Immunization of mice with a newly identified thyroid-stimulating hormone receptor splice variant induces Graves'-like disease.
Immunization of mice with a newly identified thyroid-stimulating hormone receptor splice variant induces Graves'-like disease.
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DOI：
10.1016/j.jaut.2013.02.004
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发表时间：
2013-06
影响因子：
12.8
通讯作者：
T. Endo;Teturo Kobayashi
中科院分区：
文献类型：
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作者：
T. Endo;Teturo Kobayashi
从小鼠甲状腺中克隆了促甲状腺激素受体（TSHR） cDNA。该cDNA序列显示其编码一个739个氨基酸的TSHR剪接变体，缺少外显子5 （TSHR739）。在成年小鼠甲状腺样本中，TSHR739mRNA的表达量约为全长TSHR (TSHR764) mRNA表达量的10%。ecfp标记的tshr739整合到质膜中，但缺乏TSH结合活性，并且对TSH不产生cAMP。然而，来自Graves病患者的促甲状腺抗体刺激了表达TSHR739的HEK293细胞中cAMP的产生。定量PCR结果显示，tshr739转录物水平在胎鼠甲状腺样品中较低，但随着小鼠出生和生长，tshr739转录物水平升高。我们采用质粒注射联合骨骼肌电穿孔，用TSHR739、TSHR764或对照质粒免疫BALB/c小鼠；tshr739引起甲状腺肿、高125i摄取活性、甲状腺毒症和促甲状腺抗体的产生，但TSHR764或对照组没有。这些结果表明，免疫自体TSHR抗原TSHR739可诱导小鼠Graves样疾病，TSHR739是自身免疫性甲状腺疾病的候选自身抗原。
We have cloned a thyroid-stimulating hormone receptor (TSHR) cDNA from mouse thyroid glands. The sequence of this cDNA indicated that it encoded a 739 amino acid TSHR splice variant that lacked exon 5 (TSHR739). In thyroid gland samples from adult mice, the amount of TSHR739mRNA was about 10% of the amount of full-length TSHR (TSHR764) mRNA. A eCFP-tagged TSHR739integrated into plasma membrane, but lacked TSH binding activity and it did not produce cAMP in response to TSH. However, thyroid-stimulating antibodies from patients with Graves' disease stimulated cAMP production in HEK293 cells that expressed TSHR739. Quantitative PCR revealed that TSHR739transcript levels were low in the fetal mouse thyroid samples, but TSHR739transcript levels increased after birth and as the mice grew. We used plasmid injection combined with electroporation into skeletal muscles to immunize BALB/c mice with TSHR739, TSHR764,, or control plasmid; TSHR739caused goiters, high125I uptake activity, thyrotoxicosis, and production of thyroid-stimulating antibodies, but TSHR764, or control did not. These results indicated that immunization with an autologous TSHR antigen, TSHR739, induced Graves'-like disease in mice, and that TSHR739is a candidate autoantigen in autoimmune thyroid disease.