Mycobacterium tuberculosis and Mycobacterium avium modify the composition of the phagosomal membrane in infected macrophages by selective depletion of cell surface-derived glycoconjugates.
The growth of pathogenic mycobacteria in phagosomes of the host cell correlates with their ability to prevent phagosome maturation. The underlying molecular mechanism remains elusive. In a previous study, we have shown that Mycobacterium avium depletes the phagosome membrane of cell surface-derived glycoconjugates (de Chastellier and Thilo, Eur. J. Cell Biol. 81, 17-25, 2002). We now extended these quantitative observations to the major human pathogen, Mycobacterium tuberculosis (H37Rv). At increasing times after infection of mouse bone marrow-derived macrophages, cell-surface glycoconjugates were labelled enzymatically with [3H]galactose. Subsequent endocytic membrane traffic resulted in a redistribution of this label from the cell surface to endocytic membranes, including phagosomes. The steady-state distribution was measured by quantitative autoradiography at the electron microscope level. Relative to early endosomes, with which phagosomes continued to fuse and rapidly exchange membrane constituents, the phagosome membrane was depleted about 3-fold, starting during infection and in the course of 9 days thereafter. These results were in quantitative agreement with our previous observations for Mycobacterium avium. For the latter case, we now showed by cell fractionation that the depletion was selective, mainly involving glycoproteins in the 110-210 kDa range. Together, these results indicated that pathogenic mycobacteria induced and maintained a bulk change in phagosome membrane composition that could be of special relevance for survival of pathogenic mycobacteria within phagosomes.
Pietersen
Raydean
R
Division of Medical Biochemistry, Faculty of Health Sciences, University of Cape Town, South Africa.
Thilo
Lutz
L
de Chastellier
Chantal
C
D000818:Animals; D001345:Autoradiography; D002478:Cells, Cultured; D004705:Endocytosis; D005260:Female; D006001:Glycoconjugates / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D007425:Intracellular Membranes / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D008264:Macrophages / Q000166:cytology* / Q000382:microbiology / Q000648:ultrastructure; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron; D009162:Mycobacterium avium / Q000472:pathogenicity / Q000502:physiology* / Q000648:ultrastructure; D009169:Mycobacterium tuberculosis / Q000472:pathogenicity / Q000502:physiology* / Q000648:ultrastructure; D010588:Phagosomes / Q000378:metabolism* / Q000382:microbiology* / Q000648:ultrastructure; D013997:Time Factors
D006001:Glycoconjugates; D006023:Glycoproteins