostics
2008
Foot-and-Mouth Disease Multiplexed
Nucleic Acid Assay Enhancements and
Validation
Sayed
USDA APHIS
PIADC
Bounpheng/
McIntosh
USDA/FADDL/
TVMDL
DHS S&T
Develop an optimized and validated ELISA to discriminate Ad5-FMD
vaccinated from FMD infected swine with objectives: (A) Identify the
dominant antigenic region(s) of 3D protein in swine; (B) Develop Mab(s)
In progress.
against the identified antigenic fragment/peptide of 3D; (C) Develop an
2014 end
ELISA for detection of FMDV antibodies in infected swine; (D) Evaluate
date.
the suitability of the 3D ELISA as a DIVA test for Ad5-FMDV
vaccinated/challenged swine. It was discovered that different epitopes
are recognized in swine and in cattle.
DHS S&T
In progress
The overall objective of this effort is to perform a comprehensive
feasibility study of a novel pen-side PCR system (PanNAT™) for sensitive
detection of FMD virus in the field from oral swabs of infected cattle. This
proof of principle study will be performed using samples collected from
experimentally inoculated cattle and pigs.
Bounpheng
TVMDL/ LLNL/
BIOO Sci
DHS S&T
The objective of this project is to develop a set of Mabs to FMDV using
two protein expression systems and to evaluate a bioinformatics approach
to develop FMDV cross reactive Mabs to facilitate research and
development of FMDV vaccine and diagnostics. Cross reactive Mabs will be
developed using in silico ranked linear epitopes from VP1, VP2, and VP3
FMDV sequences. Diverse specificity Mabs will be developed using
recombinant proteins from the structural proteins VP1, VP2 and VP3 of
In progress.
selected serotypes of FMDV; these recombinant proteins will be produced
2013 end
in E. coli and baculovirus expression systems. The specificity of these
date.
Mabs will be evaluated and the epitope identified. These Mabs will have
the following specificities: 1. Linear epitopes of the structural proteins
VP1, VP2 and VP3. 2. Conformational epitopes from baculovirus
expressed structural proteins VP1 of FMDV serotypes O, A and Asia1. 3.
Cross reactive Mabs between all serotypes of FMDV. 4. Serotype (s) FMDV
specific. Mabs to linear and conformational epitopes of FMDV serotype A
and O will be produced.
Beckham
(collaboration with
LLNL)
USDA APHIS
PIADC
DHS S&T
Terminated
Bench validation was completed; no further work was authorized.
FMD
FMD
FMD
FMD
FMD
Area
Diagnostics
Diagnostics
Vacccine
Vaccine
Vaccine
Y ear
2012
Project Title
Establishment of a FMD Antisera Bank
Primary
Investigator
Wu
2012
Molecular Epidemiology and Biosurveillance
of FMD In Endemic Regions of Middle East,
Southeast Asia and Africa
2012
Development of novel foot and mouth
disease virus with multiple mutations for
de los Santos, Rieder
evaluation as live attenuated DIVA vaccine
candidates
2000
2004
Development and Testing of a Subunit
Vaccine for Foot-and-Mouth Disease
Development of an Antiviral and Vaccine
Approach to Control Foot-and-Mouth
Disease
Rodriguez
Grubman
Grubman
Institution
USDA APHIS
USDA ARS
PIADC
APHIS USDA
PIADC
USDA ARS NAA
USDA ARS NAA
Funding
Agency
DHS S&T
DHS S&T
Status
In progress
Comments
A steady supply of antisera is needed for at least two of DSS’s routine
diagnostic assays, ELISA’s, and virus neutralization tests. In addition to
the diagnostic applications, the availability of a wide range of well
characterized antisera has been identified by Reagent and Vaccine
Services Section (RVSS) as a gap in the development of new diagnostic
assays and the improvement of existing ones. This was recently
experienced by the RVSS in the development of the FMD liquid phase
blocking ELISA (LPBE), and it is expected to be even more critical during
the validation phase. The validation of serologic assays, a mission
critical element for Proficiency Validation Service Section (PVSS), requires
significant amounts and varieties of FMD antisera.
Goals: 1. Gain a better understanding of the epidemiology of FMDV
strains circulating in selected endemic regions of Central and Southeast
Asia and Africa. 2. Determine the phylogenetic relationships of FMDV
In progress.
strains in endemic regions. 3. Determine the antigenic relationship
2015 end date.
