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Fig. 3 | Journal for ImmunoTherapy of Cancer

Fig. 3

From: PD1Hi CD8+ T cells correlate with exhausted signature and poor clinical outcome in hepatocellular carcinoma

Fig. 3

Assessment of pro/anti-inflammatory cytokines profiling and cytotoxic potential of tumor infiltrating PD1Hi CD8+ T cells. a-b, Representative flow cytometric plots (a) and accumulated data (b) to show the pro-inflammatory cytokines IFN-γ, IL-2, TNF-α, GM-CSF, IL-4, IL-22, IL-17a and anti-inflammatory IL-10 secreting profile of tumor infiltrating PD1Hi, PD1Int and PD1 CD8+ T cells following the stimulation of PMA, ionomycin and BFA for 5 h (n = 9). c, Representative flow cytometric overlays of intracellular Granzyme B, perforin and CD107a expression of tumor infiltrating PD1Hi, PD1Int, and PD1 CD8+ T cells. Granzyme B and perforin were detected from fresh samples (n = 11) and CD107a expression was measured after overnight stimulation of coated anti-CD3 (10 μg/mL) and soluble anti-CD28 mAb (1 μg/mL) (n = 8). d, Apoptosis quantification of HCCLM3 tumor cell line after co-culture with anti-CD3 (10 μg/mL)/CD28 mAb (1 μg/mL) stimulated CD8+ T cells subsets for 18 h and pooled statistics of viable HCCLM3 tumor cells shown in bar graph (n = 6). Significance was assessed by Wilcoxon matched-pairs signed rank test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001

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