cDNA cloning of functional T cell receptor gamma/delta chains expressed in human peripheral blood lymphocytes

We have identified in earlier studies two V delta rearrangements corresponding to a 4.5-kb Eco RI fragment detected with a V delta1 probe and to a 7-kb Eco RI band detected with a V delta2 probe. These rearrangements have been found in two human T cell clones, F6C7 and G6, displaying surface phenotypes unfrequent in human peripheral blood, namely Ti gamma A+ BB3- (F6C7) and Ti gamma A- BB3+ (G6). Herein, we report the sequences of the functional transcripts encoded by these rearranged genes and show that the 4.5- and the 7-kb Eco RI fragments correspond to V1/D3/J delta 3 and to V2/D3/J delta 3 recombinations, respectively. In addition, we have sequenced the V2/D3/J1/C delta transcripts expressed in two clones, AB12 and VTC, which have a Ti gamma A+ BB3+ surface phenotype corresponding to that of most gamma/delta peripheral lymphocytes. Analyses of the delta transcripts expressed by these four cells further strengthen the hypothesis that anti-BB3 and anti-delta-TCS-1 monoclonal antibodies recognize a V delta 2- and a V1/(D)/J delta 1-encoded epitope, respectively. Sequence of the gamma transcripts expressed by AB12 and F6C7 cells shows that they encode a V9/JP/C gamma 1 chain. Finally, we confirm that non-combinatorial diversity in the gamma and delta proteins is generated by both junctional flexibility and N-region addition without any somatic mutation.