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. 2018 Oct 1;215(10):2617–2635. doi: 10.1084/jem.20180300

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© 2018 Ito et al.

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Figure 4. — Insulin-specific CD4 T cells show FoxP3⁺ T reg cell phenotype in islets of NOD mice. (A) Representative flow-cytometric analysis of tetramer⁺ CD4⁺ T cells from pancreatic islets of 9-wk-old NOD mice labeled with FoxP3-specific or isotype control antibodies. (B) Percentage of FoxP3⁺ cells among islet-infiltrating CD4⁺ T cells from 9-wk-old NOD mice labeled with four different I-A^g7 tetramers (% FoxP3⁺ of tetramer⁺ CD4⁺ T cells; n = 8 mice/group). (C) Analysis of FoxP3 labeling for tetramer⁺ CD4 T cells from islets of control mice (C) or M98A/M98A mice (KI) at 9 wk or 12 wk of age (n = 7 to 11 mice/group). (D) Percentage of IFN-γ⁺ cells among tetramer⁺ FoxP3⁺ CD4 T cells from islets of 9-wk-old NOD mice. Cells were stimulated with PMA and ionomycin and stained with IFN-γ antibody (n = 4–5 mice/group). (E) Representative flow-cytometric analysis of GITR, Helios, and CTLA-4 expression in InsB12-20–labeled FoxP3⁺ CD4⁺ T cells from pancreatic islets of 9-wk-old NOD mice. Cell surface GITR and intracellular Helios as well as CTLA-4 were examined. (F) Summary plots of GITR⁺, Helios⁺, or CTLA-4⁺ cells for InsB12-20–labeled FoxP3⁺ CD4⁺ T cells (n = 6 to 12 mice/group). Statistical analyses were performed with the one-way ANOVA test (B) and the Mann-Whitney test (C), mean + SD (B–D), or mean – SD (F) are shown (*, 0.01 ≤ P < 0.05; **, 0.001 ≤ P < 0.01). Data represent two independent experiments.