Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1996 Oct;64(10):4054–4059. doi: 10.1128/iai.64.10.4054-4059.1996

Invasion of respiratory epithelial cells by Burkholderia (Pseudomonas) cepacia.

J L Burns 1, M Jonas 1, E Y Chi 1, D K Clark 1, A Berger 1, A Griffith 1
PMCID: PMC174336  PMID: 8926068

Abstract

Pulmonary infections caused by Burkholderia (Pseudomonas) cepacia are an important cause of morbidity and mortality in cystic fibrosis (CF) patients. Several features suggestive of cellular invasion and intracellular sequestration of B. cepacia in CF are persistence of infection in the face of antibiotic therapy to which the organism demonstrates in vitro susceptibility and a propensity to cause bacteremic infections in patients with CF. Epithelial cell invasion was demonstrated in vitro in A549 cells by a modified gentamicin protection assay. The kinetics of invasion appear to be saturable. Electron microscopy of invaded monolayers showed intracytoplasmic bacteria enclosed by membrane-bound vacuoles. No lysosomal fusion with these vacuoles was observed. Intraepithelial cell replication was suggested by electron microscopy and confirmed by both a quantitative assay and a visual assay. Cytochalasin D, but not colchicine, inhibited invasion, suggesting a role for microfilaments but not microtubules. The invasion phenotype in B. cepacia may be an important virulence factor for CF infections.

Full Text

The Full Text of this article is available as a PDF (1,008.4 KB).

