Cyril gay nfid vaccine research conference, fmd vaccines, april 2013


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  • At the end of this period all countries will have reached at least PCP Stage 2.
  • Have!
  • Cyril gay nfid vaccine research conference, fmd vaccines, april 2013

    2. 2. Development of vaccines toward the global control and eradication of Foot- and-Mouth Disease (FMD)Cyril Gerard Gay, DVM, PhD Senior National Program Leader Animal Production and Protection Agricultural Research Service
    3. 3. Presentation Outline 1. Importance of Animal Agriculture 2. Disease Threats 3. Cost of FMD 4. FMD Eradication 5. FMD Virology and Pathogenicity 6. FMD vaccines 7. Conclusions 33
    4. 4. Importance of AnimalImportance of Animal AgricultureAgriculture • FAO estimates that livestock contribute 40% of the global value of agricultural output and support the livelihoods and food security of almost 1 billion people 44
    5. 5. 2121stst Century ChallengesCentury Challenges • World population is projected to reach 9 billion • Global food production will need to double in order to meet these food demands. • 73% increase in consumption of animal protein 55 58% increase in consumption of dairy products
    6. 6. Presentation Outline 1. Importance of Animal Agriculture 2. Disease Threats 3. Cost of FMD 4. FMD Eradication 5. FMD Virology and Pathogenicity 6. FMD vaccines 7. Conclusions 66
    7. 7. List of 17 Most DamagingList of 17 Most Damaging Animal Disease ThreatsAnimal Disease Threats 1. Highly Pathogenic AI (F) 2. Foot-and-Mouth Disease 3. Rift Valley fever (F) 4. Exotic Newcastle Disease 5. Nipah and Hendra virus (F) 6. Classical swine fever 7. African swine fever 8. Bovine spongiform encephalopathy (F) 9. Rinderpest (E) 10. Japanese encephalitis (F) 11. African horse sickness 12. Venezuelan equine (F) encephalitis 13. Contagious bovine pleuropneumonia 14. Ehrlichia ruminantium (Heartwater) 15. Eastern equine encephalitis (F) 16. Coxiella burnetii (F) 17. Akabane virus F: Potentially fatal to humans Yellow text: FBI pathogens of Concern E: Eradicated H5N1 Avian Influenza Virus Source: PHIL CDC
    8. 8. Emerging Diseases (and re-emerging diseases) Human Animal • HIV/AIDS • Ebola* • Hantaan • Legionaire’s disease • BSE* • SARS* • Dengue • West Nile* • Nipah virus* • Rift Valley Fever* • Chikungunya virus • H5N1, H3N2v, H7N9* • pandemic H1N1 • BSE* • CWD • West Nile* • Foot-and-Mouth Disease • Classical Swine Fever • Blue Ear Pig Disease • Rift Valley Fever* • Avian Influenza H5N1/H7N9* • Nipah and Hendra* • Bluetongue • African Swine Fever • African Horse Sickness • pandemic H1N1** * Zoonoses **Reverse Zoonosis
    9. 9. Presentation Outline 1. Importance of Animal Agriculture 2. Disease Threats 3. Cost of FMD 4. FMD Eradication 5. FMD Virology and Pathogenicity 6. FMD vaccines 7. Conclusions 99
    10. 10. HistoryHistory • Over 100 years of research in FMDV • 1924-British Minister of Agriculture appointed a committee “to initiate, direct and conduct investigations into FMD… discovering means whereby the invasion of the new disease may be rendered less harmful to agriculture…” (from B.W. Mahy, 2005) • Successful eradication in Europe • US free since 1929 In 1898, Freidrich Loeffler and Paul Frosch showed that a virus was responsible for foot-and-mouth disease in cattle 1010
