Foreign Animal Disease Research Unit USDA/ARS
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1 Foreign Animal Disease Research Unit USDA/ARS Overview USAHA FED 2012 Agricultural Research Service, Plum Island Animal Disease Center
2 APPROVED NEW CRIS PROJECTS Intervention Strategies to Support the Global Control and Eradication of FMD Countermeasures to Control Foreign Animal Diseases of Swine: CSF/ASF Ecology and pathogenesis of Re-Emerging VSV in North America
3 Foot and Mouth Disease FMD is considered number one threat to American Agriculture and related industries (livestock, crops, farm supply, etc) FAO/OIE announced eradication from Americas by 2023 global strategy key to eradication FMD is main barrier to international trade of animal products FMD is one of the main threats to global food security Maasai Hearding; Painting By Kahare Miano Countries infected with FMD are more prone to food insecurity as a result of the impact of FMD at household level and through reduced access to local, national and international markets and of animal draught power for agriculture OIE/FAO, 2009
4 T.R. Doel / Virus Research 91 (2003) 81/99 86 Currrent vaccines
5 Concerns with FMD Vaccines SAFETY: Require adaptation and growth of large volumes of wild type virus in cells è possibility of escape of virus from manufacturing facilities EFFICACY: Narrow antigenic coverage (serotype / subtype) Short duration of immunity <6 months Vaccinated and exposed animals become carriers
6 Characteristics of an Ideal FMD Vaccine Effective, rapid and long-lasting protection with one inoculation Prevents viral transmission Allow differentiation of infected from vaccinated animals (DIVA) Safe: produced without the need for virulent FMDV No need for adaptation of field strains to cell culture Prevent development of carrier state Broad antigenic coverage Stable antigen long shelf life Long duration of immunity
7 5 NTR S IRES δ L STRUCTURAL NON-STRUCTURAL 3B 23 3 NTR 1A 1B 1C 1D 2A 2B 2C 3A 3C 3D A poly (C) RE1 RE2 3D pol ELISA 3B ELISA 2 Negative markers Targeting NSP Vaccine seed antigens Easy swap of capsid sequences Attenuating factor Deletion of Leader protein (543 bp)
8 Safety Data Cattle Bovine # a Virus Viremia, Maximum Titer. b (DPI) c Virus in Saliva, Maximum Titer b (DPI) c Fever d (DPI) e Maximum Clinical Score/ Maximum achievable f (DPI) g Neutralization Titer maximun (Starting DPI) h Shedding in air. Maximun Titer i (DPI) c 7109 A 24 WT 7.60 (3) 8.90 (3) Yes (2,3) 5/5 (7) 2.4 (5) 5.57 (5) 7110 A 24 WT 7.31 (4) (3) Yes (2-5) 5/5 (5) 2.4 (6) ND 9143 A 24 WT3B m 3D m 7.40 (4) 9.03 (5) Yes (3) 1/5 (5) (6) 9144 A 24 WT3B m 3D m 7.92 (4) 8.85 (5) Yes (4) 4/5 (7) (7) 9145 A 24 LL3B m 3D m Negative Negative No 0/5 1.5 ND j 9146 A 24 LL3B m 3D m Negative Negative No 0/5 2.4 Negative FMD-LL3B3D is fully attenuated in cattle!
9 Efficacy Data Cattle BEI-inactivated Vaccine formulated with montadine ISA 260 adjuvant (Sepic-WOW) 21 dpv challenge with FMDV-A24 N=4 N=4 N=4 N=4 Commercial Tetravalent FMDV Vaccine 1xBEI- Vx FMD-LL3D FMDV Vaccine 1 x BEI Marker virus FMD-LL3B3D FMDV Vaccine 1 x BEI Marker virus Naïve unvaccinated controls No clinical disease (0/4)* No clinical disease (0/4)* No clinical disease (0/4)* 100 % clinical disease (4/4) Inactivated vaccines prepared with FMD-LL3D and FMD-LL3B3D induced complete protection against challenge!
