Development of Th1, Tregs and Th17 cell subsets after <i>C. muridarum</i> infections.

<p>(A) and (B) C57BL/6 mice were infected with 1×10⁵<i>C. muridarum</i> i.v. or i.vag.. PmpG-1_303–311:I-A^b+ cells were enriched from spleen, DLNs and NDLNs 7 and 14 days post i.v. and i.vag. infections, respectively. (A) Histograms showing expression levels of T-bet and GATA3 on CD4⁺PmpG-1_303–311:I-A^b+ cells from naïve and infected mice. (B) FACS plots showing expression of Foxp3 on CD4+ cells. (C) RORγt-GFP reporter mice were infected i.vag. with 1×10⁵<i>C. muridarum</i>. Cells from spleen, DLNs and genital tract were isolated and stained with pooled RplF_51–59:I-A^b, Aasf_24–32:I-A^b and PmpG-1_303–311:I-A^b tetramers (R.A.P.:I- A^b). CD4⁺CD44^hi R.A.P.:I- A^b+ cells were further gated for GFP expression. RORγt-GFP⁻ mice were used as a negative control for setting the GFP⁺ gates in our analysis. (D) C57BL/6 mice were infected with 1×10⁵<i>C. muridarum</i> intravaginally. IFNγ and IL-17A production by CD4 T cells from DLNs and genital tract were detected by flow cytometry after ex vivo stimulation with phorbol 12-myristate 13-acetate and ionomycin in the presence of brefeldin A.