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. 2020 Sep 7;8(2):e001187. doi: 10.1136/jitc-2020-001187

Figure 2.

Figure 2

B cell activation capacity of follicular helper T cells (Tfh) from isotype-treated and antiprogrammed cell death protein 1 (anti-PD-1)-treated mice. (A) Quantification of interferon (IFN)γ, interleukin (IL)-4, IL-17 and IL-21 production in circulating T follicular helper cells (cTfh) and Tfh from spleen of anti-PD-1-treated and isotype-treated mice. For stimulation experiments, Tfh were isolated and pooled from isotype-treated and anti-PD-1-treated mice (n=7 and n=4, respectively). They were subsequently treated for 5 hours with PMA/ionomycin (P/Io) plus brefeldin for the last 2 hours, and cytokine production was assessed by flow cytometry. (B) Representative plots of data showed in (A). Percentages of IFNγ+, IL-4+, IL-17+ and IL-21+ subpopulations are included. (C) Histograms show CD69 expression in Tfh from different conditions stimulated as in (A) to confirm effective cell activation. (D) Quantification of IFNγ, IL-4, IL-17 and IL-21 expression in P/Io-stimulated and control naïve CD4+ T lymphocytes from peripheral blood and spleen of anti-PD-1-treated and isotype-treated mice. Naïve T lymphocytes were isolated and pooled from seven isotype-treated mice and four anti-PD-1-treated mice, respectively. (E) Histograms show CD69 expression in T-naïve cells from different conditions stimulated as in (A) to confirm effective cell activation. *P<0.05; ****p<0.0001.