2010 Annual Report 1a.Objectives (from AD-416) To improve Foot-and-Mouth Disease Virus (FMDV) vaccine potency and duration of immunity, we will study the cellular immune response to infection and the ability to refine the killed virus vaccine for FMDV or the recombinant empty capsid vaccine. The objective of this agreement is to analyze T cell responses to FMDV infection in swine and cattle. 1b.Approach (from AD-416) The state of the art for analysis of the cellular immune response of T lymphocytes to vaccination is: identification of T cell epitopes, mapping of those epitopes to the presenting transplantation antigens known as histocompatibility complex (MHC) molecules and developing MHC tetramers for tracking T cell responses. This technology is used extensively in laboratory mice but is expensive in a genetically diverse human population. Even given the expense, the technology is being applied in the clinic more and more often. Our preliminary studies indicate common cattle breeds, such as Holstein, can be analyzed using this technology. The University of Copenhagen will generate the information required to design and test MHC tetramers for analysis of cellular immune reponse to Foot-and-Mouth Disease Virus (FMDV). ARS, PIADC will utilize this information to analyze responses of cattle and swine to vaccination and FMDV infection. 3.Progress Report The objectives of this collaboration are to improve Foot-and-Mouth Disease Virus (FMDV) vaccine potency and duration of immunity through the study of the cellular immune response to infection and the response to the killed virus vaccine for FMDV or the recombinant empty capsid vaccine in swine and cattle. Specifically, the mapping and identification of potential T cell epitopes capable of being bound by the presenting major histocompatability complex (MHC) molecules will lead to development of MHC tetramers for tracking T cell responses. Generated recombinant porcine beta 2 microglobulin , and the recombinant porcine MHC’s; SLA-1*0401, SLA-2*04:01 and SLA-3*04:01. Porcine beta 2 microglobulin, SLA-1*04:01, and SLA-2*0401 have successfully been used to generate functional peptide binding assays and are the subject of paper in preparation. We have validated that our recombinant SLA-3*0401 molecules are functional and stable, and at the same time identified its specificity. This information can now be used to search for SLA-3*04:01 restricted porcine T cell responses. The major accomplishments over the life of this project include: 1. Establishment of functional porcine MHC class I molecules and analysis of their specificity. 2. Establishment of predictors covering porcine MHC class I molecules. 3. Establishment of recombinant reagents suitable for tetramer staining of porcine T cells. The following technologies have been transferred in FY 2010: 1. The information and reagents have been transferred from Copenhagen to ARS, PIADC. 2. The ability to generate tetramers has been transferred from Copenhagen to ARS, PIADC. This project was monitored through email and telephone exchange as well as site visits to ARS, PIADC. In addition, a University of Copenhagen PhD student is currently conducting work at ARS, PIADC on this project.