10 Characteristics of an Ideal FMD Vaccine Effective, rapid and long-lasting protection with one inoculation Prevents viral transmission Allow differentiation of infected from vaccinated animals (DIVA) Safe: produced without the need for virulent FMDV No need for adaptation of field strains to cell culture Prevent development of carrier state Broad antigenic coverage Stable antigen long shelf life Long duration of immunity
11 Novel Ad5-FMD Vaccine A novel FMD vaccine was developed by ARS scientists under the leadership of Dr. Marvin Grubman This vaccine utilizes a defective human adenovirus vector to deliver genes coding for FMDV structural proteins Human Defective Adenovirus 5 vector - Lacks necessary proteins for growth - Delivers and expresses transgenes in target cells M. Grubman
12 First Licensed Molecular FMD Vaccine N=6 1 o Vx N=6 N=6 1 o Vx 1 o Naive N=4 2 o Vx N=4 2 o Naive N=4 2 o Naive No clinical disease 100% protection (10/10) No clinical disease 100% protection (10/10) 100 % clinical disease (10/10) DHS- TAD Group
13 Characteristics of an Ideal FMD Vaccine Effective, rapid and long-lasting protection with one inoculation Prevents viral transmission Allow differentiation of infected from vaccinated animals (DIVA) Safe: produced without FMDV!! No need for adaptation of field strains to cell culture Prevent development of carrier state Broad antigenic coverage Stable antigen long shelf life Long duration of immunity
14 Implications What does this vaccines add for US preparedness? Possibility of domestic production Rapid response to new strains Opened regulatory path to other molecular vaccines Implications to the hemisphere? Potential application in final stages of eradication?
15 APPROVED NEW CRIS PROJECTS Intervention Strategies to Support the Global Control and Eradication of FMD Countermeasures to Control Foreign Animal Diseases of Swine: CSF/ASF Ecology and pathogenesis of Re-Emerging VSV in North America
16 ASF On The Move 2012 Beginning of 2007 in R. of Georgia and has since spread to the neighboring countries of Armenia, Azerbaijan, Ukraine and Russia
17 ASFV Research Program Develop intervention strategies to control ASF virus by identifying virus-host determinants of virulence and transmission and by developing technologies to enable the development of ASF vaccines that are efficacious against the most prevalent ASF strains. Development and standardization of a challenge model to assess ASFV experimental immunogens Comparative studies of early pathogenesis events in swine during infection with highly virulent and attenuated ASFV strains using a natural route of infection. Identification of immune mechanisms mediating protection induced by experimental live attenuated vaccine strains Functional Genomics and development of ASFV experimental vaccines
18 Challenge model Hypothesis: Oronasal (simulated natural) inoculation of pigs will allow elucidation of critical virus-host interactions which may be targeted by novel countermeasures Standardized intraoral deposition of ASFV under sedation Dr. Jonathan Arzt
19 Functional genomics LVR CVR RVR Structural proteins: p30, p72, p54 Immune response modulation: 5EL (IkB), 8CR (lectin), 8DR (CD2) Prevention of apoptotic cell death: 5HL (Bcl2), 4CL (iap) Host range and virulence associated genes NL, UK, 9GL, TK Multigene family (MGF) 360 genes Multigene family (MGF) 530 genes Dr. Manuel Borca
20 Genetic engineering of ASFV ASFV 0kB X X 180kB PCR Amplify left flank Amplify right flank INSERT REPORTER GENE P72 GUS Transfer Vector (TV) INFECTION RECOMBINATION Macrophages x ASFV TRANSFECTION TV
21 9GL confers strain-specific protection ASFV challenge (1,000-10,000 LD 100 ) Immunization MAL 9GL TEN 9GL Pr4 9GL Protection in pig MAL (Malawi 83) YES NO NO TEN (Tengani 54) NO YES NO RSA1 (Pr4 96) NO NO YES RSA2 (Pr5 96) YES RSA3 (CR1 96) YES RSA4 (CR3 96) YES RSA5 (O1 96) YES
22 Production of recombinant Georgia ASFV ΔCD-2 like Δ ASFV genes 9GL NL UK MGF CD-2 like
23 What next Comparative studies of early pathogenesis events in swine during infection with highly virulent and attenuated ASFV strains using a natural route of infection. Determine protection induced by experimental live attenuated vaccine strains Identification of immune mechanisms mediating protection induced by experimental live attenuated vaccine strains Expression of multiple ASFV immunogens in replicating viral vectors (e.g. vaccinia) to assess immune response and protection from challenge
24 Strategic partnerships Government: DHS, USDA/APHIS, EPA, DOD, DOS, DOE Stake holder: National Pork Board (>$2 M in last 4 years) Academia: UConn, Kansas State, U.Georgia, Stonybrook, U. Wisconsin, U. Vermont, Texas A&M Industry: Pfizer, Merial, Intervet, GenVec
25 GLOBAL PARTNERSHIPS To establish and sustain global research partnerships to generate scientific knowledge and discover the tools to successfully prevent and control FADs Countries with current research collaborations India Australia Spain Pakistan Vietnam South Africa UK Argentina Kenya Cameroon Russia R. Georgia S.Korea Japan Mexico Uganda Israel Denmark
26
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