DOI QR코드

DOI QR Code

Production of Monoclonal Antibodies Specific to FimA of Porphyromonas gingivalis and Their Inhibitory Activity on Bacterial Binding

  • Koh, Eun-Mi (Division of Biological Sciences and The Institute for Molecular Biology and Genetics, Chonbuk National University) ;
  • Kim, Ju (Jeonju Biomaterials Institute) ;
  • Lee, Jin-Yong (Department of Dental Microbiology, Kyunghee University Dental School) ;
  • Kim, Tae-Geum (Division of Biological Sciences and The Institute for Molecular Biology and Genetics, Chonbuk National University)
  • Received : 2009.09.10
  • Accepted : 2009.09.28
  • Published : 2009.10.31

Abstract

Background: The FimA of Porphyromonas gingivalis is a crucial pathogenic component of the bacteria and has been implicated as a target for vaccine development against the periodontal diseases. Methods: In this study, the purified fimbriae (FimA subunit polymers) protein was used for immunization in their native form and B hybridoma clones producing antibodies specific to FimA were established. Results: The monoclonal antibodies prepared from selected two clones, designated #123 (IgG2b/ kappa) and #265 (IgG1/kappa), displayed different patterns of binding activity against the cognate antigen. Both antibodies reacted with conformational epitopes expressed by partially dissociated oligomers, but not with monomer as elucidated by Western blot analysis. Ascites fluid containing the monoclonal antibodies showed the inhibitory activity against P. gingivalis to saliva-coated hydroxyapatite beads, an in vitro model for the pellicle-coated tooth surface. Conclusion: These results suggest that the monoclonal antibodies could be used as vaccine material against the periodontal diseases through passive immunization.

References

  1. Andrian E, Grenier D, Rouabhia M: Porphyromonas gingivalis-epithelial cell interactions in periodontitis. J Dent Res 85;392-403, 2006 https://doi.org/10.1177/154405910608500502
  2. Griffen AL, Becker MR, Lyons SR, Moeschberger ML, Leys EJ: Prevalence of porphyromonas gingivalis and periodontal health status. J Clin Microbiol 36;3239-3242, 1998
  3. Amano A: Molecular interaction of Porphyromonas gingivalis with host cells: implication for the microbial pathogenesis of periodontal disease. J Periodontol 74;90-96, 2003 https://doi.org/10.1902/jop.2003.74.1.90
  4. Ezzo PJ, Cutler CW: Microorganisms as risk indicators for periodontal disease. Periodontol 2000 32;24-35, 2003 https://doi.org/10.1046/j.0906-6713.2003.03203.x
  5. Shibata Y, Hosogi Y, Hayakawa M, Hori N, Kamada M, Abiko Y: Construction of novel human monoclonal antibodies neutralizing Porphyromonas gingivalis hemagglutination activity using transgenic mice expressing human Ig loci. Vaccine 23;3850-3856, 2005 https://doi.org/10.1016/j.vaccine.2005.01.159
  6. Jotwani R, Cutler CW: Fimbriated porphyromonas gingivalis is more efficient than fimbria-deficient P. gingivalis in entering human dendritic cells in vitro and induces an inflammatory Th1 effector response. Infect Immun 72;1725-1732, 2004 https://doi.org/10.1128/IAI.72.3.1725-1732.2004
  7. Sharma A, Honma K, Evans RT, Hruby DE, Genco RJ: Oral immunization with recombinant streptococcus gordonii expressing porphyromonas gingivalis FimA domains. Infect Immun 69;2928-2934, 2001 https://doi.org/10.1128/IAI.69.5.2928-2934.2001
  8. Shin EA, Lee JY, Kim TG, Park YK, Langridge WH: Synthesis and assembly of an adjuvanted Porphyromonas gingivalis fimbrial antigen fusion protein in plants. Protein Expr Purif 47;99-109, 2006 https://doi.org/10.1016/j.pep.2005.09.005
  9. Takahashi Y, Kumada H, Hamada N, Haishima Y, Ozono S, Isaka M, Yasuda Y, Tochikubo K, Umemoto T: Induction of immune responses and prevention of alveolar bone loss by intranasal administration of mice with Porphyromonas gingivalis fimbriae and recombinant cholera toxin B subunit. Oral Microbiol Immunol 22;374-380, 2007 https://doi.org/10.1111/j.1399-302X.2007.00373.x
  10. Evans RT, Klausen B, Genco RJ: Immunization with fimbrial protein and peptide protects against Porphyromonas gingivalis-induced periodontal tissue destruction. Adv Exp Med Biol 327;255-262, 1992
  11. Evans RT, Klausen B, Sojar HT, Bedi GS, Sfintescu C, Ramamurthy NS, Golub LM, Genco RJ: Immunization with Porphyromonas (Bacteroides) gingivalis fimbriae protects against periodontal destruction. Infect Immun 60;2926-2935, 1992
  12. Hamada N, Watanabe K, Sasakawa C, Yoshikawa M, Yoshimura F, Umemoto T: Construction and characterization of a fimA mutant of Porphyromonas gingivalis. Infect Immun 62;1696-1704, 1994
  13. 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 33;479-485, 1988 https://doi.org/10.1016/0003-9969(88)90028-3
  14. Casadevall A: Passive antibody administration (immediate immunity) as a specific defense against biological weapons. Emerg Infect Dis 8;833-841, 2002 https://doi.org/10.3201/eid0808.010516
  15. Ma JK, Hunjan M, Smith R, Kelly C, Lehner T: An investigation into the mechanism of protection by local passive immunization with monoclonal antibodies against Streptococcus mutans. Infect Immun 58;3407-3414, 1990
  16. Lee JY, Sojar HT, Amano A, Genco RJ: Purification of major fimbrial proteins of Porphyromonas gingivalis. Protein Expr Purif 6;496-500, 1995 https://doi.org/10.1006/prep.1995.1066
  17. Lee JY, Sojar HT, Bedi GS, Genco RJ: Synthetic peptides analogous to the fimbrillin sequence inhibit adherence of Porphyromonas gingivalis. Infect Immun 60;1662-1670, 1992
  18. Kirakodu SS, Govindaswami M, Novak MJ, Ebersole JL, Novak KF: Optimizing qPCR for the quantification of periodontal pathogens in a complex plaque biofilm. Open Dent J 2;49-55, 2008 https://doi.org/10.2174/1874210600802010049
  19. Hosogi Y, Duncan MJ: Gene expression in Porphyromonas gingivalis after contact with human epithelial cells. Infect Immun 73;2327-2335, 2005 https://doi.org/10.1128/IAI.73.4.2327-2335.2005
  20. Dickinson DP, Kubiniec MA, Yoshimura F, Genco RJ: Molecular cloning and sequencing of the gene encoding the fimbrial subunit protein of Bacteroides gingivalis. J Bacteriol 170;1658-1665, 1988
  21. Ito HO, Nakashima T, So T, Hirata M, Inoue M: Immunodominance of conformation-dependent B-cell epitopes of protein antigens. Biochem Biophys Res Commun 308;770-776, 2003 https://doi.org/10.1016/S0006-291X(03)01466-9
  22. Yoshimura F, Sugano T, Kawanami M, Kato H, Suzuki T. Detection of specific antibodies against fimbriae and membrane proteins from the oral anaerobe Bacteroides gingivalis in patients with periodontal diseases. Microbiol Immunol 31;935-941, 1987 https://doi.org/10.1111/j.1348-0421.1987.tb03154.x