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

• Project/Area Number: 15390639
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Orthodontic/Pediatric dentistry
• Research Institution: Nagasaki University
• Principal Investigator: FUJIWARA Taku Nagasaki University, Graduate School of Biomedical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (00228975)
• Co-Investigator(Kenkyū-buntansha): FUKUMOTO Satoshi Kyushu University, Graduate School of Dental Science, Associate Professor, 歯学研究院, 助教授 (30264253) ; HOSHINO Tomonori Nagasaki University, Graduate School of Biomedical Sciences, Instructor, 大学院・医歯薬学総合研究科, 助手 (00359960)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥13,700,000 (Direct Cost: ¥13,700,000); Fiscal Year 2004: ¥5,900,000 (Direct Cost: ¥5,900,000); Fiscal Year 2003: ¥7,800,000 (Direct Cost: ¥7,800,000)
• Keywords: glucosyltransferase / Streptococcus mutans / DNA vaccine / Stretococcus mutans / S.mutans

Research Abstract

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.

Research Products

• [Journal Article] PCR detection and identification of oral streptococci in saliva samples using gtf genes (2005)
  • Author(s): Hoshino, T., Kawaguchi, M., Shimizu, N., Hoshino, N., Ooshima T., Fujiwara, T.
  • Journal Title: Diagnostic Microbiology and Infectious Disease 48 Pages: 159-199
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Method for rapid identification of oral streptococci by PCR using 16S-23S ribosomal RNA intergenic spacer gene (2005)
  • Author(s): Hoshino, T., Izumi, T., Ooshima, T., Fujiwara, T.
  • Journal Title: Pediatric Dental Journal (IN PRESS)
  • NAID: 130004721778
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] IMMUNOGENICITY OF GTF DOMAINS FROM STR. MUTANS FOR DNA VACCINE CANDIDATE (2005)
  • Author(s): FUJIWARA, T.*, HOSHINO, T., KONISHI, I., FUKUMOTO.S.
  • Journal Title: The 4^<th> Conference of Pediatric Dentistry Association of Asia Pages: 89-89
• [Journal Article] Immunogenicity of gtf domains from Str.mutans for DNA vaccine candidate (2004)
  • Author(s): Fujiwara, T., Hoshino, T., Konisi, I., Fukumoto, S.
  • Journal Title: The 4^<th> Conference of Pediatric Dentistry Association of Asia Pages: 89-89
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Immunogenicity of gtf domains from Str.mutans for DNA vaccine candidate (2004)
  • Author(s): Fujiwara, T., Hoshino, T., Konisi, I., Fukumoto, S.
  • Journal Title: The 4^<th> Conference of Pediatric Dentistry Association of Asia 89
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] PCR detection and identification of oral streptococci in saliva samples using gtf genes (2004)
  • Author(s): Hoshino, T., Kawaguchi, M., Shimizu, N., Hoshino, N., Ooshima T., Fujiwara, T.
  • Journal Title: Diag Microbiol Infect Dis 48 Pages: 195-199
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Method for rapid identification of oral streptococci by PCR using 16S-23S ribosomal RNA intergenic spacer gene
  • Author(s): Hoshino, T., Izumi, T., Ooshima, T., Fujiwara, T.
  • Journal Title: Pediatric Dental Journal (in press)
  • NAID: 130004721778
  • Description: 「研究成果報告書概要(欧文)」より