Fig 2: CD86 expression on CD8+ T cells in GILs(A) Representative histogram of CD86 expression on CD8+ T cells of Naïve spleen cells (SPCs), postoperative day (POD)14 SPCs, and POD14 graft-infiltrating lymphocytes (GILs). Data are representative of four independent experiments.(B) Quantitative data of the median fluorescence intensity (MFI) of CD86 expression on CD8+ T cells of Naïve SPCs, POD14 SPCs and POD14 GILs, as evaluated by flow cytometry (n = 4 for each group). Statistical analysis by one-way ANOVA followed by a post hoc test. ∗∗∗∗p < 0.0001. Values are shown as the mean ± SEM.(C) The gating strategy of CD86low, CD86mid, and CD86high CD8+ T cell subsets based on the CD86 expression level (Left). The proportion of CD86low, CD86mid, and CD86high CD8+ T cell subsets among CD8+ T cells of Naïve SPCs, POD14 SPCs, and POD14 GILs, as evaluated by flow cytometry (n = 4 for each group) (Right). ∗∗∗∗p < 0.0001 by one-way ANOVA with Tukey’s post hoc test vs. POD14 SPCs. Values are shown as the mean ± SEM.(D) Representative immunofluorescence staining of CD86, CD8β, and PD-1 in a hepatic allograft on POD14. Image data are representative of three independent experiments. Scale bar = 10 μm.(E) Representative histogram of PD-1, TIM-3, and CD39 expression on CD86low, CD86mid, and CD86high CD8+ T cell subsets of Naïve SPCs, POD14 SPCs, and POD14 GILs. Data are representative of four independent experiments.(F) Quantitative data of the MFI of PD-1, TIM-3, and CD39 expression on CD86low, CD86mid, and CD86high CD8+ T cell subsets of Naïve SPCs, POD14 SPCs, and POD14 GILs, as evaluated by flow cytometry (n = 4 for each group). Statistical analysis by one-way ANOVA followed by a post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Values are shown as the mean ± SEM.(G) The mRNA expression level of CD86 in CD8+ T cells of Naïve SPCs, POD14 SPCs, and POD14 GILs. Statistical analysis by one-way ANOVA followed by a post hoc test. ∗∗∗∗p < 0.0001. Values are shown as the mean ± SEM.(H) The representative flow cytometry analysis of CD86 and CTLA-4 expression on CD8+ T cells of POD14 GILs. Data are representative of three independent experiments.