Differential expression of a panel of optimized melanoma-specific TCRs in peripheral blood T cells. (A) Jurkat/MA cells were transduced with titrated aliquots (Figure S1) of viral supernatants of vectors encoding the gene-optimized Mart-1–specific TCRs 1D3, 2C2, and DMF4 or the Gp100-specific TCR R6C12. Transduced cells were stained with anti-TCRαβ 4 days after transduction. Histograms show levels of TCR expression for the different TCRs. (B) Retroviral aliquots as used in panel A were used to transduce human peripheral blood T cells. TCR expression was determined by staining with anti-CD8 and A2.1-Mart-1_{(26-35, 27 A>L)} or A2.1-Gp100_{(209-217, 210 T>M)} tetramers. The numbers in the upper-right and lower-right corners indicate the percentage of tetramer⁺ CD8⁺ and tetramer⁺ CD8⁻ cells, respectively.