Figure 5.

Tetramer binding of WT and ΔTCR–transduced T cells. (A) vα3_WT/vβ3_WT (bold line) and vα3_c-90/vβ3_WT (dashed line) TCR–transduced CD4⁺ T cells were stained with p53_264-272Tet–PE at 37°C at the indicated tetramer concentrations. (B) vα3_WT/vβ3_WT (□) and vα3_c-90/vβ3_WT (▪) TCR–transduced CD4⁺ T cells were tested for binding to p53_264-272Tet–PE after 60 min by flow cytometry as indicated in A, and the avidity (K_D) of tetramer binding to T cells was determined by Scatchard analysis, with the 95% confidence band indicated by thin dashed lines. p53_264-272Tet–PE binding to vα3_WT/vβ3_WT (□) and vα3_c-90/vβ3_WT (▪) TCR–transduced (C) CD8⁺ and (D) CD4⁺ T cells was determined at 37°C at the indicated time points by flow cytometry, and t_1/2 for half maximal binding was determined. After p53_264-272Tet–PE binding to vα3_WT/vβ3_WT (□) and vα3_c-90/vβ3_WT (▪) TCR–transduced (E) CD8⁺ and (F) CD4⁺ T cells, the cells were washed and incubated with the HLA class I blocking antibody W6/32 at 37°C. The percent inhibition of tetramer binding and t_1/2 for inhibition were calculated. All data are representative of at least two independent experiments.