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Staphylococcus aureus adherence to nasal epithelial cells in a physiological in vitro model

  • Infectious Disease/Cellular Pathology
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Summary

Nasal carriage of Staphylococcus aureus represents a risk factor for subsequent invasive infections and interpatient transmission of strains. No physiological in vitro model of nasal epithelial cells is available to study both patient- and bacteria-related characteristics and their interaction, leading to adherence and colonization. Starting with tissues from human nasal polyps, a confluent, squamous, nonkeratinized epithelium in collagen-coated 96-well microtiter plates was obtained after 14 d. This in vitro cell-layer was characterized histologically, ultrastructurally, and immunohistochemically and showed features that were indistinguishable from those observed in the squamous nonkeratinized epithelium found in the posterior part of the vestibulum nasi. Adherence experiments were performed with four different 3H-thymidine-labeled Staphylococcus aureus strains. The effect of bacterial inoculum size, temperature of incubation, and incubation medium were studied. The adherence results were found to be reproducible, reliable and sensitive, allowing detection of small quantitative differences in adherence between the Staphylococcus aureus strains. There was no significant difference in adherence at 23° C and 37° C, nor between the incubation medium M199 and phosphate-buffered saline. Plastic adherence could be reduced and standardized with use of siliconized tips and a constant bacterial inoculum volume of 100 µl/well. This physiological and reliable in vitro cell-culture model offers a unique opportunity to study Staphylococcus aureus adherence to squamous, nonkeratinized nasal epithelial cells and both patient and bacterial characteristics involved in this interaction.

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References

  1. Aly, M. Y. Histology of the human nasopharyngeal mucosa. J. Anat. 99:657–672; 1965.

    Google Scholar 

  2. Aly, R.; Shinefield, H. R.; Strauss, W. G.; Maibach, H. I. Bacterial adherence to nasal mucosal cells. Infect. Immun. 17:546–549; 1977.

    PubMed  CAS  Google Scholar 

  3. Anderson, M. P.; Welsh, M. J. Calcium and cAMP activate different chloride channels in the apical membrane of normal and cystic fibrosis epithelia. Proc. Natl. Acad. Sci. USA 88:6003–6007; 1991.

    Article  PubMed  CAS  Google Scholar 

  4. Becker, S.; Koren, H. S.; Henke, D. C. Interleukin-8 expression in normal nasal epithelium and its modulation by infection with respiratory syncitial virus and cytokines tumor necrosis factor, interleukin-1, and interleukin-6. Am. J. Respir. Cell. Mol. Biol. 8:20–27; 1993.

    PubMed  CAS  Google Scholar 

  5. Bibel, D. J.; Aly, R.; Shinefield, H. R.; Maibach, H. I.; Strauss, W. G. Importance of the keratinized epithelial cells in bacterial adherence. J. Investig. Dermatol. 79:250–253; 1982.

    Article  PubMed  CAS  Google Scholar 

  6. Boysen, M. The surface structure of the human nasal mucosa. Ciliated and metaplastic epithelium in normal individuals. A correlated study by scanning/transmission electron and light microscopy. Virchows Arch. B. Cell. Pathol. Incl. Mol. Pathol. 40:279–294; 1982.

    PubMed  CAS  Google Scholar 

  7. Brezillon, S.; Dupuit, F.; Hinnrasky, J.; Marchand, V.; Kalin, N.; Tummler, B.; Puchelle, E. Decreased expression of the CFTR protein in remodeled human nasal epithelium from non-cystic fibrosis patients. Lab. Invest. 72:191–200; 1995.

    PubMed  CAS  Google Scholar 

  8. Chandrakasan, G.; Torchia, D. A.; Piez, K. A. Preparation of intact monomeric collagen from rat tail tendon and skin and the structure of the nonhelical ends in solution. J. Biol. Chem. 251:6062–6067; 1976.

    PubMed  CAS  Google Scholar 

  9. Cheung, A. L.; Koomey, J. M.; Lee, S.; Jaffe, E. A.; Fischetti, V. A. Recombinant human tumor necrosis factor alpha promotes adherence to Staphylococcus aureus to cultured human endothelial cells. Infect. Immun. 59:3827–3831; 1991.

