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. 1997 Sep;65(9):3759–3767. doi: 10.1128/iai.65.9.3759-3767.1997

Interaction of Mycobacterium avium with environmental amoebae enhances virulence.

J D Cirillo 1, S Falkow 1, L S Tompkins 1, L E Bermudez 1
PMCID: PMC175536  PMID: 9284149

Abstract

Environmental mycobacteria are a common cause of human infections. Recently, contaminated domestic water supplies have been suggested as a potential environmental source of several mycobacterial diseases. Since many of these mycobacterial species replicate best intracellularly, environmental hosts have been sought. In the present study, we examined the interaction of Mycobacterium avium with a potential protozoan host, the water-borne amoeba Acanthamoeba castellanii. We found that M. avium enters and replicates in A. castellanii. In addition, similar to that shown for mycobacteria within macrophages, M. avium inhibits lysosomal fusion and replicates in vacuoles that are tightly juxtaposed to the bacterial surfaces within amoebae. In order to determine whether growth of M. avium in amoebae plays a role in human infections, we tested the effects of this growth condition on virulence. We found that growth of M. avium in amoebae enhances both entry and intracellular replication compared to growth of bacteria in broth. Furthermore, amoeba-grown M. avium was also more virulent in the beige mouse model of infection. These data suggest a role for protozoa present in water environments as hosts for pathogenic mycobacteria, particularly M. avium.

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Selected References

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  1. Allen P. G., Dawidowicz E. A. Phagocytosis in Acanthamoeba: I. A mannose receptor is responsible for the binding and phagocytosis of yeast. J Cell Physiol. 1990 Dec;145(3):508–513. doi: 10.1002/jcp.1041450317. [DOI] [PubMed] [Google Scholar]
  2. Allen P. G., Dawidowicz E. A. Phagocytosis in Acanthamoeba: II. Soluble and insoluble mannose-rich ligands stimulate phosphoinositide metabolism. J Cell Physiol. 1990 Dec;145(3):514–521. doi: 10.1002/jcp.1041450318. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Armstrong J. A., Hart P. D. Response of cultured macrophages to Mycobacterium tuberculosis, with observations on fusion of lysosomes with phagosomes. J Exp Med. 1971 Sep 1;134(3 Pt 1):713–740. doi: 10.1084/jem.134.3.713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Arnow P. M., Chou T., Weil D., Shapiro E. N., Kretzschmar C. Nosocomial Legionnaires' disease caused by aerosolized tap water from respiratory devices. J Infect Dis. 1982 Oct;146(4):460–467. doi: 10.1093/infdis/146.4.460. [DOI] [PubMed] [Google Scholar]
  6. Barker J., Brown M. R., Collier P. J., Farrell I., Gilbert P. Relationship between Legionella pneumophila and Acanthamoeba polyphaga: physiological status and susceptibility to chemical inactivation. Appl Environ Microbiol. 1992 Aug;58(8):2420–2425. doi: 10.1128/aem.58.8.2420-2425.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Barker J., Brown M. R. Trojan horses of the microbial world: protozoa and the survival of bacterial pathogens in the environment. Microbiology. 1994 Jun;140(Pt 6):1253–1259. doi: 10.1099/00221287-140-6-1253. [DOI] [PubMed] [Google Scholar]
  8. Barker J., Lambert P. A., Brown M. R. Influence of intra-amoebic and other growth conditions on the surface properties of Legionella pneumophila. Infect Immun. 1993 Aug;61(8):3503–3510. doi: 10.1128/iai.61.8.3503-3510.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Benson C. A. Disease due to the Mycobacterium avium complex in patients with AIDS: epidemiology and clinical syndrome. Clin Infect Dis. 1994 Apr;18 (Suppl 3):S218–S222. doi: 10.1093/clinids/18.supplement_3.s218. [DOI] [PubMed] [Google Scholar]
