Analyses of antigen-specific T cells in systemic types of autoimmune disorders.

抗原特异性T细胞参与系统性自身免疫性疾病的发病机制仍有争议。从本质上讲，活化的抗原特异性T细胞应该在淋巴细胞群中形成累积克隆。因此，检测这些T细胞以了解抗原特异性免疫应答在疾病中的贡献是至关重要的。然而，可用于分析T细胞克隆性的方法仍然有限。在这方面，我们已经建立了一种新的方法，使用相结合的逆转录酶-聚合酶链反应的T细胞受体β链转录本和单链构象多态性（SSCP）。利用这种方法，可以监测抗原刺激过程中T细胞克隆反应的动态变化。对几种自身免疫性疾病，包括类风湿性关节炎、系统性红斑狼疮及其动物模型的分析揭示了抗原特异性T细胞免疫应答的参与。此外，利用SSCP凝胶中条带的可重复迁移性，我们现在能够比较不同样品中积累的T细胞克隆的身份，而不需要对每个克隆进行核苷酸测序。因此，这些信息可以阐明患者中均匀或稳定的免疫反应的发生，并且还表明这些反应在发病机制中起重要作用。开发这个系统，我们也建立了一个新的策略，以确定这些在体内积累的T细胞克隆的抗原特异性。

The involvement of antigen-specific T cells in the pathogenesis of systemic types of autoimmune disorders is still controversial. Essentially, activated antigen-specific T cells should form accumulating clones among the lymphocyte population. Therefore, it is crucial to detect such T cells in order to know the contribution of antigen -specific immune responses in the diseases. However, the methods available to analyze T cell clonality are still limited. In this respect, we have established a novel method using a combination of reverse transcriptase-polymerase chain reaction of T cell receptor beta chain transcripts and single strand conformation polymorphism (SSCP). Using this method, the dynamic changes of T cell clonal responses during an antigenic stimulation could be monitored. Analyses of several autoimmune disorders, including rheumatoid arthritis, systemic lupus erythematosus and their animal models, revealed the involvement of antigen specific T cell immune responses. Furthermore, taking advantage of the reproducible mobility of a band in SSCP gel, we are now able to compare identities of the accumulated T cell clones in different samples Without the need for nucleotide sequencing of each clone. Such information can thus elucidate the occurrence of uniform or stable immunological reactions in the patients and also suggests that these reactions play an important role in the pathogenesis. Developing this system, we are also establishing a new strategy to determine the antigen specificities of these in vivo accumulating T cell clones.