| Project/Area Number |
15590354
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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 |
Experimental pathology
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| Research Institution | Hoshi University Faculty of Pharmaceutical Science |
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Principal Investigator |
YOSHIDA Tadashi Hoshi University Faculty of Pharmaceutical Science, Dept of Pathophysiology, Professor, 薬学部, 教授 (00182767)
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| Co-Investigator(Kenkyū-buntansha) |
MUTO Akihiro Hoshi University Faculty of Pharmaceutical Science, Dept of Pathophysiology, Assistant Professor, 薬学部, 講師 (90239468)
KANEDA Toshio Hoshi University Faculty of Pharmaceutical Science, Dept of Pathophysiology, Instructor, 薬学部, 助手 (70339521)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | TSH receptor / Anti-TSH receptor antibody / Graves' disease / Animal model / Transgenic mice / エレクトロポレーション |
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| Research Abstract |
In Graves' disease, autoantibody directed against TSH receptor (TSHR) mimic the action of TSH and are therefore called thyroid-stimulating antibody (TSAb). TSAb caused overstimulation of the thyroid gland and hyperthyroidism. Animal models are very useful tools to clarify the pathophysiology of autoimmune thyroid disease.
Recently, several animal models of Graves' disease have been developed, some of which employed genetic immunization. A transgenic mouse line that expresses Cre recombinase under control of the human keratinocyte gene promoter was established. The activity and specificity of the keratin-driven Cre recombinase were examined by using Northern blotting. This keratin-Cre transgenic mouse was specifically delete loxP-inserted lac Z gene in keratinocytes and induced TSHR gene during development and/or in adult keratinocytes. However, we could not detect the activities of anti-TSHR antibody and high levels of serum thyroid hormone.
We then used the technique of electroporation (EP) for establishment of a new model of Graves' disease in expectation of greatly enhanced hTSHR expression in vivo. Among non-viral techniques for gene transfer by plasmid vector in vivo, the method with direct injection of plasmid DNA into muscles is simple, while is restricted by the relatively low expression levels of the transferred gene.
Moreover, we established hTSHR expressing stable clone (AHK12), having functional response to bTSH, for the detection of biological activity of autoantibody and ELISA for the detection of autoantibody against rhTSHR-289His, corresponding to the extracellular A subunit TSHR.
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