    PubMed  CAS  Google Scholar 

  10. Cole, G. W.; Silverberg, N. L. The adherence of Staphylococcus aureus to human corneocytes. Arch. Dermatol. 122:166–169; 1986.

    Article  PubMed  CAS  Google Scholar 

  11. Dunkle, L. M.; Blair, L. L.; Fortune, K. P. Transformation of a plasmid encoding an adhesin of Staphylococcus aureus into a non-adherent staphylococcal strain. J. Infect. Dis. 153:670–675; 1986.

    PubMed  CAS  Google Scholar 

  12. Emura, M.; Riebe, M.; Ochiai, A.; Aufderheide, M.; Germann, P.; Mohr, U. New functional cell-culture approach to pulmonary carcinogenesis and toxicology. J. Cancer Res. Clin. Oncol. 116:557–562; 1990.

    Article  PubMed  CAS  Google Scholar 

  13. Foster, T. J.; McDevitt, D. Surface-associated proteins of Staphylococcus aureus: their possible roles in virulence. FEMS Microbiol. Lett. 118:199–206; 1994.

    Article  PubMed  CAS  Google Scholar 

  14. Greunert, D. C.; Finkbeiner, W. E.; Widdicombe J. H. Culture and transformation of human airway epithelial cells. Am. J. Physiol. 268 (Lung Cell Mol. Physiol. 12):L347-L360; 1995.

    Google Scholar 

  15. Gulisano, M.; Pacini, S.; Ruggiero, M.; Pacini, A.; Delrio, A. N.; Pacini, P. In vitro effects of some differentiation inductors in metaplastic epithelium of the human nasal cavity. Cell. Tissue Res. 285:119–125; 1996.

    Article  PubMed  CAS  Google Scholar 

  16. Haagen, I. A.; Heezius, H. C.; Verkooyen, R. P.; Verhoef, J.; Verbrugh, H. A. Adherence of peritonitis-causing staphylococci to human peritoneal mesothelial cell monolayers. J. Infect. Dis. 161:266–273; 1990.

    PubMed  CAS  Google Scholar 

  17. Hoefnagels-Schuermans, A.; Peetermans, W. E.; Struelens, M. J.; Van Lierde, S.; Van Eldere, J. Clonal analysis and identification of epidemic strains of methicillin-resistant Staphylococcus aureus by antibiotyping, and determination of protein A gene and coagulase gene polymorphism. J. Clin. Microbiol. 35:2514–2520; 1997.

    PubMed  CAS  Google Scholar 

  18. Hynes, R. O. Alteration of cell surface proteins by viral transformation and by proteolysis. Proc. Natl. Acad. Sci. USA 70:3170–3174; 1973.

    Article  PubMed  CAS  Google Scholar 

  19. Jetten, A. M. Multistep process of squamous differentiation in tracheobronchial epithelial cells in vitro: analogy with epidermal differentiation. Environ. Health Perspect. 80:149–160; 1989.

    Article  PubMed  CAS  Google Scholar 

  20. Jordens, J. Z. Adherence of S. aureus to Hep2 monolayers and fibronectin. In: Wadström, T.; Eliasson, I.; Holder, I.; Ljungh, A., ed. Pathogenesis of wound- and biomaterial-associated infection. London: Springer-Verlag; 1990:325–327.

    Google Scholar 

  21. Jorissen, M.; Van der Schueren, B.; Van den Berghe, H.; Cassiman, J. J. The preservation and regeneration of cilia on human nasal epithelial cells cultured in vitro. Arch. Otorhinolaryngol. 246:308–314; 1989.

    Article  PubMed  CAS  Google Scholar 

  22. Kluytmans, J. A. J. W.; Mouton, J. W.; Ijzerman, E. P. F.; Vandenbroucke-Grauls, C. M. J. E.; Maat, A. W. P. M.; Wagenvoort, J. H. T.; Verbrugh, H. A. Nasal carriage of Staphylococcus aureus as a major risk factor for wound infections after cardiac surgery. J. Infect. Dis. 171:216–219; 1995.

