DHS S&T works with many partners to develop vaccines

In vitro FMD VLP Vaccine Production

DEVELOPMENT OF NEXT-GENERATION VACCINES AND DIAGNOSTICS FOR TRANSBOUNDARY ANIMAL DISEASE PREPAREDNESS

Roxann Motroni1, John Neilan2, Max Rasmussen2, Chungwon Chung2, Michael Puckette2, David Brake3, Barbara Kamicker4 1U.S. Department of Homeland Security, Science and Technology Directorate, Chemical and Biological Defense Division, Washington, D.C., USA

2 U.S. Department of Homeland Security, Science and Technology Directorate, Plum Island Animal Disease Center, Greenport, NY, USA 3 BioQuest Associates, LLC, Plum Island Animal Disease Center, Greenport, NY, USA 4 Leidos, Plum Island Animal Disease Center, Greenport, NY, USA

Funding for this program is provided by Department of Homeland Security (DHS) Science and Technology Directorate (S&T) Homeland Security Advanced Research Projects Agency (HSARPA) Chemical and Biological Defense Division. Funding for Plum Island Animal Disease Center is provided by DHS S&T, Research and Development Partnerships (RDP), National Bio and Agrodefense Facility (NBAF) Program Executive Office.

ACKNOWLEDGEMENTS

DHS S&T and Plum Island Animal Disease Center

Target FMD Vaccine Product Profile

ABSTRACT As part of its mission to accelerate the development of countermeasures to mitigate the consequences of the introduction of emerging and/or transboundary animal diseases into the U.S., the Department of Homeland Security Science and Technology Directorate’s Homeland Security Advanced Research and Projects Agency in conjunction with Plum Island Animal Disease Center funded the research, development, test and evaluation of several next-generation, rapid-response vaccine platforms and diagnostic kits for use in a transboundary animal disease outbreak. The first licensed Foot and Mouth Disease vaccine that allows differentiation between infected and vaccinated animals and can be produced on the U.S. mainland was produced through this program. Ten additional U.S. Department of Agriculture Center for Veterinary Biologics approved master seed viruses for vaccine production have been produced. These vaccines have shown greater than 80 percent efficacy using OIE challenge models following a single vaccination and are being transferred to a commercial partner. The program also licensed the first diagnostic kit for FMD that can be produced on the U.S. mainland and provides a result in less than three hours. Through the use of extensive public-private partnerships, this program continues to work to produce next-generation vaccines and diagnostics that suit the needs of both the developing and developed world. Focused on vaccine vector improvement, testing of adjuvants, and development of novel rapid response platforms that can be adapted for emerging or re-emerging threats, this program works to decrease the biological threat from transboundary animal diseases, improve animal health and increase economic security.

Figure 1. Depiction of the development of the Adeno-FMD vaccine and the creation of FMD protein. The Adenovirus is produced on a cell line that expresses replication-essential gene products of the adenovirus. The virus is purified and formulated in adjuvant. Once injected in the host, the recombinant virus infects host cells and can not replicate but expresses FMD capsid proteins. C. Schutta et al. Vaccine. 2016. 34(27): 3214-3220.

First conditionally licensed Adeno-FMD Vaccine in the U.S. for a single serotype that is also DIVA compatible

- Demonstration of > 500% reduction in cost of raw goods for commercial viability - Production of 10 USDA Center for Veterinary Biologics approved Master Seed Viruses (MSVs) against the high and medium priority strains of FMD that show >80% efficacy in challenge studies. These have transitioned to USDA and a commercial partner for further development. Discovery and patent filing for a novel 3C protease mutant that decreases the toxicity

to cells and maintains processing activity allowing for generation of VLPs in bacterial and mammalian cells.

Seroconversion in guinea pig and cattle models using in vitro produced VLPs. -Puckette et al Journal of Virology 2017 Sep 6. doi: 10.1128/JVI.00924-17

First licensed rapid-response (three-hour) FMD Diagnostic kit that can be manufactured in U.S. Currently funding Nipah oral vaccine efficacy trial in pigs in collaboration with U.S. National Institute of

Health/NIAID Rocky Mountain Labs and Canadian Food Inspection Agency.

Commercial Partners

DHS targeted funding to TAD product candidates that met the following requirements: • Efficacy: >80% (Most >90%) by OIE Challenge Standards • Safety: No adverse and minimal local reactions in food producing animals • Singe Dose Vaccination • Rapid Onset of Immunity- serum virus neutralization titers in >50% and >80% of vaccinates- 1 & 2

weeks post vaccination • Duration of Immunity: ≥ 6 months • Route/Ease of Administration- SQ, IM, ID • Commercial Feasibility- Low Cost of Goods • Stable over wide variety of storage temperatures and time • Rapid Response Platforms • Multivalent and Panvalent Technologies

The U.S. Department of Homeland Security (DHS) Science and Technology Directorate (S&T) Chemical and Biological Defense Division (CBD) works to strengthen preparedness against chemical, biological, and agricultural threats through improved threat awareness, enhanced surveillance, advanced detection and diagnostic technologies, and robust response and recovery countermeasures. The agricultural biodefense mission seeks to enhance current capabilities and develop state-of-the-art countermeasures for high-priority transboundary animal diseases (TAD). This mission is accomplished by short- and long-term research, development, test and evaluation of vaccines and diagnostics in conjunction with Plum Island Animal Disease Center (PIADC). Technology gaps are identified in coordination with internal and external stakeholders such as USDA, the livestock industry and animal health pharmaceutical industry.

Adenovirus-vectored FMD Subunit Vaccine

Next Generation FMD Vaccines are needed Foot-and-mouth disease (FMD) is a highly contagious disease affecting cloven hoofed animals (e.g. cattle, swine, sheep) and threatens worldwide food security, production and trade. Approximately 2.35 billion doses of FMD vaccines are administered yearly in endemic areas • Inactivated virus used for monovalent and multivalent vaccines

• Carries risk of accidental outbreaks due to release from high containment manufacturing • Difficult to differentiate infected versus vaccinated animals (DIVA) • Can not be produced in U.S. mainland

To create safer and more efficacious vaccines, molecular FMD vaccines express only the FMDV proteins necessary for assembly of virus-like particles (VLPs) are being developed.

• No risk of accidental outbreak • DIVA potential

DHS bridges the “valley of death” by supporting test and evaluation research of product candidates that have shown promise in target host proof-of-concept studies and de-risks the early and late stage development costs for industry partners. DHS’s extensive experience in public-private partnerships and interagency engagement contribute to the success of the program. Multiple mechanisms exist to allow further collaboration, including material transfer agreements (MTAs), cooperative research and development agreements (CRADAs), interagency agreements and contracts.

Figure 2. A. FMDV VLP production in bacterial cells seen as crystalline array structures. B. Representation of an FMDV capsid using electron microscopy images of FMDV virus-like particles produced in bacterial and mammalian cells. C. FMDV VLP production observed in a mammalian cell line. M. Puckette et al. J. Virol. 2017. Sep 6. doi: 10.1128/JVI.00924-17.

DHS advances transboundary animal disease countermeasures

3B ELISA Diagnostic Kit licensed for production in U.S. DHS in collaboration with USDA Agricultural Research Service (ARS) and Animal Plant Heath Inspection Service (APHIS), the Institute for Infectious Animal Diseases (IIAD) and VMRD recently licensed a commercial serological diagnostic kit for FMD. It has several advantages over other commercially available kits.

3-hour turn around time Able to be produced on U.S. mainland Licensed for use in cattle, pigs and sheep serum samples High sensitivity and specificity

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B. C.