Fig. 5
CCR2 is required for AM localization within TB granulomas. a CD11c+ cells were purified from lungs of 30 dpi HN878-infected CCR2-GFP(+/KI) or CCR2-GFP(KI/KI) mice and 106 cells were IT transferred into B6 HN878-infected mice (n = 5) at 30 dpi. a–c Lungs were harvested at 50 dpi and examined for localization of SiglecF+ GFP+ cells within TB granulomas using the morphometric tool of the Zeiss Axioplan microscope. d CCR2-GFP(+/KI) or CCR2-GFP(KI/KI) BMDMs were stimulated in vitro with 20 µg/mL irradiated Mtb HN878 for 24 h. Migration towards uninfected, H37Rv- or HN878-infected epithelial cell supernatants was analyzed via transwell chemotaxis assays and flow cytometry (n = 3). e Ccl2 mRNA localization was determined within FFPE lung sections from B6 and CCR2−/− HN878-infected using RNAScope in situ hybridization (ISH). Arrows point to Ccl2 mRNA localization (brown). f B6 and CCR2−/− mice (n = 5) were infected with HN878 and percentage of AMs with airway label CD45.2 delivered IT was calculated on 14 dpi by flow cytometry. Grav gravity control, Un. uninfected, AMs alveolar macrophages. n = 5. b, c Student’s t-test. n = 3. d 2-Way ANOVA with Bonferroni post test. f Student’s t-test
