| Project/Area Number |
07557033
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Immunology
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| Research Institution | Juntendo University School of Medicine, Department of Immunology |
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Principal Investigator |
RA Chisei Juntendo University School of Medicine, Associate Professor, 医学部, 講師 (60230851)
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| Co-Investigator(Kenkyū-buntansha) |
YAGI Shintaro Tonen Corporation, Research & Development Laboratory Senior Investigator, 総合研究所, 主任研究員
NAITO Koji The Grean Cross Corpration, Basic Research Laboratory Senior Investigator, 中央研究所, 主任研究員
TAKAI Toshiro Asahi Breweries, LTD., Bioscience Research & Development Laboratory Senior Inves, 主任研究員
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| Project Period (FY) |
1995 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥12,000,000 (Direct Cost: ¥12,000,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1996: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1995: ¥6,000,000 (Direct Cost: ¥6,000,000)
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| Keywords | High-affinity IgE receptor (FcepsilonRI) / IgE / Mast Cell / Allergy / aniti-FcepsilonRIAb / IgE-binding inhibition / Humanizaed antibody |
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| Research Abstract |
1.We established a mouse model for IgE-mediated allergy by transplanting a IgE-producing hybridoma under the back skin. We analyzed allergic inflammation induced by IgE in this mouse model and confirmed this allergic inflammation was prevented by soluble human FcepsilonRIalpha ectodomain generated by gene-engineering.
2.We established mouse monoclonal antibodies which recognize human FcepsilonRIalpha ectodomain. Of these antibodies we picked up the one (CRA2) that strongly inhibits IgE binding to FcepsilonRI and tried to humanize this CRA2by CDR-grafting for a specific anti-allergic reagent. The humanized CRA2 (hu CRA2) was as active as the original mouse CRA2 (mo CRA2) and the Fab fragment of hu CRA2 inhibited IgE binding to and histamine release from human basophils. In in vivo experiments with rhesus monkeys, the hu CRA2 exhibited almost no antigenicity and efficiently suppressed allergic inflammation induced by IgE.We are now trying to increase the affinity of this Ab to FcepsilonRI by amino-acid exchange and to develop a specific anti-allergic reagent.
3.We have been performing screening of a variety of substances which can inhibit IgE binding to FcepsilonRI and found out several peomising substances. We are now trying to define the chemical structures of these substances.
4.In order to clarify tertiary structure of the IgE binding site on the FcepsilonRIalpha, we have tried to crystalize the human FcepsilonRIalpha ectodomain and succeeded in generating a partial crystal. Still now we are trying to produce a complete crystal of the FcepsilonRIalpha for X-ray crystallography.
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