E2 induces IL-17–producing γδ+ T cells in the FGT

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E2 induces IL-17–producing γδ+ T cells in the FGT E2 induces IL-17–producing γδ+ T cells in the FGT. Vaginal cells from OVX WT (C57BL/6) mice treated with E2 or placebo (mock) pellets (n = 5) were isolated, pooled, and stimulated with a cell stimulation mixture containing Golgi inhibitors and PMA plus ionomycin for 18 h. E2 induces IL-17–producing γδ+ T cells in the FGT. Vaginal cells from OVX WT (C57BL/6) mice treated with E2 or placebo (mock) pellets (n = 5) were isolated, pooled, and stimulated with a cell stimulation mixture containing Golgi inhibitors and PMA plus ionomycin for 18 h. (A) Cells were stained with Abs against CD3, CD4, and IL-17 and analyzed by flow cytometry. ERKO mice were used as additional controls. Dead cells were excluded, and total IL-17+ cells were gated. (B) The differences in percentage and (C) MFI of IL-17+ cells were compared between E2 and mock mice from three independent experiments. Data were analyzed using the unpaired t test with 95% confidence interval (*p < 0.05, **p < 0.01). (D) Subsets of IL-17+ cells were further identified based on CD3 and CD4 expression. (E) The distributions of the cell subsets among all IL-17–producing cells comparing E2-treated and mock-treated mice. (F) The difference in percentage of IL-17–producing γδ+ cells was compared between E2 and mock mice from three independent experiments. Data are mean ± SEM, and significance was calculated using an unpaired t test with 95% confidence interval (*p < 0.05). Data are representative of three independent experiments with similar results. Varun C. Anipindi et al. ImmunoHorizons 2019;3:317-330 Copyright © 2019 The Authors