(vaccine matching) of FMDV strains circulating in selected regions with
currently available vaccine strains.
National Pork
Board
The objective of this project is to improve current strategies to generate
FMDV containing specific attenuating factors and antigenic markers and
In Progress, 1 evaluate their potential as vaccine candidates. This proposal builds upon
yr timeline
previous and ongoing research where mutations are being introduced into
the leader protein of FMDV in order to attenuate the virus and derive virus
strains that could be used as vaccine candidates
National Pork
Board
Completed
We have tested the potency and efficacy of replication-defective
recombinant human adenoviruses containing the capsid and 3C proteinase
coding regions of foot-and-mouth disease virus (FMDV) as vaccine
candidates in swine. This vaccine, which lacks the coding region of several
FMDV nonstructural proteins, has a number of advantages over the current
whole virus inactivated vaccine including the ability to readily distinguish
vaccinated from infected animals using current technology. Inoculation of
swine with this vaccine resulted in either complete protection from
disease after virulent homologous virus challenge or significant reduction
in clinical signs as compared to co-housed un-inoculated control animals.
The optimal vaccine regimen tested was a low dose initial inoculation
followed by a high dose boost. All animals inoculated by this regimen
were completely free of disease after challenge. Increased efficacy of the
two-dose regimen was associated with heightened levels of FMDV specific
IgG1 and IgG2 antibodies. Expression and processing of the FMDV capsid
precursor protein, requiring a biologically active 3C proteinase, was
essential for induction of FMDV-specific neutralizing antibodies and
protection from challenge. This vaccine is safe since there is no evidence
of sero-conversion of co-housed unvaccinated control animals. These
experimental results suggest that the Ad5-FMDV virus vector is a
promising vaccine candidate against FMD and additional efforts to improve
its potency and efficacy are underway.
Completed
In this study we have demonstrated that delivery of the combination of
our FMD subunit vaccine and interferon alpha can enhance the long-term
protection afforded swine with the vaccine alone. Furthermore, we are
developing additional antiviral reagents, i.e.., interferon beta, that may
allow us to supplement our current approach as well as develop a more
comprehensive understanding of the interactions between FMDV and its
host.
National Pork
Board
Y ear
Project Title
Primary
Investigator
Institution
Funding
Agency
Status
Comments
Brough and Butman
GenVec
DHS S&T
Research milestones are associated with the construction, in vitro and in
vivo (cattle focus). Several AdFMD vaccine candidates have been made
and tested in cattle. Development milestones are associated with
In progress.
manufacturing process improvements and stability and potency
2013 end date.
assessment of AdFMD vaccine candidates producted under improved
manufacturing processes. Development milestones are aligned with DHS
S&T Vx and Dx CRADA with Merial.
Massare
Novavax
DHS S&T
Goals is construction and testing (cattle focus) of virus-like particle
In progress.
recombinant, subunit FMDV vaccine produced in insect cells. POC cattle
2014 end date.
study planned for Spring 2013.
Kern
Inovio
DHS S&T
(CRADA/No
Direct
Funding)
Goal is construction and testing (swine focus) of FMD DNA vaccines based
In progress.
on Inovio's SynCon technology and Cellectra delivery (electroporation)
End date TBD.
device. POC swine study planned for Spring 2013.
Merial
DHS S&T
(CRADA/No
Direct
Funding)
Goal is testing and evaluation of (3) AdFMD vaccine candidates in swine,
production of master seed vacine viruses, and improvement of vaccine
In progress.
manufacturing process. One POC efficacy study in swine has been
End date TBD.
completed and target efficacy level was not achieved. Efforts are ongoing
to improve vaccine potency.
FMD
Vaccine
2010
Research and Development of Molecular
FMD Vaccines (historically cattle focused)
FMD
Vaccine
2011
Development of VLP Vaccine as a
Countermeasure for FMD
FMD
Vaccine
2011
FMD DNA Vaccine
2011
Testing and Evaluation of Human
Adenovirus Replication Deficient Vectored
FMD Vaccines (swine focus)
Miller
Benchmark
Biolabs
DHS S&T
In progress.
2014 end date
(if all option
years awarded)
Mittal
Purdue
University
DHS S&T
Goal is construction and testing (cattle focus) of replication competent
In progress.
and replication deficient bovine adenovirus vectored FMD vaccine
2014 end date.
candidates.
