FIG 2.

Host response to H. ohiense, but not H. capsulatum, is associated with a significant influx of neutrophils and a concomitant depletion of alveolar macrophages. Lung cells from C57BL/6J mice were analyzed at the indicated times after intranasal inoculation with 2,500 H. capsulatum or H. ohiense. (A to C) Concatenated live, CD45⁺ lung cells were visualized 8 days (A), 12 days (B), or 16 days (C) after Histoplasma infection compared to control mice using a combination of tSNE (see Fig. S2A in the supplemental material) and a sequential gating scheme (Fig. S2B). Concatenated events from mice at each time point depict color-coded cell populations derived from the sequential gating scheme and tSNE expression profiles (Fig. S2C) that were overlaid onto each tSNE plot. Plots to the right include events that were derived from either mock-infected, H. capsulatum-, or H. ohiense-inoculated mice, and populations of interest include neutrophils (orange), Ly6C^hi MHC-II⁻ monocytes (purple), Ly6C^hi MHC-II⁺ inflammatory monocytes (magenta), and alveolar macrophages (cyan). Numbers associated with colored clusters indicate the average percentage among CD45⁺ cells. Each plot includes 20,000 to 40,000 pooled events (10,000 events per mouse with two mock-infected mice per time point or four Histoplasma-infected mice per time point). cDC, conventional dendritic cells. (D to F) Bars represent the mean total number (±SD) of neutrophils (D), inflammatory monocytes (E), and alveolar macrophages (F) recovered from the lungs of mice infected with either H. capsulatum (blue bars) or H. ohiense (red bars) (n = 4 per time point). Horizontal dotted lines indicate the mean number of cells recovered from mock-infected mice (n = 10, 1 or 2 mice were analyzed at each time point). Data are representative of two separate time course experiments, and statistical significance was evaluated using nonparametric Mann-Whitney tests; *, P < 0.05.