Trafficking and release of mycobacterial lipids from infected macrophages

Wandy L. Beatty, Elizabeth R. Rhoades, Heinz Joachim Ullrich, Delphi Chatterjee, John E. Heuser, David G. Russell

Research output: Contribution to journalArticle

276 Scopus citations

Abstract

Analysis of infected macrophages revealed that lipid-containing moieties of the mycobacterial cell wall are actively trafficked out of the mycobacterial vacuole. To facilitate the analysis of vesicular trafficking from mycobacteria-containing phagosomes, surface-exposed carbohydrates were labeled with hydrazide-tagged markers. The distribution of labeled carbohydrate/lipid moieties and subsequent interaction with cellular compartments were analyzed by immunoelectron microscopy and by fluorescence microscopy of live cells. The released mycobacterial constituents were associated with several intracellular organelles and were enriched strikingly in tubular endocytic compartments. Subcellular fractionation of infected macrophages by density gradient electrophoresis showed temporal movement of labeled bacterial constituents through early and late endosomes. Thin layer chromatography analysis of these subcellular fractions confirmed their lipid nature and revealed five dominant bacteria-derived species. These mycobacterial lipids were also found in extracellular vesicles isolated from the medium and could be observed in un-infected 'bystander' cells. Their transfer to bystander cells could expand the bacteria's sphere of influence beyond the immediate confines of the host cell.

Original languageEnglish
Pages (from-to)235-247
Number of pages13
JournalTraffic (Copenhagen, Denmark)
Volume1
Issue number3
DOIs
StatePublished - Mar 2000

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