  10. Bermudez L. E., Parker A., Goodman J. R. Growth within macrophages increases the efficiency of Mycobacterium avium in invading other macrophages by a complement receptor-independent pathway. Infect Immun. 1997 May;65(5):1916–1925. doi: 10.1128/iai.65.5.1916-1925.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Bermudez L. E., Petrofsky M., Kolonoski P., Young L. S. An animal model of Mycobacterium avium complex disseminated infection after colonization of the intestinal tract. J Infect Dis. 1992 Jan;165(1):75–79. doi: 10.1093/infdis/165.1.75. [DOI] [PubMed] [Google Scholar]
  12. Bermudez L. E., Shelton K., Young L. S. Comparison of the ability of Mycobacterium avium, M. smegmatis and M. tuberculosis to invade and replicate within HEp-2 epithelial cells. Tuber Lung Dis. 1995 Jun;76(3):240–247. doi: 10.1016/s0962-8479(05)80012-7. [DOI] [PubMed] [Google Scholar]
  13. Bermudez L. E., Young L. S. Factors affecting invasion of HT-29 and HEp-2 epithelial cells by organisms of the Mycobacterium avium complex. Infect Immun. 1994 May;62(5):2021–2026. doi: 10.1128/iai.62.5.2021-2026.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Bermudez L. E., Young L. S. Tumor necrosis factor, alone or in combination with IL-2, but not IFN-gamma, is associated with macrophage killing of Mycobacterium avium complex. J Immunol. 1988 May 1;140(9):3006–3013. [PubMed] [Google Scholar]
  15. Bertram M. A., Inderlied C. B., Yadegar S., Kolanoski P., Yamada J. K., Young L. S. Confirmation of the beige mouse model for study of disseminated infection with Mycobacterium avium complex. J Infect Dis. 1986 Jul;154(1):194–195. doi: 10.1093/infdis/154.1.194. [DOI] [PubMed] [Google Scholar]
  16. Bowers B., Korn E. D. The fine structure of Acanthamoeba castellanii. I. The trophozoite. J Cell Biol. 1968 Oct;39(1):95–111. doi: 10.1083/jcb.39.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Brown R. C., Bass H., Coombs J. P. Carbohydrate binding proteins involved in phagocytosis by Acanthamoeba. Nature. 1975 Apr 3;254(5499):434–435. doi: 10.1038/254434a0. [DOI] [PubMed] [Google Scholar]
  18. CLARK H. F., SHEPARD C. C. EFFECT OF ENVIRONMENTAL TEMPERATURES ON INFECTION WITH MYCOBACTERIUM MARINUM (BALNEI) OF MICE AND A NUMBER OF POIKILOTHERMIC SPECIES. J Bacteriol. 1963 Nov;86:1057–1069. doi: 10.1128/jb.86.5.1057-1069.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Carter R. F., Cullity G. J., Ojeda V. J., Silberstein P., Willaert E. A fatal case of meningoencephalitis due to a free-living amoeba of uncertain identity--probably acanthamoeba sp. Pathology. 1981 Jan;13(1):51–68. doi: 10.3109/00313028109086829. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Colville A., Crowley J., Dearden D., Slack R. C., Lee J. V. Outbreak of Legionnaires' disease at University Hospital, Nottingham. Epidemiology, microbiology and control. Epidemiol Infect. 1993 Feb;110(1):105–116. doi: 10.1017/s0950268800050731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Costerton J. W., Cheng K. J., Geesey G. G., Ladd T. I., Nickel J. C., Dasgupta M., Marrie T. J. Bacterial biofilms in nature and disease. Annu Rev Microbiol. 1987;41:435–464. doi: 10.1146/annurev.mi.41.100187.002251. [DOI] [PubMed] [Google Scholar]
  23. Crowle A. J., Tsang A. Y., Vatter A. E., May M. H. Comparison of 15 laboratory and patient-derived strains of Mycobacterium avium for ability to infect and multiply in cultured human macrophages. J Clin Microbiol. 1986 Nov;24(5):812–821. doi: 10.1128/jcm.24.5.812-821.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Dillon J., Millson C., Morris I. Mycobacterium kansasii infection in the wrist and hand. Br J Rheumatol. 1990 Apr;29(2):150–153. doi: 10.1093/rheumatology/29.2.150. [DOI] [PubMed] [Google Scholar]
  25. Essig A., Heinemann M., Simnacher U., Marre R. Infection of Acanthamoeba castellanii by Chlamydia pneumoniae. Appl Environ Microbiol. 1997 Apr;63(4):1396–1399. doi: 10.1128/aem.63.4.1396-1399.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Frehel C., de Chastellier C., Lang T., Rastogi N. Evidence for inhibition of fusion of lysosomal and prelysosomal compartments with phagosomes in macrophages infected with pathogenic Mycobacterium avium. Infect Immun. 1986 Apr;52(1):252–262. doi: 10.1128/iai.52.1.252-262.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Grange J. M. Environmental mycobacteria and human disease. Lepr Rev. 1991 Dec;62(4):353–361. doi: 10.5935/0305-7518.19910041. [DOI] [PubMed] [Google Scholar]
