Identification of virulent factors related to rearrangement of skeleton Proteins in mycobacterial infections.

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To identify new virulent factors of intracellular bacteria, such as Mycobacterium tuberculosis and Rhodococcus equi, murine macrophage-like J774. 1 cells were infected with virulent or avirulent R. equi and observed by using immunocytological methods. Observations by light microscopy and transmission electron microscopy showed that the virulent bacteria increased into single membrane vacuoles. Observations by confocal fluorescence microscopy with an endosomal marker and lysosomal markers revealed that most phagosomes containing virulent bacteria were fused with lysosomes. To monitor acidification of phagosomes containing R. equi, the cells infected with R. equi were stained with acidotropic probe. The phagosomes containing avirulent R. equi became acidic. In contrast, virulent R. eqiu did not change pH. The medium of virulent R. equi was suppressed acidification of acidic compartments in macrophages. These results suggested that virulent R. equi can survive phagolysosomal environment in macrophages by secreting substance which suppress acidification of phagolysosome.