Development of Anti-Dental Caries Vaccine Using Recombinant Lactic Streptococci

利用重组乳酸链球菌开发抗龋齿疫苗

• 批准号: 63870086
• 负责人: KOGA Toshihiko
• 金额: $ 7.94万
• 依托单位: National Institute of Health
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B).
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63870086/
• 关键词: Dental Caries; Streptococcus mutans; Streptococcus sobrinus; Lactic Streptococci; Vaccine; DNA組換え; う蝕

Streptococcus mutans has been implicated as a causative organism of human dental caries. S. Mutans produces a cell surface protein antigen (PAc) with a molecular mass of 190 kilodaltons. In this study, we constructed a recombinant lactic streptococcus with carries th pac gene of S. Mutans for development of an oral vaccine against dental caries.1. We cloned the pac gene of S. Mutans MT8148 (serotype c) and determined the complete nucleotide sequence of the gene. The pac gene consisted of 4,695 base pairs. The initial gene product contained 1,565 amino acids and had a molecular weight of 170,773.2. To make clear the biological function of PAc, we constructed genetically several PAc-defective transformants fo S. mutans. The cell hydrophobicity of PAc-defective transformants was markedly lower than that of the parent strain. Resting cells of the hydrophilic strains attached in lower number to saliva-coated hydroxyapatite than did the parent strain. These findings suggest that cell surface PAc of S. Mutans participates in attachment of the streptococcal cell to aquired pellicles on tooth surfaces.3. Homology between PAc of S. Mutans and a 210-kilodalton protein antigen of Streptococcus sobrinus was high in the central regions.4. The pac gene from S. mutans was joined to shuttle vector pSA3. The chimeric plasmid pSM1 was successfully transformed into Streptococcus lactis. The recombinant S. Lactis produced a small amount of PAc. Significant salivary immunoglobulin A and serum immunoglobulin G responses to PAc were induced in mice immunized orally with the recombinant S. lactis.

变形链球菌被认为是人类龋齿的致病微生物。 S. Mutans 产生分子量为 190 千道尔顿的细胞表面蛋白抗原 (PAc)。本研究构建了携带变形链球菌th pac基因的重组乳酸链球菌，用于口腔龋齿疫苗的开发。 1．我们克隆了变形链球菌MT8148（血清型c）的pac基因，并确定了该基因的完整核苷酸序列。 pac 基因由 4,695 个碱基对组成。最初的基因产物含有1,565个氨基酸，分子量为170,773.2。为了明确PAc的生物学功能，我们从基因角度构建了几种PAc缺陷型变形链球菌转化体。 PAc缺陷型转化子的细胞疏水性明显低于亲本菌株。亲水菌株的静息细胞比亲本菌株附着在唾液包被的羟基磷灰石上的数量更少。这些结果表明，变形链球菌细胞表面的PAc参与了链球菌细胞与牙齿表面后天菌膜的附着。3．变异链球菌PAc与远缘链球菌210kD蛋白抗原的同源性在中部区域较高。 4．将来自变形链球菌的 pac 基因连接至穿梭载体 pSA3。嵌合质粒pSM1成功转化乳酸链球菌。重组乳链球菌产生少量的PAc。口服重组乳酸链球菌免疫小鼠，诱导出对 PAc 的显着唾液免疫球蛋白 A 和血清免疫球蛋白 G 反应。

项目成果

N. Okahashi et al: "Molecular characterization of a surface protein antigen gene from serotype c Streptococcus mutans, implicated in dental caries." Molecular Microbiology. 3. 673-678 (1989)

N. Okahashi 等人：“c 血清型变形链球菌表面蛋白抗原基因的分子特征，与龋齿有关。”

岡橋暢夫: 微生物. 4. 3-12 (1988)

冈桥伸夫：微生物。4. 3-12 (1988)

I.Takahashi et al.: "Homology between surface protein antigen genes of Streptococcus sobrinus and Streptococcus mutans" FEBS Letters. 249. 383-388 (1989)

I.Takahashi 等人：“远缘链球菌和变形链球菌表面蛋白抗原基因之间的同源性”FEBS Letters。

M.Iwaki et al.: "Oral immunization with recombinant Streptococcus lactis carring the Streptococcus mutans surface protein antigen gene" Infection and Immunity. 58. 2929-2934 (1990)

M.Iwaki 等人：“携带变形链球菌表面蛋白抗原基因的重组乳酸链球菌的口服免疫”感染和免疫。

I.Takahashi et al.: "Intranasal immunization with recombinant protein antigen of serotype c <Streptococcus>___ー <mutans>___ー and chorea toxin B subunit" Archives of Oral Biology. 35. 475-477 (1990)

I. Takahashi 等人：“用血清型 c <链球菌>___<变异体>____ 和舞蹈病毒素 B 亚基的重组蛋白抗原进行鼻内免疫”口腔生物学档案 35. 475-477 (1990)。