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. 1997 Jul;65(7):2861–2867. doi: 10.1128/iai.65.7.2861-2867.1997

Epithelial cell polarity affects susceptibility to Pseudomonas aeruginosa invasion and cytotoxicity.

S M Fleiszig 1, D J Evans 1, N Do 1, V Vallas 1, S Shin 1, K E Mostov 1
PMCID: PMC175402  PMID: 9199460

Abstract

Intact tissues are relatively resistant to Pseudomonas aeruginosa-induced disease, and injury predisposes tissue to infection. Intact epithelia contain polarized cells that have distinct apical and basolateral membranes with unique lipids and proteins. In this study, the role of cell polarity in epithelial cell susceptibility to P. aeruginosa virulence mechanisms was tested. Madin-Darby canine kidney (MDCK) cells, human corneal epithelial cells, and primary cultures of two different types of airway epithelial cells were grown on Transwell filters or in plastic tissue culture wells. P. aeruginosa invasion of cells was quantified by gentamicin survival assays with two isolates that invade epithelial cells (6294 and PAO1). Cytotoxic activity was assessed by trypan blue exclusion assays with two cytotoxic strains (6206 and PA103). Basolateral surfaces of cells were exposed by one of two methods: EGTA pretreatment of epithelial cells or growth of cells in low-calcium medium. Both methods of exposing basolateral membranes increased epithelial cell susceptibility to P. aeruginosa invasion and cytotoxicity. Migrating cells were also found to be more susceptible to P. aeruginosa invasion than confluent monolayers that had established membrane polarity. Monolayers of MDCK cells that had been selected for resistance to killing by concanavalin A were resistant to both cytotoxicity and invasion by P. aeruginosa because they were more efficiently polarized for their susceptibility to P. aeruginosa virulence factors than regular MDCK cells and not because they were defective in glycosylation. These results suggest that there are factors on the basolateral surfaces of epithelial cells that promote interaction with P. aeruginosa or that there are inhibitory factors on the apical cell surface. Thus, cell polarity of intact epithelia is likely to contribute to defense against P. aeruginosa infection.

