Location: Foreign Animal Disease Research
2012 Annual Report
During FY 2012, the analysis of bovine leukocyte antigens (BoLA) Class I expression in 81 Holstein cattle from Nordic Farm, a UVM Dairy Center of Excellence Research herd, has identified 9 dominant alleles that are expressed in 23 to 44 percent of animals tested. Expression of these alleles has been confirmed by cloning and sequenceing of BoLA class I genes in 17 animals. Overall 99 percent of animals tested are positive for at least one of these alleles. Further patterns of allelic expression are being identified in individual animals contributing to our understanding of class I gene expression. We have also completed class I expression typing for 15 bovine primary fibroblast cell lines and 4 bovine immortalized epithelial cell lines. We have synthesized 6 class I molecules and an additional 5 molecules are in the pipeline for synthesis. At least 4 of these BoLA class I molecules will be used in peptide binding and cytotoxic lymphocyte (CTL) response assays. This panel of 11 BoLA class I molecules will be made available as a resource to other scientists.
We have successfully transferred reagents and technologies and adapted new technologies for the development of key tools to be used in immune response assays. Specifically we have transferred the gene constructs and applied methods for recombinant protein expression of a panel of bovine cytokines using an adenovirus expression system. We have demonstrated biological activity of these cytokines and established a pipeline for production of monoclonal antibodies that recognize these lymphocyte markers.
During FY 2012 the recombinant replication defective adenoviral vector expression systems have been transferred from ARS, PIADC to University of Vermont. MHC/peptide binding analysis has been transferred from the collaborators at University of Copenhagen to University of Vermont.
No publications have been produced during FY 2012.