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Development of Diagnosis and Vaccine for Lyme Borreliosis in Japan.

Research Project

Project/Area Number 03807025
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 細菌学
Research InstitutionUniversity of Shizuoka

Principal Investigator

IWAMOTO Yoshihisa  University of Shizuoka, Dept. of microbiology, Professor, 薬学部, 助教授 (60046290)

Co-Investigator(Kenkyū-buntansha) FUKUNAGA Masahito  Fukuyama Universuty, Dept of Microbiology, Professor, 薬学部, 教授 (20132483)
MASUZAWA Toshiyuki  University of Shizuoka, Dept. of Microbiology, Research Associate, 薬学部, 助手 (10181645)
柳原 保武  静岡県立大学, 薬学部, 教授 (30046255)
Project Period (FY) 1991 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1993: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1991: ¥700,000 (Direct Cost: ¥700,000)
KeywordsBorrelia burgdorferi / Lyme borreliosis / PCR / RFLP / Vaccine / Osp A / Osp B / Flagellin / 単クローン抗体 / 抗原構造 / シュルツェマダニ / ヤマトマダニ / 単クロ-ン抗体 / Ixodes persulcatus / Ixodes ovatus
Research Abstract

Development of Gene diagnosis for Lyme Disease : Primer pairs targeting to flagellin and outer surface protein A (Osp A) and Osp B gene were chemically synthesized. Flagellin primer set could amplified the gene with all strains used in this study isolated in Japan, North Americand Europe. The amplified DNA fragment was detected in sample including one cell by second PCR using the amplified DNA fragment from first PCR as template. The DNA was amplified in samples of urine and ear punch from mice experimentally infected with Borrelia burgdorferi. Restriction fragment length polymorphism (RFLP) on an fragment amplified with Osp A-Osp B gene primer set gave the information for genospecies determination in B.burgdorferi sensulato.
Development of Vaccine ageinst Lyme Disease : Outbred ddy mice inoculated with live cells of B.burgdorferi into hind footpad displayd swelling of the footpad at days 5 to 11 after inoculation. The mice inoculated with neither heat-inactivated cells at 56 C for 30 m … More in nor medium show any significant histopathological change. B.burgdorferi was cultured from bladder and heart of the mice at day 14 after inoculation. The outbred ddy mice model is useful for evaluating the effectiveness of vaccination against Lyme disease. Cross-protective activity of B.burgdorferi sensu lato was examined in outbred ddy mice. Isolates from Japan, North American and Europe belonging to four genospecies, B.burgdorferi sensu stricto, B.garinil, B.afzelii and B.japonica used in this study were further classified into Osp A serotypes depend on the reactivity with monoclonal antibody to Osp A.The mice were protected by active immunization with monoclonal antibody to Osp A.The mice were protected by active immunization with the vaccine prepared from the challenge strain, but no crossprotection was observed by immunization with vaccine prepared from borrelial culture belonging to heterologous serotypes. The mice were partially protected by vaccination with the Borrelia belonging in the same serotype as challeng strain. No cross-protection was observed among the four genospecies.
These results suggested that protective antigens besides Osp A are present in B.burgdorferi sensu lato and polyvalent vaccine is necessary for prophylaxis of Lyme disease. Less

