Figure 5

Tumor cell recognition by CMV tetramer⁺ T cells. TIL 5 TCR-transduced and untransduced CMV-stimulated bulk cultures were cocultured in the presence of 10 μg/ml brefeldin A for 5 hours in a 1:1 ratio with A2⁺, MART⁺ tumor cells (SK23 MEL, 1300 MEL) or A2⁺, MART-1^- tumor cells (UOK131 RCC). Cells were then collected, stained with PE-conjugated HLA-A2 MHC CMV tetramer, fixed, permeabilized, and stained with FITC-conjugated anti-IFN-γ mAb. Samples were analyzed using two-color flow cytometry. The percentage of dual positive staining cells (upper right quadrant) is as indicated.