  28. Grange J. M., Yates M. D., Boughton E. The avian tubercle bacillus and its relatives. J Appl Bacteriol. 1990 May;68(5):411–431. doi: 10.1111/j.1365-2672.1990.tb02892.x. [DOI] [PubMed] [Google Scholar]
  29. Heineman H. S., Spitzer S., Pianphongsant T. Fish tank granuloma. A hobby hazard. Arch Intern Med. 1972 Jul;130(1):121–123. [PubMed] [Google Scholar]
  30. 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]
  31. Horsburgh C. R., Jr Mycobacterium avium complex infection in the acquired immunodeficiency syndrome. N Engl J Med. 1991 May 9;324(19):1332–1338. doi: 10.1056/NEJM199105093241906. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Isberg R. R., Falkow S. A single genetic locus encoded by Yersinia pseudotuberculosis permits invasion of cultured animal cells by Escherichia coli K-12. Nature. 1985 Sep 19;317(6034):262–264. doi: 10.1038/317262a0. [DOI] [PubMed] [Google Scholar]
  34. Kelly P. J., Karlson A. G., Weed L. A., Lipscomb P. R. Infection of synovial tissues by mycobacteria other than Mycobacterium tuberculosis. J Bone Joint Surg Am. 1967 Dec;49(8):1521–1530. [PubMed] [Google Scholar]
  35. Kiely J. L., O'Riordan D. M., Sheehan S., Curtin J., Hogan J., Bredin C. P. Tenosynovitis due to mycobacteria other than tuberculosis: a hazard of water sports and hobbies. Respir Med. 1995 Jan;89(1):69–71. doi: 10.1016/0954-6111(95)90077-2. [DOI] [PubMed] [Google Scholar]
  36. King C. H., Fields B. S., Shotts E. B., Jr, White E. H. Effects of cytochalasin D and methylamine on intracellular growth of Legionella pneumophila in amoebae and human monocyte-like cells. Infect Immun. 1991 Mar;59(3):758–763. doi: 10.1128/iai.59.3.758-763.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. LeChevallier M. W., Cawthon C. D., Lee R. G. Factors promoting survival of bacteria in chlorinated water supplies. Appl Environ Microbiol. 1988 Mar;54(3):649–654. doi: 10.1128/aem.54.3.649-654.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ly T. M., Müller H. E. Ingested Listeria monocytogenes survive and multiply in protozoa. J Med Microbiol. 1990 Sep;33(1):51–54. doi: 10.1099/00222615-33-1-51. [DOI] [PubMed] [Google Scholar]
  39. Maniar A. C., Vanbuckenhout L. R. Mycobacterium kansasii from an environmental source. Can J Public Health. 1976 Jan-Feb;67(1):59–60. [PubMed] [Google Scholar]
  40. Martínez A. J. Free-living amebic meningoencephalitides: comparative study. Neurol Neurocir Psiquiatr. 1977;18(2-3):391–401. [PubMed] [Google Scholar]
  41. Meenhorst P. L., Reingold A. L., Groothuis D. G., Gorman G. W., Wilkinson H. W., McKinney R. M., Feeley J. C., Brenner D. J., van Furth R. Water-related nosocomial pneumonia caused by Legionella pneumophila serogroups 1 and 10. J Infect Dis. 1985 Aug;152(2):356–364. doi: 10.1093/infdis/152.2.356. [DOI] [PubMed] [Google Scholar]
  42. Moffat J. F., Edelstein P. H., Regula D. P., Jr, Cirillo J. D., Tompkins L. S. Effects of an isogenic Zn-metalloprotease-deficient mutant of Legionella pneumophila in a guinea-pig pneumonia model. Mol Microbiol. 1994 Jun;12(5):693–705. doi: 10.1111/j.1365-2958.1994.tb01057.x. [DOI] [PubMed] [Google Scholar]
  43. Moffat J. F., Tompkins L. S. A quantitative model of intracellular growth of Legionella pneumophila in Acanthamoeba castellanii. Infect Immun. 1992 Jan;60(1):296–301. doi: 10.1128/iai.60.1.296-301.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Montecalvo M. A., Forester G., Tsang A. Y., du Moulin G., Wormser G. P. Colonisation of potable water with Mycobacterium avium complex in homes of HIV-infected patients. Lancet. 1994 Jun 25;343(8913):1639–1639. doi: 10.1016/s0140-6736(94)93093-7. [DOI] [PubMed] [Google Scholar]
