1887

Abstract

has emerged as a crucial periodontal pathogen with possible implications for systemic disease. The aim of this study was to isolate the S-layer from and examine its virulence potential as a part of efforts to characterize virulence factors of . The role of the S-layer in the haemagglutinating and adherent/invasive activities was evaluated. It was observed that the S-layer alone was able to mediate haemagglutination. In adherent and invasive studies, transmission electron microscopy clearly revealed that cells were able to attach to and invade KB cells, showing the formation of a microvillus-like extension around adherent and intracellular bacteria. The quantitative analysis showed that five different strains exhibited attachment (1·9–2·3 %) and invasion (0·4–0·7 %) capabilities. It was also observed through antibody inhibition assays that adherent/invasive activities of are mediated by the S-layer. Furthermore, an immunization study adopting a murine abscess model was used to prove that the S-layer is involved in the infectious process of abscess formation. While mice immunized with purified S-layer and whole cells did not develop any abscesses when challenged with viable cells, unimmunized mice developed abscesses. Collectively, the data obtained from these studies indicate that the S-layer of is a virulence factor.

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2003-12-01
2019-11-14
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References

  1. Adlerberth, I., Hanson, L. A., Svanborg, C., Svennerholm, A. M., Nordgren, S. & Wold, A. E. ( 1995; ). Adhesins of Escherichia coli associated with extra-intestinal pathogenicity confer binding to colonic epithelial cells. Microb Pathog 18, 373–385.[CrossRef]
    [Google Scholar]
  2. Arakawa, S., Nakajima, T., Ishikura, H., Ichinose, S., Ishikawa, I. & Tsuchida, N. ( 2000; ). Novel apoptosis-inducing activity in Bacteroides forsythus: a comparative study with three serotypes of Actinobacillus actinomycetemcomitans. Infect Immun 68, 4611–4615.[CrossRef]
    [Google Scholar]
  3. Baker, P. J., Evans, R. T. & Roopenian, D. C. ( 1994; ). Oral infection with Porphyromonas gingivalis and induced alveolar bone loss in immunocompetent and severe combined immunodeficient mice. Arch Oral Biol 39, 1035–1040.[CrossRef]
    [Google Scholar]
  4. Beveridge, T. J., Pouwels, P. H., Sara, M. & 22 other authors ( 1997; ). Functions of S-layers. FEMS Microbiol Rev 20, 99–149.[CrossRef]
    [Google Scholar]
  5. Bird, P. S., Shakibaie, F., Gemmell, E., Polak, B. & Seymour, G. J. ( 2001; ). Immune response to Bacteroides forsythus in a murine model. Oral Microbiol Immunol 16, 311–315.[CrossRef]
    [Google Scholar]
  6. Blaser, M. J. & Pei, Z. ( 1993; ). Pathogenesis of Campylobacter fetus infections: critical role of high-molecular-weight S-layer proteins in virulence. J Infect Dis 167, 372–377.[CrossRef]
    [Google Scholar]
  7. Booth, B. A., Boesman-Finkelstein, M. & Finkelstein, R. A. ( 1984; ). Vibrio cholerae hemagglutinin/protease nicks cholera enterotoxin. Infect Immun 45, 558–560.
    [Google Scholar]
  8. Borinski, R. & Holt, S. C. ( 1990; ). Surface characteristics of Wolinella recta ATCC 33238 and human clinical isolates: correlation of structure with function. Infect Immun 58, 2770–2776.
    [Google Scholar]
  9. Califano, J. V., Gunsolley, J. C., Schenkein, H. A. & Tew, J. G. ( 1997; ). A comparison of IgG antibody reactive with Bacteroides forsythus and Porphyromonas gingivalis in adult and early-onset periodontitis. J Periodontol 68, 734–738.[CrossRef]
    [Google Scholar]
  10. Chandad, F., Guillot, E. & Mouton, C. ( 1997; ). Detection of Bacteroides forsythus by immunomagnetic capture and a polymerase chain reaction-DNA probe assay. Oral Microbiol Immunol 12, 311–317.[CrossRef]
    [Google Scholar]
  11. Coleman, S. E., Brady, L. J. & Boyle, M. D. ( 1990; ). Colloidal gold immunolabeling of immunoglobulin-binding sites and beta antigen in group B streptococci. Infect Immun 58, 332–340.
