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. 2017 Jun 30;8:768. doi: 10.3389/fimmu.2017.00768

Figure 2.

Figure 2

GrB-producing B cells from healthy individuals inhibited Th1 and Th17 cell responses. Purified peripheral blood CD19+ B cells and CD4+CD25− T cells from healthy individuals (n = 10) were cocultured with anti-CD3 antibody (3 µg/ml), anti-CD28 antibody (3 µg/ml), CpG stimulation (10 µg/ml), recombinant human IL-21 (50 ng/ml), and anti-B-cell receptor (10 µg/ml) in the presence of anti-GrB antibody or isotype control. After 3 days, the cells were stained with anti-CD4 antibody, anti-IFN-gamma antibody, anti-IL-17A antibody, anti-TCR-zeta antibody, annexin V, and 7-AAD and analyzed by FACS. (A) The representative frequencies of Th1 cells measured by FACS were showed, which were obviously increased in the presence of anti-GrB antibody. (B) The frequencies of Th17 cells under the blockade of GrB were detected by FACS. The frequencies of TCR zeta+ T cells (C) and apoptotic T cells (D) were detected by flow cytometry and the representative flowcharts as well as the statistical results were shown. *P < 0.05, **P < 0.01, ***P < 0.001, by Wilcoxon signed-rank test. GrB, granzyme B.