Study on the immunological mechanism of anti-TSH receptor antibody production by using transgenic mice

转基因小鼠产生抗TSH受体抗体的免疫学机制研究

• 批准号: 15590354
• 负责人: YOSHIDA Tadashi
• 金额: $ 2.24万
• 依托单位: Hoshi University Faculty of Pharmaceutical Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590354/
• 关键词: TSH receptor; Anti-TSH receptor antibody; Graves' disease; Animal model; Transgenic mice; エレクトロポレーション

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.

在Graves病中，针对TSH受体的自身抗体(TSHR)模拟TSH的作用，因此被称为甲状腺刺激抗体(TSAb)。TSAb可引起甲状腺过度刺激和甲状腺机能亢进。动物模型是阐明自身免疫性甲状腺疾病的病理生理机制的非常有用的工具。最近，人们已经建立了几种Graves病的动物模型，其中一些采用了基因免疫。建立了人角质形成细胞基因启动子控制下表达Cre重组酶的转基因小鼠系。Northern印迹法检测角蛋白驱动的Cre重组酶的活性和特异性。该角蛋白-Cre转基因小鼠在角质形成细胞中特异性地删除了loxP插入的lac Z基因，并在发育过程中和/或在成年角质形成细胞中诱导了TSHR基因。然而，我们无法检测到抗TSHR抗体的活性和血清中高水平的甲状腺激素，因此我们利用电穿孔(EP)技术建立了一种新的Graves病模型，以期在体内大大增强hTSHR的表达。在非病毒载体体内基因转移技术中，直接肌肉注射法简单，但受基因表达水平较低的限制，我们建立了对bTSH有功能反应的hTSHR稳定表达克隆(AHK12)，用于检测自身抗体的生物学活性；建立了检测针对胞外A亚单位TSHR的自身抗体的ELISA方法。

项目成果

MTX療法におけるアフター性口内炎に対するマレイン酸イルソグラジンの効果、リウマチ科

马来酸伊索拉定对MTX治疗后遗症性口腔炎的影响，风湿科

• 发表时间: 2004
• 期刊: リウマチ科 32
• 作者: 吉田正;Yoshida T.;吉田 正;吉田正;吉田正
• 通讯作者: 吉田正

骨・関節の疾患、薬と疾病、II

骨关节疾病、药物与疾病II

• 发表时间: 2005
• 作者: 吉田正;Yoshida T.;吉田 正;吉田正;吉田正;Yoshida T.;吉田 正;Yoshida T. et al.;Yoshida T. et al.;吉田正;Yoshida T. et al.;Yoshida T.;吉田正;吉田正;吉田正;吉田 正
• 通讯作者: 吉田 正

ファーマシューティカルノート

医药笔记

• 发表时间: 2003
• 作者: 吉田正;Yoshida T.;吉田 正;吉田正;吉田正;Yoshida T.;吉田 正;Yoshida T. et al.;Yoshida T. et al.;吉田正;Yoshida T. et al.;Yoshida T.;吉田正;吉田正;吉田正;吉田 正;吉田 正;吉田正;吉田 正;吉田正;吉田正
• 通讯作者: 吉田正

Prospective study of high-dose intravenous immunoglobulin for the treatment of steroid-resistant polymyositis and dermatomyositis.

大剂量静脉注射免疫球蛋白治疗类固醇抵抗性多发性肌炎和皮肌炎的前瞻性研究。

• 发表时间: 2003
• 期刊: Mod Rheumatol 13
• 作者: 吉田正;Yoshida T.;吉田 正;吉田正;吉田正;Yoshida T.;吉田 正;Yoshida T. et al.;Yoshida T. et al.
• 通讯作者: Yoshida T. et al.

慢性疾患薬物療法のツボ : 関節リウマチ

慢性病药物治疗要点：类风湿性关节炎

• 发表时间: 2005
• 作者: 吉田正;Yoshida T.;吉田 正;吉田正;吉田正;Yoshida T.;吉田 正;Yoshida T. et al.;Yoshida T. et al.;吉田正;Yoshida T. et al.;Yoshida T.;吉田正;吉田正;吉田正
• 通讯作者: 吉田正