    [Google Scholar]
  12. Dorn, B. R., Burks, J. N., Seifert, K. N. & Progulske-Fox, A. ( 2000; ). Invasion of endothelial and epithelial cells by strains of Porphyromonas gingivalis. FEMS Microbiol Lett 187, 139–144.[CrossRef]
    [Google Scholar]
  13. Duguid, J. P., Clegg, S. & Wilson, M. I. ( 1979; ). The fimbrial and non-fimbrial haemagglutinins of Escherichia coli. J Med Microbiol 12, 213–227.[CrossRef]
    [Google Scholar]
  14. Dzink, J. L., Socransky, S. S. & Haffajee, A. D. ( 1988; ). The predominant cultivable microbiota of active and inactive lesions of destructive periodontal diseases. J Clin Periodontol 15, 316–323.[CrossRef]
    [Google Scholar]
  15. Falkler, W. A., Jr & Hawley, C. E. ( 1977; ). Hemagglutinating activity of Fusobacterium nucleatum. Infect Immun 15, 230–238.
    [Google Scholar]
  16. Falkow, S., Isberg, R. R. & Portnoy, D. A. ( 1992; ). The interaction of bacteria with mammalian cells. Annu Rev Cell Biol 8, 333–363.[CrossRef]
    [Google Scholar]
  17. Finlay, B. B. & Falkow, S. ( 1997; ). Common themes in microbial pathogenicity revisited. Microbiol Mol Biol Rev 61, 136–169.
    [Google Scholar]
  18. Garduno, R. A., Moore, A. R., Olivier, G., Lizama, A. L., Garduno, E. & Kay, W. W. ( 2000; ). Host cell invasion and intracellular residence by Aeromonas salmonicida: role of the S-layer. Can J Microbiol 46, 660–668.[CrossRef]
    [Google Scholar]
  19. Han, Y. W., Shi, W., Huang, G. T., Kinder, H. S., Park, N. H., Kuramitsu, H. & Genco, R. J. ( 2000; ). Interactions between periodontal bacteria and human oral epithelial cells: Fusobacterium nucleatum adheres to and invades epithelial cells. Infect Immun 68, 3140–3146.[CrossRef]
    [Google Scholar]
  20. Higuchi, N., Murakami, Y., Moriguchi, K., Ohno, N., Nakamura, H. & Yoshimura, F. ( 2000; ). Localization of major, high molecular weight proteins in Bacteroides forsythus. Microbiol Immunol 44, 777–780.[CrossRef]
    [Google Scholar]
  21. Honma, K., Kuramitsu, H. K., Genco, R. J. & Sharma, A. ( 2001; ). Development of a gene inactivation system for Bacteroides forsythus: construction and characterization of a BspA mutant. Infect Immun 69, 4686–4690.[CrossRef]
    [Google Scholar]
  22. Ishiguro, E. E., Kay, W. W., Ainsworth, T., Chamberlain, J. B., Austen, R. A., Buckley, J. T. & Trust, T. J. ( 1981; ). Loss of virulence during culture of Aeromonas salmonicida at high temperature. J Bacteriol 148, 333–340.
    [Google Scholar]
  23. Katz, J., Yang, Q. B., Zhang, P., Potempa, J., Travis, J., Michalek, S. M. & Balkovetz, D. F. ( 2002; ). Hydrolysis of epithelial junctional proteins by Porphyromonas gingivalis gingipains. Infect Immun 70, 2512–2518.[CrossRef]
    [Google Scholar]
  24. Kerosuo, E. ( 1998; ). Ultrastructure of the cell envelope of Bacteroides forsythus strain ATCC 43037T. Oral Microbiol Immunol 3, 134–137.
    [Google Scholar]
  25. Kroes, I., Lepp, P. W. & Relman, D. A. ( 1999; ). Bacterial diversity within the human subgingival crevice. Proc Natl Acad Sci U S A 96, 14547–14552.[CrossRef]
    [Google Scholar]
  26. Laemmli, U. K. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.[CrossRef]
    [Google Scholar]
  27. Lai, C. H., Listgarten, M. A., Shirakawa, M. & Slots, J. ( 1987; ). Bacteroides forsythus in adult gingivitis and periodontitis. Oral Microbiol Immunol 2, 152–157.[CrossRef]
    [Google Scholar]
  28. Lamont, R. J., Chan, A., Belton, C. M., Izutsu, K. T., Vasel, D. & Weinberg, A. ( 1995; ). Porphyromonas gingivalis invasion of gingival epithelial cells. Infect Immun 63, 3878–3885.
