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.

来自链球菌突变的葡萄糖基转移酶（GTF）合成蔗糖的粘合剂葡萄糖，是龋齿的重要毒力因子。这项研究的目的是：1）确定GTF结构域的免疫原性以选择抗帕里斯DNA疫苗的靶区域，2）构建针对候选结构域的DNA疫苗，3）分析DNA疫苗的抗钙化效应的分析，通常具有三个可变（可变）（可变的，型，型，型，glucanting和glucan-inding）。通过Paker的方法预测了每个结构域的抗原性，然后构建了携带来自S.Mutans MT8148的GTF域的重组质粒。蛋白质印迹分析表明，催化和葡萄糖结合区域的免疫原性很低。另一方面，可变区域被认为具有免疫原性。DNA疫苗质粒遏制GTF的可变区域是使用携带Adeno病毒启动子的PAFC3构建的。将疫苗质粒转染到293个细胞中，并通过Northen印迹分析确认mRNA的表达。在下一步中，我们将在体外和体内（动物）实验中确定DNA疫苗的抗癌活性。

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.