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Fig. 4 | Journal for ImmunoTherapy of Cancer

Fig. 4

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

Fig. 4

Increased infiltration of exhausted PD1Hi CD8+ T cells in HCC tumor tissues revealed by multiplex immunohistochemistry. a, Representative multiplex immunofluorescence images to show the distribution of CD8+ T cell subsets in peri-tumor and tumor: red arrows (CD8+TIM3+PD1Hi), purple arrow (CD8+TIM3PD1Hi), green arrow (CD8+PD1Int) and cyan arrow (CD8+PD1). Tissue slides were stained by TSA method and scanned at 20x by Vectra3.0 Automated Imaging System. Scale bar, 50 μm. b, t-SNE analysis of CD8+ T cells from paired peri-tumor and tumor tissues validated the distinct classification of four CD8+ T cell subsets. c, Relative distribution analysis of total CD3+ T cells in paired peri-tumor and tumor tissues by dividing the total CD3+ T cells into CD8+ (cytotoxic T cells) and other cells (upper panel). CD8+ T cells were further split into CD8+PD1, CD8+PD1Int and CD8+PD1Hi subpopulations (middle panel); and finally CD8+PD1+ T cells were separated into CD8+TIM3+PD1Hi and CD8+TIM3PD1Int basing on the TIM3 expression (lower panel). d-h, Comparisons of the proportions of CD8+ among CD3+ T cells (d) and CD8+PD1Hi (e), CD8+PD1Int (f), CD8+TIM3+PD1Hi (g) and CD8+TIM3PD1Hi (h) among CD8+PD1+ T cells between paired peri-tumor and tumor tissues in training cohort (n = 358). Error bars indicated median with interquartile range. 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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