Low-dose immunization with adenovirus expressing the thyroid-stimulating hormone receptor A-subunit deviates the antibody response toward that of autoantibodies in human Graves' disease
Low-dose immunization with adenovirus expressing the thyroid-stimulating hormone receptor A-subunit deviates the antibody response toward that of autoantibodies in human Graves' disease
复制标题
DOI：
10.1210/en.2003-1134
复制
发表时间：
2004-01-01
期刊：
影响因子：
4.8
通讯作者：
Rapoport, B
中科院分区：
文献类型：
--
作者：
Chen, CR;Pichurin, P;Rapoport, B
使用表达 TSH 受体 (TSHR) 的腺病毒进行免疫会导致 25-50% 的小鼠出现甲状腺功能亢进症。更有效的是使用 TSHR A 亚基腺病毒进行免疫（65-84% 甲状腺功能亢进）。然而，这些小鼠的 TSHR 抗体特征并不能准确模拟典型格雷夫斯患者的自身抗体特征，具有明显的 TSH 阻断抗体反应。我们假设这种次优抗体反应是由于标准剂量的 TSHR 腺病毒提供了太大的免疫刺激所致。为了检验这一假设，我们比较了用通常数量（10（11））和少得多的病毒颗粒（10（9）和10（7））免疫的BALB/c小鼠。无论病毒剂量如何，小鼠患甲状腺功能亢进的比例相似（68-80%）。然后我们检查了每组中 TSHR 抗体的定性。第二次免疫后，所有小鼠的血清均具有 TSH 结合抑制（TBI）活性，TBI 值与病毒剂量成正比。第三次注射后，所有组的 TBI 值都接近最大。值得注意的是，为了证实我们的假设，用逐渐降低的病毒剂量进行免疫产生的 TSHR 抗体接近人类格雷夫斯病自身抗体的特征，如下：1) ELISA 上的 TSHR 抗体滴度较低，2) 较低的 TSH 阻断抗体活性，但不降低甲状腺刺激抗体活性。总之，用表达游离TSHR A亚基的腺病毒进行低剂量免疫提供了甲亢患病率高的诱导动物模型，并且TSHR抗体更类似于格雷夫斯病中的自身抗体。
Immunization with adenovirus expressing the TSH receptor (TSHR) induces hyperthyroidism in 25-50% of mice. Even more effective is immunization with a TSHR A-subunit adenovirus (65-84% hyperthyroidism). Nevertheless, TSHR antibody characteristics in these mice do not mimic accurately those of autoantibodies in typical Graves' patients, with a marked TSH-blocking antibody response. We hypothesized that this suboptimal antibody response was consequent to the standard dose of TSHR-adenovirus providing too great an immune stimulus. To test this hypothesis, we compared BALB/c mice immunized with the usual number (10(11)) and with far fewer viral particles (10(9) and 10(7)). Regardless of viral dose, hyperthyroidism developed in a similar proportion (68-80%) of mice. We then examined the qualitative nature of TSHR antibodies in each group. Sera from all mice had TSH binding-inhibitory (TBI) activity after the second immunization, with TBI values in proportion to the viral dose. After the third injection, all groups had near-maximal TBI values. Remarkably, in confirmation of our hypothesis, immunization with progressively lower viral doses generated TSHR antibodies approaching the characteristics of autoantibodies in human Graves' disease as follows: 1) lower TSHR antibody titers on ELISA and 2) lower TSH-blocking antibody activity without decrease in thyroid-stimulating antibody activity. In summary, low-dose immunization with adenovirus expressing the free TSHR A-subunit provides an induced animal model with a high prevalence of hyperthyroidism as well as TSHR antibodies more closely resembling autoantibodies in Graves' disease.