Monoclonal Thyroid Stimulating Antibodies

促甲状腺激素（TSH）受体（TSHR）是Graves病的主要抗原和甲状腺刺激的靶点 TSHR抗体（TSHR-Ab）在这些患者的血清中发现。TSHR-Ab的特点是其长期作用 并且最初被称为长效甲状腺刺激剂（LATS）。在本申请中，我们寻求继续我们的 成功克隆和鉴定TSH受体的单克隆抗体，这导致了第一个 具有刺激活性的单克隆抗体。特别地，单克隆抗体MS-1已经显示出有效的TSHR 当用稳定表达hTSHR的CHO细胞测试时，刺激活性明显降低至2ng/ml的IgG。 本次竞争性更新的具体目标是：目标1：确定额外的单克隆甲状腺刺激 抗体的我们将首先继续建立在我们的成功，发展扩大小组的单克隆抗体 促甲状腺激素受体具有甲状腺刺激活性。目的2：确定TSHR-AB表位。我们将描述 衍生的TSH受体抗体的线性和非线性抗原表位，以确定主要的免疫原性 '区（MIRs）。目的3：定义抗体介导的TSHR翻译后调节 修改.我们将在CHO细胞中共表达TSHRcpp和TSHRmycconstructs，并检查TSHRcpp和TSHRmycconstructs是否占据了CHO细胞中的TSHRcpp和TSHRmycconstructs。 通过TSHR-Abs对受体分子的作用将调节FRET指数，正如我们在TSH配体中所看到的那样。另外我们有 最近开发了一种独特的测定TSHR裂解，并将检查TSHR-Ab对 受体裂解，目的是深入了解这种裂解在甲状腺自身免疫中的功能作用。 目的4：研究TSHR抗体的生物学功能及诱导的信号转导。我们会研究 TSHR-Ab在其生长刺激/抑制潜力及其活化/灭活方面特异性 分化的甲状腺细胞功能。 与TSHR-Ab具有相似特征的抗TSH抗体的分离和表征， 格雷夫斯病患者的血清将为他们的作用机制提供新的见解，并提供新的 甲状腺功能紊乱的治疗方法

The thyrotropin (TSH) receptor (TSHR) is the primary antigen of Graves' disease and the target for thyroid stimulating 'TSHR antibodies (TSHR-Ab)found in the serum of such patients. TSHR-Ab are characterized by their prolonged action and were originally called Long Acting Thyroid Stimulators (LATS). In this application, we seek to continue our successful cloning and characterization of monoclonal antibodies to the TSH receptor which have resulted in the first monoclonal antibody with stimulating activity. In particular, monoclonal antibody MS-1 has shown potent TSHR stimulating activity when tested with CHO cells stably expressing hTSHR with activity evident down to 2ng/ml ofIgG. The specific aims of this competitive renewal are: Aim 1: Identify additional monoclonal thyroid-stimulating antibodies. We will first continue to build on our success by developing an expanded panel of monoclonal antibodies to the TSH receptor with thyroid stimulating activity. Aim 2: Define TSHR-AB epitopes. We will characterize the linear and non-linear antigenic epitopes for the derived TSH receptor antibodies to define the major immunogenic 'regions (MIRs) on the TSH receptor. Aim 3: Defining antibody-mediated regulation of TSHR post-translational modifications. We will co-express TSHRcpp TSHRmycconstructs in CHO cells and examine whether occupation of the receptor molecule by TSHR-Abs will modulate the FRET index as we have seen for TSH ligand. In addition, we have recently developed a unique assay for measuring TSHR cleavage and will examine the influence of TSHR-Abs on receptor cleavage with the aim of gaining insight into the functional role of such cleavage in thyroid autoimmunity. Aim 4: Examine the biologic function and signal transduction induced by TSHR-Abs. We will examine the specificity of TSHR-Abs with regard to their growth stimulating/inhibiting potential and their activarion/inacrivation of differentiated thyroid cell function. The isolation and characterization of antibodies to the TSH with similar characteristics to the TSHR-Abs found in the serum of Graves' disease patients will provide major new insight into their mechanisms of action and offer novel approaches to the control of thyroid dysfunction.