Characterization of T cell receptor (TCR) of neurological autoimmune disease-inducing T cells and TCR-based immunotherapy with DNA vaccines

在本研究中，我们改进并应用CDR3分型分析这一检测T细胞克隆性增殖的敏感方法来鉴定携带自身免疫病相关T细胞(TCR)的T细胞。根据分析结果，我们制作了编码脑源性T细胞TCR的Vbeta链的DNA疫苗，并试图保护神经抗原+完全弗氏佐剂免疫的动物免受实验性自身免疫性脑脊髓炎(EAE)的发生。DNA疫苗治疗的基本原理如下。当DNA疫苗注射到动物身上时，TCR基因被转录并翻译成TCR oritubs。然后，这些蛋白在免疫动物体内过度表达，诱导抗TcR抗体和/或调节性T细胞，从而抑制携带相同TCR的脑源性T细胞的增殖。在动物模型中建立的TcR诊断和DNA疫苗治疗将为t…提供有用的信息在Lewiw大鼠诱导的EAE模型中，我们发现：1)Vβ8.2克隆性扩增存在于侵入中枢神经系统的T细胞中；2)这一发现不仅在中枢神经系统中观察到，而且在外周血淋巴细胞中也可观察到，这使得利用外周血来诊断该病成为可能；3)当携带Vβ8.2的主要脑源性T细胞被相应的抗体耗尽时，携带Vβ10的第二脑源性T细胞被新激活。这些发现表明，基于TCR的免疫治疗应该同时针对这些T细胞，以获得足够的疾病抑制。在我们最近的研究中，我们发现了CDR3谱型未知的、通过注射相应的TCR DNA疫苗而成功预防疾病的自身免疫性心脏炎诱发的T细胞(J.Invol，2000)。然而，疫苗在抑制EAE方面的效果是不够的。通过改进疫苗结构和确定最有效的疫苗给药方案，应该建立可靠的DNA疫苗接种。我们的最终目标是开发基于TCR的免疫疗法，用于治疗人类神经自身免疫性疾病。较少

In this study, we have modified and applied CDR3 spectratyping analysis which is a sensitive method to detect T cell clonal expansion to identify T cells bearing autoimmune disease-associated T cells (TCR). Based on the results obtained by the analysis, we have made DNA vaccines i.e. expression vectors encoding the Vbeta chain of TCR of encepahlotogenic T cells, and tried to protect animals immunized with neuroantigen plus complete Freund adjuvant from the development of experimental autoimmune encephalomyelitis(EAE). The basic principle of DNA vaccination therapy is as follows. When DNA vaccines are injected in animals, TCR genes are transcribed and translated into TCR oriteubs. Then, these proteins overexpressed in vaccinated animals induce anti-TCR antibodies and/or regulatory T cells, which in tern inhibit the proliferation of encephalitogenic T cells bearing the same TCR.The TCR duagbisus abd DNA vaccination therapy established in animal models will provide useful information to t … More he development of TCR analysis and TCR-based immunotherapy in human diseases in the near future.In EAE induced in Lewiw rats, we found that 1)Vβ8.2 clonal expansion was observed in T cells infiltrating the central nevous system(CNS), 2)this finding was observed not only in the CNS, but also in peripheral blood lymphocyts, enabling the diagnosis of the disease satus using the peripheral blood, and that 3)when the major encephalitogenic T cells bearing Vβ8.2 were depleted with the corresponding antibody, second encephalitogenic T cells bearing Vβ10 were newly activated. These findings suggest that TCR-based immunotherapy should be directed at these T cells simultaneously to obtain sufficient desease suppression.In our recent studies, we identified autoimmune cardititis-inducing T cells that had been not known by CDR3 spectratyping and seceeded in preventing the disease by injection of the corresponding TCR DNA vaccine(J.Immunol., 2000). However, the effects of vaccines on EAE suppression is not sufficient. By improving the vaccine constructs and determining the most effective protocols for vaccine administration, reliable DNA vaccination should be established. Our final goal is to develop TCR-based immunotherapy for human neurological autoimmune diseases. Less