FIGURE 5.
Ag-specific TEFF exhibit reduced responsiveness to anti–TIM-3 Ab compared with TEM. Ag-specific CD8 T cells were cultured as in Fig. 1 (standard) or stimulated an additional three times with aAPC loaded with MART1 peptide (terminal), as described in Materials and Methods, and analyzed by flow cytometry for expression of (A) CCR7 versus CD45RA and (B) CD45RO, TIM-3, CD127, and CD25. Cells stimulated with standard or terminal protocol were treated with anti–TIM-3 mAb (black bars) or isotype control (gray bars), and analyzed for (C) expression of CCR7 versus CD45RA by flow cytometry and (D) cytokine production. Data shown are representative of four to six biological replicates per treatment condition performed in three independent experiments.