Hayward Sarah L, Suzuki Kunimasa, Elliott John F
Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta, Canada.
J Immunol Methods. 2007 Jun 30;323(2):114-22. doi: 10.1016/j.jim.2007.04.002. Epub 2007 Apr 30.
Autoimmune (Hashimoto's) thyroiditis is a chronic inflammatory disease which affects >3% of the population and shows an increasing prevalence with increasing age. Anti-thyroid autoantibodies, particularly against thyroperoxidase (also known as thyroid peroxidase or TPO), occur commonly in humans with autoimmune thyroid disease, and assays for anti-TPO autoantibodies are used in clinical diagnosis. In contrast anti-TPO autoantibodies have not been observed in classical mouse models of autoimmune thyroiditis, except in cases where mice were deliberately immunized with TPO. In the past, detection of anti-TPO autoantibodies in mice has relied on an indirect immunofluorescence assay (iIFA) which screens for thyroid follicle membrane staining in frozen sections of mouse thyroid glands. Since recent transgenic mouse models of autoimmune thyroiditis spontaneously develop anti-TPO autoantibodies, an assay other than serial dilution and iIFA would be useful to detect and quantify these autoantibodies. In this paper we describe such an assay, based on the capacity of autoimmune mouse sera to bind to the extracellular domain of mouse TPO which was produced in a radioactively labeled form using a coupled in vitro transcription/translation system. The same approach, using human TPO, could provide a highly sensitive method to detect anti-TPO autoantibodies in humans.
自身免疫性(桥本氏)甲状腺炎是一种慢性炎症性疾病,影响着超过3%的人群,且患病率随年龄增长而上升。抗甲状腺自身抗体,尤其是抗甲状腺过氧化物酶(也称为甲状腺过氧化物酶或TPO)的抗体,在患有自身免疫性甲状腺疾病的人群中普遍存在,抗TPO自身抗体检测用于临床诊断。相比之下,在自身免疫性甲状腺炎的经典小鼠模型中未观察到抗TPO自身抗体,除非小鼠被特意用TPO免疫。过去,小鼠中抗TPO自身抗体的检测依赖于间接免疫荧光分析(iIFA),该方法通过检测小鼠甲状腺冰冻切片中的甲状腺滤泡膜染色来进行筛选。由于最近的自身免疫性甲状腺炎转基因小鼠模型会自发产生抗TPO自身抗体,因此除了系列稀释和iIFA之外的检测方法对于检测和定量这些自身抗体将很有用。在本文中,我们描述了这样一种检测方法,该方法基于自身免疫小鼠血清与小鼠TPO细胞外结构域结合的能力,小鼠TPO细胞外结构域是使用耦合的体外转录/翻译系统以放射性标记形式产生的。使用人TPO的相同方法可以提供一种高灵敏度的方法来检测人血清中的抗TPO自身抗体。