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. 1995 Apr;63(4):1541–1551. doi: 10.1128/iai.63.4.1541-1551.1995

Characterization of Pseudomonas aeruginosa-induced MDCK cell injury: glycosylation-defective host cells are resistant to bacterial killing.

G Apodaca 1, M Bomsel 1, R Lindstedt 1, J Engel 1, D Frank 1, K E Mostov 1, J Wiener-Kronish 1
PMCID: PMC173187  PMID: 7890421

Abstract

As a model for bacterium-induced epithelial cell injury, we have studied the interaction of Pseudomonas aeruginosa with polarized Madin-Darby canine kidney (MDCK) cells grown on filters. Following an initial period of bacterial adhesion, foci of injured host cells, which consisted of a central region of cell debris, surrounded by cells that were permeable and apparently necrotic, were formed. Host cell death was quantified by measuring the increased permeability of the monolayer to the macromolecular tracer [14C]inulin. Using this MDCK model system, we have identified bacterial and host cell factors necessary for the host cell damage. The ability of P. aeruginosa to cause MDCK cell damage was independent of elastase or exotoxin A production. In contrast, bacteria with a mutation in the regulatory locus exsA (which are deficient in exoenzyme S production) neither bound to nor caused host cell injury. MDCK cells with defects in cell surface glycosylation were resistant to cell injury, indicating that bacteria may require host cell glycolipids and/or glycoproteins as points of adhesion to cause subsequent host cell injury.

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Selected References

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  1. Andersson B., Dahmén J., Frejd T., Leffler H., Magnusson G., Noori G., Edén C. S. Identification of an active disaccharide unit of a glycoconjugate receptor for pneumococci attaching to human pharyngeal epithelial cells. J Exp Med. 1983 Aug 1;158(2):559–570. doi: 10.1084/jem.158.2.559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Apodaca G., Katz L. A., Mostov K. E. Receptor-mediated transcytosis of IgA in MDCK cells is via apical recycling endosomes. J Cell Biol. 1994 Apr;125(1):67–86. doi: 10.1083/jcb.125.1.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker N., Hansson G. C., Leffler H., Riise G., Svanborg-Edén C. Glycosphingolipid receptors for Pseudomonas aeruginosa. Infect Immun. 1990 Jul;58(7):2361–2366. doi: 10.1128/iai.58.7.2361-2366.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baltimore R. S., Christie C. D., Smith G. J. Immunohistopathologic localization of Pseudomonas aeruginosa in lungs from patients with cystic fibrosis. Implications for the pathogenesis of progressive lung deterioration. Am Rev Respir Dis. 1989 Dec;140(6):1650–1661. doi: 10.1164/ajrccm/140.6.1650. [DOI] [PubMed] [Google Scholar]
  5. Breitfeld P. P., Casanova J. E., Harris J. M., Simister N. E., Mostov K. E. Expression and analysis of the polymeric immunoglobulin receptor in Madin-Darby canine kidney cells using retroviral vectors. Methods Cell Biol. 1989;32:329–337. doi: 10.1016/s0091-679x(08)61178-4. [DOI] [PubMed] [Google Scholar]
  6. Bryan C. S., Reynolds K. L. Bacteremic nosocomial pneumonia. Analysis of 172 episodes from a single metropolitan area. Am Rev Respir Dis. 1984 May;129(5):668–671. doi: 10.1164/arrd.1984.129.5.668. [DOI] [PubMed] [Google Scholar]
  7. Brändli A. W., Hansson G. C., Rodriguez-Boulan E., Simons K. A polarized epithelial cell mutant deficient in translocation of UDP-galactose into the Golgi complex. J Biol Chem. 1988 Nov 5;263(31):16283–16290. [PubMed] [Google Scholar]
  8. Ceri H., McArthur H. A., Whitfield C. Association of alginate from Pseudomonas aeruginosa with two forms of heparin-binding lectin isolated from rat lung. Infect Immun. 1986 Jan;51(1):1–5. doi: 10.1128/iai.51.1.1-5.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Coburn J., Gill D. M. ADP-ribosylation of p21ras and related proteins by Pseudomonas aeruginosa exoenzyme S. Infect Immun. 1991 Nov;59(11):4259–4262. doi: 10.1128/iai.59.11.4259-4262.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Coburn J., Kane A. V., Feig L., Gill D. M. Pseudomonas aeruginosa exoenzyme S requires a eukaryotic protein for ADP-ribosyltransferase activity. J Biol Chem. 1991 Apr 5;266(10):6438–6446. [PubMed] [Google Scholar]
  12. Coburn J., Wyatt R. T., Iglewski B. H., Gill D. M. Several GTP-binding proteins, including p21c-H-ras, are preferred substrates of Pseudomonas aeruginosa exoenzyme S. J Biol Chem. 1989 May 25;264(15):9004–9008. [PubMed] [Google Scholar]
  13. Doig P., Smith N. R., Todd T., Irvin R. T. Characterization of the binding of Pseudomonas aeruginosa alginate to human epithelial cells. Infect Immun. 1987 Jun;55(6):1517–1522. doi: 10.1128/iai.55.6.1517-1522.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Frank D. W., Nair G., Schweizer H. P. Construction and characterization of chromosomal insertional mutations of the Pseudomonas aeruginosa exoenzyme S trans-regulatory locus. Infect Immun. 1994 Feb;62(2):554–563. doi: 10.1128/iai.62.2.554-563.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grant M. M., Niederman M. S., Poehlman M. A., Fein A. M. Characterization of Pseudomonas aeruginosa adherence to cultured hamster tracheal epithelial cells. Am J Respir Cell Mol Biol. 1991 Dec;5(6):563–570. doi: 10.1165/ajrcmb/5.6.563. [DOI] [PubMed] [Google Scholar]
  16. Gumbiner B., Simons K. A functional assay for proteins involved in establishing an epithelial occluding barrier: identification of a uvomorulin-like polypeptide. J Cell Biol. 1986 Feb;102(2):457–468. doi: 10.1083/jcb.102.2.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hannan L. A., Lisanti M. P., Rodriguez-Boulan E., Edidin M. Correctly sorted molecules of a GPI-anchored protein are clustered and immobile when they arrive at the apical surface of MDCK cells. J Cell Biol. 1993 Jan;120(2):353–358. doi: 10.1083/jcb.120.2.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hata J. S., Fick R. B., Jr Airway adherence of Pseudomonas aeruginosa: mucoexopolysaccharide binding to human and bovine airway proteins. J Lab Clin Med. 1991 May;117(5):410–422. [PubMed] [Google Scholar]
  19. Johnson J., Warren R. L., Branstrom A. A. Effects of FP2 and a mercury resistance plasmid from Pseudomonas aeruginosa PA103 on exoenzyme production. J Clin Microbiol. 1991 May;29(5):940–944. doi: 10.1128/jcm.29.5.940-944.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Krivan H. C., Ginsburg V., Roberts D. D. Pseudomonas aeruginosa and Pseudomonas cepacia isolated from cystic fibrosis patients bind specifically to gangliotetraosylceramide (asialo GM1) and gangliotriaosylceramide (asialo GM2). Arch Biochem Biophys. 1988 Jan;260(1):493–496. doi: 10.1016/0003-9861(88)90473-0. [DOI] [PubMed] [Google Scholar]
  21. Lam J., Chan R., Lam K., Costerton J. W. Production of mucoid microcolonies by Pseudomonas aeruginosa within infected lungs in cystic fibrosis. Infect Immun. 1980 May;28(2):546–556. doi: 10.1128/iai.28.2.546-556.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Le Bivic A., Garcia M., Rodriguez-Boulan E. Ricin-resistant Madin-Darby canine kidney cells missort a major endogenous apical sialoglycoprotein. J Biol Chem. 1993 Apr 5;268(10):6909–6916. [PubMed] [Google Scholar]