    11. 11. The Cost of FMDThe Cost of FMD • Total loss of 2001 outbreak in the United Kingdom was estimated to be between $12.3 and $15 billion (US$) • 36% was lost tourism , Slaughter of 6.5 million livestock • $4.2 billion paid by government in compensation to the agriculture and food industry • Social effects, Human cost (suicides) • For some countries mass slaughter is NO LONGER an option!! (e.g. S. Korea, 2011)
    12. 12. The Cost of FMDThe Cost of FMD • On the global scale FMD causes damage and hampers development : Cost USD 5 billion per year • Outbreaks in FMD-free countries worldwide costs USD 1 billion/year • The world is a global village; risks for FMD-free countries will only increase • Fighting the disease at source should be part of the prevention strategy of FMD-free countries(Rushton, 2012)
    13. 13. Presentation Outline 1. Importance of Animal Agriculture 2. Disease Threats 3. Cost of FMD 4. FMD Eradication 5. FMD Virology and Pathogenicity 6. FMD vaccines 7. Conclusions 1313
    14. 14. Pool 7 O, A Pool 5 O, A, SAT 1, 2 Pool 6 SAT 1, 2, 3 Pool 4 A, O, SAT 1, 2, 3 Pool 2 O, A, Asia 1 Pool 1 O, A, Asia 1 Pool 3 O, A, Asia 1 Endemic Free with vaccination Intermediate, sporadic Countries with multiple zones: FMD-free, free with vaccination or not free Free. Virus present in game parks Free Pool positions are approximate and colours indicate that there are three principal pools, two of which can be subdivided into overlapping areas Status of FMD showing approximate distribution of regional virus endemic pools ®
    15. 15. Website: FAO/OIE Progressive Control Pathway for the control of FMD
    16. 16. Within a 15-year period: 1) countries that are currently in PCP Stages 0 and 1 will have progressed at least two stages along the PCP 2) countries in PCP Stages 2 or 3 should also move up two stages, but the final objective will depend on a country’s decision based on cost-effectiveness studies 3) countries or zones that already have an OIE-recognized FMD-free status maintain this status or further improve it (i.e. go from FMD-free with vaccination to FMD-free without vaccination) Objectives of FMD Control Strategy
    17. 17. - Cost of national FMD programs for 79 initial 0-2 Stage countries: 68 M - Vaccination cost for 45 initial 1-3 Stage countries (excluding India and China): 694 M - Regional level (ref. lab and epidemiology support and networks) 47 M - Global level (coordination, evaluation) 11 M Financial implications (first 5 years) (in USD as calculated by the World Bank)
    18. 18. Presentation Outline 1. Importance of Animal Agriculture 2. Disease Threats 3. Cost of FMD 4. FMD Eradication 5. FMD Virology and Pathogenicity 6. FMD vaccines 7. Conclusions 1818
    19. 19. Features of FMDV 25 nm • Family Picornaviridae, genus aphtovirus • Positive sense RNA Approximately 8.2 kb • Seven serotypes: A, O, C, Asia, Sat1, Sat2, Sat3
    20. 20. P1 P2 poly(A) 3'UTR 3B IRES S 5'UTR PKs crepoly(C) 3B1231A Lpro 1B 1C 1D 2A 2B 2C 3A 3Cpro 3Dpol P3 ** Protease Cleavage Sites Lpro unknown 2A3Cpro 2 in-frame AUGs ** FMDV genome VP0 (1AB) 3AB123 structural proteins nonstructural proteins partialpartial cleavagecleavage productsproducts P1/2A P2BC P3 3B123CD1ABC 3A L VP1 VP3 2C2B 3A 3B1 3B2 3B3 3C 3CD 3D
    21. 21. FMDV-Key Information • Systemic disease of domestic and wild cloven-hooved animals • Acute disease characterized by fever, lameness, and vesicular lesions on the feet, tongue, snout, and teats • FMD is considered to be one of the most contagious infectious disease known 2121
