1a.Objectives (from AD-416):
To improve Foot-and-Mouth Disease Virus (FMDV) vaccine potency and duration of immunity, ARS, PIADC and the University of Vermont (UV) 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):
ARS, PIADC in cooperation with the University of Copenhagen is developing state of the art technology for analysis of FMDV vaccine performance in cattle. The product of this collaboration, MHC tetramers for tracking T cell responses to vaccination, will be used to analyze genetically defined cattle. The University of Vermont collaborators maintain a genetically homogenous herd of Holstein cattle that make it possible to use the tetramer technology to analyze the response to vaccination with FMDV new vaccines. ARS, PIADC will collaborate with UV faculty to analyze responses to infection of these cattle with FMDV and mastitis.
During FY 2013 we conducted the following activities:
1: Analysis of bovine lymphycote (BoLA) Class I expression in 118 Holstein cattle has identified 10 dominant BoLA class I alleles which are expressed in greater than 90% of animals tested. We have selected 9 alleles which occur at population level frequencies of greater than 20% for further analysis. Patterns of allelic expression are being identified in individual animals contributing to our understanding of class I and II gene expression and the potential relationship between class I and class II expression.
2: A total of 15 class I molecules have been synthesized by collaborators from the University of Copenhagen. Thirteen of these molecules were found to be functional in peptide binding assays. Among the 9 high-frequency target molecules described under objective 1, only one molecule, 2*01601,was found to be non-functional in the peptide binding assays. This leaves 8 target molecules (expressed in cattle at UVM) where peptide-binding motifs have been determined by positional scanning combinatorial peptide libraries (PSCPL). Binding affinity and stability were determined using a standardized pool of peptides.
This work will allow us to apply the netMHCPan algorithm to identify poteintial T cell epitopes of FMDV that will be induced in these cattle. This will allow us to generate new reagents for the rapid and accurate assessment of T cell responses to FMDV vaccination and infection.
No technologies were transferred and no publications were produced during this period.