1. Roxann Motroni, DVM, PhD AAAS Fellow Agriculture Defense Branch Chemical and Biological Defense Division Science and Technology Directorate International Foot-and-Mouth Disease Vaccine and Diagnostic Field Trial Global Foot and Mouth Disease Research Alliance Scientific Meeting October 2015

2. Agricultural Defense Mission The Agricultural Defense mission is to enhance current capabilities and develop state-of-the-art countermeasures for high-priority foreign animal diseases (FAD). This includes near- and long-term research and development for vaccines and diagnostics, in coordination with internal and external stakeholders. HSPD-9 Paragraph 23: The Secretaries of [DHS, USDA, HHS, EPA…] in consultation with the Director of the Office of Science and Technology Policy, will accelerate and expand development of current and new countermeasures against the intentional introduction or natural occurrence of catastrophic animal, plant, and zoonotic diseases. The Secretary of Homeland Security will coordinate these activities. 2

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4. FMD Virus Vaccine, Serotype A24, Live Adenovirus Vector (USDA Code 1FM1.R0) 4 4 Conditional license granted for use in cattle on May 31, 2012 Recently renewed to May 6, 2016 Met the expiration potency after three years in storage at -18+/-5 o C. Monovalent vaccine Public-private partnership between DHS, USDA and industry Replication deficient viral vector Does not contain full FMDv genome, so can be produced at BSL-2 Recent PIADC studies Cattle - Demonstrated 6 mo. duration of immunity(experimental challenge) Swine - Demonstrated proof-of-concept efficacy (experimental challenge at 2 weeks post-vaccination)

5. 1.10X dose (calf)  Acceptable - no local or systemic reactions 2.Backpassage/Reversion to Virulence (cattle)  Inoculation of master seed virus (MSV) into 10 cattle showed no reversion to virulence  Only a small amount of vaccine virus inoculum was isolated from nasal swabs at 1-2 days post-administration in 4/10 animals  No amplification of vaccine virus was evident 3.Shed-spread (cattle, swine)  MSV could not be isolated from either 10 cattle or 10 swine after intramuscular administration.  Naive cattle and swine co-mingled with vaccinated cattle or swine do not seroconvert to adenovirus or FMD.  Portion of study published in Grubman, MJ, et. al. 2010. Future Virol. 5(1):51- 64. FMD Virus Vaccine, Serotype A24, Live Adenovirus Vector: USDA CVB Licensing Studies- Safety 5 5

6. 4.Field Safety (beef, dairy)  500 total 4 geographic, commercial sites Both sexes Young (19-22 weeks) and older (> 2yrs) Includes 100 lactating dairy; 58 pregnant (all trimesters)  96% experienced no recorded adverse events of any kind during the 3 week post-vaccination observation period.  In the remaining 4%, there were no recorded instances of serious, permanent or systemic adverse reaction attributable to vaccination.  Study conclusion: Vaccine is safe for use in healthy cattle when used per proposed label recommendations FMD Virus Vaccine, Serotype A24, Live Adenovirus Vector: USDA CVB Licensing Studies- Safety 6

7. 5.Supplemental Safety Studies Milk safety  10 lactating cows and nursing calves; milk collected daily on days 0-4 and days 7, 10, and 14  None of the milk samples from any of the vaccinated cattle tested positive for the adenovirus vector  Study conclusion: Vaccine is not shed in the mammary tissue of cattle after IM inoculation Vaccine biodistribution and persistence in cattle  Very low level of vaccine vector detected at inoculation site and draining lymph node up to 72 hrs post- inoculation  Results published:  Montiel, NA. et. al. 2013. Vet. Immunol. Immunopathol. 151(1-2):37-48  Montiel, NA. et al. 2012. Vaccine. 30(9):1690-1701. FMD Virus Vaccine, Serotype A24, Live Adenovirus Vector: USDA CVB Licensing Studies- Safety 7

