| Project/Area Number |
60480412
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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 |
Conservative dentistry
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| Research Institution | Osaka University, Faculty of Dentistry |
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Principal Investigator |
EBISU Shigeyuki Osaka University, Faculty of Dentistry, Associate Professor, 歯学部, 助教授 (50116000)
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| Co-Investigator(Kenkyū-buntansha) |
KIMURA Shigeyuki Osaka University, Faculty of Dentistry. Assistant Professor, 歯学部附属病院, 講師 (10177917)
FUKUHARA Hironobu Osaka University, Faculty of Dentistry, Assistant Professor, 歯学部附属病院, 講師 (20144503)
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| Project Period (FY) |
1985 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1986: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1985: ¥4,100,000 (Direct Cost: ¥4,100,000)
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| Keywords | Eikenella corrodens / Bacterial lectin / Monoclonal antibody / Oral colonization / 定着阻害 / 菌体間凝集 |
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| Research Abstract |
A bacterial lectin-like substance (LS), which is considered to participate in the adherence of Eikenella corrodens to various host cells, was purified from E. corrodens cells. The LS was extracted in 1% Triton X-100 with sonication from the cell envelope of E. corrodens 1073, and partially purified by galactosamine affinity chromatography and gel filtration chromatography. The LS was purified about 256-fold as evaluated by its specific hemagglutinating activity.
A method to produce monoclonal antibody against the LS (moAnti-LS) was investigated. Splenocytes of BALB/c mice immunized with purified LS and myeloma cells (SP-2) were fused, and hybridomas were screened by ELISA. Four hybridoma clones were selected. Two of them were of the IgG1 isotype and the others were IgG2b isotype. Immunoelectron microscopic study revealed that LS was found to localize in cell wall or capsule of E. corrodens cells.
E. corrodens 1073 was found to coaggregate with 8 strains of total of 42 oral bacterial strains examined, and the study on mechanisms of the coaggregation reactions of E. corrodens 1073 with Actinomyces viscosus ATCC19246, A. viscosus T14AV, Streptococcus sanguis 34, and S. sanguis ST160R indicates that the coaggregations are mediated by a system of specific cell surface interaction between the LS on E. corrodens and sugar receptors on the partner.
The moAnti-LS completely inhibited the coaggregation of E. corrodens with 2 strains of A. viscosus and 2 strains of S. sanguis. The moAnti-LS also inhibited hemagglutination of E. corrodens. These results suggested that colonization of E. corrodens in the mouth of experimental animal may be specifically inhibited by the moAnti-LS. Indeed, the establishment of E. corrodens 1073 in the periodontal pocket and the adsorption of the alveolar bone of SPF rats in vivo were both inhibited by administration of N-acetyl-D-galactosamine, a specific inhibitor of the LS.
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