    22. 22. FMDV-Key Information • Multiple subtypes reflect significant genetic and antigenic variability • Some strains of the virus and some host species show minimal or no signs of disease • The emergence of new variants of FMDV is common • Fifty percent of infected cattle become carriers • The pathogenesis of FMDV, including mechanisms of viral transmission and the carrier state, are not fully understood • The early detection of FMDV is paramount to stop the spread of the virus and disease and reduce economic impact 2222
    23. 23. Aerosol Inoculation ModelAerosol Inoculation Model The nebulizer consists of a commercially available aerosol delivery system that will produce an average of particles of 5 microns (Hess et al., 1996) and a large equine mask. Entire respiratory tract is exposed to virus. Pacheco et al, 2008 Dose: FMDV 107 TCID50 2323
    24. 24. Summary of FMD Early Pathogenesis in Cattle V V V V V V V V V V V V Systemic Circulation Aerosol Exposure T = 0.1 HPA T = 3 – 6 HPA T = 12 HPA T = 24 HPA T = 48 HPA 24
    25. 25. FMD-Bovine Mouth 25
    26. 26. FMD-Bovine Hoof 26
    27. 27. Presentation Outline 1. Importance of Animal Agriculture 2. Disease Threats 3. Cost of FMD 4. FMD Eradication 5. FMD Virology and Pathogenicity 6. FMD vaccines 7. Conclusions 2727
    28. 28. FMDV- Commercial Vaccines • Conventional inactivated vaccines have been successfully used in disease eradication programs in endemic areas of Africa, South America, and Europe • Requires adaptation of wild type virus to cell culture • Virulent virus grown on BHK cells • Production of large volume requires BSL-3 facilities • Virus yield sometimes low for hard-to-adapt viruses • Inactivated with binary ethyleneimine (BEI) • Non-structural proteins removed • Non-formulated bulk fluids are stored frozen for stockpiling • Adjuvanted with alum or oil emulsion • Vaccines provide fail to induce long lasting immunity 2828
    29. 29. T.R. Doel / Virus Research 91 (2003) 81/99 86 Current Vaccines 29 Inactivated Vaccine
    30. 30. FMDV- Vaccine Information • FMDV Serotype O is less immunogenic • FMDV Serotype O is more prevalent in South America • Vaccines for FMDV Serotype O need a higher payload than Serotypes A, C, Asia, or SAT • FMDV Serotypes SAT1, SAT2, SAT3 antigens are less stable • FMDV Serotypes A and SAT 2 are more hypervariable than other serotypes 3030
    31. 31. Risk of Vaccine Production withRisk of Vaccine Production with Virulent FMDVVirulent FMDV On Friday August 3, 2007 FMD was detected in a farm in Southern England located within 6 miles of the Pirbright Laboratory site Outbreak resulted in trade barriers and billions $$ loses 3131
    32. 32. Concerns with FMD Vaccines in Disease-Free Countries • Require adaptation and growth of large volumes of wild type virus in cells • Escape of virus from manufacturing facilities • Require banking of multiple antigen concentrates • Some antigens lack stability (low potency/short shelf life) • Onset of protection 7-14 days • Short duration of immunity <6 months • Difficult to differentiate vaccinated from infected animals (DIVA) due to presence of NS proteins • Vaccinated and exposed animals become carriers 3232
    33. 33. Characteristics of the “Ideal” FMD Vaccine • Effective, rapid and long-lasting protection with one inoculation • Prevents viral transmission • Allow differentiation of infected from vaccinated animals (DIVA) • Produced without the need for virulent FMDV • Prevent development of carrier state • Protection against multiple serotypes • Stable antigen – long shelf life • Reasonable cost to enable eradication programs