FMD
Vaccine
FMD
Vaccine
2012
Acquisition of Master Seed Virus (MSV) and
Working Seed Virus (WSV) Lots for FMD
Molecular Vaccine Countermeasures
FMD
Vaccine
2012
Bovine Adenovirus Vector-Based Vaccine
for FMD
FMD
Vaccine
2012
Construction and Evaluation of
Recombinant MVA-BN FMDV Candidates
FMD
Vaccine
2012
Replicon-based Rapid Response Vaccines
for FMD
2010
Production of FMD Virus Pseudovirion
Vaccine Candidates Using a Plant Transient
System
2011
and Improved Challenge Systems Improved Challenge Systems for FMD
Vaccine and Biotherapeutics Testing in
Cattle and Pigs
FMD
FMD
Area
Vaccine
Vaccine
David
optimal vector parameters, ii) construction and evaluaton of two
recombinatn MVA-BN-FMDV constructs expressing 3C in trans, and iii)
grade).
Weinberger
Bavarian Nordic
DHS S&T
In progress. optimal vector parameters, ii) construction and evaluaton of two
2014 end date. recombinatn MVA-BN-FMDV constructs expressing 3C in trans, and iii)
grade).
Kamrud
Harrisvaccines
DHS S&T
In progress. Goals is rapid, large-scale (50-100K doses) of VEE replicon particle FMDV
2014 end date. vaccine candidate (cattle focus).
Hiatt
Kentucky
BioProcessing
DHS S&T
Vx/Dx
Arzt
USDA ARS PIADC
DHS S&T
In progress.
The key goal of the project is to produce FMDV pseudovirion structures in
plants and develop scaleable manners for purification and vaccine
production. To date, efforts to produce pseudovirion structures in tobacco
plants have failed. Modified SOW is in progress to use an alernative
strategy to improve capsid production and solubility.
The primary goal of this proposed research is the optimization of new,
simulated natural systems for challenging cattle and pigs with foot-andmouth disease virus (FMDV). These novel systems will be optimized in
In progress.
studies using naïve animals and subsequently validated in vaccinate2014 end date.
challenge experiments. Additionally, the novel systems will be used to
assess pigs’ and steers’ ability to transmit FMDV in the pre-clinical phase
of disease.
FMD
FMD
Area
Vaccine
Vaccine
Y ear
Project Title
Primary
Investigator
Institution
Funding
Agency
Status
Comments
2011
Rational design of attenuated foot-andmouth disease virus strains for
development of improved disease
countermeasures
de los Santos
USDA ARS NAA
National Pork
Board
Completed
Our goal is to develop alternative control strategies that could improve current
FMD countermeasure programs. An attenuated vaccine is expected to elicit more
rapid innate immunity and a long lived adaptive immunity to effectively control
disease. Moreover, induction of innate immunity could result in early protection
against multiple FMDV serotypes. Importantly, attenuated strains are excellent
new tools to study the interactions between FMDV and the host immune system
and ultimately could lead to the development of novel strategies to counteract
FMD. During the past year and with the support of NPB we have successfully
derived a mutant strain of FMDV that did not cause disease in swine (FMDV-SAP
mutant). Interestingly inoculation of swine with this mutant strain induced a strong
immune response that protected animals against infection with the parental (wild
type) virus, as early as two days post vaccination. Studies in animals and in
cultured swine cells demonstrated that, in contrast to the parental wild type virus,
the mutant variant was unable to block some inflammatory responses thus limiting
dissemination of the virus beyond the original site of inoculation. Furthermore we
have started studies to add more mutations to this virus aiming to increase the
stability of the original mutations therefore decreasing the probability of reversion
to virulence. Our results indicated that manipulation of the viral genome in the
region that encodes for the leader protein is a viable alternative to derive less
pathogenic FMDV strains that could be used as the basis for live attenuated
vaccines against FMD or as seeds to grow the virus for manufacturing safer
inactivated vaccines.
2012
Exploiting the potential of leader
proteinase coding sequence of foot-andmouth disease virus to derive attenuated
strains suitable for pathogenesis studies
and development of improved
countermeasures.
de los Santos
USDA ARS NAA
National Pork
Board
In progress
Goal is to develop alternative control strategies that could improve
current FMD countermeasure programs. An attenuated vaccine is expected
to elicit more rapid innate immunity and a long lived adaptive immunity to
effectively control disease.
