1995 Fiscal Year Final Research Report Summary
Immunoresponsive Analysis of Protection against Periodontopathogen
| Project/Area Number |
05454512
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
病態科学系歯学(含放射線系歯学)
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| Research Institution | TOKYO DENTAL COLLEGE |
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Principal Investigator |
OKUDA Katsuji Tokyo Dental College, Microbiology, Professor, 歯学部, 教授 (40085741)
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| Co-Investigator(Kenkyū-buntansha) |
YAMANAKA Ayumi Tokyo Dental College, Microbiology, Assistant, 歯学部, 助手 (40231667)
KATO Tetsuo Tokyo Dental College, Microbiology, Asspcoate Professor, 歯学部, 助教授 (00159253)
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| Project Period (FY) |
1993 – 1995
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| Keywords | Periodontal disease / Gram-negative rod / A.actinomycetemcomitans / P.gingivalis / Campylobacter rectus / Protection / IgG response / vaccination |
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| Research Abstract |
Specific gram-negative bacteria including Actinobacillus actinomycetemcomitans, Prophyromonas gingivalis and Campylobacter restus are periodontopathogen. The purpose of this study is to critify the importance of the fimbriae attachment factor of A.actinomycetemcomitants and the significance of anti-fimbrial antibody functions as an attachment inhibitor. Fimbrial antigen was prepared from the A.actinomycetemcomitans 310-a strain. Oligopeptides were synthesized according to the amino acid sequence of the fimbrial protein. The peptide antigen was conjugated with branched lysine polymer resin beads. The peptide antigen was suspended in PBS emulsified with incomplete Freund's adjuvant and used to immunize rabbits. A rabbit antiserum reacted with an approximately 54kDa protein of the fimbriae protein from A.actinomycetemcomitans 310-a and with those of other fimbriated strains. This antiserum atrongly inhibited the attachment of fimbriated A.actinomycetemcomitans strains to saliva-coated hyd
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roxyapatite beads, buccal epithelial cells, and a fibroblast cell line, Gin-1.
The relative avidity of serum immunoglobulin G antibody for the fimbria antigens from A.actinomycetemcomitans and P.gingivalis was assessed by diethylamine dissociation enzyme-linked immunosorbent assay in patients with adult periodontitis. High-titer sera from patients not harboring had significantly higher avidities for the fimbria antigen than did high-titer sera from patients with the infections in their periodontal pockets. The elicited antibodies against the fimbria antigen may afford protection against A.actinomycetemcomitans and P.gingivalis infections.
Rabbit polyclonal antiserum to whole cells of C.restus ATCC33238 and monoclonal mouse antibodies against surface antigens of the microorganisms were prepared. Monoclonal antibody against 150KDa S-layr protein from C.rectus ATCC33238 reacted with 4 out of 9 C.rectus strains. The monoclonal antibody recognized 150KDa S-layr inhibited dose dependently the C.rectus attachment to human gingival fibroblast, G1 cells, in vitro. Phagocytic killing of C.rectus ATCC33238 by peritoneal leukocytes prepared from guinea pig was significantly enhanced by the rabbit antiserum and moderately enhanced by monoclonal antibody against S-layr, but was not enhanced monoclonal antibody to lipopolysaccharide.
Electron micrographs of leukocytes exposed to C.rectus cells on the absence of antibody showed injury, indicating that the microorganisms possesses a leukocyte toxic activity. Less
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