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. Author manuscript; available in PMC: 2019 Dec 15.
Published in final edited form as: Clin Cancer Res. 2018 Aug 21;24(24):6236–6247. doi: 10.1158/1078-0432.CCR-18-1100

Figure 3. PD-1 Blockade Increases Expression of IL-10.

Figure 3.

(A) CD3+ T-cells were isolated by negative magnetic separation from baseline patient PBMC, cultured for two hours with IgG or αPD-1 (5ug/mL) and then activated with CD3/CD28 Dynabeads for 72 hours. Supernatant IL-10 levels were assessed in a Luminex assay. (B) T-cells were treated with IgG or αPD-1 (5ug/mL), and stattic (200nM) or DMSO control. IL-10 production was assessed by Luminex. (C-D) Isolated CD3+ T-cells were evaluated by intracellular flow cytometry for IL-10 expression after treatment with IgG or αPD-1 (5ug/mL) (C) without or (D) with the addition of CD3/CD28 Dynabeads. Indicated cell populations were gated on and percentage of IL-10 positive cells determined. All bar graphs illustrate means +/− SEM for triplicates/group. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.