Figure 2
Lung γδ+T cells are the predominant leukocyte population expressing IL-17A during pulmonary infection with C. neoformans strain H99γ during neutrophil depletion. BALB/c mice received an intranasal inoculum of 1 × 104 CFU of C. neoformans strain H99γ in 50 μl of sterile PBS. Mice were treated with isotype control antibody (white bars) or with anti-1A8 antibody (gray bars). The lungs were excised at day 7 post-inoculation and a single cell suspension generated using enzymatic digestion. The leukocytes were stained with anti-mouse antibodies (CD45, 1A8 (Neut), CD4, CD8, F4/80 (Mac), CD11b/CD11c (DC), CD4/Fox3p (Treg), CD3/NKp46 (NKT), NKp46/CD45 (NK), γδ/CD45 (γδ+ T cells), CD19 (B cell), SiglecF/CD11b (Eosinophil), fixed, permeabilized, and incubated with anti-IL-17A antibodies and evaluated by flow cytometry. Flow cytometry data are cumulative results of three independent experiments using pooled leukocytes from 4 mice per group per experiment. Results shown are the percentage of leukocytes expressing the indicated surface markers and IL-17A. Asterisks (*) indicate where significant differences (P < 0.05) were observed between isotype control-treated and anti-1A8-treated mice infected with C. neoformans strain H99γ.