Molecular cloning of eye muscle autoantigen in thyroid-associated ophthalmopathy.

甲状腺相关眼病眼肌自身抗原的分子克隆。

• 批准号: 05670889
• 负责人: HIROMATSU Yuji
• 金额: $ 1.34万
• 依托单位: Kurume University School of Medcine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670889/
• 关键词: Graves'ophthalmopathy; Anti-eye muscle antibody; cDNA cloning; バセドウ病; ophthalmopathy; cDNA cloning

There is considerable evidence that thyroid-associated ophthalmopathy is an autoimmune disorder. However, orbital autoantigen (s) has not been fully elucidated. We have previously reported the presence of autoantibodies against eye muscle membrane 64 kDa proteins. In the present study we attempted to clone and characterized the eye muscle autoantigen (s) in TAO.We screened human skeletal muscle cDNA library with positive sera from patients with severe TAO and human eye muscle cDNA library with the rabbit sera immunized with porcine eye muscle 64 kDa proteins.No positive clone was obtained from human skeletal muscle library. In contrast, 4 positive clones (200-600bp) were obtained from human eye muscle library. One of them (S13) was sequenced by automatic DNA sequencer and identified as an IgE binding protein. Dot blot study showed that 11 out of 20 sera from patients with TAO reacted with the fusion protein, whereas only one of 6 controls did.The relevance of S13 clone in TAO is not known. Recent reports on the presence of IgE in eye muscle tissue from patients with TAO and the exacerbation of Graves' disease after attack of allergic rhinitis support the notion that IgE binding protein may play some role in the development of TAO.

有相当多的证据表明甲状腺相关眼病是一种自身免疫性疾病。然而，眼眶自身抗原尚未完全阐明。我们以前曾报道过存在抗眼肌膜64 kDa蛋白的自身抗体。本研究对TAO眼肌自身抗原进行了克隆和鉴定，用TAO患者的阳性血清筛选人骨骼肌cDNA文库，用猪眼肌64 kDa蛋白免疫的兔血清筛选人眼肌cDNA文库，均未获得阳性克隆。从人眼肌文库中获得4个200- 600 bp的阳性克隆。其中一个（S13）经全自动DNA测序仪测序鉴定为IgE结合蛋白。斑点杂交结果显示，20例TAO患者血清中有11例与融合蛋白反应，而6例正常人血清中只有1例与融合蛋白反应。最近关于TAO患者眼肌组织中IgE的存在以及过敏性鼻炎发作后Graves病加重的报道支持IgE结合蛋白可能在TAO的发展中起一定作用的观点。