    34. 34. Adenovirus-Vectored FMD Vaccine Expressing Empty Viral Capsids • Contains all protective epitopes present on current inactivated virus vaccine but lacks infectious viral nucleic acid and non-structural protein (NSP) • Allows to “clearly” distinguish vaccinated from infected animals using 3D and other NSP diagnostic tests • Can be safely produced in the United States “Left-out” proteins can be used for DIVA tests Processed products display epitopes resembling intact capsid. 3D FMDV Empty Capsid Vaccine P1 2A 2B’ 3B’ 3C VP0 VP3 VP 1 3C L P1 2A 2B 2C 3A 3B 3C Remove regions unnecessary for capsid formation. 3D DHS TAD Program Licensed by CVB-APHIS
    35. 35. 2 Negative markers: DIVA tests 3Dpol ELISA 3B ELISA 3B23 3’NTR IRES S 1A 1B 1C 1D 2A 2B 2C 3A 3C 3D A poly(C) NON-STRUCTURALSTRUCTURAL δ L 5’NTR RE1 RE2 Easy swap of capsid sequences Vaccine seed antigens Deletion of Leader protein (543 bp) Attenuating factor FMD-LL3B3D: A Safe Platform For FMD Vaccine Production With Built-In DIVA Markers Key Features 36
    36. 36. FMD Vaccine Product Profiles :FMD Vaccine Product Profiles : Current Inactivated versusCurrent Inactivated versus Inactivated FMD-LL3B3DInactivated FMD-LL3B3D NoNoProvides cross-serotype protection NoCompatible with “vaccinate to live” strategy NoNoReadily deployable (ready to use) NoNoLong duration of immunity PossibleNoDomestic production (USA) +/-Marked vaccine (DIVA capable) Early onset of immunity (7 DPV) Prevents viral transmission MOLECULAR INACTIVATED CURRENT INACTIVATED PRODUCT PROFILE      Possible
    37. 37. ProteinaseProteinase domain/De-domain/De- ubiquitinaseubiquitinase Topology,Topology, DNA binding,DNA binding, transcriptiontranscription regulationregulation Topology,Topology, interaction withinteraction with phosphoproteins,phosphoproteins, signalingsignaling interferenceinterference InteractionInteraction with eIF4Gwith eIF4G SAP FHA-likeProteinase 1 29 75 112 167 183 201 N C Lab Lb Bioinformatics analysis suggests the presence of multiple domains (by Dr. James Zhu)
    38. 38. Disruption of the L protein SAP domain results in attenuation in vitro SAP domains are conserved protein domains present in eukaryotic nuclear proteins involved in chromosomal organization and repression of transcription. Double mutation of FMDV L protein SAP domain results in: Altered L protein sub-cellular distribution: L SAP mutant localizes only to the cytoplasm of infected cells by 6 hpi while L wild type is in the cytoplasm and nucleus. L SAP mutant is unable to cause degradation of NF-κB inducing higher levels of IFN, inflammatory cytokines and chemokines in comparison to WT. (de los Santos et al., 2009)
    39. 39. 00 11 22 33 44 55 66 77dpidpi Virus intradermal inoculation inVirus intradermal inoculation in right rear foot-padright rear foot-pad Temperature Serum Nasal Swabs Temperature Plasma/Serum Clinical signs Nasal Swabs 1414 2121 x3x3 x3x3 x3x3xx 33 x3x3 Group #1:Group #1: Group #2:Group #2: Group #3:Group #3: Group #4:Group #4: Group #5:Group #5: FMDV A12-WT 1x105 pfu/pig FMDV A12-WT 1x106 pfu/pig FMDV A12-SAP 1x105 pfu/pig FMDV A12-SAP 1x106 pfu/pig FMDV A12-SAP 1x107 pfu/pig Serum Does disruption of the L protein SAPDoes disruption of the L protein SAP domain results in attenuationdomain results in attenuation in vivo?in vivo?
