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. 2018 Dec 18;8:632. doi: 10.3389/fonc.2018.00632

Figure 2.

Figure 2

PD-L1 is involved in the MALT1 protease activity-mediated immunosuppressive property of ABC-DLBCL cells. (A) Flow cytometric analysis of DLBCL cell lines for PD-L1. One representative experiment of three is depicted. The black curve in each histogram show staining by specific mAbs, and isotype control results are shown in gray. (B) DLBCL cells were pretreated with vehicle or VRPR for 12 h prior to exposure to CFSE-labeled Vγ9Vδ2 T lymphocytes for 6 h. Proportions of PD-L1+ DLBCL cells within DLBCL cells were detected by flow cytometry (n = 5). (C) Cytotoxicity of Vγ9Vδ2 T lymphocytes toward ABC-DLBCL cells pretreated with vehicle, anti-PD-L1 antibody, or anti-PD-L1 antibody + VRPR (α-PD-L1+ VRPR) for 12 h (n = 5). Western blot to detect Bcl-10 cleavage products; β-actin was used as the loading control. (D) Flow cytometric analysis of Vγ9Vδ2 T lymphocytes for PD-1 (n = 10). The black curve in each histogram show staining by specific mAbs, and isotype control results are shown in gray. (E) CFSE-labeled DLBCL cells were pretreated with vehicle or VRPR for 12 h prior to exposure to Vγ9Vδ2 T lymphocytes for 6 h. Expression of PD-1 on Vγ9Vδ2 T lymphocytes was detected by flow cytometry (n = 4). (F) Proportion of PD-1+ Vγ9Vδ2 T lymphocytes from the graph in (E) are shown (n = 4). Data are shown as the means ± SD, **p < 0.01; ***p < 0.001.