Abstract
The potential role of inhaled protozoa as a niche for intrapulmonary replication of Legionella pneumophila was investigated in vivo with mutant strains of L. pneumophila which have reduced virulence for the amoeba Hartmannella vermiformis. L. pneumophila AA488 and AA502 were derived from wild-type strain AA100 after transposon mutagenesis. These mutants have reduced virulence for H. vermiformis but are fully virulent for mononuclear phagocytic cells. A/J mice, which are susceptible to replicative L. pneumophila lung infections, were inoculated intratracheally with L. pneumophila AA100, AA488, or AA502 (10[6] bacteria per mouse) or were coinoculated with one of the L. pneumophila strains (10[6] bacteria per mouse) and uninfected H. vermiformis (10[6] amoebae per mouse). The effect of coinoculation with H. vermiformis on intrapulmonary growth of each L. pneumophila strain was subsequently assessed. In agreement with our previous studies, coinoculation with H. vermiformis significantly enhanced intrapulmonary growth of the parent L. pneumophila strain (AA100). In contrast, intrapulmonary growth of L. pneumophila AA488 or AA502 was not significantly enhanced by coinoculation of mice with H. vermiformis. These studies demonstrate that L. pneumophila virulence for amoebae is required for maximal intrapulmonary growth of the bacteria in mice coinoculated with H. vermiformis and support the hypothesis that inhaled amoebae may potentiate intrapulmonary growth of L. pneumophila by providing a niche for bacterial replication.
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- Abu Kwaik Y. The phagosome containing Legionella pneumophila within the protozoan Hartmannella vermiformis is surrounded by the rough endoplasmic reticulum. Appl Environ Microbiol. 1996 Jun;62(6):2022–2028. doi: 10.1128/aem.62.6.2022-2028.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Anand C. M., Skinner A. R., Malic A., Kurtz J. B. Interaction of L. pneumophilia and a free living amoeba (Acanthamoeba palestinensis). J Hyg (Lond) 1983 Oct;91(2):167–178. doi: 10.1017/s0022172400060174. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barbaree J. M., Fields B. S., Feeley J. C., Gorman G. W., Martin W. T. Isolation of protozoa from water associated with a legionellosis outbreak and demonstration of intracellular multiplication of Legionella pneumophila. Appl Environ Microbiol. 1986 Feb;51(2):422–424. doi: 10.1128/aem.51.2.422-424.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Breiman R. F., Fields B. S., Sanden G. N., Volmer L., Meier A., Spika J. S. Association of shower use with Legionnaires' disease. Possible role of amoebae. JAMA. 1990 Jun 6;263(21):2924–2926. [PubMed] [Google Scholar]
- Brieland J. K., Remick D. G., Freeman P. T., Hurley M. C., Fantone J. C., Engleberg N. C. In vivo regulation of replicative Legionella pneumophila lung infection by endogenous tumor necrosis factor alpha and nitric oxide. Infect Immun. 1995 Sep;63(9):3253–3258. doi: 10.1128/iai.63.9.3253-3258.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brieland J., Freeman P., Kunkel R., Chrisp C., Hurley M., Fantone J., Engleberg C. Replicative Legionella pneumophila lung infection in intratracheally inoculated A/J mice. A murine model of human Legionnaires' disease. Am J Pathol. 1994 Dec;145(6):1537–1546. [PMC free article] [PubMed] [Google Scholar]
- Brieland J., McClain M., Heath L., Chrisp C., Huffnagle G., LeGendre M., Hurley M., Fantone J., Engleberg C. Coinoculation with Hartmannella vermiformis enhances replicative Legionella pneumophila lung infection in a murine model of Legionnaires' disease. Infect Immun. 1996 Jul;64(7):2449–2456. doi: 10.1128/iai.64.7.2449-2456.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cianciotto N. P., Eisenstein B. I., Mody C. H., Toews G. B., Engleberg N. C. A Legionella pneumophila gene encoding a species-specific surface protein potentiates initiation of intracellular infection. Infect Immun. 1989 Apr;57(4):1255–1262. doi: 10.1128/iai.57.4.1255-1262.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cirillo J. D., Falkow S., Tompkins L. S. Growth of Legionella pneumophila in Acanthamoeba castellanii enhances invasion. Infect Immun. 1994 Aug;62(8):3254–3261. doi: 10.1128/iai.62.8.3254-3261.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Edelstein P. H. Improved semiselective medium for isolation of Legionella pneumophila from contaminated clinical and environmental specimens. J Clin Microbiol. 1981 Sep;14(3):298–303. doi: 10.1128/jcm.14.3.298-303.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feeley J. C., Gibson R. J., Gorman G. W., Langford N. C., Rasheed J. K., Mackel D. C., Baine W. B. Charcoal-yeast extract agar: primary isolation medium for Legionella pneumophila. J Clin Microbiol. 1979 Oct;10(4):437–441. doi: 10.1128/jcm.10.4.437-441.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fields B. S., Fields S. R., Loy J. N., White E. H., Steffens W. L., Shotts E. B. Attachment and entry of Legionella pneumophila in Hartmannella vermiformis. J Infect Dis. 1993 May;167(5):1146–1150. doi: 10.1093/infdis/167.5.1146. [DOI] [PubMed] [Google Scholar]