8. Minimum Protective Dose (MPD) Studies: 14 days post single dose vaccination (n=34 vaccinates) 94 % SVN positive 97% protection against generalized disease 91% protection against plasma viremia (VI, rRT-PCR) 7 days post single dose vaccination (n=10 vaccinates) 50% SVN positive 100% protection against generalized disease 90% protection against plasma virema (VI, rRT-PCR) Product release dose (‘normal potency’) is 15.8 X MPD Supplemental Information: FMD Virus Vaccine, Serotype A24, Live Adenovirus Vector: USDA CVB Licensing Studies- Efficacy 8 100% (12/12) protected against generalized disease and viremia following direct contact challenge at 1 week post- single dose vaccination (Grubman, MJ, et. al. 2010. Future Virol. 5(1):51-64.) 88% (91/104) of the vaccinated animals tested from the Field Safety Study had positive FMDV SVN titers (>0.9 log10) at approximately 12 months post-single dose vaccine administration

9. Ad5FMD Vaccine Candidate Pipeline 1.14 AdFMD monovalent vaccine candidates efficacy tested 2.9 AdFMD vaccine candidates have been identified for master seed virus production 4 MSVs have CVB approval, 1 is very near approval and 4 will be submitted in the next quarter for approval. 3.AdFMD candidates for 6 of 7 major serotypes Multiple, serotype A candidates Multiple serotype O candidates Includes vaccine candidates for most recent outbreaks in S. Korea (O/South Korea/2010) and Egypt (SAT2/Egypt/2012) 9

10. 10 The question remains, how will this vaccine and companion diagnostic platform perform in an endemic and/or outbreak situation?

11. International Vaccine and Diagnostic Trial Objectives  Demonstrate effectiveness of Ad5 vectored, multivalent developmental vaccine in an outbreak and/or endemic situation  Critical to establishing efficacy and promoting commercial potential.  Demonstrate process for using vaccine and companion Differentiating Infected from Vaccinated Animals (DIVA) diagnostic in a disease situation  Build international partnerships to facilitate future trials in endemic settings 11

12. Overarching Goal Identify and foster the development of collaborative opportunities for international partnerships to further advance the state of the art for FAD vaccines and diagnostics 12 Build enduring partnerships for the future

13. Tasks Year 1Year 2Year 3Year 4 13 1 - Workshop(s) to identify partner countries and detailed plan for case-control study and validating 3B DIVA diagnostic assay in endemic countries 2 - Production of additional master seed viruses needed for the bi- or tri-valent vaccine Analysis of data and final report Procurement of required vaccine and diagnostic kits Begin field study, initial study period of 6 months If vaccine efficacious at 6 months, continue study to 12 and 18 mo. time points to obtain information related to duration of immunity, effect of boostering, and serological response to vaccine Future possibility of additional trials with African or Classical Swine Fever vaccines, or additional FMD vector platforms in other countries Begin in country planning for vaccine trial

14.  A series of meetings/workshops to establish a common understanding of DHS goals and objectives  Discuss logistical considerations and how to best succeed at the execution of an international with Adenovirus Vectored vaccine and the companion 3B ELISA (short term)  Establish general recommendations for international field trials  Identify potential field trial locations and partners for current and future collaborations (Notice of Intent to CRADA)  Release an “Expression of Interest” for potential partners to respond to once the study requirements and design are established Vaccine and Diagnostic Trial Approach 14

15.  Defining Requirements for International Field Trials for Conventional and Next-Generation Foot and Mouth Disease Virus (FMDV) Vaccines and Diagnostics  Washington, DC June 10-12th, 2014  The project working groups have reevaluated all of the recommendations made at the workshop and made revisions where necessary.  Preference for a cluster randomized trial and baseline epidemiologic study in partner country prior to the field trial to better inform the study design.  Finalized the criteria for country selection and developed a country outreach strategy. Also, developing the expression of interest to go to partner countries.  Starting to work through regulatory requirements but will need more input when a partner country is selected. Project Progress to Date 15

16. Questions? 16 If interested in additional information please contact; Roxann Motroni DVM, PhD AAAS Fellow, Agricultural Defense Chemical and Biological Defense Division Homeland Security Advanced Research Projects Agency Science and Technology Directorate Roxann.b.Motroni@associates.HQ.DHS.GOV

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