Fig. 6

Association of tumor-associated macrophages infiltration with PD-1^Int and TIM3⁺PD1^Hi CD8⁺ TILs. a-c, Correlation analysis between the proportion of tumor infiltrating CD8⁺TIM3⁺PD1^Hi (a), CD8⁺TIM3⁻PD1^Hi (b) and CD8⁺PD1^Int (c) among CD8⁺PD1⁺ TILs and the proportion of PDL1⁺ TAMs within CD68⁺ macrophages per core respectively. Correlation is evaluated by the Spearman correlation coefficient. d, Representative multiplex immune-fluorescence image to show the staining for CD8 (yellow), PD1 (green), TIM3 (red), CD68 (cyan), PDL1 (magenta) in the HCC tumor tissue. Cellular phenotype of the fluorescence image depicted the spatial location of CD68⁺PDL1⁺ (blue dots), CD8⁺TIM3⁺PD1^Hi (red dots), CD8⁺PD1^Int (green dots) in the situ tumor tissue. Solid plots and dash line connected the nearest cells within 20 μm from the CD8⁺TIM3⁺PD1^Hi and CD8⁺PD1^Int to the CD68⁺PDL1⁺ respectively. Scale bar, 200 μm. (e) The infiltrating density of PDL1⁺ TAMs within the indicated hierarchy distances of CD8⁺TIM3⁺PD1^Hi and CD8⁺PD1^Int in the HCC tumor tissues, respectively. Error bars indicated median with interquartile range. Significance was assessed by Wilcoxon matched-pairs signed rank test. ****, P < 0.0001. TAMs: tumor associated macrophages