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. 2018 Oct 1;215(10):2520–2535. doi: 10.1084/jem.20180684

Figure 4.

Figure 4.

CXCR5+ TIM-3– CD8+ T cells are efficient in rapidly producing effector molecules. (A–D) Sorted CXCR5+ TIM-3–, CXCR5– TIM-3–, and CXCR5– TIM-3+ CD8+ TILs were stimulated for 3 h with PMA/ionomycin, and CD107a expression and cytokine production was determined by flow cytometry (n = 7; all performed in independent experiments). (A) Representative dot plots showing the analysis of IL-2, TNF, and IFNγ production by NSCLC tumor-infiltrating CXCR5+ TIM-3–, CXCR5– TIM-3–, and CXCR5– TIM-3+ CD8+ T cells following short-term PMA/ionomycin stimulation. Numbers in the dot plots indicate the percentage of cells identified by the gates. (B) Representative histograms showing the expression of CD107a by the indicated subsets after short-term PMA/ionomycin stimulation. Numbers in the histograms indicate the percentage of cells identified by the gates. (C) Pie charts representing the proportion of CXCR5+ TIM-3–, CXCR5– TIM-3–, and CXCR5– TIM-3+ CD8+ TILs expressing and producing different combinations of CD107a, IL-2, IFNγ, and TNF after PMA/ionomycin stimulation (n = 7). Frequencies were corrected by background subtraction as determined in unstimulated controls. **, P < 0.01; ***, P < 0.001; partial permutation tests, using SPICE software. (D) Frequency of CXCR5+ TIM-3–, CXCR5– TIM-3–, and CXCR5– TIM-3+ CD8+ TILs expressing and producing different combinations of CD107a, IL-2, IFNγ, and TNF after PMA/ionomycin stimulation (n = 7). Data are given as mean ± SEM. *, P < 0.05 versus CXCR5+ TIM-3– CD8+ TILs; +, P < 0.05 versus CXCR5– TIM-3– CD8+ TILs, paired Wilcoxon t test.