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.