Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1996 Nov;62(11):3933–3938. doi: 10.1128/aem.62.11.3933-3938.1996

Expression of functional Porphyromonas gingivalis fimbrillin polypeptide domains on the surface of Streptococcus gordonii.

A Sharma 1, H Nagata 1, N Hamada 1, H T Sojar 1, D E Hruby 1, H K Kuramitsu 1, R J Genco 1
PMCID: PMC168210  PMID: 8899979

Abstract

Genetically engineering bacteria to express surface proteins which can antagonize the colonization of other microorganisms is a promising strategy for altering bacterial environments. The fimbriae of Porphyromonas gingivalis play an important role in the pathogenesis of periodontal diseases. A structural subunit of the P. gingivalis fimbriae, fimbrillin, has been shown to be an important virulence factor, which likely promotes adherence of the bacterium to saliva-coated oral surfaces and induces host responses. Immunization of gnotobiotic rats with synthetic peptides based on the predicted amino acid sequence of fimbrillin has also been shown to elicit a specific immune response and protection against P. gingivalis-associated periodontal destruction. In this study we engineered the human oral commensal organism Streptococcus gordonii to surface express subdomains of the fimbrillin polypeptide fused to the anchor region of streptococcal M6 protein. The resulting recombinant S. gordonii strains expressing P. gingivalis fimbrillin bound saliva-coated hydroxyapatite in a concentration-dependent manner and inhibited binding of P. gingivalis to saliva-coated hydroxyapatite. Moreover, the recombinant S. gordonii strains were capable of eliciting a P. gingivalis fimbrillin-specific immune response in rabbits. These results show that functional and immunologically reactive P. gingivalis fimbrillin polypeptides can be expressed on the surface of S. gordonii. The recombinant fimbrillin-expressing S. gordonii strains may provide an effective vaccine or a vehicle for replacement therapy against P. gingivalis. These experiments demonstrated the feasibility of expressing biologically active agents (antigens or adhesin molecules) by genetically engineered streptococci. Such genetically engineered organisms can be utilized to modulate the microenvironment of the oral cavity.

Full Text

The Full Text of this article is available as a PDF (283.1 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amano A., Sharma A., Lee J. Y., Sojar H. T., Raj P. A., Genco R. J. Structural domains of Porphyromonas gingivalis recombinant fimbrillin that mediate binding to salivary proline-rich protein and statherin. Infect Immun. 1996 May;64(5):1631–1637. doi: 10.1128/iai.64.5.1631-1637.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boyd J., McBride B. C. Fractionation of hemagglutinating and bacterial binding adhesins of Bacteroides gingivalis. Infect Immun. 1984 Aug;45(2):403–409. doi: 10.1128/iai.45.2.403-409.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. De Nardin A. M., Sojar H. T., Grossi S. G., Christersson L. A., Genco R. J. Humoral immunity of older adults with periodontal disease to Porphyromonas gingivalis. Infect Immun. 1991 Dec;59(12):4363–4370. doi: 10.1128/iai.59.12.4363-4370.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Deslauriers M., Haque S., Flood P. M. Identification of murine protective epitopes on the Porphyromonas gingivalis fimbrillin molecule. Infect Immun. 1996 Feb;64(2):434–440. doi: 10.1128/iai.64.2.434-440.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dickinson D. P., Kubiniec M. A., Yoshimura F., Genco R. J. Molecular cloning and sequencing of the gene encoding the fimbrial subunit protein of Bacteroides gingivalis. J Bacteriol. 1988 Apr;170(4):1658–1665. doi: 10.1128/jb.170.4.1658-1665.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Evans R. T., Klausen B., Genco R. J. Immunization with fimbrial protein and peptide protects against Porphyromonas gingivalis-induced periodontal tissue destruction. Adv Exp Med Biol. 1992;327:255–262. doi: 10.1007/978-1-4615-3410-5_27. [DOI] [PubMed] [Google Scholar]
  7. Gibbons R. J., Hay D. I., Schlesinger D. H. Delineation of a segment of adsorbed salivary acidic proline-rich proteins which promotes adhesion of Streptococcus gordonii to apatitic surfaces. Infect Immun. 1991 Sep;59(9):2948–2954. doi: 10.1128/iai.59.9.2948-2954.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hanazawa S., Murakami Y., Hirose K., Amano S., Ohmori Y., Higuchi H., Kitano S. Bacteroides (Porphyromonas) gingivalis fimbriae activate mouse peritoneal macrophages and induce gene expression and production of interleukin-1. Infect Immun. 1991 Jun;59(6):1972–1977. doi: 10.1128/iai.59.6.1972-1977.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Isogai H., Isogai E., Yoshimura F., Suzuki T., Kagota W., Takano K. Specific inhibition of adherence of an oral strain of Bacteroides gingivalis 381 to epithelial cells by monoclonal antibodies against the bacterial fimbriae. Arch Oral Biol. 1988;33(7):479–485. doi: 10.1016/0003-9969(88)90028-3. [DOI] [PubMed] [Google Scholar]
  10. Kawata Y., Hanazawa S., Amano S., Murakami Y., Matsumoto T., Nishida K., Kitano S. Porphyromonas gingivalis fimbriae stimulate bone resorption in vitro. Infect Immun. 1994 Jul;62(7):3012–3016. doi: 10.1128/iai.62.7.3012-3016.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kubo S., Kubota H., Ohnishi Y., Morita T., Matsuya T., Matsushiro A. Expression and secretion of an Arthrobacter dextranase in the oral bacterium Streptococcus gordonii. Infect Immun. 1993 Oct;61(10):4375–4381. doi: 10.1128/iai.61.10.4375-4381.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lamont R. J., Bevan C. A., Gil S., Persson R. E., Rosan B. Involvement of Porphyromonas gingivalis fimbriae in adherence to Streptococcus gordonii. Oral Microbiol Immunol. 1993 Oct;8(5):272–276. doi: 10.1111/j.1399-302x.1993.tb00573.x. [DOI] [PubMed] [Google Scholar]
  13. Lamont R. J., Hsiao G. W., Gil S. Identification of a molecule of Porphyromonas gingivalis that binds to Streptococcus gordonii. Microb Pathog. 1994 Nov;17(5):355–360. doi: 10.1006/mpat.1994.1081. [DOI] [PubMed] [Google Scholar]
  14. Lee J. Y., Sojar H. T., Bedi G. S., Genco R. J. Synthetic peptides analogous to the fimbrillin sequence inhibit adherence of Porphyromonas gingivalis. Infect Immun. 1992 Apr;60(4):1662–1670. doi: 10.1128/iai.60.4.1662-1670.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lee J. Y., Sojar H. T., Sharma A., Bedi G. S., Genco R. J. Active domains of fimbrillin involved in adherence of Porphyromonas gingivalis. J Periodontal Res. 1993 Nov;28(6 Pt 2):470–472. doi: 10.1111/j.1600-0765.1993.tb02105.x. [DOI] [PubMed] [Google Scholar]
  16. Malek R., Fisher J. G., Caleca A., Stinson M., van Oss C. J., Lee J. Y., Cho M. I., Genco R. J., Evans R. T., Dyer D. W. Inactivation of the Porphyromonas gingivalis fimA gene blocks periodontal damage in gnotobiotic rats. J Bacteriol. 1994 Feb;176(4):1052–1059. doi: 10.1128/jb.176.4.1052-1059.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Medaglini D., Pozzi G., King T. P., Fischetti V. A. Mucosal and systemic immune responses to a recombinant protein expressed on the surface of the oral commensal bacterium Streptococcus gordonii after oral colonization. Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):6868–6872. doi: 10.1073/pnas.92.15.6868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Murray P. A., Levine M. J., Tabak L. A., Reddy M. S. Specificity of salivary-bacterial interactions: II. Evidence for a lectin on Streptococcus sanguis with specificity for a NeuAc alpha 2, 3Ga1 beta 1, 3Ga1NAc sequence. Biochem Biophys Res Commun. 1982 May 31;106(2):390–396. doi: 10.1016/0006-291x(82)91122-6. [DOI] [PubMed] [Google Scholar]
  19. Naito Y., Gibbons R. J. Attachment of Bacteroides gingivalis to collagenous substrata. J Dent Res. 1988 Aug;67(8):1075–1080. doi: 10.1177/00220345880670080301. [DOI] [PubMed] [Google Scholar]
  20. Ogawa T., Ogo H., Uchida H., Hamada S. Humoral and cellular immune responses to the fimbriae of Porphyromonas gingivalis and their synthetic peptides. J Med Microbiol. 1994 Jun;40(6):397–402. doi: 10.1099/00222615-40-6-397. [DOI] [PubMed] [Google Scholar]
  21. Ogawa T. The potential protective immune responses to synthetic peptides containing conserved epitopes of Porphyromonas gingivalis fimbrial protein. J Med Microbiol. 1994 Nov;41(5):349–358. doi: 10.1099/00222615-41-5-349. [DOI] [PubMed] [Google Scholar]
  22. Ogawa T., Uchida H., Hamada S. Porphyromonas gingivalis fimbriae and their synthetic peptides induce proinflammatory cytokines in human peripheral blood monocyte cultures. FEMS Microbiol Lett. 1994 Feb 15;116(2):237–242. doi: 10.1111/j.1574-6968.1994.tb06707.x. [DOI] [PubMed] [Google Scholar]
  23. Oggioni M. R., Pozzi G. A host-vector system for heterologous gene expression in Streptococcus gordonii. Gene. 1996 Feb 22;169(1):85–90. doi: 10.1016/0378-1119(95)00775-x. [DOI] [PubMed] [Google Scholar]
  24. Pozzi G., Contorni M., Oggioni M. R., Manganelli R., Tommasino M., Cavalieri F., Fischetti V. A. Delivery and expression of a heterologous antigen on the surface of streptococci. Infect Immun. 1992 May;60(5):1902–1907. doi: 10.1128/iai.60.5.1902-1907.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Scannapieco F. A., Bergey E. J., Reddy M. S., Levine M. J. Characterization of salivary alpha-amylase binding to Streptococcus sanguis. Infect Immun. 1989 Sep;57(9):2853–2863. doi: 10.1128/iai.57.9.2853-2863.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sharma A., Sojar H. T., Lee J. Y., Genco R. J. Expression of a functional Porphyromonas gingivalis fimbrillin polypeptide in Escherichia coli: purification, physicochemical and immunochemical characterization, and binding characteristics. Infect Immun. 1993 Aug;61(8):3570–3573. doi: 10.1128/iai.61.8.3570-3573.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shiroza T., Kuramitsu H. K. Construction of a model secretion system for oral streptococci. Infect Immun. 1993 Sep;61(9):3745–3755. doi: 10.1128/iai.61.9.3745-3755.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shiroza T., Kuramitsu H. K. Development of a heterodimer plasmid system for the introduction of heterologous genes into streptococci. Plasmid. 1995 Sep;34(2):85–95. doi: 10.1006/plas.1995.9998. [DOI] [PubMed] [Google Scholar]
  29. Sojar H. T., Lee J. Y., Genco R. J. Fibronectin binding domain of P. gingivalis fimbriae. Biochem Biophys Res Commun. 1995 Nov 22;216(3):785–792. doi: 10.1006/bbrc.1995.2690. [DOI] [PubMed] [Google Scholar]
  30. Stinson M. W., Levine M. J., Cavese J. M., Prakobphol A., Murray P. A., Tabak L. A., Reddy M. S. Adherence of Streptococcus sanguis to salivary mucin bound to glass. J Dent Res. 1982 Dec;61(12):1390–1393. doi: 10.1177/00220345820610120101. [DOI] [PubMed] [Google Scholar]
  31. Stinson M. W., Safulko K., Levine M. J. Adherence of Porphyromonas (Bacteroides) gingivalis to Streptococcus sanguis in vitro. Infect Immun. 1991 Jan;59(1):102–108. doi: 10.1128/iai.59.1.102-108.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES