| Project/Area Number |
08671150
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
内分泌・代謝学
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
AKAMIZU Takashi Kyoto Univ. Graduate.Sch.of Med.Assist., 医学研究科, 助手 (20231813)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUDA Fumihiko Kyoto Univ. Graduate.Sch.of Med.Assist., 医学研究科, 助手 (50212220)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1996: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Graves'disease, / TSH receptor, / Ig genes / autoantibody, / autoimmune thyroid disease |
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| Research Abstract |
Anti-thyrotropin (TSH) receptor autoantibodies (TSHRAbs) are known to be involved in Geaves' disease. To elucidate the molecular mechanism of the pathogenesis of Graves' disease, we previously isolated and reconstituted the immunoglobulin genes of two B cell clones prcducing a monoclonal thyroid stimulating antibody (TSAb), a stimulating type of TSHRAb, obtainedfrom patients with Graves' disease. In the present study, we produced a large amount of recombinant monoclonal TSAbs in eukariotic cells using these genes and applied them to severral further investigations. First, we tried to identify their epitopes in the TSHR by using a panel of mutants of the extracellular domain of TSHR.Both antibodies abrogated or reduced TSAb activities in mutants of the N-terminal but not C-terminal region of the extracellular domain of TSHR,while TSH had opposite effect. Secondly, purified antibodies were radiolabeled and tested for binding to cells expressing high levels of TSHR.Although their affinities were lower than that of TSH,their bindings were not displaced by TSH.The antibody bindings were not mutually competitive, either. The finding indicates that these antibodies interact with the multiple conformational arrays of receptor determinants in the N-terminus and transduce a signal through binding sites different from TSH.
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