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The Journal of Immunology, 1998, 160: 1290-1296.
Copyright © 1998 by The American Association of Immunologists

Cytokine Activation Leads to Acidification and Increases Maturation of Mycobacterium avium-Containing Phagosomes in Murine Macrophages1

Ulrich E. Schaible2,*, Sheila Sturgill-Koszycki*, Paul H. Schlesinger{dagger} and David G. Russell3,*

* Departments of Molecular Microbiology and {dagger} Physiology and Cell Biology, Washington University, School of Medicine, St. Louis, MO 63110

Mycobacterium avium (MAC) organisms multiply in phagosomes that have restricted fusigenicity with lysosomes, do not acidify due to a paucity of vacuolar proton-ATPases, yet remain accessible to recycling endosomes. During the course of mycobacterial infections, IFN-{gamma}-mediated activation of host and bystander macrophages is a key mechanism in the regulation of bacterial growth. Here we demonstrate that in keeping with earlier studies, cytokine activation of host macrophages leads to a decrease in MAC viability, demonstrable by bacterial esterase staining with fluorescein diacetate as well as colony-forming unit counts from infected cells. Analysis of the pH of MAC phagosomes demonstrated that the vacuoles in activated macrophages equilibrate to pH 5.2, in contrast to pH 6.3 in resting phagocytes. Biochemical analysis of MAC phagosomes from both resting and activated macrophages confirmed that the lower intraphagosomal pH correlated with an increased accumulation of proton-ATPases. Furthermore, the lower pH is reflected in the transition of MAC phagosomes to a point no longer accessible to transferrin, a marker of the recycling endosomal system. These alterations parallel the coalescence of bacterial vacuoles from individual bacilli in single vacuoles to communal vacuoles with multiple bacilli. These data demonstrate that bacteriostatic and bactericidal activities of activated macrophages are concomitant with alterations in the physiology of the mycobacterial phagosome.




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G. St. John, N. Brot, J. Ruan, H. Erdjument-Bromage, P. Tempst, H. Weissbach, and C. Nathan
Peptide methionine sulfoxide reductase from Escherichia coli and Mycobacterium tuberculosis protects bacteria against oxidative damage from reactive nitrogen intermediates
PNAS, August 14, 2001; 98(17): 9901 - 9906.
[Abstract] [Full Text] [PDF]


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