Multiple
Agents
Multiple
Agents
Multiple
Agents
Multiple
Agents
Multiple
Agents
Area
Diagnostics
Diagnostics
Diagnostics
Diagnostics
Diagnostics
Y ear
2008
2011
2012
Project Title
Enhancements of High-Throughput
Diagnosis for FAD
Development of a multiplex RT-qPCR
assay for the simultaneous detection of
endemic and foreign animal diseases in
oral fluid samples.
Assessment of Two Methods for Nucleic
Acid Stability and Viral Inactivation of
FAD Viruses Collected from Oral Fluid
Rope Samples
2009
Vesicular Disease Diagnostic Reagent
Production
2011
The Matrix-Chaperone: Ambient
Temperature Biospecimen Collection,
Transport & Banking For Simplified
Animal Disease Screening (swine and
cattle).
Primary
Institution
Investigator
McIntosh
Beckham/
McIntosh/
Bounpheng
Mayr
Jia
Hogan
USDA
APHIS
PIADC
TMVDL/FAD
DL
USDA
APHIS
USDA
APHIS
PIADC
IntegenX
Funding
Agency
Status
Comments
DHS S&T
Vx/Dx
Construction and select agent certification of the highthroughput lab that will allow diagnostic work using
Completed
PCR and serology was completed. Validation of the lab
is underway.
DHS S&T
The objective of this effort is to develop an optimized
oral fluid nucleic acid purification method and a
multiplex reverse transcriptase quantitative PCR assay
(mpRT-qPCR) that can be used to simultaneously
detect several economically important swine endemic
In progress and foreign animal diseases from the same sample.
APHIS will conduct bench validation of a multiplex RTqPCR assay for the simultaneous detection of endemic
and foreign animal diseases in oral fluid samples. The
multiplex is currently under development by FAZD under
a separate agreement
DHS S&T
Two (or more) methods for the preservation of nucleic
acid while neutralizing virus infectivity will be
examined. Whitman FTA paper and PrimeStore MTM
media will be examined to determine if they are
capable of stabilizing viral nucleic acids, RNA and DNA,
In progress from rope oral fluid samples while at the same time
inactivating/neutralizing any viable virus present in the
oral fluid so as to make transport of the samples to
laboratories outside of the outbreak zone risk free.
The recent approval to fill this position will result in the
job posting within the month.
DHS S&T
Vx/Dx
DHS S&T
USDA APHIS FADDL established an in-house reagent
production capacity to produce large quantities of
highly specific rabbit and guinea pig antisera against
homologous FMDV, SVDV, and VSV to be used in the
In Progress. vesicular disease Ag ELISA, as well as to fulfill the
2013 end diagnostic surge capacity required in the event of a
vesicular disease outbreak. A result of this effort is
date.
that this lab was recognized as a FMD Reference lab by
FAO and the World Organization for Animal Health,
resulting in significantly less reliance on foreign labs.
This project will develop and validate a new technology
for the collection, transport and banking of
In progress biospecimens for animal disease screening that is
based on ambient-temperature, dry-state sample
preservation
Area
Y ear
Multiple
Agents
Diagnostics
2011 &
2008
Multiple
Agents
Post-Harvest
2012
Project Title
Development of Pan-viral DNA
Microarrays for the Detection of
Emerging and Foreign Animal Diseases
Disinfection of foreign animal disease
viruses on surfaces relevant to the Pork
Packing Industry
Primary
Institution
Investigator
McIntosh
Rodriguez and
Krug
USDA
APHIS
PIADC
USDA
APHIS
PIADC
Funding
Agency
Status
Comments
DHS S&T
Vx/Dx
An enhanced version of the microarray platform is being
Completed validated that will detect more viruses isolated from a
variety of organisms.
National Pork
Board
The main objective of this proposal is to determine the
efficacy of chemical disinfectants against foreign
animal disease (FAD) viruses dried on surfaces relevant
to the Pork Packing industry. A second objective of this
Completed project is to ensure that necessary surface-specific
changes are made to existing protocols to ensure valid
results are generated. A third objective of this proposal
is to validate the use of non-FAD viruses as surrogates
for FAD viruses in disinfection assays.
Page 33 of 32

Strategic Planning for Swine Disease Research

Transcription

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