Pichurin P, Yan X M, Farilla L, Guo J, Chazenbalk G D, Rapoport B, McLachlan S M
Autoimmune Disease Unit, Cedars-Sinai Research Institute and UCLA School of Medicine, Los Angeles, California 90048, USA.
Endocrinology. 2001 Aug;142(8):3530-6. doi: 10.1210/endo.142.8.8301.
Two approaches have been developed to induce TSH receptor antibodies in mice with properties resembling those in Graves' disease, the Shimojo model of injecting live fibroblasts coexpressing the TSH receptor and major histocompatibility complex antigen Class II, and TSH receptor-DNA vaccination. Thyroid-stimulating antibodies appear to occur less commonly after DNA vaccination, but there has been no direct comparison of these models. We performed a three-way comparison of 1) AKR/N and 2) BALB/c mice vaccinated with TSH receptor-DNA and 3) AKR/N mice injected with fibroblasts expressing the TSH receptor and the major histocompatibility complex antigen class II of AKR/N mice. TSH receptor-DNA vaccinated mice had low or undetectable levels of TSH receptor antibodies determined by ELISA or flow cytometry. Nonspecific binding precluded comparisons with sera from Shimojo mice by these assays. TSH binding inhibition and thyroid-stimulating antibody were undetectable in TSH receptor-DNA vaccinated mice. In Shimojo mice, TSH binding inhibition was positive in approximately 60%, and thyroid-stimulating antibodies were positive in hyperthyroid animals. Unlike the negative antibody data, splenocytes from TSH receptor-vaccinated (but not Shimojo) mice proliferated and produced the Th1 cytokine interferon-gamma in response to TSH receptor antigen. In conclusion, DNA vaccination is less effective at inducing TSH receptor antibodies than the Shimojo approach, but it permits the future characterization of TSH receptor-specific T cells generated without adjuvant.
已经开发出两种方法来在小鼠中诱导出具有类似于格雷夫斯病患者特性的促甲状腺激素（TSH）受体抗体，一种是岛尾模型，即注射共表达TSH受体和主要组织相容性复合体抗原II类的活成纤维细胞，另一种是TSH受体DNA疫苗接种。DNA疫苗接种后促甲状腺激素刺激抗体出现的频率似乎较低，但尚未对这些模型进行直接比较。我们对以下三组进行了三向比较：1）接种TSH受体DNA的AKR/N小鼠和2）BALB/c小鼠，以及3）注射表达AKR/N小鼠TSH受体和主要组织相容性复合体抗原II类的成纤维细胞的AKR/N小鼠。通过酶联免疫吸附测定（ELISA）或流式细胞术测定，接种TSH受体DNA的小鼠的TSH受体抗体水平较低或检测不到。由于非特异性结合，这些检测方法无法将其与岛尾小鼠的血清进行比较。在接种TSH受体DNA的小鼠中未检测到TSH结合抑制和促甲状腺激素刺激抗体。在岛尾小鼠中，约60%的TSH结合抑制呈阳性，甲状腺功能亢进的动物中促甲状腺激素刺激抗体呈阳性。与阴性抗体数据不同的是，接种TSH受体疫苗（而非岛尾模型）的小鼠的脾细胞在接触TSH受体抗原后会增殖并产生Th1细胞因子干扰素-γ。总之，DNA疫苗接种在诱导TSH受体抗体方面比岛尾方法效果差，但它有助于未来对无佐剂情况下产生的TSH受体特异性T细胞进行特性分析。
