1887

Abstract

The major extracellular enzyme of , 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 multiplying intracellularly. In this study, was demonstrated to produce protease in guinea-pig and human alveolar macrophages infected . After 24 h infection, approximately 0.1 to 0.2 μg of protease per 10bacteria 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.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-138-8-1671
1992-08-01
2024-05-04
Loading full text...

Full text loading...

/deliver/fulltext/micro/138/8/mic-138-8-1671.html?itemId=/content/journal/micro/10.1099/00221287-138-8-1671&mimeType=html&fmt=ahah

References

  1. Baskerville A., Conlan J. W., Ashworth L. A. E., Dowsett A. B. 1986; Pulmonary damage caused by a protease from Legionella pneumophila . British Journal of Experimental Pathology 67:527–536
    [Google Scholar]
  2. Belyi I. F., Vertiev I. V., Tartakovskii I. S., Ezepchuck I. V., Prosorovskii S. V. 1988; Characteristics of the cytolysin of Legionella pneumophila . Zhurnal Mikrobiologii, Epidemiologii i Immunobiologii 2:4–7
    [Google Scholar]
  3. Blander S. J., Horwitz M. A. 1989; Vaccination with the major secretory protein of Legionella pneumophila induces cell-mediated and protective immunity in a guinea pig model of Legionnaires’ disease. Journal of Experimental Medicine 169:691–705
    [Google Scholar]
  4. Blander S. J., Szeto L., Shuman H. A., Horwitz M. A. 1990; An immunoprotective molecule, the major secretory protein of Legionella pneumophila, is not a virulence factor in a guinea pig model of Legionnaires’ disease. Journal of Clinical Investigation 86:817–824
    [Google Scholar]
  5. Conlan J. W., Baskerville A., Ashworth L. A. E. 1986; Separation of Legionella pneumophila proteases and purification of a protease which produces lesions like those of Legionnaires’ disease in guinea pig lung. Journal of General Microbiology 132:1565–1574
    [Google Scholar]
  6. Conlan J. W., Williams A., Ashworth L. A. E. 1988; In vivo production of a tissue-destructive protease by Legionella pneumophila in the lungs of experimentally infected guinea pigs. Journal of General Microbiology 134:143–149
    [Google Scholar]
  7. Diamant M., Rechnitzer C., Christensen L. D., Pedersen B. K. 1991; Cleavage of the CD4 molecular complex by Legionella pneumophila protease. Immunology and Infectious Diseases 1:167–171
    [Google Scholar]
  8. Dreyfus L. A., Iglewski B. H. 1986; Purification and characterization of an extracellular protease of Legionella pneumophila . Infection and Immunity 51:736–743
    [Google Scholar]
  9. Elliott J. A., Winn W. C. Jr 1986; Treatment of alveolar macrophages with cytochalasin D inhibits uptake and subsequent growth of Legionella pneumophila . Infection and Immunity 51:31–36
    [Google Scholar]
  10. Fitzgeorge R. B. 1985; The effect of antibiotics on the growth of Legionella pneumophila in guinea pig alveolar phagocytes infected in vivo by an aerosol. Journal of Infection 10:189–193
    [Google Scholar]
  11. Fitzgeorge R. B., Featherstone A. S. R., Baskerville A. 1988; Effects of polymorphonuclear leucocyte depletion on experimental Legionnaires’ disease. British Journal of Experimental Pathology 69:105–112
    [Google Scholar]
  12. Horwitz M. A., Silverstein S. C. 1980; Legionnaires’ disease bacterium (Legionella pneumophila) multiplies intracellularly in human monocytes. Journal of Clinical Investigation 66:441–450
    [Google Scholar]
  13. Jepras R. I., Fitzgeorge R. B., Baskerville A. 1985; A comparison of virulence of two strains of Legionella pneumophila based on experimental aerosol infection of guinea pigs. Journal of Hygiene 95:29–38
    [Google Scholar]
  14. Kishimoto R. A., White J. D., Shirey F. G., McGann V. G., Berendt R. F., Larson E. W., Hedlund K. W. 1981; In vitro response of guinea pig peritoneal macrophages to Legionella pneumophila . Infection and Immunity 31:1209–1213
    [Google Scholar]
  15. Mintz C. S., Chen J., Shuman H. A. 1988; Isolation and characterization of auxotrophic mutants of Legionella pneumophila that fail to multiply in human monocytes. Infection and Immunity 56:1449–1455
    [Google Scholar]
  16. Nash T. W., Libby D. M., Horwitz M. A. 1984; Interaction between the Legionnaires’ disease bacterium (Legionella pneumophila) and human alveolar macrophages. Influence of antibody, lymphokines, and hydrocortisone. Journal of Clinical Investigation 74:771–782
    [Google Scholar]
  17. Rajagopalan P., Dournon E., Vilde J.-L., Pocidalo J.-J. 1987; Direct activation of human monocyte-derived macrophages by a bacterial glycoprotein extract inhibits the intracellular multiplication of virulent Legionella pneumophila serogroup 1. Infection and Immunity 55:2234–2239
    [Google Scholar]
  18. Rechnitzer C., Kharazmi A. 1992; Effect of Legionella pneumophila cytotoxic protease on human neutrophil and monocyte function. Microbial Pathogenesis 12:115–125
    [Google Scholar]
  19. Rechnitzer C., Tvede M., Doring G. 1989a; A rapid method for purification of homogeneous Legionella pneumophila protease using fast protein liquid chromatography. FEMS Microbiology Letters 59:39–44
    [Google Scholar]
  20. Rechnitzer C., Diamant M., Pedersen B. K. 1989b; Inhibition of human natural killer cell activity by Legionella pneumophila protease. European Journal of Clinical Microbiology and Infectious Diseases 8:989–992
    [Google Scholar]
  21. Rosenfeld J. S., Kueppers F., Newkirk T., Tamada R., Meissler J. J. Jr, Eisenstein T. K. 1986; A protease from Legionella pneumophila with cytotoxic and dermal ulcerative activity. FEMS Microbiology Letters 37:51–58
    [Google Scholar]
  22. Szeto L., Shuman H. A. 1990; The Legionella pneumophila major secretory protein, a protease, is not required for intracellular growth or cell killing. Infection and Immunity 58:2585–2592
    [Google Scholar]
  23. Williams A., Baskerville A., Dowsett A. B., Conlan J. W. 1987; Immunocytochemical demonstration of the association between Legionella pneumophila, its tissue-destructive protease, and pulmonary lesions in experimental Legionnaires’ disease. Journal of Pathology 153:257–264
    [Google Scholar]
  24. Yamamoto Y., Klein T. W., Newton C. A., Widen R., Friedman H. 1987; Differential growth of Legionella pneumophila in guinea pig versus mouse macrophage culture. Infection and Immunity 55:1369–1374
    [Google Scholar]
  25. Yoshida S., Mizuguchi Y., Nikaido Y., Mitsuyama M., Nomoto K. 1987; Fate of Legionella pneumophila, Philadelphia-1 strain in resident, elicited, activated and immune peritoneal macrophages of guinea pigs. Infection and Immunity 55:2477–2482
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-138-8-1671
Loading
/content/journal/micro/10.1099/00221287-138-8-1671
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error