Human leukocyte antigen (HLA) genes are the most polymorphic genes in the human genome. Knowledge about HLA polymorphism in relation to possible peptide-based, T-cell-restricted vaccination protocols is important for understanding the physiology of T-cell recognition and to improve strategies of T-cell antigen-specific vaccination. During the 2-year project, the HLA Laboratory has developed and perfected techniques for high-resolution typing of HLA class I and class II molecules using polymerase chain reaction (PCR) techniques. This technique was compared for yield and accuracy of information to other techniques, including directed heteroduplex analysis and automated sequencing of genomic DNA. Multiple new HLA alleles were discovered in the HLA Laboratory last year by combining these strategies. It was concluded that because of the ever-increasing number of HLA alleles, the only definitive strategy for typing is sequencing of HLA alleles. This could be achieved by using a high throughput sequencer such as the 3700 ABI. The next year will be devoted to the development of high throughput HLA sequencing strategies. In addition, the HLA Laboratory has developed a comprehensive method for typing T-cell receptor (TCR) based on Vb-specific, PCR-based amplification and directed heteroduplex analysis. Most recently, this laboratory has perfected soluble, epitope-restricted HLA tetramer technology and the measurement of gene expression. These techniques allow high-resolution/high efficiency characterization of TCR responses to HLA class I restricted vaccines. These techniques are currently being utilized for the immunologic monitoring of patients undergoing peptide-based vaccination. - HLA, Immunogenetics, vaccination, TCR, polymorphism,
