T CELL RECEPTOR GENE USAGE IN EAE

本建议的目的是为了更清楚地了解 自身免疫性疾病的潜在机制，实验性过敏 脑脊髓炎（EAE）。 EAE被用作模型，希望 所获得的知识可以应用于其他自身免疫性疾病。 EAE是一个 中枢神经系统的麻痹性和炎性疾病， 在许多方面类似于最普通的人类的病理学， 脱髓鞘疾病、多发性硬化（MS）。 的新进展 在分子水平上对T细胞受体（TCR）的分析揭示了 自身反应性T细胞克隆只表达有限数量的 T细胞受体特异性。 这让人产生了希望， 某些自身免疫性疾病的调节可以通过 对TCR本身的操纵。 然而，在小鼠模型中，分析 到目前为止，只有三种菌株，或者严格地说，两种 单倍型，因为三个菌株中的两个携带相同的主要 组织相容性复合体基因和识别相同的髓鞘基本 蛋白（MBP）T细胞表位。 在大鼠中，只有刘易斯菌株 详细分析。 人类的研究还相当初级。 到 概括TCR基因使用中有限异质性的概念， 自身反应性T细胞，这项研究将分析两个TCR特异性 遗传多样的小鼠品系，C57 BI/6和BALB/C。 这两种菌株 在通过过继转移诱导EAE方面是独特， 受体小鼠的疾病迹象。 EAE仅在 接受者在稍后阶段用MBP激发。 调查 在这些“相对耐药”的细胞中表达的TCR基因的性质 因此，研究TCR基因与免疫机制的关系具有重要意义 使用. 为此，来自B6和BALB/C的MBP特异性T细胞克隆将被克隆。 生成的. 将对克隆进行致脑炎性和MBP检测 它们识别的表位。 TCR基因产物的细胞表面表达将 用特异性单克隆抗体进行监测。 在分子水平上， 已知V区基因特异性DNA探针将用于定量RNA 在致脑炎克隆中表达。 这样的分析也将提供 关于特定V区基因V-β- 8.2疾病的表现。 由于B6受体小鼠不会发生疾病， 这为进一步研究MBP的作用机制提供了一个理想的体系。 转移细胞的体内运输模式与其 EAE诱导中的“相对抗性”。 通过使用 Thy-1遗传标记，供体细胞的存在可以在各种 受体的组织，特别是大脑和脊髓。 采取 总之，这些研究将提供一个更好的评价， EAE和MS的潜在机制。

The objective of this proposal is to develop a clearer understanding of the underlying mechanisms of the autoimmune disease, experimental allergic encephalomyelitis (EAE). EAE is used as a model with the hope that the knowledge gained can be applied to other autoimmune diseases. EAE is a paralytic and inflammatory disease of the central nervous system and in many respects resembles the pathology of the most common human demyelinating disease, multiple sclerosis (MS). Recent development in the analysis of the T cell receptor (TCR) at the molecular level has revealed that clones of autoreactive T cells only express a restrict number of the available T cell receptor specificities. This raises the hope that perhaps modulation of certain autoimmune diseases can be achieved through manipulation of the TCR itself. However, in the murine model, the analysis has so far been made only in three strains or strictly speaking, in two haplotypes because two of the three strains carry identical major histocompatibility complex genes and recognize the same myelin basic protein (MBP) T cell epitope. In rat, only the Lewis strain has been analysed in detail. Human study is still rather preliminary. To generalize the concept of limited heterogeneity in TCR gene usage by autoreactive T cells, this study will analyse the TCR specificities of two genetically diverse strains of mice, C57BI/6 and BALB/C. These two strains are unique in that induction of EAE by adoptive transfer fail to generate signs of disease in the recipient mice. EAE is induced only after the recipients are challenged at a later stage with MBP. The investigation of the nature of the TCR genes expressed in these "relatively resistant" strains will be of value to correlate immune mechanisms with TCR gene usage. To this end, MBP-specific T cell clones from B6 and BALB/C will be generated. Clones will be tested for encephalitogenicity and the MBP epitope they recognize. Cell surface expresion of TCR gene products will be monitored with specific monoclonal antibodies. At the molecular level, DNA probes specific for known V-region genes will be used to quantify RNA expression in encephalitogenic clones. Such analysis will also provide information regarding the prevalence of a specific V-region gene, V-beta- 8.2, in disease expression. Since B6 recipient mice do not develop disease until after challenged with MBP, this provides an ideal system to study the in vivo trafficking patterns of the transferred cells in relation to their "relative resistance" in EAE induction. By using the expression of the Thy-1 genetic marker, the presence of donor cells can be traced in various tissues of the recipient, especially in the brain and spinal cord. Taken together, these studies will provide a better appreciation of the underlying mechanisms of EAE and of MS.