Isolation and characterization of the mycobacterial phagosome: segregation from the endosomal/lysosomal pathway

Molecular Microbiology
Z HasanJean Pieters

Abstract

Mycobacteria have the ability to persist within host phagocytes, and their success as intracellular pathogens is thought to be related to the ability to modify their intracellular environment. After entry into phagocytes, mycobacteria-containing phagosomes acquire markers for the endosomal pathway, but do not fuse with lysosomes. The molecular machinery that is involved in the entry and survival of mycobacteria in host cells is poorly characterized. Here we describe the use of organelle electrophoresis to study the uptake of Mycobacterium bovis bacille Calmette Guerin (BCG) into murine macrophages. We demonstrate that live, but not dead, mycobacteria occupy a phagosome that can be physically separated from endosomal/lysosomal compartments. Biochemical analysis of purified mycobacterial phagosomes revealed the absence of endosomal/lysosomal markers LAMP-1 and beta-hexosaminidase. Combining subcellular fractionation with two-dimensional gel electrophoresis, we found that a set of host proteins was present in phagosomes that were absent from endosomal/lysosomal compartments. The residence of mycobacteria in compartments outside the endosomal/lysosomal system may explain their persistence inside host cells and their sequestration f...Continue Reading

Citations

Aug 27, 2009·Archivum Immunologiae Et Therapiae Experimentalis·Rogelio Hernandez-PandoDiana Aguilar
Jul 22, 1998·Journal of Immunological Methods·L RamachandraC V Harding
Apr 5, 2003·Comparative Immunology, Microbiology and Infectious Diseases·J HostetterN Cheville
May 19, 2001·Microbes and Infection·J Pieters
May 17, 2006·Veterinary Immunology and Immunopathology·Dirk WerlingTracey J Coffey
Nov 17, 2010·Proteomics·Qingbo LiGiovanni Lostumbo
Apr 1, 2011·Veterinary Immunology and Immunopathology·Richard J WhittingtonAuriol C Purdie
Jun 11, 2008·Cell Host & Microbe·Jean Pieters

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