Differences between Mycobacterium-Host Cell Relationships in Latent Tuberculous Infection of Mice Ex Vivo and Mycobacterial Infection of Mouse Cells In Vitro

Figure 6

((a)–(c)) The cells with IFNγ in the fragments of mouse splenic granulomas. The scale bars are 10 μm each. ((a), (c)) Colocalization of the markers on the confocal images of cells (yellow signal). ((a), (c)-(d)) Nuclei stained by DAPI (blue signal). (a) Confocal immunofluorescent localization of IFNγ (green signal) and CD80 (red signal) in granuloma macrophages (Mph) and a fibroblast at the center of the images. In the right panel, the same fragment as in the other panels restained for acid-fast BCG-mycobacteria by the ZN method. An IFNγ-producing macrophage with BCG-mycobacteria reproducing in it is indicated by the white arrows on the fluorescent images and the black arrow on the ZN image. (b) Immunochemical localization of IFNγ in granuloma cells and control mouse peritoneal macrophages. The brown color indicates the presence of IFNγ in these cells. (c) Confocal immunofluorescent localization of IFNγ (green signal) and CD11b (red signal) in granuloma macrophages, with a multinucleate Langhans giant cell indicated by the white arrows in the three lower panels. (d) Bone marrow macrophages following infection with the BCG vaccine in vitro and after culture for several hours not stained by the IFNγ- (green signal) and IL-1α-specific antibodies (red signal) on the confocal fluorescent images. Abbreviations as in Figure 1.

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