1887

Abstract

strains W50 and ATCC 33277 were shown to bind to cultured human fibroblast (MRC-5) cells using flow cytometry. As the concentration of strain W50 cells was increased relative to the concentration of MRC-5 cells, the number of W50 cells bound per MRC-5 cell increased, as did the percentage of MRC-5 cells with bacteria bound. However, this relationship was only seen for strain ATCC 33277 at low cell concentrations: at high bacterial cell concentrations strain ATCC 33277 auto-aggregated and binding to the MRC-5 cells decreased. Strain W50 was therefore chosen to study the role of the surface proteinase–adhesin complexes (RgpA–Kgp complexes) in binding to MRC-5 cells. W50 cells treated with an inhibitor of the RgpA–Kgp complexes exhibited reduced binding to MRC-5 cells. The purified active and proteinase-inactive RgpA–Kgp complexes competitively inhibited binding of W50 to MRC-5 cells, and isogenic mutants of W50 lacking RgpA/B and Kgp displayed reduced binding. W50 mutant cells lacking Kgp exhibited the lowest binding to MRC-5 cells, suggesting an important role for this proteinase and its associated adhesins in binding to fibroblasts.

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2008-10-01
2020-01-23
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References

  1. Bieber, D., Ramer, S. W., Wu, C. Y., Murray, W. J., Tobe, T., Fernandez, R. & Schoolnik, G. K. ( 1998; ). Type IV pili, transient bacterial aggregates, and virulence of enteropathogenic Escherichia coli. Science 280, 2114–2118.[CrossRef]
    [Google Scholar]
  2. Chen, T. & Duncan, M. J. ( 2004; ). Gingipain adhesin domains mediate Porphyromonas gingivalis adherence to epithelial cells. Microb Pathog 36, 205–209.[CrossRef]
    [Google Scholar]
  3. Cohen, J. ( 1969; ). Statistical Power Analysis for the Behavioral Sciences. New York: Academic Press.
  4. Hamada, N., Watanabe, K., Sasakawa, C., Yoshikawa, M., Yoshimura, F. & Umemoto, T. ( 1994; ). Construction and characterization of a fimA mutant of Porphyromonas gingivalis. Infect Immun 62, 1696–1704.
    [Google Scholar]
  5. Hassell, T. M. ( 1993; ). Tissues and cells of the periodontium. Periodontol 2000 3, 9–38.[CrossRef]
    [Google Scholar]
  6. Knutton, S., Shaw, R. K., Anantha, R. P., Donnenberg, M. S. & Zorgani, A. A. ( 1999; ). The type IV bundle-forming pilus of enteropathogenic Escherichia coli undergoes dramatic alterations in structure associated with bacterial adherence, aggregation and dispersal. Mol Microbiol 33, 499–509.[CrossRef]
    [Google Scholar]
  7. Kontani, M., Ono, H., Shibata, H., Okamura, Y., Tanaka, T., Fujiwara, T., Kimura, S. & Hamada, S. ( 1996; ). Cysteine protease of Porphyromonas gingivalis 381 enhances binding of fimbriae to cultured human fibroblasts and matrix proteins. Infect Immun 64, 756–762.
    [Google Scholar]
  8. Kontani, M., Kimura, S., Nakagawa, I. & Hamada, S. ( 1997; ). Adherence of Porphyromonas gingivalis to matrix proteins via a fimbrial cryptic receptor exposed by its own arginine-specific protease. Mol Microbiol 24, 1179–1187.[CrossRef]
    [Google Scholar]
  9. Kuramitsu, H., Tokuda, M., Yoneda, M., Duncan, M. & Cho, M. I. ( 1997; ). Multiple colonization defects in a cysteine protease mutant of Porphyromonas gingivalis. J Periodontal Res 32, 140–142.[CrossRef]
    [Google Scholar]
  10. Lamont, R. J. & Jenkinson, H. F. ( 1998; ). Life below the gum line: pathogenic mechanisms of Porphyromonas gingivalis. Microbiol Mol Biol Rev 62, 1244–1263.
    [Google Scholar]
  11. McCulloch, C. A. & Bordin, S. ( 1991; ). Role of fibroblast subpopulations in periodontal physiology and pathology. J Periodontal Res 26, 144–154.[CrossRef]
    [Google Scholar]
