Fig. 2
AMs are preferentially infected with HN878 and CCR2 expression on AMs is Mtb strain-dependent. a The flow cytometry gating strategy for myeloid populations. Briefly, AMs were defined as CD11b−CD11c+SiglecF+ cells. mDCs were defined as CD11b+CD11c+ cells. Neutrophils were defined as CD11b+CD11c−Gr-1hi cells, monocytes were defined as CD11b+CD11c−Gr-1lo cells, and recruited macrophages were defined as CD11b+CD11c−Gr-1− cells. Mtb-GFP+ cells were gated from individual subsets. b B6 mice were aerosol-infected with ~100 CFU of H37Rv-GFP or HN878-GFP and myeloid cell subsets infected with Mtb-GFP were determined by flow cytometry on 30 dpi (n = 5). c, d B6 mice (n = 5) were infected with aerosolized H37Rv or HN878 and the total number (c) and percentage (d) of CCR2+ myeloid lung cell populations were determined by flow cytometry and compared to uninfected controls. e Representative histograms of each subset displaying CCR2 expression by antibody staining compared to CCR2−/−. f Mean fluorescence intensity (MFI) of CCR2 expression on myeloid populations was normalized relative to MFI of unstained, uninfected controls. Un. uninfected, AMs alveolar macrophages, Monos monocytes, RMs recruited macrophages, Neuts neutrophils, mDCs myeloid dendritic cells. a–d 2-Way ANOVA with Bonferroni post test was used. f Student’s t-test
