Comprehensive analysis of the functional TCR repertoire at the single-cell level

A Vβ TCR repertoire is analyzed for understanding the T-cell population in the immune response. However, the TCR repertoire of the Vα–Vβ pair is difficult to analyze because no suitable analytical method is available. Here, we have applied the single-cell 5′-RACE method for amplifying TCR cDNAs from single T-cells and analyzed the repertoire of Vα–Vβ pairs in human T-cells that responded to a superantigen, SEB. We found that the TCR Vβ profile of the SEB-stimulated CD4+ T-cells was in accordance with the previous reports, that the TCR Vα profile also exhibited a prominent difference, and that the TCR Vα–Vβ pairs of the SEB-responding T-cells were promiscuous. We have also found a significant dual TCRα expression in single T-cells. This is the first report of a comprehensive analysis of the functional repertoire of Vα–Vβ pairs at the single T-cell level. This novel method may contribute to TCR-based immunotherapeutics.