Studies on the pathogenesis of Graves' disease and new treatment using our recently established mouse model.

1. CD 4 ^+ CD 25 ^+调节性T细胞（Treg）对疾病发展的抑制作用：抗CD 25抗体耗竭Treg可诱导30%的抗性C57 BL/6小鼠出现Graves'甲亢，并使敏感的BALB/c小鼠血清T_4水平升高200%。这种效应归因于刺激性抗体滴度降低和阻断性抗体滴度增加。此外，在向未处理野生型小鼠的转移实验中，来自Graves小鼠的脾细胞诱导很少的抗TSHR抗体，但是CD 25去除的脾细胞产生低但显著水平的抗体。通过调节性细胞因子抑制疾病发展：构建表达IL-10或TGF-β（两者都是调节性细胞因子）的腺病毒，并与编码TSHR的腺病毒（Ad-TSHR）一起给予小鼠。只有Ad-IL-10能显著抑制甲亢.通过诱导凋亡的Fas配体（FasL）抑制疾病的发展：构建表达FasL的腺病毒，并使用GMCSF和IL-4与Ad-TSHR一起感染来自骨髓细胞的树突状细胞（DC）。这些树突状细胞可抑制肌内注射Ad-TSHR诱导的抗TSHR免疫应答和Graves病的发生。TSHR特异性T细胞系目前正在通过TSHR蛋白和三种肽建立，这些肽在回忆试验中诱导IFN γ释放。将研究单个克隆产生细胞因子和诱导甲状腺功能亢进的能力以及T细胞受体的氨基酸序列。

1. Suppression of disease development by CD4^+CD25^+ regulatory T cells (Treg) : Depletion of Treg by anti-CD25 antibody induced Graves' hyperthyroidism in 30 % of resistant C57BL/6 mice, and increased serum T_4 levels by 200 % in susceptible BALB/c mice. This effect was attributed to decreased stimulatory antibody titers and increased blocking antibody titers. Furthermore, in transfer experiments to naive wt mice, splenocytes from Graves' mice induced little anti-TSHR antibody, but CD25-depleted splenocytes did low but significant levels of antibody.2. Suppression of disease development by regulatory cytokines : adenovirus expressing IL-10 or TGF-beta (both are regulatory cytokines) were constructed, and administered to mice together with adenovirus coding the TSHR (Ad-TSHR). Only Ad-IL-10 significantly inhibited hyperthyroidism.3. Suppression of disease development by apoptosis-inducing Fas ligand (FasL) : Adenovirus expressing FasL was constructed, and infected into dendritic cells (DCs) derived from bone-marrow cells by using GMCSF and IL-4 together with Ad-TSHR. These DCs inhibited anti-TSHR immune response and development of Graves' disease induced by intramuscular injection of Ad-TSHR.4. TSHR-specific T cell lines are now being established by the TSHR protein and three : peptides which induced IFNgamma release in recall assay. Abilities of individual clones to produce cytokine(s) and to induce hyperthyroidism as well as amino acid sequences of T cell receptors will be studied.