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. 2021 Feb 26;12:577766. doi: 10.3389/fimmu.2021.577766

Figure 6.

Figure 6

Flow cytometry analysis of nivolumab effect on PD-1 expression in sarcoma-specific CD8+ T cells activated by PMA/IM. SaOS-2 and SYO-1 cell lines were left untreated or treated with sunitinib for 24 h at concentrations of 20 μM and 15 μM, respectively, and used for monocyte-derived DC pulsing for another 24 h at a ratio of 1:2 (DC:sarcoma cell). DCs loaded with untreated SaOS-2/SYO-1 (- SUN) or sunitinib-treated SaOS-2/SYO-1 cells (+ SUN) were then used to stimulate and restimulate autologous CD3+ T cells at a ratio of 1:10 (DC:T cell). 24 h after restimulation, the stimulated CD3+ effector T cells were activated by PMA/IM in the presence or absence of nivolumab (20 µg/ml) for 4 h and the reaction was stopped by adding Brefeldin A. The percentages of IFN-γ-producing CD3/CD8+ T cells were evaluated by flow cytometry. The values are represented as mean ± SEM of 3 independent experiments; *p < 0.05; **p < 0.01.