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1997 Fiscal Year Final Research Report Summary

Molecular Analysis of Infectious Mechanisin of A.actinomycelemcomitans

Research Project

Project/Area Number 08457484
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Morphological basic dentistry
Research InstitutionTOKYO DENTAL COLLEGE

Principal Investigator

OKUDA Katuji  TOKYO DENTAL COLLEGE,DEPARTMENT OF DENTISTRY,PROFESSOR, 歯学部, 教授 (40085741)

Co-Investigator(Kenkyū-buntansha) YAMANAKA Ayumi  TOKYO DENTAL COLLEGE,DEPARTMENT OF DENTISTRY,ASSISTANT, 歯学部, 助手 (40231667)
MIURA Tadashi  TOKYO DENTAL COLLEGE,DEPARTMENT OF DENTISTRY,ASSISTANT, 歯学部, 助手 (10266570)
ISHIHARA Kazuyuki  TOKYO DENTAL COLLEGE,DEPARTMENT OF DENTISTRY,LECTURER, 歯学部, 講師 (00212910)
KATO Tetsuo  TOKYO DENTAL COLLEGE,DEPARTMENT OF DENTISTRY,ASSISTANT PROFESSOR, 歯学部, 助教授 (00159253)
Project Period (FY) 1996 – 1997
KeywordsPeriodontal disease / Antibody / Attachment factor / fop gene / cloning / A.actinomycetemcomitans / fimbrial / vaccination
Research Abstract

Fundamental advances to maintain oral health such as preventing periodontal disease are required in the elderly society. Actinobacillus actinomycetemcomitans is considered to be one of the most important periodontopathic bacteria. We found that sera from A.actinomycetemcomitans culture-negative periodontitis patients with elevated titers of IgG against fimbriae had significantly higher avidity for the fimbrial antigen than did sera from culture-positive patients with elevated titers. We extracted A.actinomycetemcomitans fimbrial antigen and determined the N-terminal amino acid sequence. We prepared primers according to the N-terminal aminoacid sequence and fimbrial-associated protein was cloned. The plasmid obtained was found to harbor a 1.7kb fragment which contained a 228bp open reading frame encoding 75 amino acids. The fimbriae associated protein gene (fap) was strongly expressed in fimbriated A.actinomycetemcomitans 310a but not in non-fimbriated strains. We sunthesized an oligope … More ptide, according to the amino acid sequence and the synthetic peptide was conjugated by the branched lysine polymer resin beads method, which has the advantage of Increasing molecular size and immunogenicity. An synthetic peptide with the sequence of V-A-V-F-Y-S-N-N-G-F-I-A-N-L-Q-S-K-F-F-S-L produced highest anti-fimbrial antibody levels in the ELISA test. We found that rabbit antiserum against a synthetic peptide inhibited the attachment of A.actinomycetemcomitans strains to saliva-coated hydroxyapatite beads, human buccal epithelial cells, and cultured gingival fibroblasts in vitro. These findings suggest that the synthetic fimbrial antigen acts as a vaccine for inducing an antibody response that inhibits A.actinomycetemcomitans colonization.
We demonstrated the genetic regulation of immune responses to the synthetic A.actinomycetemcomitans fimbrial antigen using various inbred strains of mice. We examined the effective adjuvant activity and immunization route to produce s-IgA against the synthetic A.actinomycetemcomitansfimbrial antigen in saliva using BALB/c mice. The synthetic peptide antigen suspended in PBS was injected with Freund adjuvant Ribi adjuvant, liposome, plasmid DNA coding mouse IL-4, or/and cholera toxin. BALB/c mice were immunized through subcutaneous, nasal mucosa or oral route. We found that a group of mice immunized through nasal mucosa with a mixture of liposomes, the plasmid DNA and cholera toxin. Mice immunized through nasal mucosal route resulted in higher s-IgA production against the synthetic antigen, and delayd type hypersensitivity was also significantly enhanced in the group. We are now studying the role of s-IgA antibody against the fimbrial antigen in colonization inhibition by A.actinomycetemcomitans. Less

  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] Kimizuka, R.et al: "Characterization of Actinobacillus actinomycetemcomitans hemolysin" Microbiology and Immunology. 40. 717-723 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ishihara, K.et al: "Characterization of the Treponemadenticola prt p gene encoding a prolyl Phenylalanine Specific protease (dentilisin)" Infection and Immunity. 64. 5178-5186 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kato, T.et al: "Isolation and characterization of hemolysin genes from Actinobacillis actinomy cetemcomitans" FEMS Microbiol.Lett. 143. 217-222 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Honma, K.et al: "Genetic Control of immune responses to a synthetic fimbrial antigen of Actinobacillus actinomycetemcomitans" Microbiology and Immunity. 41. 609-614 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ishihara, K.et al: "Cloning and sequence analysis of fimbriac associated (fap) gene from Actinobacillus actinomycetemcomitans" Microbial Pathogenesis. 23. 63-69 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Okuda, K.et al: "Role for the S-layer of Campylobacterrectus ATCC 33238 in complement mediated killing and phagocytic killing by leukocytes from guinea pig and human peripheral blood" Oral Disenses. 3. 113-120 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 奥田 克爾: "口腔の感染症とアレルギー" 一世印刷出版, 1-315 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 奥田 克爾: "歯周病学" 永末書店, 29-39 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kimizuka R., Miura T.and Okuda K.: "Characterization of Actinobacillus actinomycetemcomitans hemolysin" Microbiol.Immunol. 40. 717-723 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kato T., Kimizuka R.and Okuda K.: "Isolation and characterization of hemolysin genes from Actinobacillus actinomycetemcomitans" FEMS Microbiol.Lett.143. 217-221 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ishihara K., Miura T., Kuramitsu, H.K.and Okuda K.: "Characterization of the Treponema denticola prtP gene encoding a prolylphenylalanine specific protease (dentilisin)" Infect Immun. 64. 5178-5186 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Honma K,Ishii N,Kato T,Ishihara K,Okuda K.and Okuda K.: "Genetic control of immune responses to a synthetic fimbrial antigen of Actinobacillus actinomycetemcomitans" Microbiol.Immun.41. 609-614 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ishihara K,Honma K,Miura T.Kato T.and Okuda K.: "Cloning and sequence analysis of fimbriae associated (fap) gene from Actinobacillus actinomycetemcomitans" Microbial Pathogen. 23. 63-69 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ishihara K,Miura T,Kimizuka R,Ebihara Y,Mizuno Y.and Okuda K.: "Oral bacterial inhibit the Helicobacter pylori growth" FEMS Microbial.Lett.159. 355-361 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Okuda K,Kigure T,Yamada S,Kaneko T,Ishihara K,Miura T,Kato T.and Takazoe I.: "Role for the S-layr of Campylobacter rectus ATCC33238 in complement mediated killing and phagocytic killing by leukocytes from guinea pig and human peripheral blood" Oral Disease. 3. 113-120 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Saito T,Takatsuka T,Kato T,Ishihara K.and Okuda K.: "Adherence of oral streptococci to an immobilized antimicrobial agent" Arch.oral Biol. 42. 539-545 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Saito T,Ishihara K,Kato T and Okuda K.: "Cloning expression, and sequencing of a protease gene from Bacteroides forsythus" Infect.Immun.65. 4888-4891 (1997)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1999-03-16  

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