Figure 3

Higher Th17 and Th1 expansion in the spleen and lung tissues of IL-10 KO mice than WT mice after Cm infection. Single cell suspensions of spleen and lungs of IL-10 KO and WT mice were prepared before infection (A–C) or at d 7 after intranasal Cm infection (D-I), as described in Materials and Methods. Intracellular cytokine staining was performed to detect the percentage of IL-17- and IFNε-producing CD4⁺ T cells by using fluorescence-conjugated antibodies, including FITC-CD3ε, PE-CD4 and allophycocyanin conjugated IL-17A or IFNγ. (A) Representative gating strategy was shown. Cells were gated on CD3γ⁺CD4⁺cells. (B, C) Representative data of intracellular IL-17 and IFNγ staining of CD4⁺T cells from naive IL-10 KO or WT mice. (D, G) Representative data of intracellular IL-17 and IFNγ staining of CD4⁺ T cells from infected mice. (E, H) Summary of the percentage of Th17 or Th1 cells in the spleen and lungs. (F, I) Spleen (7.5 × 10⁵) and lung cells (5 × 10⁵) were cultured for 72 h, and the culture supernatants were tested for IL-17 and IFNγ protein by ELISA. Data are shown as mean ± SD (n = 4). *p < 0.05; **p < 0.01. Representative data of three independent experiments are shown.