  12. Nakagawa, I., Amano, A., Kuboniwa, M., Nakamura, T., Kawabata, S. & Hamada, S. ( 2002; ). Functional differences among FimA variants of Porphyromonas gingivalis and their effects on adhesion to and invasion of human epithelial cells. Infect Immun 70, 277–285.[CrossRef]
    [Google Scholar]
  13. Neiders, M. E., Chen, P. B., Suido, H., Reynolds, H. S., Zambon, J. J., Shlossman, M. & Genco, R. J. ( 1989; ). Heterogeneity of virulence among strains of Bacteroides gingivalis. J Periodontal Res 24, 192–198.[CrossRef]
    [Google Scholar]
  14. O'Brien-Simpson, N. M., Paolini, R. A., Hoffmann, B., Slakeski, N., Dashper, S. G. & Reynolds, E. C. ( 2001; ). Role of RgpA, RgpB, and Kgp proteinases in virulence of Porphyromonas gingivalis W50 in a murine lesion model. Infect Immun 69, 7527–7534.[CrossRef]
    [Google Scholar]
  15. O'Brien-Simpson, N. M., Veith, P. D., Dashper, S. G. & Reynolds, E. C. ( 2003; ). Porphyromonas gingivalis gingipains: the molecular teeth of a microbial vampire. Curr Protein Pept Sci 4, 409–426.[CrossRef]
    [Google Scholar]
  16. O'Brien-Simpson, N. M., Veith, P. D., Dashper, S. G. & Reynolds, E. C. ( 2004; ). Antigens of bacteria associated with periodontitis. Periodontol 2000 35, 101–134.[CrossRef]
    [Google Scholar]
  17. O'Brien-Simpson, N. M., Pathirana, R. D., Paolini, R. A., Chen, Y. Y., Veith, P. D., Tam, V., Ally, N., Pike, R. N. & Reynolds, E. C. ( 2005; ). An immune response directed to proteinase and adhesin functional epitopes protects against Porphyromonas gingivalis-induced periodontal bone loss. J Immunol 175, 3980–3989.[CrossRef]
    [Google Scholar]
  18. Ogawa, T., Ogo, H. & Kinoshita, A. ( 1997; ). Antagonistic effect of synthetic peptides corresponding to the binding regions within fimbrial subunit protein from Porphyromonas gingivalis to human gingival fibroblasts. Vaccine 15, 230–236.[CrossRef]
    [Google Scholar]
  19. Pathirana, R. D., O'Brien-Simpson, N. M., Veith, P. D., Riley, P. F. & Reynolds, E. C. ( 2006; ). Characterization of proteinase-adhesin complexes of Porphyromonas gingivalis. Microbiology 152, 2381–2394.[CrossRef]
    [Google Scholar]
  20. Pathirana, R. D., O'Brien-Simpson, N. M., Visvanathan, K., Hamilton, J. A. & Reynolds, E. C. ( 2007; ). Flow cytometric analysis of adherence of Porphyromonas gingivalis to oral epithelial cells. Infect Immun 75, 2484–2492.[CrossRef]
    [Google Scholar]
  21. Pike, R. N., Potempa, J., McGraw, W., Coetzer, T. H. & Travis, J. ( 1996; ). Characterization of the binding activities of proteinase-adhesin complexes from Porphyromonas gingivalis. J Bacteriol 178, 2876–2882.
    [Google Scholar]
  22. Sheikh, J., Czeczulin, J. R., Harrington, S., Hicks, S., Henderson, I. R., Le Bouguenec, C., Gounon, P., Phillips, A. & Nataro, J. P. ( 2002; ). A novel dispersin protein in enteroaggregative Escherichia coli. J Clin Invest 110, 1329–1337.[CrossRef]
    [Google Scholar]
  23. Socransky, S. S. & Haffajee, A. D. ( 2002; ). Dental biofilms: difficult therapeutic targets. Periodontol 2000 28, 12–55.[CrossRef]
    [Google Scholar]
  24. Socransky, S. S., Haffajee, A. D., Cugini, M. A., Smith, C. & Kent, R. L., Jr ( 1998; ). Microbial complexes in subgingival plaque. J Clin Periodontol 25, 134–144.[CrossRef]
    [Google Scholar]
  25. van Steenbergen, T. J., Kastelein, P., Touw, J. J. & de Graaff, J. ( 1982; ). Virulence of black-pigmented Bacteroides strains from periodontal pockets and other sites in experimentally induced skin lesions in mice. J Periodontal Res 17, 41–49.[CrossRef]
    [Google Scholar]
  26. Veith, P. D., Talbo, G. H., Slakeski, N., Dashper, S. G., Moore, C., Paolini, R. A. & Reynolds, E. C. ( 2002; ). Major outer membrane proteins and proteolytic processing of RgpA and Kgp of Porphyromonas gingivalis W50. Biochem J 363, 105–115.[CrossRef]
    [Google Scholar]
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