Selected References

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

  1. Cheng H. P., Lessie T. G. Multiple replicons constituting the genome of Pseudomonas cepacia 17616. J Bacteriol. 1994 Jul;176(13):4034–4042. doi: 10.1128/jb.176.13.4034-4042.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chi E., Mehl T., Nunn D., Lory S. Interaction of Pseudomonas aeruginosa with A549 pneumocyte cells. Infect Immun. 1991 Mar;59(3):822–828. doi: 10.1128/iai.59.3.822-828.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ewanowich C. A., Sherburne R. K., Man S. F., Peppler M. S. Bordetella parapertussis invasion of HeLa 229 cells and human respiratory epithelial cells in primary culture. Infect Immun. 1989 Apr;57(4):1240–1247. doi: 10.1128/iai.57.4.1240-1247.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Finlay B. B., Falkow S. Comparison of the invasion strategies used by Salmonella cholerae-suis, Shigella flexneri and Yersinia enterocolitica to enter cultured animal cells: endosome acidification is not required for bacterial invasion or intracellular replication. Biochimie. 1988 Aug;70(8):1089–1099. doi: 10.1016/0300-9084(88)90271-4. [DOI] [PubMed] [Google Scholar]
  5. Finlay B. B., Falkow S. Salmonella interactions with polarized human intestinal Caco-2 epithelial cells. J Infect Dis. 1990 Nov;162(5):1096–1106. doi: 10.1093/infdis/162.5.1096. [DOI] [PubMed] [Google Scholar]
  6. Finlay B. B., Ruschkowski S., Dedhar S. Cytoskeletal rearrangements accompanying salmonella entry into epithelial cells. J Cell Sci. 1991 Jun;99(Pt 2):283–296. doi: 10.1242/jcs.99.2.283. [DOI] [PubMed] [Google Scholar]
  7. Fleiszig S. M., Zaidi T. S., Fletcher E. L., Preston M. J., Pier G. B. Pseudomonas aeruginosa invades corneal epithelial cells during experimental infection. Infect Immun. 1994 Aug;62(8):3485–3493. doi: 10.1128/iai.62.8.3485-3493.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gahring L. C., Heffron F., Finlay B. B., Falkow S. Invasion and replication of Salmonella typhimurium in animal cells. Infect Immun. 1990 Feb;58(2):443–448. doi: 10.1128/iai.58.2.443-448.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gaillard J. L., Berche P., Mounier J., Richard S., Sansonetti P. In vitro model of penetration and intracellular growth of Listeria monocytogenes in the human enterocyte-like cell line Caco-2. Infect Immun. 1987 Nov;55(11):2822–2829. doi: 10.1128/iai.55.11.2822-2829.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gold R., Jin E., Levison H., Isles A., Fleming P. C. Ceftazidime alone and in combination in patients with cystic fibrosis: lack of efficacy in treatment of severe respiratory infections caused by Pseudomonas cepacia. J Antimicrob Chemother. 1983 Jul;12 (Suppl A):331–336. doi: 10.1093/jac/12.suppl_a.331. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Jørgensen S., Skov K. W., Diderichsen B. Cloning, sequence, and expression of a lipase gene from Pseudomonas cepacia: lipase production in heterologous hosts requires two Pseudomonas genes. J Bacteriol. 1991 Jan;173(2):559–567. doi: 10.1128/jb.173.2.559-567.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Konkel M. E., Mead D. J., Hayes S. F., Cieplak W., Jr Translocation of Campylobacter jejuni across human polarized epithelial cell monolayer cultures. J Infect Dis. 1992 Aug;166(2):308–315. doi: 10.1093/infdis/166.2.308. [DOI] [PubMed] [Google Scholar]
  14. Kooi C., Cox A., Darling P., Sokol P. A. Neutralizing monoclonal antibodies to an extracellular Pseudomonas cepacia protease. Infect Immun. 1994 Jul;62(7):2811–2817. doi: 10.1128/iai.62.7.2811-2817.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lonon M. K., Woods D. E., Straus D. C. Production of lipase by clinical isolates of Pseudomonas cepacia. J Clin Microbiol. 1988 May;26(5):979–984. doi: 10.1128/jcm.26.5.979-984.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McKevitt A. I., Woods D. E. Characterization of Pseudomonas cepacia isolates from patients with cystic fibrosis. J Clin Microbiol. 1984 Feb;19(2):291–293. doi: 10.1128/jcm.19.2.291-293.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Miliotis M. D. Acridine orange stain for determining intracellular enteropathogens in HeLa cells. J Clin Microbiol. 1991 Apr;29(4):830–831. doi: 10.1128/jcm.29.4.830-831.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nakazawa T., Yamada Y., Ishibashi M. Characterization of hemolysin in extracellular products of Pseudomonas cepacia. J Clin Microbiol. 1987 Feb;25(2):195–198. doi: 10.1128/jcm.25.2.195-198.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pier G. B., Grout M., Zaidi T. S., Olsen J. C., Johnson L. G., Yankaskas J. R., Goldberg J. B. Role of mutant CFTR in hypersusceptibility of cystic fibrosis patients to lung infections. Science. 1996 Jan 5;271(5245):64–67. doi: 10.1126/science.271.5245.64. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rosenstein B. J., Hall D. E. Pneumonia and septicemia due to Pseudomonas cepacia in a patient with cystic fibrosis. Johns Hopkins Med J. 1980 Nov;147(5):188–189. [PubMed] [Google Scholar]
  21. Rubens C. E., Raff H. V., Jackson J. C., Chi E. Y., Bielitzki J. T., Hillier S. L. Pathophysiology and histopathology of group B streptococcal sepsis in Macaca nemestrina primates induced after intraamniotic inoculation: evidence for bacterial cellular invasion. J Infect Dis. 1991 Aug;164(2):320–330. doi: 10.1093/infdis/164.2.320. [DOI] [PubMed] [Google Scholar]
  22. Rubens C. E., Smith S., Hulse M., Chi E. Y., van Belle G. Respiratory epithelial cell invasion by group B streptococci. Infect Immun. 1992 Dec;60(12):5157–5163. doi: 10.1128/iai.60.12.5157-5163.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sajjan U. S., Sun L., Goldstein R., Forstner J. F. Cable (cbl) type II pili of cystic fibrosis-associated Burkholderia (Pseudomonas) cepacia: nucleotide sequence of the cblA major subunit pilin gene and novel morphology of the assembled appendage fibers. J Bacteriol. 1995 Feb;177(4):1030–1038. doi: 10.1128/jb.177.4.1030-1038.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Vasil M. L., Krieg D. P., Kuhns J. S., Ogle J. W., Shortridge V. D., Ostroff R. M., Vasil A. I. Molecular analysis of hemolytic and phospholipase C activities of Pseudomonas cepacia. Infect Immun. 1990 Dec;58(12):4020–4029. doi: 10.1128/iai.58.12.4020-4029.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wilson C. B., Jacobs R. F., Smith A. L. Cellular antibiotic pharmacology. Semin Perinatol. 1982 Apr;6(2):205–213. [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES