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. 1994 Dec;145(6):1537–1546.

Replicative Legionella pneumophila lung infection in intratracheally inoculated A/J mice. A murine model of human Legionnaires' disease.

J Brieland 1, P Freeman 1, R Kunkel 1, C Chrisp 1, M Hurley 1, J Fantone 1, C Engleberg 1
PMCID: PMC1887509  PMID: 7992856

Abstract

The role of host immune responses in the pathogenesis of Legionnaires' disease is incompletely understood, due in part to the current lack of an animal model that is both susceptible to replicative Legionella pneumophila-induced lung infection and for which species-specific immunological reagents are available. We have developed a model of replicative L. pneumophila lung infection in intratracheally inoculated A/J mice. L. pneumophila was obtained in the exponential growth phase and inoculated into the trachea of 6- to 8-week-old female A/J mice. Microbiological and histopathological evidence of infection was demonstrated in mice inoculated with 10(6) colony-forming units. Development of an acute pneumonia that resembled human Legionnaires' disease coincided with exponential growth of the bacteria in the lung 24 to 48 hours after intratracheal inoculation of L. pneumophila. This was associated with increased plasma levels of interferon-gamma at 24 hours after inoculation. After 48 hours, the bacteria were gradually eliminated from the lung over the next 5 days, corresponding with resolution of the inflammatory response in the lung, thereby mimicking the outcome frequently seen in the immunocompetent human host. Treatment of animals with anti-interferon-gamma antibody enhanced bacterial replication and disease progression, indicating an important role of host immune response in resolution of the infection. Because of the availability of murine-specific reagents, this model of replicative L. pneumophila lung infection in A/J mice after intrapulmonary inoculation of L. pneumophila potentially provides an important tool for future studies investigating the role of host immune responses in the pathogenesis of Legionnaires' disease in the immunocompetent host.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abu Kwaik Y., Eisenstein B. I., Engleberg N. C. Phenotypic modulation by Legionella pneumophila upon infection of macrophages. Infect Immun. 1993 Apr;61(4):1320–1329. doi: 10.1128/iai.61.4.1320-1329.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baskerville A., Dowsett A. B., Fitzgeorge R. B., Hambleton P., Broster M. Ultrastructure of pulmonary alveoli and macrophages in experimental Legionnaires' disease. J Pathol. 1983 Jun;140(2):77–90. doi: 10.1002/path.1711400202. [DOI] [PubMed] [Google Scholar]
  3. Bhardwaj N., Nash T. W., Horwitz M. A. Interferon-gamma-activated human monocytes inhibit the intracellular multiplication of Legionella pneumophila. J Immunol. 1986 Oct 15;137(8):2662–2669. [PubMed] [Google Scholar]
  4. Blanchard D. K., Djeu J. Y., Klein T. W., Friedman H., Stewart W. E., 2nd Protective effects of tumor necrosis factor in experimental Legionella pneumophila infections of mice via activation of PMN function. J Leukoc Biol. 1988 May;43(5):429–435. doi: 10.1002/jlb.43.5.429. [DOI] [PubMed] [Google Scholar]
  5. Blanchard D. K., Stewart W. E., 2nd, Klein T. W., Friedman H., Djeu J. Y. Cytolytic activity of human peripheral blood leukocytes against Legionella pneumophila-infected monocytes: characterization of the effector cell and augmentation by interleukin 2. J Immunol. 1987 Jul 15;139(2):551–556. [PubMed] [Google Scholar]
  6. Buchmeier N. A., Schreiber R. D. Requirement of endogenous interferon-gamma production for resolution of Listeria monocytogenes infection. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7404–7408. doi: 10.1073/pnas.82.21.7404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Byrd T. F., Horwitz M. A. Interferon gamma-activated human monocytes downregulate transferrin receptors and inhibit the intracellular multiplication of Legionella pneumophila by limiting the availability of iron. J Clin Invest. 1989 May;83(5):1457–1465. doi: 10.1172/JCI114038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cianciotto N., Eisenstein B. I., Engleberg N. C., Shuman H. Genetics and molecular pathogenesis of Legionella pneumophila, an intracellular parasite of macrophages. Mol Biol Med. 1989 Oct;6(5):409–424. [PubMed] [Google Scholar]
  9. Davis G. S., Winn W. C., Jr, Beaty H. N. Legionnaires Disease. Infections caused by Legionella pneumophilia and Legionella-like organisms. Clin Chest Med. 1981 Jan;2(1):145–166. [PubMed] [Google Scholar]
  10. Davis G. S., Winn W. C., Jr, Gump D. W., Craighead J. E., Beaty H. N. Legionnaires' pneumonia after aerosol exposure in guinea pigs and rats. Am Rev Respir Dis. 1982 Dec;126(6):1050–1057. doi: 10.1164/arrd.1982.126.6.1050. [DOI] [PubMed] [Google Scholar]
  11. Dondero T. J., Jr, Rendtorff R. C., Mallison G. F., Weeks R. M., Levy J. S., Wong E. W., Schaffner W. An outbreak of Legionnaires' disease associated with a contaminated air-conditioning cooling tower. N Engl J Med. 1980 Feb 14;302(7):365–370. doi: 10.1056/NEJM198002143020703. [DOI] [PubMed] [Google Scholar]
  12. Engleberg N. C., Carter C., Demarsh P., Drutz D. J., Eisenstein B. I. A Legionella-specific DNA probe detects organisms in lung tissue homogenates from intranasally inoculated mice. Isr J Med Sci. 1986 Oct;22(10):703–705. [PubMed] [Google Scholar]
  13. Friedman H., Klein T. W., Widen R., Newton C., Blanchard D. K., Yamamoto Y. Legionella pneumophila immunity and immunomodulation: nature and mechanisms. Adv Exp Med Biol. 1988;239:327–341. doi: 10.1007/978-1-4757-5421-6_32. [DOI] [PubMed] [Google Scholar]
  14. Fujio H., Yoshida S., Miyamoto H., Mitsuyama M., Mizuguchi Y. Investigation of the role of macrophages and endogenous interferon-gamma in natural resistance of mice against Legionella pneumophila infection. FEMS Microbiol Immunol. 1992 Apr;4(4):183–191. doi: 10.1111/j.1574-6968.1992.tb04993.x. [DOI] [PubMed] [Google Scholar]
  15. Horwitz M. A., Silverstein S. C. Activated human monocytes inhibit the intracellular multiplication of Legionnaires' disease bacteria. J Exp Med. 1981 Nov 1;154(5):1618–1635. doi: 10.1084/jem.154.5.1618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Horwitz M. A., Silverstein S. C. Legionnaires' disease bacterium (Legionella pneumophila) multiples intracellularly in human monocytes. J Clin Invest. 1980 Sep;66(3):441–450. doi: 10.1172/JCI109874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jensen W. A., Rose R. M., Wasserman A. S., Kalb T. H., Anton K., Remold H. G. In vitro activation of the antibacterial activity of human pulmonary macrophages by recombinant gamma interferon. J Infect Dis. 1987 Mar;155(3):574–577. doi: 10.1093/infdis/155.3.574. [DOI] [PubMed] [Google Scholar]
  18. Katz S. M., Hashemi S. Electron microscopic examination of the inflammatory response to Legionella pneumophila in guinea pigs. Lab Invest. 1982 Jan;46(1):24–32. [PubMed] [Google Scholar]
  19. Klein T. W., Yamamoto Y., Brown H. K., Friedman H. Interferon-gamma induced resistance to Legionella pneumophila in susceptible A/J mouse macrophages. J Leukoc Biol. 1991 Jan;49(1):98–103. doi: 10.1002/jlb.49.1.98. [DOI] [PubMed] [Google Scholar]
  20. Levi M. H., Pasculle A. W., Dowling J. N. Role of the alveolar macrophage in host defense and immunity to Legionella micdadei pneumonia in the guinea pig. Microb Pathog. 1987 Apr;2(4):269–282. doi: 10.1016/0882-4010(87)90125-2. [DOI] [PubMed] [Google Scholar]
