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. 2020 Jan 28;11:52. doi: 10.3389/fimmu.2020.00052

Figure 3.

Figure 3

Foxp3+ Tregs and IL-10 production are cell-intrinsically controlled by PTPN22 in steady state and LIP. (A) Ab-mediated depletion efficacy of CD4+Foxp3+ Tregs and CD4+Foxp3− conventional CD4 T cells 3 days post anti-CD4 and anti-CD8 injection. Data shown as mean with SD with N = 7 animals per group. Pooled data of two independent experiments. (B) Proportion and absolute numbers of regulatory T cells 2 weeks after T cell depletion in combination with IL-7R blockade and (C) 8 weeks post depletion. Each dot represents one animal. In (B) N = 7–8 animals per group, shown data is pooled of two independent experiments. In (C) N = 4–6 animals per group, one representative experiment out of three independent experiments. Two-way ANOVA with Sidak's multiple comparison test. (D) IL-10 production by CD4 T cells, CD4+CD25+ Tregs and CD4+CD25− conventional T cells 2 weeks post depletion. To analyze IL-10 production cells were re-stimulated in presence of Brefeldin A ex vivo. Each dot represents one animal. Each dot represents one animal. N = 6–8 animals per group, shown data is pooled of two independent experiments. Two-way ANOVA with Sidak's multiple comparison test. (E) Bone marrow from CD45.1 PTPN22 WT and CD45.2 PTPN22 KO donor animals was mixed 1:1 and transplanted in into fully irradiated CD45.1/2 PTPN22 WT recipient mice. The Foxp3+ frequency within PTPN22 WT CD45.1+CD4+ or PTPN22 KO CD45.2+CD4+ cell pool was assessed 15 weeks post transfer. Each dot represents one animal (N = 11), data is pooled of two independent experiments. A paired t-test was performed to compare PTPN22 WT and KO cells within the same recipient. (F,G) Phenotypical analysis of Foxp3+ expressing PTPN22 WT CD45.1+CD4+ and PTPN22 KO CD45.2+CD4+ cells from the experiment outlined in (E). Each dot represents one animal (N = 11), data is pooled of two independent experiments. A paired t-test was performed to compare PTPN22 WT and KO cells within the same recipient. A p < 0.05 was considered significant *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.