title: Lipopolysaccharide of Legionella pneumophila shed in a liquid culture as a nonvesicular fraction arrests phagosome maturation in amoeba and monocytic host cells.
abstract: After uptake by susceptible host cells, Legionella pneumophila displays the ability to arrest phagolysosome fusion. To elucidate the role of lipopolysaccharide (LPS) in this mechanism, we investigated its influence on Acanthamoeba castellanii, A/J mouse macrophages and human monocytes. For this, legionellae were cultured in broth to the replicative, noninfectious phase or to the infectious phase expressing virulence traits. Shed LPS-enriched outer membrane vesicles (OMV) and LPS species <300 kDa were obtained from L. pneumophila Corby strains possessing the virulence-associated LPS epitope recognized by monoclonal antibody (MAb) 3/1 and its mutant TF 3/1, which has lost this epitope due to a mutation in the lag-1 gene. The shed LPS components were attached by specific antibodies to latex beads and added to the host cells for phagocytosis. We demonstrated for the first time that evasion of lysosomal degradation of phagosomes for up to 5 h can also be set off by LPS that is not tied up in OMV. Moreover, our cell culture models showed that the influence of MAb 3/1-positive and -negative LPS was identical. Our data clearly substantiate that LPS is an independent factor for evading lysosomal degradation, which is independent of the bacterial expression of known virulence traits.
authors.affiliation: Institute of Medical Microbiology and Hygiene, Medical Faculty of the Technical University Dresden, Dresden, Germany.
authors.forename: Eva M
authors.initials: EM
authors.lastname: Seeger
authors.forename: Maria
authors.initials: M
authors.lastname: Thuma
authors.forename: Esteban
authors.initials: E
authors.lastname: Fernandez-Moreira
authors.forename: Enno
authors.initials: E
authors.lastname: Jacobs
authors.forename: Marc
authors.initials: M
authors.lastname: Schmitz
authors.forename: Jürgen H
authors.initials: JH
authors.lastname: Helbig
mesh_terms: D048248:Acanthamoeba castellanii / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D054884:Host-Pathogen Interactions; D006801:Humans; D016952:Legionella pneumophila / Q000737:chemistry*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008247:Lysosomes / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D009000:Monocytes / Q000378:metabolism; D010587:Phagocytosis / Q000187:drug effects; D010588:Phagosomes / Q000187:drug effects* / Q000378:metabolism*
chemical_list: D008070:Lipopolysaccharides