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

Figure 7

((a)–(c)) Bacterial lipid- and glycolipid-presenting molecule CD1d in the cells in the fragments of mouse splenic granulomas. The scale bars are ((a, central right panel), (b)–(d)) 10 μm each and (a, other panels) 20 μm each. ((b)-(c)) Colocalization of the markers on the confocal images of cells (yellow signal). ((b)–(d)) Nuclei stained by DAPI (blue signal). (a) Immunochemical localization of CD1d in granuloma cells, control mouse peritoneal macrophages (Mph), and fibroblasts (Fibr). The brown color indicates the presence of CD1d in these cells. ((b)-(c)) Confocal immunofluorescent localization of CD1d (green signal) and IL-1α (red signal) in granuloma macrophages and fibroblasts. (b) CD1d- and IL-1α-producing fibroblasts are indicated by the white arrows on the fluorescent images and the black arrows on the 3D image. (c) In the right panel, the same fragment as in the left panel restained for acid-fast BCG-mycobacteria by the ZN method. A CD1d- and IL-1α-producing macrophage with BCG-mycobacteria reproducing in it is indicated by the white arrow on the fluorescent image and the black arrow on the ZN image. (d) Confocal immunofluorescent localization of CD1d (red signal) in the cells in the cultures of bone marrow macrophages following infection with the BCG vaccine in vitro and after culture for several hours. The macrophage with CD1d in it is indicated by the red arrow, and a megakaryocyte without the antigen is indicated by the white arrow. Abbreviations as in Figure 1.

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