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

1.CD_4~+CD25~+调节性T细胞(Treg)抑制疾病的发生：抗CD25抗体去除Treg后，30%耐药的C57BL/6小鼠出现Graves‘甲亢，易感BALB/c小鼠血清T_4水平升高200%。这种效果归因于刺激性抗体效价的降低和阻断抗体效价的增加。此外，在对幼小鼠的移植实验中，来自Graves‘s小鼠的脾细胞诱导了少量的抗TSHR抗体，但CD25耗竭的脾细胞产生了低水平但显著的抗体水平。调节性细胞因子抑制疾病进展：构建表达IL-10或TGF-β的腺病毒(两者均为调节性细胞因子)，并与编码TSHR的腺病毒(Ad-TSHR)一起接种于小鼠。仅Ad-IL-10能显著抑制甲亢。通过凋亡诱导Fas配体(FasL)抑制疾病进展：构建表达FasL的腺病毒，并用GMCSF和IL-4联合Ad-TSHR感染骨髓来源的树突状细胞(DC)。这些树突状细胞抑制肌肉注射Ad-TSHR诱导的抗TSHR免疫应答和Graves病的发生。TSHR特异的T细胞系目前正在由TSHR蛋白和三种多肽建立：在Recall实验中诱导IFNGamma释放的多肽。将研究单个克隆产生细胞因子(S)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.