Flow cytometric analysis of perforin, granzyme B, IFN-g, TNF-a, IL-2, and PD-1 expression by CD8+ T cells in PBMCs and tumor tissues.

Freshly isolated PBMCs and digested tumor tissues from 10 paired patients were stained by flow cytometry. Flow cytometric plots were pregated on TCRab+CD8+cells, excluding dead cells and naïve T cells (CD45RA+CCR7+ T cells). (A) Flow cytometric plots of perforin and granzyme B intracellular staining among CD8+ T cells are shown. (B) The frequencies of the expression levels of perforin and granzyme B in CD8+ T cells from paired PBMCs and tumor tissues are shown (n = 10). (C) Flow cytometric plots of IFN-g, TNF-a, and IL-2 intracellular staining among CD8+ T cells are shown. (D) The frequencies of the expression levels of IFN-g, TNF-a, and IL-2 in CD8+ T cells from paired PBMCs and tumor tissues are shown (n = 10). (E) Representative flow cytometric plots of PD-1 expression among CD8+ T cells are shown. (F) The frequencies of the expression levels of PD-1 in CD8+ T cells from paired PBMCs and tumor tissues are shown (n = 10). P-values were calculated by the Wilcoxon signed-rank test between paired PBMCs and tumor tissues. A single asterisk indicates P P < 0.01.