Figure 5.

Polyclonal CD8⁺ and CD4⁺ T cells from a HLA-A*0201⁺ patient with metastatic rectal cancer transduced with retroviruses encoding the α and β chains of a HLA-A*0201-restricted, NY-ESO-1_157-165-specific TCR recognize DAC-treated HLA-A*0201⁺ CRC cells. PBMC collected from the patient prior to the receipt of adjuvant chemotherapy were transduced with retroviruses encoding the wild type 1G4 NY-ESO-1_157-165-specific TCR or its α95:LY or β51:AI variants, then analyzed by flow cytometry with a NY-ESO-1_157-165/HLA-A*0201 tetramer and anti-CD4 or anti-CD8 mAbs, and tested for cytolytic activity against CRC targets. (A) Flow cytometric analysis of a natively CD8⁺ NY-ESO-1_157-165-specific CTL clone and polyclonal CD8⁺ T cells from the CRC patient transduced with retroviruses encoding the wild type or variant 1G4 TCRs. (B) Cytolytic activity of patient T cells transduced with the wild type 1G4 TCR (black) or its α95:LY (dark gray) or β51:AI (light gray) variants against saline- or DAC-treated SW480 cells. (C) Cytolytic activity of CD4⁺ (light gray) or CD8⁺ (black) 1G4 α95:LY TCR-transduced patient T cells against control (−) or DAC-treated (+) HLA-A*0201 dermal fibroblasts and CRC cells at E:T 5:1. (D) Cytolytic activity of polyclonal CD4⁺ (gray) or CD8⁺ (black) T cells that were isolated from the same rectal cancer patient after 7 months of adjuvant chemotherapy, then transduced with the 1G4 α95:LY variant TCR, against control (broken lines) or DAC-treated (solid lines) SW480 cells. Data shown are mean values +/− SEM.