Fig 5. Neutrophil depletion is associated with decreased intracellular cytokine levels and changes in transcription factor mRNA expression related to T helper subsets during the chronic stages of P. brasiliensis infection.

BALB/c mice infected with 1.5x10⁶ P. brasiliensis yeast cells and treated with an isotype control Ab or anti-Ly6G mAb at 4 weeks post-infected and analyzed at 12 weeks post-infection. Flow cytometry was performed to identify the following lymphocytes subsets: A) Th1 (LT CD4⁺IFNγ⁺), B) Th2 (LT CD4⁺IL-4⁺), C) Th17 (LT CD4⁺IL-17⁺), D) Th22 (LT CD4⁺IL-22⁺), and E) Treg cells (LT CD4⁺CD25⁺ FoxP3⁺). Data shown represent median and IQR (n = 4–5 mice/group; representative of two independent experiments). Relative quantification of the mRNA expression for T-bet (F), GATA-3 (G), RORc (H), Ahr (I), FoxP3 (J), iNOS2 (K) and Arginase-1 (L) were measured in lungs of mice. Results are expressed as mean ± SEM (n = 4–5 mice/group; representative of two independent experiments). *, P<0.05 comparing infected untreated mice versus control mice, or comparing infected-anti-Ly6G mAb treated mice versus infected-untreated mice. PBS, control mice; PBS + Iso, control mice treated with isotype control Ab (clone: 2A3); PBS + anti-neutrophils, control mice treated with anti-Ly6G mAb (clone: 1A8); Pb18, infected-untreated mice; Pb18 + Iso, infected mice treated with isotype control Ab; Pb18 + anti-neutrophils, infected mice treated with anti-Ly6G mAb.