ANIMAL MODELS OF HYPERTHYROIDISM

Graves病是一种常见的人类自身免疫性疾病，由刺激TSH受体的自身抗体引起 (TSHR)。目前还没有这种疾病的自发动物模型。我们将使用两种诱导小鼠模型：“裸鼠”。 TSHR-DNA疫苗接种和“Shimojo”方法(注射表达TSHR的成纤维细胞)。有了这些型号，我们将 分析针对TSHR的免疫反应中的以下问题： 1.微生物的影响或缺乏自我耐受性：我们将研究TSHR的效价和功能活性 TSHR-DNA疫苗联合微生物制剂诱导的抗体以及TSHR基因敲除小鼠的抗体 无法获得对TSHR的自我容忍的人。 2.细胞因子和细胞相互作用：野生型小鼠和B细胞敲除小鼠将被用来确定(A) 细胞因子是由TSHR蛋白攻击的脾细胞产生的；(B)CD4+或CD8+T细胞是否参与 (C)如果需要B细胞来诱导TSHR特有的记忆T细胞， 3、TSHR特异性T细胞识别的表位：合成的TSHR多肽将用于确定表位 被T细胞识别：(A)克隆自TSHR-DNA疫苗的BALB/c小鼠；(B)缺乏耐受性的TSHR基因敲除小鼠 对TSHR；(C)易患(DR3)或预防(DQ6)Graves‘病的转HL A基因小鼠。自然而然 还将研究3个TSHR多肽， 3.共刺激信号：共刺激分子的作用将在体外通过使用抗体来阻断 CD40/CD40-1配体和CD28/B7-1/2相互作用；(B)在体内使用CD28基因被破坏的小鼠， CD40、B7-1或B7-2；(C)体内注射抗B7-1抗体(或对照)和TSHR-成纤维细胞(表达 B7-1)。 5.TSHR卵裂和脱落的作用：我们 高度糖基化的A亚单位起着一定的作用 不劈开或脱落的TSHR；(B)检查 甘露糖受体的抗原(如TPO)

Graves' disease is a common human autoimmune disorder caused by autoantibodies that stimulate the TSH receptor (TSHR). There are no spontaneous animal models of the disease. We will use two induced mouse models; "naked" TSHR-DNA vaccination, and the "Shimojo" approach (injecting TSHR-expressing fibroblasts). With these models, we will analyze the following issues in the immune response to the TSHR:- 1. Influence of micro-organisms or lack of self tolerance: We will study the titers and functional activity of TSHR antibodies induced by TSHR-DNA vaccination in combination with microbial products, as well as in TSHR-knockout mice that cannot acquire self tolerance to the TSHR. 2. CYtokine and cellular interactions: Wild type mice and B-cell knockout mice will be used to determine (a) which cytokines are produced by splenocytes challenged with TSHR-protein; (b) whether CD4+ or CD8+ T cells are involved in the response; (c) if B cells are required to induce memory T cells specific for the TSHR, 3, Epitopes recognized by TSHR-specific T cells: Synthetic TSHR peptides will be used to determine the epitopes recognized by T cells (a) cloned from TSHR-DNA vaccinated BALB/c mice; (b) TSHR knockout mice that lack tolerance to the TSHR; (c) mice transgenic for HLA that predispose (DR3) or protect against (DQ6) Graves' disease. Naturally 3rocessed TSHR peptides will also be studied, 3. Co-stimulatory signals: The role of co-stimutatory molecules will be explored (a) in vitro by using antibodies to block CD40/CD40-1igand and CD28/B7-1/2 interactions; (b) in vivo using mice with disrupted genes ("knockouts") for CD28, CD40, B7-1 or B7-2; (c) in vivo by injecting mice with anti-B7-1 (or control) together with TSHR -fibroblasts (that express B7-1). 5. Role of TSHR cleavage and shedding: We heavily glycosylated A subunit plays a role non-cleaving or shedding TSHR; (b) examining antigens (such as TPO) to the mannose receptor