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Analysis of the population of human T cell receptor gamma and delta chain variable region subfamilies by reverse dot blot hybridization.

We have developed a simple method to analyze the population of T cell receptor (TCR) gamma and delta chain variable (V) region subfamilies by the application of reverse dot blot hybridization, which was originally developed for the analysis of human HLA-DR polymorphism. The four oligonucleotides corresponding to each TCR-gamma V region subfamily and the six oligonucleotides to each TCR-delta V region gene were synthesized, and tailed with dTTP. The cDNA was amplified by ligation-mediated PCR in the presence of biotinylated deoxynucleotides. Hybridization between immobilized specific oligoprobes and biotinylated target DNA was nonradioactively detected by a reaction using alkaline phosphatase. The population of the V region subfamilies of TCR-gamma and -delta in PBL analyzed by reverse dot blot hybridization described here showed good correlations with the result of colony hybridization.