T CELL RECEPTOR GENE USAGE IN EAE

这项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，实验性变态反应 脑脊髓炎(EAE)。EAE被用作一个模型，希望 所获得的知识可以应用于其他自身免疫性疾病。EAE是一种 中枢神经系统的麻痹和炎症性疾病 许多方面都与最常见的人类的病理相似 脱髓鞘疾病，多发性硬化(MS)。中国的最新发展 对T细胞受体(TCR)的分子水平分析揭示了 自身反应性T细胞的克隆只表达有限数量的 可获得的T细胞受体特异性。这让人们产生了这样的希望：或许 某些自身免疫性疾病的调节可以通过 操纵TCR本身。然而，在小鼠模型中，分析 到目前为止只有三个品系或者严格地说是两个品系 单倍型，因为三个菌株中有两个携带相同的主要基因 组织相容性复合体基因和识别相同的髓鞘基础 蛋白质(MBP)T细胞表位。在老鼠身上，只有刘易斯菌株 对此进行了详细的分析。人体研究仍处于相当初步的阶段。至 推广TCR基因使用中的有限异质性的概念 自身反应性T细胞，这项研究将分析两种TCR的特异性 C57BI/6和BALB/C这两个品系的小鼠的遗传多样性 其独特之处在于通过不能产生收养转移而诱导EAE 受体小鼠有疾病的迹象。只有在以下情况下才会诱发EAE 接受者在稍后阶段接受MBP的挑战。对…的调查 在这些“相对抗性”中表达的TCR基因的性质 菌株将有价值与TCR基因相关的免疫机制 用法。为此，来自B6和BALB/C的MBP特异性T细胞克隆将是 已生成。克隆人将接受脑原性和MBP的测试 他们识别的表位。TCR基因产物在细胞表面的表达 用特定的单抗进行监测。在分子水平上， 针对已知V区基因的DNA探针将用于量化RNA 在脑源性克隆中的表达。这种分析还将提供 关于特定V区基因V-β-的流行情况的信息- 8.2，在疾病表现上。因为B6受体小鼠不会得病 在受到MBP挑战之前，这提供了一个理想的系统来研究 转移的细胞在体内的转运模式与其 EAE诱导中的“相对阻力”。通过使用 THY-1基因标记，供体细胞的存在可以在各种不同的 受赠者的组织，尤指大脑和脊髓。已被占用 综上所述，这些研究将更好地了解 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.