title: Bacterial Membrane Vesicles Mediate the Release of Mycobacterium tuberculosis Lipoglycans and Lipoproteins from Infected Macrophages.
abstract: Mycobacterium tuberculosis is an intracellular pathogen that infects lung macrophages and releases microbial factors that regulate host defense. M. tuberculosis lipoproteins and lipoglycans block phagosome maturation, inhibit class II MHC Ag presentation, and modulate TLR2-dependent cytokine production, but the mechanisms for their release during infection are poorly defined. Furthermore, these molecules are thought to be incorporated into host membranes and released from infected macrophages within exosomes, 40-150-nm extracellular vesicles that derive from multivesicular endosomes. However, our studies revealed that extracellular vesicles released from infected macrophages include two distinct, largely nonoverlapping populations: one containing host cell markers of exosomes (CD9, CD63) and the other containing M. tuberculosis molecules (lipoglycans, lipoproteins). These vesicle populations are similar in size but have distinct densities, as determined by separation on sucrose gradients. Release of lipoglycans and lipoproteins from infected macrophages was dependent on bacterial viability, implicating active bacterial mechanisms in their secretion. Consistent with recent reports of extracellular vesicle production by bacteria (including M. tuberculosis), we propose that bacterial membrane vesicles are secreted by M. tuberculosis within infected macrophages and subsequently are released into the extracellular environment. Furthermore, extracellular vesicles released from M. tuberculosis-infected cells activate TLR2 and induce cytokine responses by uninfected macrophages. We demonstrate that these activities derive from the bacterial membrane vesicles rather than exosomes. Our findings suggest that bacterial membrane vesicles are the primary means by which M. tuberculosis exports lipoglycans and lipoproteins to impair effector functions of infected macrophages and circulate bacterial components beyond the site of infection to regulate immune responses by uninfected cells.
authors.affiliation: Department of Pathology, Case Western Reserve University/University Hospitals Case Medical Center, Cleveland, OH;
authors.forename: Jaffre J
authors.initials: JJ
authors.lastname: Athman
authors.affiliation: Department of Pathology, Case Western Reserve University/University Hospitals Case Medical Center, Cleveland, OH;
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Molecular Biology and Microbiology, Case Western Reserve University/University Hospitals Case Medical Center, Cleveland, OH; Center for AIDS Research, Case Western Reserve University/University Hospitals Case Medical Center, Cleveland, OH; and.
authors.forename: David J
authors.initials: DJ
authors.lastname: McDonald
authors.affiliation: Center for AIDS Research, Case Western Reserve University/University Hospitals Case Medical Center, Cleveland, OH; and Division of Infectious Diseases, Case Western Reserve University/University Hospitals Case Medical Center, Cleveland, OH.
authors.forename: W Henry
authors.initials: WH
authors.lastname: Boom
authors.affiliation: Department of Pathology, Case Western Reserve University/University Hospitals Case Medical Center, Cleveland, OH; Center for AIDS Research, Case Western Reserve University/University Hospitals Case Medical Center, Cleveland, OH; and.
authors.forename: Clifford V
authors.initials: CV
authors.lastname: Harding
authors.affiliation: Department of Pathology, Case Western Reserve University/University Hospitals Case Medical Center, Cleveland, OH; Center for AIDS Research, Case Western Reserve University/University Hospitals Case Medical Center, Cleveland, OH; and paw28@case.edu.
authors.forename: Pamela A
authors.initials: PA
authors.lastname: Wearsch
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D055354:Exosomes / Q000276:immunology / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000276:immunology; D008074:Lipoproteins / Q000276:immunology; D008168:Lung / Q000166:cytology / Q000276:immunology; D016676:Macrophages, Alveolar / Q000276:immunology* / Q000382:microbiology; D051379:Mice; D008810:Mice, Inbred C57BL; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D022142:Secretory Vesicles / Q000276:immunology*; D051195:Toll-Like Receptor 2 / Q000378:metabolism; D014397:Tuberculosis, Pulmonary / Q000276:immunology* / Q000382:microbiology
chemical_list: D008070:Lipopolysaccharides; D008074:Lipoproteins; C493486:Tlr2 protein, mouse; D051195:Toll-Like Receptor 2