| Project/Area Number | 03807025 |
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
細菌学
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| Research Institution | University of Shizuoka |
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Principal Investigator |
IWAMOTO Yoshihisa University of Shizuoka, Dept. of microbiology, Professor, 薬学部, 助教授 (60046290)
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| 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)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed(Fiscal Year 1993)
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| 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)
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| Keywords | Borrelia 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 |
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| 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
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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
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