    40. 40. Clinical Score pfu/ml 0 2 4 6 8 10 12 14 16 18 0dpc 1dpi 2dpi 3dpi 4dpi 5dpi 6dpi 7dpi 10e5 A12-SAP 10e5 A12-WT 10e6 A12-WT 10e6 A12-SAP 10e7 A12-SAP Viremia Nasal swabs 0dpi 1dpi 2dpi 3dpi 4dpi 5dpi 6dpi 1.0E+00 1.0E+01 1.0E+02 1.0E+03pfu/ml 0.0E+00 4.0E+02 8.0E+02 1.2E+03 1.6E+03 0dpi 1dpi 2dpi 3dpi 4dpi 5dpi 6dpi 7dpi A12-SAP mutant is attenuatedA12-SAP mutant is attenuated in vivoin vivoScore(max.17)
    41. 41. Animals inoculated with A12-SAP areAnimals inoculated with A12-SAP are completely protected when challengedcompletely protected when challenged with WT FMDV at 21 dayswith WT FMDV at 21 days Group Animal Challenge virus at 21dpi Dose Viremia (dpc, day of onset, duration) PFU in nasal swabs (dpc, day of onset, duration) Neutralizing antibodies PRN70 7dpc A12-SAP 1x105 pfu/ pig 90 A12-WT 1x105 Neg. Neg. >3.1 91 Neg. Neg. >3.1 92 Neg. Neg. >3.1 A12-SAP 1x106 pfu/ pig 93 A12-WT 1x105 Neg. Neg. >3.1 94 Neg. Neg. >3.1 95 Neg. Neg. >3.1 A12-SAP 1x107 pfu/ pig 96 A12-WT 1x105 Neg. Neg. 3.0 97 Neg. Neg. 3.1 98 Neg. Neg. >3.1
    42. 42. x3x3 x3x3 x3x3 FMDV A12-SAP 1x106 pfu/pig FMDV A12-SAP 1x106 pfu/pig FMDV A12-SAP 1x106 pfu/pig CONTROL PBS x3x3 x3x3 FMDV A12-SAP 1x106 pfu/pig VACCINATION DOSE TIME OF CHALLENGE CHALLENGE DOSE 14dpv14dpv 7dpv7dpv 4dpv4dpv 2dpv2dpv 14dpi14dpi FMDV A12-WT 5x105 pfu/pig FMDV A12-WT 5x105 pfu/pig FMDV A12-WT 5x105 pfu/pig FMDV A12-WT 5x105 pfu/pig FMDV A12-WT 5x105 pfu/pig SAP mutant vaccination experiment withSAP mutant vaccination experiment with early challenge in swineearly challenge in swine
    43. 43. Inoculation with FMDV A12-SAP confersInoculation with FMDV A12-SAP confers protection as early as 2 days postprotection as early as 2 days post vaccinationvaccination Virusinserumornasalswabs (pfu/ml) ClinicalScore 1.0E+00 1.0E+01 1.0E+02 1.0E+03 1.0E+04 1.0E+05 1.0E+06 0dpc 1dpc 2dpc 3dpc 4dpc 5dpc 6dpc 7dpc 0 2 4 6 8 10 12 14 16 18 14 dpv 4 dpv 7 dpv 2 dpv control viremia NScs
    44. 44. SummarySummary A12-SAP mutant is avirulent in swine but induces a strong neutralizing antibody response In vivo attenuation correlates with increased levels of pro-inflammatory cytokines whose transcription depends on NF-κB Vaccination of swine with A12-SAP results in complete protection against homologous challenge as early as 2 days post-inoculation, when no adaptive immune response is detectable
    45. 45. Conclusions 1. Importance of Animal Agriculture 2. Disease Threats 3. Cost of FMD 4. FMD Eradication 5. FMD Virology and Pathogenicity 6. FMD vaccines 4646 Publications
    46. 46. Global Foot-and-Mouth Disease Research Alliance 4747
    47. 47. AcknowledgementAcknowledgement • Luis Rodriguez • Marvin Grubman • Jonathan Arzt • Juan Pacheco • Elizabeth Rieder • James Zhu • Teresa de los Santos • Bill Golde 4848
    48. 48. Thank you!
    49. 49. Thank you !!!