LYMPHOCYTES THAT SUPPRESS EAE

调节性(抑制)淋巴细胞被认为起着关键作用。 在许多现象中，从自身免疫到移植 宽容。然而，他们的行动机制在很大程度上仍然 由于所研究系统的复杂性质，这是未知的。至 简化问题，我们生成了一个最小的鼠标模型 实验性自身免疫性脑脊髓炎(EAE) 多发性硬化症。我们已经培育出转基因小鼠(T/R+)，用于 编码T细胞受体α链和β链的基因 来自脑脊髓性T细胞克隆。大多数人 T/R+小鼠不会发生EAE。相反，当T/R+小鼠 与RAG-1 KO小鼠杂交产生T/R小鼠，100% T/R子代自发发育为EAE。因为这块破布- 1基因突变阻止T/R小鼠产生成熟的B和T 淋巴细胞，这些小鼠唯一含有的淋巴细胞是抗MBP 转基因T细胞。相比之下，T/R+小鼠除了 数量大致相同的抗MBP T细胞，有些是非转基因的 (内源性)具有不同谱系的Alphabeta T细胞，以及 Gammadelta T细胞..因此，T/R-小鼠构成了单克隆性 可以向其添加定义的蜂窝组件及其 评估了EAE法规的重要性。我们的目标是了解 这种调控究竟是如何在细胞和分子中发生的 级别。我们想要鉴定淋巴细胞亚群 通过将T/R+小鼠与小鼠杂交来对抗EAE 仅对T/R缺乏的一个淋巴间隔进行敲除- 小鼠，并将纯化(分类)的细胞亚群注射到 T/R-小鼠在EAE发病前后。另外，我们知道 T/R+小鼠的EAE抵抗不是由两者中的任何一个失败引起的 在胸腺对MBP特异性T细胞的阳性选择中，也没有一个 增加了对这些细胞的负面选择。此外，反- 两型猪外周淋巴器官中的MBP T细胞 老鼠并不是无能的。为了更好地定义 调节细胞上的抗MBP细胞，我们还将比较抗MBP细胞 T/R-小鼠晚期MBP细胞与T/R+小鼠MBP细胞的比较 T细胞的反应，即在“关闭”期间，通过激活诱导 细胞死亡或其他抗炎机制。最后， 调节细胞在EAE控制中的作用的证据也是 通过过继转移抗MBP诱导EAE提供 CD4+TH2细胞。MBP特异性TH2细胞向TH2细胞的转移 将细胞导入RAG-1 KO或TCRαKO小鼠导致EAE 然而，发育正常和TCR Delta KO小鼠 接受者是有抵抗力的。这强烈地表明， Alphabeta T细胞可保护受者免受EAE的侵袭。我们计划 定义这些单元格对以下方面的监管要求 特定的单元格类型、所需的最小单元格数量 调节，以及这类细胞的特异性。增加的 对此负向调控的认识 定义的系统可能不仅适用于多个 硬化症，但也与其他自身免疫性疾病和 关于宽容的一般知识。

Regulatory (suppressor) lymphocytes are assumed to play a key role in a number of phenomena, from autoimmunity to transplantation tolerance. However, their mechanism of action remains largely unknown due to the complex nature of the systems studied. To simplify matters, we generated a minimal mouse model of experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis. We have generated transgenic mice (T/R+) for the genes encoding the alpha and beta chains of the T cell receptor from an encephalomyelitogenic T cell clone. The majority of the T/R+ mice do not develop EAE. In contrast, when T/R+ mice were crossed with RAG-1 KO mice to generate T/R-mice, 100% of the T/R- progeny developed EAE spontaneously. Because the RAG- 1 mutation prevents T/R- mice from generating mature B and T lymphocytes, the only lymphocytes these mice contain are anti-MBP transgenic T-cells. In contrast, T/R+ mice have, in addition to roughly similar numbers of anti-MBP T-cell, some non transgenic (endogenous) alphabeta T-cells with diverse repertoires, as well as gammadelta T-cells.. Thus, T/R- mice constitute a monoclonal system to which defined cellular components can be added and their importance in EAE regulation assessed. Our goal is to understand precisely how this regulation occurs at the cellular and molecular levels. We would like to identify the lymphocyte subpopulation responsible for EAE resistance by crossing T/R+ mice with mice knockout for only one of the lymphoid compartments lacking in T/R- mice, and by injecting purified (sorted) cell subpopulations into T/R-mice before and after the onset of EAE. In addition, we know that the EAE resistance of T/R+ mice is caused by neither a failure in thymic positive selection of MBP specific T cells, nor an increased negative selection of these same cells. Furthermore, anti- MBP T cells in the peripheral lymphoid organs of both types of mice are not anergic. In order to better define the action of regulatory cells on anti-MBP cells, we will also compare the anti- MBP cells of T/R- mice to those of T/R+ mice in the later stages of T cell response, i.e., during "shut off," by activation induced cell death or other anti-inflammatory mechanisms. Finally, evidence for a role of regulatory cells in EAE control is also provided by EAE induction by adoptive transfer of anti- MBP CD4+ TH2 cells. Transfer of MBP specific TH2 cells into TH2 cells into RAG-1 KO or TCR alpha KO mice results in EAE development, however, both normal and TCR delta KO mouse recipients are resistant. This strongly suggests that the presence of alphabeta T cells protects recipients against EAE. We plan to define the requirements of these cells for regulation with regard to the specific cell type, the minimum number of cells necessary for regulation, and the specificity of such cells. The increased understanding of negative regulation and control provided by this defined system is potentially applicable not only to multiple sclerosis, but also to other autoimmune diseases and to the knowledge of tolerance in general.