Inhibition of dental plaque formation and determination of anti-cariogenic effect by DNA vaccine

DNA疫苗抑制牙菌斑形成及防龋效果测定

• 批准号: 15390639
• 负责人: FUJIWARA Taku
• 金额: $ 8.77万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15390639/
• 关键词: glucosyltransferase; Streptococcus mutans; DNA vaccine; Stretococcus mutans; S.mutans

Glucosyltransferases(GTFs) from Streptococcus mutans synthesis the adhesive glucans from sucrose and are important virulence factor of dental caries. The purposes of this investigation are : 1)Determination of the immunogenicity of GTF domains to select target region of anti-caries DNA vaccine, 2)Construct DNA vaccine against the candidate domain, 3)Analysis of anti-cariogenic effect of the DNA vaccine.GTFs commonly possesses three (variable, catalytic, and glucan-binding) domains. Antigenicity of each domain was predicted by Paker's method, then the recombinant plasmids carrying these domains of GTFs from S.mutans MT8148 were constructed. Western blot analysis revealed that immunogenicity of the catalytic and glucan-binding regions was low. On the other hand, the variable region was thought to possess immunogenicity.DNA vaccine plasmid containig the variable region of GTF was constructed using pAFC3 which was carrying the adeno virus promoter. The vaccine plasmid was transfected into 293 cells and the expression of mRNA was confirmed by Northen blot analysis.To the next step, we will determine the anti-cariogenic activity of the DNA vaccine in in vitro and in vivo (animal) experiments.

变形链球菌的葡萄糖基转移酶（GTF）从蔗糖合成粘附葡聚糖，是龋齿的重要毒力因子。本研究的目的是：1）测定GTF结构域的免疫原性，以选择抗龋DNA疫苗的靶区域；2）针对候选结构域构建DNA疫苗；3）分析DNA疫苗的抗龋效果。 GTF 通常具有三个结构域（可变结构域、催化结构域和葡聚糖结合结构域）。通过Paker法预测各结构域的抗原性，然后构建携带来自S.mutans MT8148的GTF的这些结构域的重组质粒。蛋白质印迹分析显示催化区和葡聚糖结合区的免疫原性较低。另一方面，可变区被认为具有免疫原性。利用携带腺病毒启动子的pAFC3构建了含有GTF可变区的DNA疫苗质粒。将疫苗质粒转染293细胞，并通过Northen blot分析证实mRNA的表达。下一步，我们将通过体外和体内（动物）实验测定DNA疫苗的抗龋齿活性。

项目成果

Method for rapid identification of oral streptococci by PCR using 16S-23S ribosomal RNA intergenic spacer gene

• DOI: 10.1016/s0917-2394(05)70051-3
• 发表时间: 2005-01-01
• 期刊: PEDIATRIC DENTAL JOURNAL
• 影响因子: 0.8
• 作者: Hoshino, Tomonori;Izumi, Tomohiro;Fujiwara, Taku
• 通讯作者: Fujiwara, Taku

PCR detection and identification of oral streptococci in saliva samples using gtf genes

• DOI: 10.1016/j.diagmicrobio.2003.10.002
• 发表时间: 2004-03-01
• 期刊: DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE
• 影响因子: 2.9
• 作者: Hoshino, T;Kawaguchi, M;Fujiwara, T
• 通讯作者: Fujiwara, T

IMMUNOGENICITY OF GTF DOMAINS FROM STR. MUTANS FOR DNA VACCINE CANDIDATE

STR. GTF 结构域的免疫原性

• 发表时间: 2005
• 期刊: The 4^<th> Conference of Pediatric Dentistry Association of Asia
• 作者: FUJIWARA;T.*;HOSHINO;T.;KONISHI;I.;FUKUMOTO.S.
• 通讯作者: FUKUMOTO.S.

Immunogenicity of gtf domains from Str.mutans for DNA vaccine candidate

候选 DNA 疫苗中变形链球菌 gtf 结构域的免疫原性

• 发表时间: 2004
• 期刊: The 4^<th> Conference of Pediatric Dentistry Association of Asia
• 作者: Fujiwara;T.;Hoshino;T.;Konisi;I.;Fukumoto;S.
• 通讯作者: S.