ANIMAL MODELS OF HYPERTHYROIDISM

甲亢动物模型

• 批准号: 6700253
• 负责人: Sandra M McLachlan
• 金额: $ 35.96万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-15 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6700253
• 关键词: ANIMAL; MODELS; HYPERTHYROIDISM

DESCRIPTION (provided by applicant): 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 processed TSHR peptides will also be studied, 4. Co-stimulatory signals: The role of co-stimulatory 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 will test the hypothesis that TSHR cleavage followed by shedding of the heavily glycosylated A subunit plays a role in the immune response to the TSHR:- (a) using DNA encoding a non-cleaving or shedding TSHR; (b) examining binding of the heavily glycosylated TSHR A-subunit vs less glycosylated antigens (such as TPO) to the mannose receptor that captures glycosylated organisms for presentation to T cells.

描述（由申请人提供）：Graves病是一种常见的人类自身免疫性疾病，由刺激TSH受体（TSHR）的自身抗体引起。没有自发的疾病动物模型。我们将使用两种诱导的小鼠模型：“裸”TSHR-DNA疫苗接种和“Shimojo”方法（注射表达TSHR-DNA的成纤维细胞）。通过这些模型，我们将分析TSHR免疫反应中的以下问题： 1.微生物的影响或缺乏自身耐受性：我们将研究TSHR-DNA疫苗接种与微生物产品组合诱导的TSHR抗体的滴度和功能活性，以及不能获得对TSHR的自身耐受性的TSHR敲除小鼠。 2.细胞因子和细胞相互作用：将使用野生型小鼠和B细胞敲除小鼠来确定（a）哪些细胞因子由用TSHR-蛋白质激发的脾细胞产生;（B）CD 4+或CD 8 + T细胞是否参与应答;（c）如果需要B细胞来诱导TSHR特异性记忆T细胞， 3.由TSHR特异性T细胞识别的表位：合成的TSHR肽将用于确定由T细胞识别的表位：（a）从TSHR-DNA接种的BALB/c小鼠克隆的T细胞;（B）缺乏对TSHR的耐受性的TSHR敲除小鼠;（c）易感（DR 3）或防止（DQ 6）格雷夫斯病的HLA转基因小鼠。还将研究天然加工的TSHR肽， 4.共刺激信号：共刺激分子的作用将通过以下方法进行探索：（a）体外使用抗体阻断CD 40/CD 40 - 1配体和CD 28/B7-1/2相互作用;（B）体内使用基因被破坏的小鼠（c）通过向小鼠注射抗B7 -1（或对照）以及TSHR -成纤维细胞（表达B7-1）进行体内。 5. TSHR切割和脱落的作用：我们将测试以下假设：TSHR切割，然后是高度糖基化的A亚基的脱落，在对TSHR的免疫应答中起作用：-（a）使用编码非切割或脱落TSHR的DNA;（B）检查高度糖基化的TSHR A亚基相对于较少糖基化的抗原（如TPO）与甘露糖受体的结合，所述甘露糖受体捕获糖基化的生物体以呈递给T细胞。