- Fields B. S., Nerad T. A., Sawyer T. K., King C. H., Barbaree J. M., Martin W. T., Morrill W. E., Sanden G. N. Characterization of an axenic strain of Hartmannella vermiformis obtained from an investigation of nosocomial legionellosis. J Protozool. 1990 Nov-Dec;37(6):581–583. doi: 10.1111/j.1550-7408.1990.tb01269.x. [DOI] [PubMed] [Google Scholar]
- Fields B. S., Shotts E. B., Jr, Feeley J. C., Gorman G. W., Martin W. T. Proliferation of Legionella pneumophila as an intracellular parasite of the ciliated protozoan Tetrahymena pyriformis. Appl Environ Microbiol. 1984 Mar;47(3):467–471. doi: 10.1128/aem.47.3.467-471.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Henke M., Seidel K. M. Association between Legionella pneumophila and amoebae in water. Isr J Med Sci. 1986 Sep;22(9):690–695. [PubMed] [Google Scholar]
- Holden E. P., Winkler H. H., Wood D. O., Leinbach E. D. Intracellular growth of Legionella pneumophila within Acanthamoeba castellanii Neff. Infect Immun. 1984 Jul;45(1):18–24. doi: 10.1128/iai.45.1.18-24.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Horwitz M. A. The Legionnaires' disease bacterium (Legionella pneumophila) inhibits phagosome-lysosome fusion in human monocytes. J Exp Med. 1983 Dec 1;158(6):2108–2126. doi: 10.1084/jem.158.6.2108. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McClain M. S., Engleberg N. C. Construction of an alkaline phosphatase fusion-generating transposon, mTn10phoA. Gene. 1996 Apr 17;170(1):147–148. doi: 10.1016/0378-1119(95)00856-x. [DOI] [PubMed] [Google Scholar]
- 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]
- Meduri G. U., Kohler G., Headley S., Tolley E., Stentz F., Postlethwaite A. Inflammatory cytokines in the BAL of patients with ARDS. Persistent elevation over time predicts poor outcome. Chest. 1995 Nov;108(5):1303–1314. doi: 10.1378/chest.108.5.1303. [DOI] [PubMed] [Google Scholar]
- 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]
- Newsome A. L., Baker R. L., Miller R. D., Arnold R. R. Interactions between Naegleria fowleri and Legionella pneumophila. Infect Immun. 1985 Nov;50(2):449–452. doi: 10.1128/iai.50.2.449-452.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pearlman E., Jiwa A. H., Engleberg N. C., Eisenstein B. I. Growth of Legionella pneumophila in a human macrophage-like (U937) cell line. Microb Pathog. 1988 Aug;5(2):87–95. doi: 10.1016/0882-4010(88)90011-3. [DOI] [PubMed] [Google Scholar]
- Rowbotham T. J. Current views on the relationships between amoebae, legionellae and man. Isr J Med Sci. 1986 Sep;22(9):678–689. [PubMed] [Google Scholar]
- Rowbotham T. J. Preliminary report on the pathogenicity of Legionella pneumophila for freshwater and soil amoebae. J Clin Pathol. 1980 Dec;33(12):1179–1183. doi: 10.1136/jcp.33.12.1179. [DOI] [PMC free article] [PubMed] [Google Scholar]
- St John R. C., Dorinsky P. M. Immunologic therapy for ARDS, septic shock, and multiple-organ failure. Chest. 1993 Mar;103(3):932–943. doi: 10.1378/chest.103.3.932. [DOI] [PubMed] [Google Scholar]
- 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]
- Swanson M. S., Isberg R. R. Association of Legionella pneumophila with the macrophage endoplasmic reticulum. Infect Immun. 1995 Sep;63(9):3609–3620. doi: 10.1128/iai.63.9.3609-3620.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Swanson M. S., Isberg R. R. Identification of Legionella pneumophila mutants that have aberrant intracellular fates. Infect Immun. 1996 Jul;64(7):2585–2594. doi: 10.1128/iai.64.7.2585-2594.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tyndall R. L., Domingue E. L. Cocultivation of Legionella pneumophila and free-living amoebae. Appl Environ Microbiol. 1982 Oct;44(4):954–959. doi: 10.1128/aem.44.4.954-959.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wadowsky R. M., Butler L. J., Cook M. K., Verma S. M., Paul M. A., Fields B. S., Keleti G., Sykora J. L., Yee R. B. Growth-supporting activity for Legionella pneumophila in tap water cultures and implication of hartmannellid amoebae as growth factors. Appl Environ Microbiol. 1988 Nov;54(11):2677–2682. doi: 10.1128/aem.54.11.2677-2682.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]