Figure 3.

mTOR signaling regulates the effector function of CD8+ T cells. Purified CD8+ T cells were stimulated with αCD3 antibody (5 μg/mL) with or without R848 (10 μg/mL) for 48 h in the presence of rapamycin (2 μM) or Akti (1 μM). (A) mTOR and phosphorylated mTOR in CD8+ T cells were detected by western blotting. The level of phosphorylated mTOR and Akt were further determined by flow cytometry. (B) The activation of CD8+ T cells was assessed by staining with αCD25, αCD44, and αCD69 antibodies. The expression of CD25, CD44, and CD69 in the αCD3+R848 stimulated cells were expressed as fold changes compared to the αCD3 stimulated cells in the corresponding treatment of rapamycin/Akti-1/2. (C) IFN-γ and TNF-α secretion were detected by specific ELISAs. (D) IRF4 expression was shown by representative dot plots and MFI in CD8+ T cells. Data are representative of three independent experiments. Data are presented as mean ± SD. The statistical relevance was determined by One-way ANOVA (B, right panel) or Two-way ANOVA (B, left panel, C,D): *p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant.