CD8 T Lymphocyte Regulation of EAE Disease Phenotype

描述（由申请人提供）：多发性硬化症及其动物模型，实验性自身免疫性脑脊髓炎（EAE），是一组异质性自身免疫性脑炎性疾病。拟议研究的长期目标是开发改进的动物模型，以了解和治疗疾病亚型，特别是那些严重的亚型。具体目的是评估CD 8 T淋巴细胞在介导EAE疾病表型严重程度中的作用。假设是，在自身反应性Th 1 CD 4 T淋巴细胞引发中枢神经系统（CNS）炎症后，随后在特定的关键时期实质募集CD 8 T淋巴细胞导致更严重的临床疾病。为了检验这一假设，在（C57BL6xB10.PL）F1小鼠中开发了EAE的过继转移模型。初步数据表明，与B10.PL小鼠相比，髓磷脂碱性蛋白（MBP）特异性（Acl-11）Th 1 CD 4 T淋巴细胞的过继转移可在免疫活性（C57BL6xB10.PL）F1小鼠中诱导严重的EAE。组织学表征、基因表达分析和CD 8耗竭揭示了（C57BL6xB10.PL）F1小鼠中CD 8细胞毒性T淋巴细胞（CTL）的致病作用。这些结果表明，依赖于遗传背景，CD 8 T淋巴细胞可以介导疾病的严重程度。我们的假设将在三个具体目标中得到进一步检验。在目的1中，我们计划使用CD 8缺陷和CTL效应缺陷小鼠来阐明CD 4和CD 8 T淋巴细胞在急性病变中的致病作用。目的2：明确重症EAE早期CD 8 T淋巴细胞进入的关键期及相关的中枢神经系统微环境变化。在目标3中，我们提出分离CNS特异性CD 8 T淋巴细胞克隆，在体外对其进行表征，并评估其诱导CNS炎性疾病的能力。这些研究将提供一个更好的了解炎症，脱髓鞘疾病的中枢神经系统中的CTL介导的损伤。此外，EAE模型具有临床意义，因为它更准确地反映了自身反应性CD 4和CD 8 T淋巴细胞在多发性硬化急性脱髓鞘病变发病机制中的复杂作用。由于CTL可在急性MS病变中引发轴突损伤，从而导致慢性、进行性临床缺陷，因此该模型也可用于筛选抑制CD 4 T淋巴细胞引发的炎症以及CTL效应机制的潜在MS治疗剂。

DESCRIPTION (provided by applicant): Multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis (EAE), are a heterogeneous group of autoimmune, inflammatory diseases of the brain. The long-term goal of the proposed studies is to develop improved animal models to understand and treat disease subtypes, particularly those that are severe. The specific objective is to assess the role of CD8 T lymphocytes in mediating severity of EAE disease phenotype. The hypothesis is that, following initiation of central nervous system (CNS) inflammation by auto-reactive Th1 CD4 T lymphocytes, the subsequent parenchymal recruitment of CD8 T lymphocytes during a specific critical period leads to more severe clinical disease. To test this hypothesis, an adoptive transfer model of EAE was developed in (C57BL6xB10.PL) F1 mice. Preliminary data demonstrate that adoptive transfer of myelin basic protein (MBP)-specific (Acl-11) Th1 CD4 T lymphocytes can induce severe EAE in immune competent (C57BL6xB10.PL) F1 mice as compared to B10.PL mice. Histologic characterization, gene expression analysis, and CD8 depletion reveal a pathogenic role for CD8 cytotoxic T lymphocytes (CTL) in (C57BL6xB10.PL) F1 mice. These results suggest that, dependent on genetic background, CD8 T lymphocytes can mediate disease severity. Our hypothesis will be tested further in three Specific Aims. In Aim 1, we plan to use CD8-deficient and CTL effector-deficient mice to clarify the pathogenic roles of CD4 and CD8 T lymphocytes in acute lesions. Aim 2 focuses on defining the critical period of entry of CD8 T lymphocytes and associated CNS microenvironment changes in the early development of severe EAE. In Aim 3, we propose to isolate CNS-specific CD8 T lymphocyte clones, characterize them in vitro, and assess their ability to induce CNS inflammatory disease. These studies will provide a better understanding of CTL-mediated injury in inflammatory, demyelinating diseases of the CNS. In addition, this EAE model has clinical relevance because it more accurately reflects the complex roles of autoreactive CD4 and CD8 T lymphocytes in the pathogenesis of acute demyelinating lesions in multiple sclerosis. Since CTL's may initiate axonal injury in acute MS lesions with resultant chronic, progressive clinical deficits, this model should also be useful in screening potential MS therapeutics that inhibit CD4 T lymphocyte-initiated inflammation as well as CTL effector mechanisms.