    PubMed  CAS  Google Scholar 

  23. Kluytmans, J.; van Belkum, A.; Verbrugh, H. Nasal carriage of Staphylococcus aureus: epidemiology, underlying mechanisms, and associated risks. Clin. Microbiol. Rev. 10:505–520; 1997.

    PubMed  CAS  Google Scholar 

  24. Luzar, M. A.; Coles, G. A.; Faller, B., et al. Staphylococcus aureus nasal carriage and infection in patients on continuous ambulatory peritoneal dialysis. N. Engl. J. Med. 322:505–509; 1990.

    Article  PubMed  CAS  Google Scholar 

  25. Meyer, D. H.; Lippmann, J. E.; Fives-Taylor, P. M. Invasion of epithelial cells by Actinobacillus actinomycetecomitans: a dynamic, multistep process. Infect. Immun. 64:2988–2997; 1996.

    PubMed  CAS  Google Scholar 

  26. Poston, S. M.; Glancey, G. R.; Wyatt, J. E.; Hogan, T.; Foster, T. J. Coelimination of mec and spa genes in Staphylococcus aureus and the effect of agr and protein A production on bacterial adherence to cell monolayers. J. Med. Microbiol. 39:422–428; 1993.

    Article  PubMed  CAS  Google Scholar 

  27. Proctor, R. A. Fibronectin: a brief overview of its structure, function and physiology. Rev. Infect. Dis. 9 (Suppl. 4):S317-S321; 1987.

    PubMed  CAS  Google Scholar 

  28. Robinson, C. B.; Wu, R. Mucin synthesis and secretion by cultured tracheal cells: effects of collagen gel substratum thickness. In Vitro Cell. Dev. Biol. 29A:469–477; 1993.

    CAS  Google Scholar 

  29. Shibl, A. M. Effect of antibiotics on adherence of microorganisms to epithelial cell surfaces. Rev. Infect. Dis. 7:51–65; 1985.

    PubMed  CAS  Google Scholar 

  30. Stosiek, P.; Kasper, M.; Moll, R. Changes in cytokeratin expression accompany squamous metaplasia of the human respiratory epithelium. Virchows Arch. Pathol. Anat. Histopathol. 421:133–141; 1992.

    Article  CAS  Google Scholar 

  31. Toppozada, H.; Toppozada, M.; El-Ghazzawi, I.; Elwany, S. The human respiratory nasal mucosa in females using contraceptive pills. An ultramicroscopic and histochemical study. J. Laryngol. Otol. 98:43–51; 1984.

    PubMed  CAS  Google Scholar 

  32. Ward, T. T. Comparison of in vitro adherence of methicillin-sensitive and methicillin-resistant Staphylococcus aureus to human nasal epithelial cells. J. Infect. Dis. 166:400–404; 1992.

    PubMed  CAS  Google Scholar 

  33. Wyatt, J. E.; Poston, S. M.; Noble, W. C. Adherence of Staphylococcus aureus to cell monolayers. J. Appl. Bacteriol. 69:834–844; 1990.

    PubMed  CAS  Google Scholar 

  34. Yankaskas, J. R.; Cotton, C. U.; Knowles, M. R.; Gatzy, J. T.; Boucher, R. C. Culture of human nasal epithelial cells on collagen matrix supports. Am. Rev. Respir. Dis. 132:1281–1287; 1985.

    PubMed  CAS  Google Scholar 

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Correspondence to A. Hoefnagels-Schuermans.

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Hoefnagels-Schuermans, A., Peetermans, W.E., Jorissen, M. et al. Staphylococcus aureus adherence to nasal epithelial cells in a physiological in vitro model. In Vitro Cell.Dev.Biol.-Animal 35, 472–480 (1999). https://doi.org/10.1007/s11626-999-0054-0

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  • DOI: https://doi.org/10.1007/s11626-999-0054-0

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