title: Demonstration of the intracellular production of tissue-destructive protease by Legionella pneumophila multiplying within guinea-pig and human alveolar macrophages.
abstract: The major extracellular enzyme of Legionella pneumophila, a metalloprotease, has been proposed as a pathogenic factor in Legionnaires' disease due to its cytotoxic, tissue-destructive, and phagocyte-inhibitory properties. The relevance of these activities depends on the production of the protease during infection, i.e. by L. pneumophila multiplying intracellularly. In this study, L. pneumophila was demonstrated to produce protease in guinea-pig and human alveolar macrophages infected in vitro. After 24 h infection, approximately 0.1 to 0.2 micrograms of protease per 10(6) bacteria was measured by ELISA in culture supernatants and lysates of the infected cells, whereas no protease could be detected immediately after infection. Immunogold labelling using anti-protease antibody showed the enzyme to be located within phagosomes and distributed throughout the macrophages. Recent observations have shown that this protease could modify host defence mechanisms through inhibition of bacterial killing by neutrophils and monocytes. The intracellular production of the enzyme in infected macrophages demonstrated here further supports a role for the protease in the pathogenesis of Legionnaires' disease.
authors.affiliation: Department of Clinical Microbiology, Rigshospitalet, Copenhagen, Denmark.
authors.forename: C
authors.initials: C
authors.lastname: Rechnitzer
authors.forename: A
authors.initials: A
authors.lastname: Williams
authors.forename: J B
authors.initials: JB
authors.lastname: Wright
authors.forename: A B
authors.initials: AB
authors.lastname: Dowsett
authors.forename: N
authors.initials: N
authors.lastname: Milman
authors.forename: R B
authors.initials: RB
authors.lastname: Fitzgeorge
mesh_terms: D000818:Animals; D001426:Bacterial Proteins; D002478:Cells, Cultured; D006168:Guinea Pigs; D006801:Humans; D007150:Immunohistochemistry; D016952:Legionella pneumophila / Q000201:enzymology* / Q000254:growth & development; D008264:Macrophages / Q000382:microbiology*; D008666:Metalloendopeptidases / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D008666:Metalloendopeptidases; C043859:pseudolysin, Pseudomonas aeruginosa