Paired cloning of the T cell receptor α and β genes from a single T cell without the establishment of a T cell clone

T cell receptors, which recognize antigen peptides on MHC molecules, are essential probes for the analysis of T cell antigen specificity. The identification of paired T cell receptor (TCR) chains, α / β or γ / δ , usually requires the establishment of T cell clones, which is not always available. In this study, we tried, as an alternative method, the paired cloning of TCR α / β genes directly from a single T cell. T cells were sorted as a single cell from which RNA was extracted. Then, TCR α / β CDR3 regions were amplified from the single cell‐derived cDNA by reverse transcriptase‐polymerase chain reaction to determine their sequences. We successfully identified pairs of TCR α / β genes, and reconstructed the TCR molecule by a bacterial expression system. This strategy makes it possible to obtain recombinant TCR molecules from a single T cell without cellular cloning and promotes the investigation of T cell antigen specificity.