  23. Marcus H., Austria A., Baker N. R. Adherence of Pseudomonas aeruginosa to tracheal epithelium. Infect Immun. 1989 Apr;57(4):1050–1053. doi: 10.1128/iai.57.4.1050-1053.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Marcus H., Baker N. R. Quantitation of adherence of mucoid and nonmucoid Pseudomonas aeruginosa to hamster tracheal epithelium. Infect Immun. 1985 Mar;47(3):723–729. doi: 10.1128/iai.47.3.723-729.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. McArthur H. A., Ceri H. Interaction of a rat lung lectin with the exopolysaccharides of Pseudomonas aeruginosa. Infect Immun. 1983 Nov;42(2):574–578. doi: 10.1128/iai.42.2.574-578.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nicas T. I., Iglewski B. H. Isolation and characterization of transposon-induced mutants of Pseudomonas aeruginosa deficient in production of exoenzyme S. Infect Immun. 1984 Aug;45(2):470–474. doi: 10.1128/iai.45.2.470-474.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pittet J. F., Wiener-Kronish J. P., McElroy M. C., Folkesson H. G., Matthay M. A. Stimulation of lung epithelial liquid clearance by endogenous release of catecholamines in septic shock in anesthetized rats. J Clin Invest. 1994 Aug;94(2):663–671. doi: 10.1172/JCI117383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Plotkowski M. C., Chevillard M., Pierrot D., Altemayer D., Zahm J. M., Colliot G., Puchelle E. Differential adhesion of Pseudomonas aeruginosa to human respiratory epithelial cells in primary culture. J Clin Invest. 1991 Jun;87(6):2018–2028. doi: 10.1172/JCI115231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Prentki P., Krisch H. M. In vitro insertional mutagenesis with a selectable DNA fragment. Gene. 1984 Sep;29(3):303–313. doi: 10.1016/0378-1119(84)90059-3. [DOI] [PubMed] [Google Scholar]
  30. Ramphal R., Guay C., Pier G. B. Pseudomonas aeruginosa adhesins for tracheobronchial mucin. Infect Immun. 1987 Mar;55(3):600–603. doi: 10.1128/iai.55.3.600-603.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ramphal R., Koo L., Ishimoto K. S., Totten P. A., Lara J. C., Lory S. Adhesion of Pseudomonas aeruginosa pilin-deficient mutants to mucin. Infect Immun. 1991 Apr;59(4):1307–1311. doi: 10.1128/iai.59.4.1307-1311.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ramphal R., Pyle M. Adherence of mucoid and nonmucoid Pseudomonas aeruginosa to acid-injured tracheal epithelium. Infect Immun. 1983 Jul;41(1):345–351. doi: 10.1128/iai.41.1.345-351.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ramphal R., Pyle M. Evidence for mucins and sialic acid as receptors for Pseudomonas aeruginosa in the lower respiratory tract. Infect Immun. 1983 Jul;41(1):339–344. doi: 10.1128/iai.41.1.339-344.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ramphal R., Sadoff J. C., Pyle M., Silipigni J. D. Role of pili in the adherence of Pseudomonas aeruginosa to injured tracheal epithelium. Infect Immun. 1984 Apr;44(1):38–40. doi: 10.1128/iai.44.1.38-40.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rosenstein I. J., Yuen C. T., Stoll M. S., Feizi T. Differences in the binding specificities of Pseudomonas aeruginosa M35 and Escherichia coli C600 for lipid-linked oligosaccharides with lactose-related core regions. Infect Immun. 1992 Dec;60(12):5078–5084. doi: 10.1128/iai.60.12.5078-5084.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rosqvist R., Magnusson K. E., Wolf-Watz H. Target cell contact triggers expression and polarized transfer of Yersinia YopE cytotoxin into mammalian cells. EMBO J. 1994 Feb 15;13(4):964–972. doi: 10.1002/j.1460-2075.1994.tb06341.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rostand K. S., Esko J. D. Cholesterol and cholesterol esters: host receptors for Pseudomonas aeruginosa adherence. J Biol Chem. 1993 Nov 15;268(32):24053–24059. [PubMed] [Google Scholar]
  38. Saiman L., Prince A. Pseudomonas aeruginosa pili bind to asialoGM1 which is increased on the surface of cystic fibrosis epithelial cells. J Clin Invest. 1993 Oct;92(4):1875–1880. doi: 10.1172/JCI116779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Simpson D. A., Ramphal R., Lory S. Genetic analysis of Pseudomonas aeruginosa adherence: distinct genetic loci control attachment to epithelial cells and mucins. Infect Immun. 1992 Sep;60(9):3771–3779. doi: 10.1128/iai.60.9.3771-3779.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wiener-Kronish J. P., Albertine K. H., Matthay M. A. Differential responses of the endothelial and epithelial barriers of the lung in sheep to Escherichia coli endotoxin. J Clin Invest. 1991 Sep;88(3):864–875. doi: 10.1172/JCI115388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wiener-Kronish J. P., Sakuma T., Kudoh I., Pittet J. F., Frank D., Dobbs L., Vasil M. L., Matthay M. A. Alveolar epithelial injury and pleural empyema in acute P. aeruginosa pneumonia in anesthetized rabbits. J Appl Physiol (1985) 1993 Oct;75(4):1661–1669. doi: 10.1152/jappl.1993.75.4.1661. [DOI] [PubMed] [Google Scholar]
  42. Woods D. E., Straus D. C., Johanson W. G., Jr, Berry V. K., Bass J. A. Role of pili in adherence of Pseudomonas aeruginosa to mammalian buccal epithelial cells. Infect Immun. 1980 Sep;29(3):1146–1151. doi: 10.1128/iai.29.3.1146-1151.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zychlinsky A. Programmed cell death in infectious diseases. Trends Microbiol. 1993 Jun;1(3):114–117. doi: 10.1016/0966-842x(93)90118-b. [DOI] [PubMed] [Google Scholar]

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