STUDIES ON THE MOLECULAR BIOLOGY OF TSH RECEPTOR AND ON THE PATHOGENETIC ROLE OF ANTI-TSH RECEPTOR ANTIBODIES IN GRAVES' DISEASE

TSH受体分子生物学及抗TSH受体抗体在Graves病中致病作用的研究

• 批准号: 62570500
• 负责人: NOBUAKI Kuzuya
• 金额: $ 1.28万
• 依托单位: UNIVERSITY OF TSUKUBA
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62570500/
• 关键词: TSH receptor; Anti-receptor antibodies; Western blotting; Graves' disease; Idiopathic myxedema; Monoclonal antibodies; サイトカイン; 甲状腺刺激抗体; 甲状腺刺激抑制抗体; Western blotting; HLA classII抗原; r-インターフェロン; 萎縮性甲状腺炎

Receptors on cell membranes generally play significant roles on the transduction of hormonal signals into cells, and abnormalities in hormone receptors closely relate to pathophysiology of several diseases. It is suggested that anti-TSH receptor antibodies play a causative role in Graves' disease, however, the molecular structure and functions of the TSH receptor have not been well clarified, making the pathogenetic significance of anti-TSH receptor antibodies still obscure. The aims of this study were as follows; 1) study on the molecular weight and the biochemical natures of the TSH receptor, 2) production of monoclonal antibodies to the TSH receptor and also of animal models of graves' disease, and 3) analysis of sequences of the cDNA of TSH receptor.We have shown that the TSH receptor is a glycoprotein, and that not only the stimulatory IgGs in patients with Graves' disease but also the blocking IgGs from patients with idiopathic myxedema react with the glycoprotein TSH receptor as well. After further purification using Graves' IgG-coupled sepharose, proteins were analyzed by Western blotting. The same 67KDa protein was stained by both the stimulatory and blocking type of IgGs suggesting that this antigen might relate to the TSH receptor.The preliminary results showed that some monoclonal antibodies against partially purified TSH receptor displaced the 125i-TSH binding to solubilized thyroid membranes stronger than others. Furthermore, we presented that the -interferon administered mice produced antithyroid antibodies in vivo, which advanced one step for making a new model of autoimmune thyroid diseases.The purpose no.3) remained undisolved in this study.

细胞膜上的受体在激素信号的传递中起着重要作用，激素受体的异常与多种疾病的病理生理密切相关。目前认为抗TSH受体抗体在Graves病的发病中起一定作用，但由于TSH受体的分子结构和功能尚未完全阐明，抗TSH受体抗体在Graves病发病中的意义仍不清楚。本研究的目的如下：1）TSH受体分子量和生化性质的研究; 2）TSH受体单克隆抗体的制备和Graves病动物模型的建立; 3）TSH受体cDNA序列的分析。不仅Graves病患者的刺激性IgG，而且特发性粘液水肿患者的阻断性IgG也与糖蛋白TSH受体反应。使用Graves IgG偶联的琼脂糖凝胶进一步纯化后，通过Western印迹分析蛋白质。刺激型和阻断型IgG均能染色同一67 KDa蛋白，提示该抗原可能与TSH受体有关，初步结果表明，某些抗部分纯化的TSH受体的单克隆抗体能较强地取代~（125）i-TSH与可溶性甲状腺膜的结合。此外，我们还发现，给予干扰素的小鼠体内产生了抗甲状腺抗体，这为建立一种新的自身免疫性甲状腺疾病模型又前进了一步。

项目成果

Y.Kawakami: The Thyroid. 71-74 (1988)

Y.Kawakami：甲状腺。

Nobuaki Kuzuya; Yasushi Kawakami; Kamejiro Yamashita: "Identification of Antigens of Abnormal Thyroid Stimulators by Western Blotting" The Thyroid 1988 (Proceedings of the International Thyroid Symposium). 185-189 (1988)

葛弥伸明；