Figure 8.

Effect MDSC depletion in the presence of pulmonary Th1, Th2, and Th17 lymphocytes of P. brasiliensis-infected mice. The presence of intracellular IFN-γ, IL-4, and IL-17 in CD4⁺ T cells in the lung infiltrating leukocytes was assessed by intracellular cytokine staining by flow cytometry. After 24 hours, 1 week, and 7 weeks of infection a group of mice received intraperitoneal injections of anti-Gr1 (clone RB6-8C5) or rat IgG2b isotype control antibody (200 µg/dose). After 96 hours, 2 weeks, and 8 weeks of infection, lung cells were stimulated in vitro with PMA/ionomycin + brefeldin A for 6 hours and subjected to intracellular staining. The lung-infiltrating lymphocytes were gated by FSC/SSC analysis. Lymphocytes were gated for CD4 expression and then for IFN-γ (A), IL-4 (B), and IL-17 (C) expression. Results are expressed in frequency and the absolute number of cells and are representative of three independent experiments. For comparisons between two groups, the mean ± SEM was obtained and analyzed by the unpaired Student’s t-test. Differences were considered significant when: *p<0.05; **p<0.01 and ***p<0.001.