Figure 6.

Eosinophils-producing IL-17A contribute for uncontrolled inflammation after acute challenge with A. fumigatus. WT and ΔdblGATA-1 mice were infected via intranasal with 40 μL of suspension containing 1 × 10⁸ conidia of A. fumigatus. BALFs were harvested at day 3 of infection, centrifuged and the cells were labeled with specific monoclonal antibodies. Samples were measured by flow cytometry and analyzed by FlowJo 7.5.3. The relevant populations were gated using morphological and surface markers approach. (A) Gating-strategy and criteria of eosinophils selection. Briefly, we excluded cellular debris and selected high complexity/granularity (SSC) cells. Eosinophils were selected as CD11c⁻ and CD11b⁺ plus Siglec F⁺ cells. After, we analyzed IL-17 production in these cells (B) Quantification of Siglec F⁺ cells. (C) Siglec F⁺IL-17A⁺ cells. (D) CD3⁺ CD4⁺ cells in lymphocytes. (E) CD3⁺ CD4⁺ RORγT⁺IL-17A⁺ cells in lymphocytes. (F) CD3⁺TCRγδ⁺ IL-17A⁺ cells in lymphocytes. Data are presented as Mean ± SD (n = 5 to 7 mice per group). *Significantly different (P < 0.05) compared WT to knockout mice group. ^#Significantly different (P < 0.05) between mice with different times of infection.