| Project/Area Number |
62570199
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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 | 国立予防衛生研究所 |
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Principal Investigator |
SATO Hiroko National Institute of Health, Senior Researcher, 体液性免疫部, 主任研究官 (80100080)
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| Co-Investigator(Kenkyū-buntansha) |
ITO Akiharu National Institute of Health, Senior Researcher, 体液性免疫部, 主任研究官 (10100067)
SATO Yuji National Institute of Health, Chief, 細菌部, 室長 (40072889)
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| Project Period (FY) |
1987 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1989: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1988: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1987: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | Pertussis Toxin / Monoclonal Antibody / Mouse-Protective Antibody / B. pertussis Mutant / Encephalopathy / Pertussis Toxin-neutralizing Antibody / Pertussis Vaccine / MouseーProtective Antibody / Pertussis Toxinーneutralizing Antibody / 百日咳毒素 / 百日咳菌変異株 / モノクローナル抗体 / 感染防御抗体 / 百日咳脳症 / トキソイド / ワクチン / PT遺伝子 / 抗百日咳毒素モノクローナル抗体 / 感染防御抗原 / 百日咳毒素部分抗原活性 |
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| Research Abstract |
Pertussis toxin (PT) having very complicated structure and activities is the most important protective antigen in a pertussis vaccine. In order to define the important structural and functional features of PT associated with its pathogenicity and immunoprotectivity, twenty monoclonal antibodies against different parts of PT were characterized. Monoclonal antibodies against subunit 1 (Sl) and subunit 2/3 (S2/3) showed high mouse protectivity in aerosol challenge system but the latter antibodies which recognize common structure between S2 and S3 showed little protection against intracerebral challenge with Bordetelia pertussis. Both protective antibodies showed a significant therapeutic effect on the mice respiratory-infected with the organisms. The other antibodies also showed diverse anti-toxic activities in vitro and in vivo assay systems. The variety of the toxin-neutralizing activities of these antibodies depends on the recognition site of each antibody on PT. Most of anti-S1 antibodies did not show so high neutralization to CHO- cell clustering or histamine-sensitizing activities of PT although anti-S2 and/or S3 did.
Using these monoclonal antibodies, protective antigenicity of various non-toxic PT produced by pertussis mutants or E. coli recombinants were analyzed. Pertussis mutants, 79G, producing stable protective antibens S234 without S1 was developed. The S1 must be unstable because of its replacement of Cys-41 with Tyr. Other genetic toxoids were also developed in E. coli and B. pertussis by change of Arg-9 to Lys and/or Glu-129 to Gly of Sl by site directed mutagenesis. The development of these mutant PT antigens suggests that S1 has the most functional structure for the toxicities of PT.
The study on mouse-encephalopathy caused with PT should be continued further more to develop an animal model for pertussis encephalopathy in human.
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