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Development of Immunological Procedure for Prevention of Dental Caries Using Fusion Protein of Colonization Factors

利用定植因子融合蛋白开发预防龋齿的免疫学方法

基本信息

批准号：
07557134
负责人：
KOGA Toshihiko
金额：
$ 6.08万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557134/
关键词：
Dental Caries Streptococcus mutans Colonization Factor Fusion Protein Passive Immunization Protein Antigen Glucosyltransferase Prevention of Dental Caries

项目摘要

Cell-surface protein antigen (PAc) and glucosyltransferases (GTFs) produced by Streptococcus mutans are considered to be major colonization factors of the organism, and the inhibition of these two factors is predicted to provide protection against dental caries. In this study, we have constructed a fusion protein PAcA : GB of the salivabinding alanine-rich region (PAcA) of PAc and the glucan-binding (GB) domain of GTF-I,an enzyme catalyzing the synthesis of water-insoluble glucan from sucrose, and a fusion protein PAcA : SB of PAcA and the sucrose-binding (SB) domain of GTF-I.The recombinant fusion proteins were purified from cell extracts of Escherichia coli harboring the fusion genes, and rabbit antibodies against these fusion proteins were prepared. The water-insoluble glucan synthesis by cell-associated and cell-free GTF preparations from S.mutans and the total glucan synthesis by GTF-I was markedly inhibited by anti-PAcA : GB IgG antibodies, but not by anti-PAcA : SB IgG antibodies. Significant inhibition of the sucrose-independent and sucrose-dependent adhesion of S.mutans to saliva-coated hydroxyapatite beads was observed when anti-PAcA : GB antibodies were added to the reaction mixture. Anti-PAcA : SB antibodies inhibited the adhesion of S.mutans to the beads in the absence of sucrose, but did not in the presence of sucrose. Immunization with the fusion protein PAcA : GB may be useful for controlling the colonization of teeth by the organism.

变形链球菌产生的细胞表面蛋白抗原（PAc）和葡糖基转移酶（GTF）被认为是生物体的主要定植因子，并且预测这两个因子的抑制提供对龋齿的保护。在这项研究中，我们构建了PAc的唾液结合丙氨酸富集区（PAc A）和GTF-1（一种催化蔗糖合成水不溶性葡聚糖的酶）的葡聚糖结合（GB）结构域的融合蛋白PAc A：GB，以及融合蛋白PAc A：PAcA的SB和GTF的蔗糖结合（SB）结构域I.从携带融合基因的大肠杆菌的细胞提取物中纯化重组融合蛋白，并制备针对这些融合蛋白的兔抗体。抗PAcA：GB IgG抗体可显著抑制变形链球菌细胞相关和无细胞GTF制剂的水不溶性葡聚糖合成，而抗PAcA：SB IgG抗体则不能显著抑制GTF-1的总葡聚糖合成。在反应混合物中加入抗-PAcA：GB抗体后，观察到变形链球菌对唾液包被的羟基磷灰石珠的蔗糖非依赖性和蔗糖依赖性粘附的显著抑制。抗-PAcA：SB抗体在不存在蔗糖的情况下抑制变形链球菌与珠的粘附，但在存在蔗糖的情况下不抑制。用融合蛋白PAcA：GB免疫可能有助于控制生物体在牙齿上的定植。