  21. McDade J. E., Shepard C. C., Fraser D. W., Tsai T. R., Redus M. A., Dowdle W. R. Legionnaires' disease: isolation of a bacterium and demonstration of its role in other respiratory disease. N Engl J Med. 1977 Dec 1;297(22):1197–1203. doi: 10.1056/NEJM197712012972202. [DOI] [PubMed] [Google Scholar]
  22. Nash T. W., Libby D. M., Horwitz M. A. IFN-gamma-activated human alveolar macrophages inhibit the intracellular multiplication of Legionella pneumophila. J Immunol. 1988 Jun 1;140(11):3978–3981. [PubMed] [Google Scholar]
  23. Nash T. W., Libby D. M., Horwitz M. A. Interaction between the legionnaires' disease bacterium (Legionella pneumophila) and human alveolar macrophages. Influence of antibody, lymphokines, and hydrocortisone. J Clin Invest. 1984 Sep;74(3):771–782. doi: 10.1172/JCI111493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schreiber R. D., Hicks L. J., Celada A., Buchmeier N. A., Gray P. W. Monoclonal antibodies to murine gamma-interferon which differentially modulate macrophage activation and antiviral activity. J Immunol. 1985 Mar;134(3):1609–1618. [PubMed] [Google Scholar]
  25. Skerrett S. J., Martin T. R. Recombinant murine interferon-gamma reversibly activates rat alveolar macrophages to kill Legionella pneumophila. J Infect Dis. 1992 Dec;166(6):1354–1361. doi: 10.1093/infdis/166.6.1354. [DOI] [PubMed] [Google Scholar]
  26. Skerrett S. J., Schmidt R. A., Martin T. R. Impaired clearance of aerosolized Legionella pneumophila in corticosteroid-treated rats: a model of Legionnaires' disease in the compromised host. J Infect Dis. 1989 Aug;160(2):261–273. doi: 10.1093/infdis/160.2.261. [DOI] [PubMed] [Google Scholar]
  27. Stein-Streilein J., Guffee J. In vivo treatment of mice and hamsters with antibodies to asialo GM1 increases morbidity and mortality to pulmonary influenza infection. J Immunol. 1986 Feb 15;136(4):1435–1441. [PubMed] [Google Scholar]
  28. Winn W. C., Jr, Davis G. S., Gump D. W., Craighead J. E., Beaty H. N. Legionnaires' pneumonia after intratracheal inoculation of guinea pigs and rats. Lab Invest. 1982 Dec;47(6):568–578. [PubMed] [Google Scholar]
  29. Winn W. C., Jr, Myerowitz R. L. The pathology of the Legionella pneumonias. A review of 74 cases and the literature. Hum Pathol. 1981 May;12(5):401–422. doi: 10.1016/s0046-8177(81)80021-4. [DOI] [PubMed] [Google Scholar]
  30. Yamamoto Y., Klein T. W., Newton C. A., Widen R., Friedman H. Differential growth of Legionella pneumophila in guinea pig versus mouse macrophage cultures. Infect Immun. 1987 Jun;55(6):1369–1374. doi: 10.1128/iai.55.6.1369-1374.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yamamoto Y., Klein T. W., Newton C. A., Widen R., Friedman H. Growth of Legionella pneumophila in thioglycolate-elicited peritoneal macrophages from A/J mice. Infect Immun. 1988 Feb;56(2):370–375. doi: 10.1128/iai.56.2.370-375.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yoshida S., Goto Y., Mizuguchi Y., Nomoto K., Skamene E. Genetic control of natural resistance in mouse macrophages regulating intracellular Legionella pneumophila multiplication in vitro. Infect Immun. 1991 Jan;59(1):428–432. doi: 10.1128/iai.59.1.428-432.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. abu Kwaik Y., Fields B. S., Engleberg N. C. Protein expression by the protozoan Hartmannella vermiformis upon contact with its bacterial parasite Legionella pneumophila. Infect Immun. 1994 May;62(5):1860–1866. doi: 10.1128/iai.62.5.1860-1866.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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