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病是由刺激TSH受体（TSHR）的自身抗体引起的常见人自身免疫性疾病。没有这种疾病的自发动物模型。我们将使用两个诱导的鼠标模型。 “ Naked” TSHR-DNA疫苗接种和“ Shimojo”方法（注射TSHR表达成纤维细胞）。通过这些模型，我们将在对TSHR的免疫反应中分析以下问题： - 1。微生物的影响或缺乏自耐力：我们将研究由TSHR-DNA疫苗接种与微生物产物结合使用的TSHR抗体的滴度和功能活性，以及​​在TSHR-KNOCKOUT小鼠中无法获得对TSHR的自耐力。 2。细胞因子和细胞相互作用：将使用野生型小鼠和B细胞基因敲除小鼠来确定（a）哪些细胞因子是由TSHR-蛋白质挑战的脾细胞产生的； （b）CD4+或CD8+ T细胞是否参与了响应； （c）如果需要B细胞诱导特定于TSHR的记忆T细胞，则 3。由TSHR特异性T细胞识别的表位：合成TSHR肽将用于确定由TSHR-DNA疫苗接种BALB/C小鼠克隆的T细胞（a）识别的表位； （b）缺乏对TSHR耐受性的TSHR敲除小鼠； （c）易感性（DR3）或预防（DQ6）坟墓疾病的HLA的小鼠。自然处理的TSHR肽也将被研究， 4。共刺激信号：通过使用抗体阻断CD40/CD40-1IGAND和CD28/B7-1/2相互作用，将在体外探索共刺激分子的作用； （b）用于CD28，CD40，B7-1或B7-2的小鼠使用小鼠的体内； （c）在体内通过将抗B7-1（或对照）与TSHR-纤维细胞（表达B7-1）一起注射抗B7-1（或对照）。 5。TSHR裂解和脱落的作用：我们将测试以下假设：TSHR裂解，然后脱落，重糖基化的亚基在对TSHR的免疫反应中起作用： - （a）使用编码非分解或脱落TSHR的DNA； （b）检查重糖基化的TSHR A-亚基与糖基化抗原（例如TPO）与甘露糖受体的结合，该抗原与甘露糖受体捕获糖基化生物，以呈现在T细胞中。