相关实验视频
Updated: Jun 15, 2025

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Generation of a Mouse Spontaneous Autoimmune Thyroiditis Model
Published on: March 17, 2023
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具有甲状腺刺激活性的甲状腺蛋白受体单克隆抗体激活NF-κB通路,诱导化学因表达
1Department of Endocrine, The second hospital & clinical medical school,Lanzhou university, Lanzhou, Gansu, China.
Journal of cellular and molecular medicine
|June 11, 2025
概括
研究人员开发了三种小鼠抗thyrotropin受体 (TSHR) 单克隆抗体 (mAbs),以了解格雷夫斯病. 这些mAbs刺激TSHR,激活NF-κB信号,并诱导化基因,提供了对自身免疫甲状腺疾病机制的见解.
科学领域:
- 免疫学 免疫学 免疫学
- 内分泌学 在内分泌学.
- 分子生物学分子生物学
背景情况:
- 甲状腺素受体 (TSHR) 的A子单元通过刺激性自身抗体参与了格雷夫斯病 (GD) 的发病.
- 病理性TSHR自身抗体反应背后的精确分子机制及其对免疫信号传递的影响仍然不完全理解.
- TSHR自身抗体可能会激活信号通路,改变化学激素配置,并加剧GD中的免疫失调.
研究的目的:
- 产生和表征小鼠抗TSHR单克隆抗体 (mAbs),以阐明TSHR自身抗体活性在格雷夫斯病中的分子基础.
- 为了研究这些mAbs对TSHR刺激,免疫细胞信号通路和人类甲状腺细胞中化学激素表达的作用.
主要方法:
- 使用昆虫百科病毒表达的TSHR 289蛋白质的生产.
- 用腺病毒表达的TSHR289对小鼠进行免疫接种,并通过混合瘤技术生成抗TSHRmAbs (1A4,7C3,22B1).
- 测试包括流式细胞计,ELISA,RT-qPCR,西部斑点测试以评估mAb活性,TSHR结合,化学激素表达 (CCL2,CCL19,CCL5) 和NF-κB通路激活.
主要成果:
- 三只小鼠的抗TSHR mAbs (1A4,7C3,22B1) 已成功生成并证明在纳米克度下具有TSHR刺激活性.
- 这些mAbs竞争性地抑制了Graves病患者血清TRAb与TSHR的结合.
- mAb刺激通过激活人类甲状腺细胞中的正规和非正规NF-κB信号来诱导化学因子CCL2,CCL19和CCL5的表达. 注射7C3增加了血清甲状腺素,导致甲状腺增生.
结论:
- 开发的抗TSHRmAbs为特征TSHR自身抗体介导病理学的分子机制提供了有价值的工具.
- 刺激性TSHR自身抗体激活NF-κB信号通路,导致诱导化学因子,这些化学因子有助于格雷夫斯病的自身免疫反应.
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