  45. Muder R. R., Yu V. L., Woo A. H. Mode of transmission of Legionella pneumophila. A critical review. Arch Intern Med. 1986 Aug;146(8):1607–1612. [PubMed] [Google Scholar]
  46. 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]
  47. Niszl I. A., Markus M. B. Processing of free-living amoebae for transmission electron microscopy. Stain Technol. 1989 Sep;64(5):259–260. doi: 10.3109/10520298909107014. [DOI] [PubMed] [Google Scholar]
  48. PHILPOTT J. A., Jr, WOODBURNE A. R., PHILPOTT O. S., SCHAEFER W. B., MOLLOHAN C. S. SWIMMING POOL GRANULOMA. A STUDY OF 290 CASES. Arch Dermatol. 1963 Aug;88:158–162. doi: 10.1001/archderm.1963.01590200046008. [DOI] [PubMed] [Google Scholar]
  49. Plum G., Clark-Curtiss J. E. Induction of Mycobacterium avium gene expression following phagocytosis by human macrophages. Infect Immun. 1994 Feb;62(2):476–483. doi: 10.1128/iai.62.2.476-483.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Ramakrishnan L., Falkow S. Mycobacterium marinum persists in cultured mammalian cells in a temperature-restricted fashion. Infect Immun. 1994 Aug;62(8):3222–3229. doi: 10.1128/iai.62.8.3222-3229.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. 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]
  52. Ryter A., Frehel C., Rastogi N., David H. L. Macrophage interaction with mycobacteria including M. leprae. Acta Leprol. 1984 Oct-Dec;2(2-4):211–226. [PubMed] [Google Scholar]
  53. SHEPARD C. C. Growth characteristics in HeLa cells of the rapidly growing acid fast bacteria, Mycobacterium fortuitum, Mycobacterium phlei, and Mycobacterium smegmatis. J Bacteriol. 1957 Jun;73(6):722–726. doi: 10.1128/jb.73.6.722-726.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. SHEPARD C. C. Growth characteristics of tubercle bacilli and certain other mycobacteria in HeLa cells. J Exp Med. 1957 Jan 1;105(1):39–48. doi: 10.1084/jem.105.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Shands K. N., Ho J. L., Meyer R. D., Gorman G. W., Edelstein P. H., Mallison G. F., Finegold S. M., Fraser D. W. Potable water as a source of Legionnaires' disease. JAMA. 1985 Mar 8;253(10):1412–1416. [PubMed] [Google Scholar]
  56. Thom S., Warhurst D., Drasar B. S. Association of Vibrio cholerae with fresh water amoebae. J Med Microbiol. 1992 May;36(5):303–306. doi: 10.1099/00222615-36-5-303. [DOI] [PubMed] [Google Scholar]
  57. 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]
  58. Wendt S. L., George K. L., Parker B. C., Gruft H., Falkinham J. O., 3rd Epidemiology of infection by nontuberculous Mycobacteria. III. Isolation of potentially pathogenic mycobacteria from aerosols. Am Rev Respir Dis. 1980 Aug;122(2):259–263. doi: 10.1164/arrd.1980.122.2.259. [DOI] [PubMed] [Google Scholar]
  59. de Chastellier C., Fréhel C., Offredo C., Skamene E. Implication of phagosome-lysosome fusion in restriction of Mycobacterium avium growth in bone marrow macrophages from genetically resistant mice. Infect Immun. 1993 Sep;61(9):3775–3784. doi: 10.1128/iai.61.9.3775-3784.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. du Moulin G. C., Stottmeier K. D., Pelletier P. A., Tsang A. Y., Hedley-Whyte J. Concentration of Mycobacterium avium by hospital hot water systems. JAMA. 1988 Sep 16;260(11):1599–1601. doi: 10.1001/jama.260.11.1599. [DOI] [PubMed] [Google Scholar]
  61. von Reyn C. F., Maslow J. N., Barber T. W., Falkinham J. O., 3rd, Arbeit R. D. Persistent colonisation of potable water as a source of Mycobacterium avium infection in AIDS. Lancet. 1994 May 7;343(8906):1137–1141. doi: 10.1016/s0140-6736(94)90239-9. [DOI] [PubMed] [Google Scholar]
  62. von Reyn C. F., Waddell R. D., Eaton T., Arbeit R. D., Maslow J. N., Barber T. W., Brindle R. J., Gilks C. F., Lumio J., Lähdevirta J. Isolation of Mycobacterium avium complex from water in the United States, Finland, Zaire, and Kenya. J Clin Microbiol. 1993 Dec;31(12):3227–3230. doi: 10.1128/jcm.31.12.3227-3230.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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