    [Google Scholar]
  29. Lee, M. D., Curtiss, R., III & Peay, T. ( 1996; ). The effect of bacterial surface structures on the pathogenesis of Salmonella typhimurium infection in chickens. Avian Dis 40, 28–36.[CrossRef]
    [Google Scholar]
  30. Liljenberg, B., Lindhe, J., Berglundh, T., Dahlen, G. & Jonsson, R. ( 1994; ). Some microbiological, histopathological and immunohistochemical characteristics of progressive periodontal disease. J Clin Periodontol 21, 720–727.[CrossRef]
    [Google Scholar]
  31. Listgarten, M. A. & Lai, C. H. ( 1999; ). Comparative microbiological characteristics of failing implants and periodontally diseased teeth. J Periodontol 70, 431–437.[CrossRef]
    [Google Scholar]
  32. Listgarten, M. A., Lai, C. H. & Young, V. ( 1993; ). Microbial composition and pattern of antibiotic resistance in subgingival microbial samples from patients with refractory periodontitis. J Periodontol 64, 155–161.[CrossRef]
    [Google Scholar]
  33. Loesche, W. J., Bretz, W. A., Kerschensteiner, D., Stoll, J., Socransky, S. S., Hujoel, P. & Lopatin, D. E. ( 1990; ). Development of a diagnostic test for anaerobic periodontal infections based on plaque hydrolysis of benzoyl-dl-arginine-naphthylamide. J Clin Microbiol 28, 1551–1559.
    [Google Scholar]
  34. Makhov, A. M., Hannah, J. H., Brennan, M. J., Trus, B. L., Kocsis, E., Conway, J. F., Wingfield, P. T., Simon, M. N. & Steven, A. C. ( 1994; ). Filamentous hemagglutinin of Bordetella pertussis. A bacterial adhesin formed as a 50-nm monomeric rigid rod based on a 19-residue repeat motif rich in beta strands and turns. J Mol Biol 241, 110–124.[CrossRef]
    [Google Scholar]
  35. Malek, R., Fisher, J. G., Caleca, A. & 7 other authors ( 1994; ). Inactivation of the Porphyromonas gingivalis fimA gene blocks periodontal damage in gnotobiotic rats. J Bacteriol 176, 1052–1059.
    [Google Scholar]
  36. Menozzi, F. D., Boucher, P. E., Riveau, G., Gantiez, C. & Locht, C. ( 1994; ). Surface-associated filamentous hemagglutinin induces autoagglutination of Bordetella pertussis. Infect Immun 62, 4261–4269.
    [Google Scholar]
  37. Meurman, J. H., Wahlfors, J., Korhonen, A., Alakuijala, P., Vaisanen, P., Torkko, H. & Janne, J. ( 1997; ). Identification of Bacteroides forsythus in subgingival dental plaque with the aid of a rapid PCR method. J Dent Res 76, 1376–1380.[CrossRef]
    [Google Scholar]
  38. Meyer, D. H., Mintz, K. P. & Fives-Taylor, P. M. ( 1997; ). Models of invasion of enteric and periodontal pathogens into epithelial cells: a comparative analysis. Crit Rev Oral Biol Med 8, 389–409.[CrossRef]
    [Google Scholar]
  39. Meyer, H. G., Wengler-Becker, U. & Gatermann, S. G. ( 1996; ). The hemagglutinin of Staphylococcus saprophyticus is a major adhesin for uroepithelial cells. Infect Immun 64, 3893–3896.
    [Google Scholar]
  40. Moncla, B. J., Braham, P. & Hillier, S. L. ( 1990; ). Sialidase (neuraminidase) activity among gram-negative anaerobic and capnophilic bacteria. J Clin Microbiol 28, 422–425.
    [Google Scholar]
  41. Moncla, B. J., Motley, S. T., Braham, P., Ewing, L., Adams, T. H. & Vermeulen, N. M. ( 1991; ). Use of synthetic oligonucleotide DNA probes for identification and direct detection of Bacteroides forsythus in plaque samples. J Clin Microbiol 29, 2158–2162.
    [Google Scholar]
  42. Moore, W. E. & Moore, L. V. ( 1994; ). The bacteria of periodontal diseases. Periodontol 2000 5, 66–77.[CrossRef]
    [Google Scholar]
  43. Munemasa, T., Takemoto, T., Dahlen, G., Hino, T., Shiba, H., Ogawa, T. & Kurihara, H. ( 2000; ). Adherence of Bacteroides forsythus to host cells. Microbios 101, 115–126.
    [Google Scholar]
  44. Murakami, Y., Higuchi, N., Nakamura, H., Yoshimura, F. & Oppenheim, F. G. ( 2002; ). Bacteroides forsythus hemagglutinin is inhibited by N-acetylneuraminyllactose. Oral Microbiol Immunol 17, 125–128.[CrossRef]
    [Google Scholar]
  45. Noack, B., Genco, R. J., Trevisan, M., Grossi, S., Zambon, J. J. & De Nardin, E. ( 2001; ). Periodontal infections contribute to elevated systemic C-reactive protein level. J Periodontol 72, 1221–1227.[CrossRef]
    [Google Scholar]
  46. Nowicki, B., Svanborg-Eden, C., Hull, R. & Hull, S. ( 1989; ). Molecular analysis and epidemiology of the Dr hemagglutinin of uropathogenic Escherichia coli. Infect Immun 57, 446–451.
    [Google Scholar]
  47. Papapanou, P. N., Sandros, J., Lindberg, K., Duncan, M. J., Niederman, R. & Nannmark, U. ( 1994; ). Porphyromonas gingivalis may multiply and advance within stratified human junctional epithelium in vitro. J Periodontal Res 29, 374–375.[CrossRef]
    [Google Scholar]
  48. Paster, B. J., Boches, S. K., Galvin, J. L., Ericson, R. E., Lau, C. N., Levanos, V. A., Sahasrabudhe, A. & Dewhirst, F. E. ( 2001; ). Bacterial diversity in human subgingival plaque. J Bacteriol 183, 3770–3783.[CrossRef]
    [Google Scholar]
  49. Persson, G. R., Schlegel-Bregenzer, B., Chung, W. O., Houston, L., Oswald, T. & Roberts, M. C. ( 2000; ). Serum antibody titers to Bacteroides forsythus in elderly subjects with gingivitis or periodontitis. J Clin Periodontol 27, 839–845.[CrossRef]
    [Google Scholar]
  50. Saito, T., Ishihara, K., Kato, T. & Okuda, K. ( 1997; ). Cloning, expression, and sequencing of a protease gene from Bacteroides forsythus ATCC 43037 in Escherichia coli. Infect Immun 65, 4888–4891.
    [Google Scholar]
  51. Sara, M. & Sleytr, U. B. ( 2000; ). S-Layer proteins. J Bacteriol 182, 859–868.[CrossRef]
    [Google Scholar]
  52. Schenkein, H. A., Barbour, S. E., Berry, C. R., Kipps, B. & Tew, J. G. ( 2000; ). Invasion of human vascular endothelial cells by Actinobacillus actinomycetemcomitans via the receptor for platelet-activating factor. Infect Immun 68, 5416–5419.[CrossRef]
    [Google Scholar]
  53. Sharma, A., Sojar, H. T., Glurich, I., Honma, K., Kuramitsu, H. K. & Genco, R. J. ( 1998; ). Cloning, expression, and sequencing of a cell surface antigen containing a leucine-rich repeat motif from Bacteroides forsythus ATCC 43037. Infect Immun 66, 5703–5710.
    [Google Scholar]
  54. Sims, T. J., Mancl, L. A., Braham, P. H., Bainbridge, B. W. & Page, R. C. ( 1998; ). Antigenic variation and cross-reactivity in Bacteroides forsythus clinical isolates detected by western blot. J Clin Immunol 18, 355–367.[CrossRef]
    [Google Scholar]
  55. Socransky, S. S. & Haffajee, A. D. ( 1994; ). Evidence of bacterial etiology: a historical perspective. Periodontol 2000 5, 7–25.[CrossRef]
    [Google Scholar]
  56. Tan, K. S., Song, K. P. & Ong, G. ( 2001; ). Bacteroides forsythus prtH genotype in periodontitis patients: occurrence and association with periodontal disease. J Periodontal Res 36, 398–403.[CrossRef]
    [Google Scholar]
  57. Tanner, A. C., Strzempko, M. N., Belsky, C. A. & McKinley, G. A. ( 1985; ). API ZYM and API An-Ident reactions of fastidious oral gram-negative species. J Clin Microbiol 22, 333–335.
    [Google Scholar]
  58. Tanner, A., Maiden, M. F., Macuch, P. J., Murray, L. L. & Kent, R. L., Jr ( 1998; ). Microbiota of health, gingivitis, and initial periodontitis. J Clin Periodontol 25, 85–98.[CrossRef]
    [Google Scholar]
  59. Towbin, H., Staehelin, T. & Gordon, J. ( 1979; ). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76, 4350–4354.[CrossRef]
    [Google Scholar]
  60. Tran, S. D., Rudney, J. D., Sparks, B. S. & Hodges, J. S. ( 2001; ). Persistent presence of Bacteroides forsythus as a risk factor for attachment loss in a population with low prevalence and severity of adult periodontitis. J Periodontol 72, 1–10.[CrossRef]
    [Google Scholar]
  61. Winkel, E. G., van Winkelhoff, A. J., Timmerman, M. F., Vangsted, T. & Van der Velden, U. ( 1997; ). Effects of metronidazole in patients with ‘refractory’ periodontitis associated with Bacteroides forsythus. J Clin Periodontol 24, 573–579.[CrossRef]
    [Google Scholar]
  62. Zambon, J. J., Grossi, S. G., Machtei, E. E., Ho, A. W., Dunford, R. & Genco, R. J. ( 1996; ). Cigarette smoking increases the risk for subgingival infection with periodontal pathogens. J Periodontol 67, 1050–1054.[CrossRef]
    [Google Scholar]
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