Kong Yi-Chi M
Wayne State University School of Medicine, Detroit, Michigan.
Curr Protoc Immunol. 2007 Aug;Chapter 15:15.7.1-15.7.21. doi: 10.1002/0471142735.im1507s78.
Experimental autoimmune thyroiditis (EAT) in mice is an excellent model for Hashimoto's thyroiditis (HT). It is induced with thyroglobulin (Tg), a known thyroid autoantigen that is common to both mouse and human and for which several conserved, thyroiditogenic epitopes have been identified. This unit describes induction and evaluation of EAT using thyroid histology and in vitro proliferative response assays. An ELISA is presented to detect the level of antibody to mouse thyroglobulin (MTg). To induce EAT, either bacterial lipopolysaccharide (LPS) or supplemented complete Freund's adjuvant (CFA) can be used as adjuvant. A support protocol for preparing MTg is included. The T cell proliferation assay can be used to examine the antigenicity of synthetic peptides derived from MTg or heterologous Tg. EAT can be adoptively transferred utilizing cells that have been expanded in vitro, as described. A protocol is provided for inducing tolerance using deaggregated MTg; induction of tolerance requires larger amounts of MTg but efficiently suppresses EAT development. Also included is a protocol to demonstrate the role of regulatory T cells in mediating tolerance. A protocol to delineate HLA association with HT is illustrated using HLA class II transgenic mice.
小鼠实验性自身免疫性甲状腺炎（EAT）是桥本甲状腺炎（HT）的优良模型。它由甲状腺球蛋白（Tg）诱导产生，Tg是一种已知的甲状腺自身抗原，在小鼠和人类中都很常见，并且已鉴定出几个保守的致甲状腺炎表位。本单元描述了使用甲状腺组织学和体外增殖反应测定法诱导和评估EAT的方法。还介绍了一种用于检测抗小鼠甲状腺球蛋白（MTg）抗体水平的ELISA方法。为诱导EAT，可使用细菌脂多糖（LPS）或补充的完全弗氏佐剂（CFA）作为佐剂。其中包括制备MTg的支持方案。T细胞增殖测定可用于检测源自MTg或异源Tg的合成肽的抗原性。如所述，EAT可利用体外扩增的细胞进行过继转移。提供了使用解聚的MTg诱导耐受的方案；诱导耐受需要大量的MTg，但能有效抑制EAT的发展。其中还包括一个证明调节性T细胞在介导耐受中作用的方案。使用HLA II类转基因小鼠说明了一种描绘HLA与HT关联的方案。