Report

(4 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • 1991 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] Toshiyuki Masuzawa: "Experimental Borrelia burgdorferi Infection of Outbred Mice." J.Clin.Microbiol.30. 3016-3018 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Masahito Fukunaga: "The 23S/5S Ribosomal RNA Genes(rrl/rrf)Are Separated from the 16S Ribosomal RNA Gene(rrs)in Borrelia burgdorferi,the Aetiological Agent of Lyme Disease." J.Gen.Microbiol.138. 871-877 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Toshiyuki Masuzawa: "Protective Activity of Antisera against Isolates of Borrelia burgdorferi from Various Geographical Origins." Microbiol.Immunol.37. 79-83 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Masahito Fukunaga: "Analysis of Borrelia Species Associated with Lyme Disease by rRNA Gene Restriction Fragment Length Polymorphism." J.Gen.Microbiol.139. 1141-1146 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 増澤俊幸: "ライム病ワクチン開発の動向." 日本細菌学雑誌. 48. 385-393 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Toshiyuki Masuzawa: "Experimental Induction of Lyme Arthritis in Outbred Mice." Microbiol.Immunol.(印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Toshiyuki Masuzawa et al.: "Experimental Borrelia bougdorferi infection of outbred mice." J Clin. Microbiol.30. 3016-3018 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Masahito Fukunaga et al.: "The 23S/5S ribosomal RNA genes (rrl/rrf) are separated from the 16S ribosomal RNA gene (rrs) in Borrelia burgdorferi, the aetiological agent of Lyme disease." J.Gen. Microbiol.138. 871-877 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Toshiyuki Masuzawa et al.: "Protective activity of antisera against isolates of Borrelia burgdorferi from various geographical origins." Microbiol. Immunol.37. 79-83 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Masahito Fukunaga et al.: "Analysis of Borrelia species associated with Lyme disease by rRNA gene restriction fragment length polymorphism." J.Gen.Microbiol.139. 1141-1146 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Toshiyuki Masuzawa.: "Current status of vaccine development of Lyme borreliosis." Jpn.J.Bacteriol.48. 385-593 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Toshiyuki Masuzawa et al.: "Experimental induction of Lyme arthritis in outbred mice." Microbiol.Immunol.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Toshiyuki Masuzawa: "Experimental Borrelia burgdorferi Infection of Outbred Moce." J.Clin.Microbiol.30. 3016-3018 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Masahito Fukunaga: "The 23S/5S Ribosomal RNA Genes (rrl/rrf) Are Separated from the 16S Ribosomal RNA Gene (rrs) in Borrelia burgdorferi,the Aetiological Agent of Lyme Disease." J.Gen.Microbiol.138. 871-877 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Toshiyuki Masuzawa: "Protective Activity of Antisera against Isolates fo Borrelia burgdorferi from Various Geographical Origins." Microbiol.Immunol.37. 79-83 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Masahito Fukunaga: "Analysis of Borrelia Species Associated with Lyme Disease by rRNA Gene Restriction Fragment Length Polymorphism." J.Gen.Microbiol.139. 1141-1146 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 増澤俊幸: "ライム病ワクチン開発の動向," 日本細菌学雑誌. 48. 385-393 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Toshiyuki Masuzawa: "Experimental Induction of Lyme Arthritis in Outbred Mice." Microbiol.Immunol.38 (印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Masuzawa,T.: "Experimental Borrelia burgdorferi infection of outbrd mice." J.Clin.Microbiol.30. 3016-3018 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Fukunaga,M.: "The 23S/5S ribosomal RNA genes (rrl/rrf) are separate from the 16S ribosomal RNA gene (rrs) in Borrelia burgdorferi,the aeti ologic agent of Lyme disease." J.Gen.Microbiol.138. 871-877 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Masuzawa,T.: "Protective activity of antisera against isolates of Borrelia burgdorferi from various geographical origins" Microbiol.Immunol.

    • Related Report
      1992 Annual Research Report
  • [Publications] Fukunaga,M.: "Analysis of Borrelia species associated with Lyme disease by rRNA gene restriction fragment length polymorphism" J.Gen.Microbiol.

    • Related Report
      1992 Annual Research Report
  • [Publications] 増沢 俊幸: "ライム病ワクチン開発の動向" 日本細菌学雑誌.

    • Related Report
      1992 Annual Research Report
  • [Publications] Masuzawa,T.: "Antigenic properties of Borrelia burgdorferi isolated from Ixodes ovatus and Ixodes persurcatus in Hokkaido,Japan" J.Clin.Microbiol.29. 1568-1573 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Isogai,E.: "Serological survey for Lyme disease in Sikadeer(Cervus nippon yesoensis)by enzymelinked Immunosorbent assay(ELISA)" Microbiol.Immunol.35. 695-703 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Masuzawa,T.: "Immunological properties of Borrelia burgdorferi isolated from the Ixodes ovatus in Shizuoka,Japan" J.Clin.Microbiol.35. 913-919 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kobayashi,Y.: "Leptospirosis" University of Tokyo Press, 534 (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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