Figure 6.

Expression of suppressive checkpoint inhibitor molecules is not enhanced by T cell activation under hyperstimulatory conditions. Human T cells and polymorphonuclear neutrophil granulocytes (PMN) were isolated from whole blood of healthy donors. PMN were incubated for 72 h in the presence or absence of nor-NOHA (1 mM). T cells were stimulated with anti-CD3/anti-CD28-tagged beads in the PMN-SN or in medium with or without the addition of 1 mM nor-NOHA for 24 h and 48 h. T cells were then harvested and stained with (A) anti-PD-1, (B) anti-TIM-3, (C) anti-LAG-3, (D) anti-CTLA-4, or (E) anti-TIGIT antibodies and analyzed by flow cytometry. A summary of expression (MFI of CD3⁺ cells) of n=3 independent experiments with blood from different donors is demonstrated. Statistical calculations were performed with one-way ANOVA and Tukey´s post hoc test (**p < 0.01, *p < 0.05).