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Selected References

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  1. Apodaca G., Bomsel M., Lindstedt R., Engel J., Frank D., Mostov K. E., Wiener-Kronish J. Characterization of Pseudomonas aeruginosa-induced MDCK cell injury: glycosylation-defective host cells are resistant to bacterial killing. Infect Immun. 1995 Apr;63(4):1541–1551. doi: 10.1128/iai.63.4.1541-1551.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Araki-Sasaki K., Ohashi Y., Sasabe T., Hayashi K., Watanabe H., Tano Y., Handa H. An SV40-immortalized human corneal epithelial cell line and its characterization. Invest Ophthalmol Vis Sci. 1995 Mar;36(3):614–621. [PubMed] [Google Scholar]
  3. Basak S., Compans R. W. Polarized entry of canine parvovirus in an epithelial cell line. J Virol. 1989 Jul;63(7):3164–3167. doi: 10.1128/jvi.63.7.3164-3167.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boyd R. L., Ramphal R., Rice R., Mangos J. A. Chronic colonization of rat airways with Pseudomonas aeruginosa. Infect Immun. 1983 Mar;39(3):1403–1410. doi: 10.1128/iai.39.3.1403-1410.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Butor C., Davoust J. Apical to basolateral surface area ratio and polarity of MDCK cells grown on different supports. Exp Cell Res. 1992 Nov;203(1):115–127. doi: 10.1016/0014-4827(92)90046-b. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Eckmann L., Kagnoff M. F., Fierer J. Epithelial cells secrete the chemokine interleukin-8 in response to bacterial entry. Infect Immun. 1993 Nov;61(11):4569–4574. doi: 10.1128/iai.61.11.4569-4574.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fleiszig S. M., Efron N., Pier G. B. Extended contact lens wear enhances Pseudomonas aeruginosa adherence to human corneal epithelium. Invest Ophthalmol Vis Sci. 1992 Sep;33(10):2908–2916. [PubMed] [Google Scholar]
  9. Fleiszig S. M., Wiener-Kronish J. P., Miyazaki H., Vallas V., Mostov K. E., Kanada D., Sawa T., Yen T. S., Frank D. W. Pseudomonas aeruginosa-mediated cytotoxicity and invasion correlate with distinct genotypes at the loci encoding exoenzyme S. Infect Immun. 1997 Feb;65(2):579–586. doi: 10.1128/iai.65.2.579-586.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Fleiszig S. M., Zaidi T. S., Pier G. B. Pseudomonas aeruginosa invasion of and multiplication within corneal epithelial cells in vitro. Infect Immun. 1995 Oct;63(10):4072–4077. doi: 10.1128/iai.63.10.4072-4077.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fleiszig S. M., Zaidi T. S., Preston M. J., Grout M., Evans D. J., Pier G. B. Relationship between cytotoxicity and corneal epithelial cell invasion by clinical isolates of Pseudomonas aeruginosa. Infect Immun. 1996 Jun;64(6):2288–2294. doi: 10.1128/iai.64.6.2288-2294.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fleiszig S. M., Zaidi T. S., Ramphal R., Pier G. B. Modulation of Pseudomonas aeruginosa adherence to the corneal surface by mucus. Infect Immun. 1994 May;62(5):1799–1804. doi: 10.1128/iai.62.5.1799-1804.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fuller S., von Bonsdorff C. H., Simons K. Vesicular stomatitis virus infects and matures only through the basolateral surface of the polarized epithelial cell line, MDCK. Cell. 1984 Aug;38(1):65–77. doi: 10.1016/0092-8674(84)90527-0. [DOI] [PubMed] [Google Scholar]
  15. Gaillard J. L., Finlay B. B. Effect of cell polarization and differentiation on entry of Listeria monocytogenes into the enterocyte-like Caco-2 cell line. Infect Immun. 1996 Apr;64(4):1299–1308. doi: 10.1128/iai.64.4.1299-1308.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gipson I. K., Yankauckas M., Spurr-Michaud S. J., Tisdale A. S., Rinehart W. Characteristics of a glycoprotein in the ocular surface glycocalyx. Invest Ophthalmol Vis Sci. 1992 Jan;33(1):218–227. [PubMed] [Google Scholar]
  17. Gonzalez-Mariscal L., Chávez de Ramírez B., Cereijido M. Tight junction formation in cultured epithelial cells (MDCK). J Membr Biol. 1985;86(2):113–125. doi: 10.1007/BF01870778. [DOI] [PubMed] [Google Scholar]
  18. Gonzalez-Mariscal L., Contreras R. G., Bolívar J. J., Ponce A., Chávez De Ramirez B., Cereijido M. Role of calcium in tight junction formation between epithelial cells. Am J Physiol. 1990 Dec;259(6 Pt 1):C978–C986. doi: 10.1152/ajpcell.1990.259.6.C978. [DOI] [PubMed] [Google Scholar]
  19. Hornick D. B., Allen B. L., Horn M. A., Clegg S. Adherence to respiratory epithelia by recombinant Escherichia coli expressing Klebsiella pneumoniae type 3 fimbrial gene products. Infect Immun. 1992 Apr;60(4):1577–1588. doi: 10.1128/iai.60.4.1577-1588.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kudoh I., Wiener-Kronish J. P., Hashimoto S., Pittet J. F., Frank D. Exoproduct secretions of Pseudomonas aeruginosa strains influence severity of alveolar epithelial injury. Am J Physiol. 1994 Nov;267(5 Pt 1):L551–L556. doi: 10.1152/ajplung.1994.267.5.L551. [DOI] [PubMed] [Google Scholar]
  21. Martinez-Palomo A., Meza I., Beaty G., Cereijido M. Experimental modulation of occluding junctions in a cultured transporting epithelium. J Cell Biol. 1980 Dec;87(3 Pt 1):736–745. doi: 10.1083/jcb.87.3.736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Meiss H. K., Green R. F., Rodriguez-Boulan E. J. Lectin-resistant mutants of polarized epithelial cells. Mol Cell Biol. 1982 Oct;2(10):1287–1294. doi: 10.1128/mcb.2.10.1287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mengaud J., Ohayon H., Gounon P., Mege R-M, Cossart P. E-cadherin is the receptor for internalin, a surface protein required for entry of L. monocytogenes into epithelial cells. Cell. 1996 Mar 22;84(6):923–932. doi: 10.1016/s0092-8674(00)81070-3. [DOI] [PubMed] [Google Scholar]
  24. Mostov K. E., Deitcher D. L. Polymeric immunoglobulin receptor expressed in MDCK cells transcytoses IgA. Cell. 1986 Aug 15;46(4):613–621. doi: 10.1016/0092-8674(86)90887-1. [DOI] [PubMed] [Google Scholar]
  25. Mounier J., Vasselon T., Hellio R., Lesourd M., Sansonetti P. J. Shigella flexneri enters human colonic Caco-2 epithelial cells through the basolateral pole. Infect Immun. 1992 Jan;60(1):237–248. doi: 10.1128/iai.60.1.237-248.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Rajasekaran A. K., Hojo M., Huima T., Rodriguez-Boulan E. Catenins and zonula occludens-1 form a complex during early stages in the assembly of tight junctions. J Cell Biol. 1996 Feb;132(3):451–463. doi: 10.1083/jcb.132.3.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rodriguez-Boulan E., Powell S. K. Polarity of epithelial and neuronal cells. Annu Rev Cell Biol. 1992;8:395–427. doi: 10.1146/annurev.cb.08.110192.002143. [DOI] [PubMed] [Google Scholar]
  29. Rodriguez D., Rodriguez J. R., Ojakian G. K., Esteban M. Vaccinia virus preferentially enters polarized epithelial cells through the basolateral surface. J Virol. 1991 Jan;65(1):494–498. doi: 10.1128/jvi.65.1.494-498.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Siliciano J. D., Goodenough D. A. Localization of the tight junction protein, ZO-1, is modulated by extracellular calcium and cell-cell contact in Madin-Darby canine kidney epithelial cells. J Cell Biol. 1988 Dec;107(6 Pt 1):2389–2399. doi: 10.1083/jcb.107.6.2389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Simons K., Fuller S. D. Cell surface polarity in epithelia. Annu Rev Cell Biol. 1985;1:243–288. doi: 10.1146/annurev.cb.01.110185.001331. [DOI] [PubMed] [Google Scholar]
  32. Topp K. S., Rothman A. L., Lavail J. H. Herpes virus infection of RPE and MDCK cells: polarity of infection. Exp Eye Res. 1997 Mar;64(3):343–354. doi: 10.1006/exer.1996.0209. [DOI] [PubMed] [Google Scholar]
  33. Tucker S. P., Thornton C. L., Wimmer E., Compans R. W. Bidirectional entry of poliovirus into polarized epithelial cells. J Virol. 1993 Jan;67(1):29–38. doi: 10.1128/jvi.67.1.29-38.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Yamaya M., Finkbeiner W. E., Chun S. Y., Widdicombe J. H. Differentiated structure and function of cultures from human tracheal epithelium. Am J Physiol. 1992 Jun;262(6 Pt 1):L713–L724. doi: 10.1152/ajplung.1992.262.6.L713. [DOI] [PubMed] [Google Scholar]

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