Elucidation of the caries-inducing mechanism by S.mutans and development of a diagnostic method to estimate caries-risk.

阐明变形链球菌诱发龋齿的机制并开发估计龋齿风险的诊断方法。

基本信息

批准号：
06671892
负责人：
FUKUSHIMA Kazuo
金额：
$ 1.34万
依托单位：
Nihon University, School of Dentistry at Matsudo
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

Recentry, we have constructed S.milleri transformants KSB8, KSC43 and NH5 which individually express S.mutans gtfB,gtfC and gtfD coding GTF-I,-SI and -S enzymes, and established hybridoma which produce monoclonal antibodies mono-specific to the three GTFs. In this study, in vitro and in vivo cariogenicities of the three transformants were compared to elucicate the caries-inducing mechanism by S.mutans. Furthermore, using the mono-specific MAbs, a high-sensitive Gross-dot method for determination of the three GTFs was developed. The results obtained were as follows :1. More stable gtfC-expressing transformant KSC43-RT was reisolated from an oral cavity of rat infected with KSC43.2. In vitro cariogenicity of the transformants was examined by a colonization test. The results showed that only KSC43-RT cells could makedly colonized on glass surfaces.3. In vivo cariogenicity of the transformants was tested by a rat model system. The results showed that the KSC43-RT cells could induced the buccal and sulcal caries, and the KSB8 cells could induced the proximal caries, but the NH5 cells did not.4. High-sensitive Gross-dot assay method was established and applied in order to evaluate the cariogenicity of S.mutans strains.5. Cell-associated GTFs from various clinical isolates of S.mutans were analyzed by the method. The results showed that the levels of three GTFs and those ratio were markedly different among the isolates.These results suggest that the gtfC product was especially important for caries development caused by S.mutans, and that the high-sensitive Gross-dot method may be useful to estimate the exact cariogenicity of individual strain of S.mutans.

最近，我们构建了S.Milleri Transforments KSB8，KSC43和NH5，它们单独表达S.Mutans GTFB，GTFC和GTFD编码GTF-I，-SI和-S酶，并建立了杂交瘤，从而产生单核抗体抗体单抗体对三个GTFS的单细胞抗体。在这项研究中，比较了这三种转化体的体外和体内致富性，以阐明S.Mutans诱导龋齿的机制。此外，使用单一特异性mAb，开发了一种高敏性总点方法来确定三个GTF。获得的结果如下：1。从感染KSC43.2的大鼠口腔中重新分离了更稳定的表达GTFC转化剂KSC43-RT。通过定殖测试检查了转化体的体外致富性。结果表明，只有KSC43-RT细胞才能在玻璃表面上植入。3。通过大鼠模型系统测试了转化体的体内创核性。结果表明，KSC43-RT细胞可能诱导颊和硫磺龋，而KSB8细胞可能诱导近端龋齿，但NH5细胞没有。4。建立并应用了高敏性的总点测定法，以评估S.Mutans菌株的致癌性。5。通过该方法分析了来自各种临床分离菌的细胞相关GTF。结果表明，在分离株之间，三个GTF和这些比率的水平明显不同。这些结果表明，GTFC产物对于由S.Mutans引起的龋齿发育尤为重要，并且高敏感的总点方法可能有助于估计S.Mutans个体菌株的精确性癌性。