Immune Regulation of Neuronal Injury and Repair

描述（由申请人提供）：我们已经建立了CD4+ T细胞介导的涉及野生型（WT）小鼠面部运动神经元（FMN）的神经保护的生理相关机制，这可能对治疗肌萎缩性侧索硬化症（ALS）这一致命的运动神经元疾病具有重要意义。在症状前阶段，在ALS转基因小鼠模型（SOD1G93A）上叠加面神经轴切术（FNA）表明，SOD1小鼠在FMN损失增加和功能恢复下降方面的行为与免疫缺陷（ID）小鼠相似，矛盾的是，其机制不是运动神经元本身固有的，而是涉及外周免疫：中枢神经系统胶质细胞相互作用的缺陷。本项目的长期目标是利用我们的FNA后免疫介导神经保护的WT小鼠模型作为模板，阐明外周免疫系统功能异常如何导致SOD1小鼠ALS模型中的运动神经元细胞丢失。我们的中心假设是CD4+效应T细胞通过调节中枢小胶质细胞和星形胶质细胞对周围神经损伤的反应，为运动神经元提供神经保护。有4个具体目标旨在：1)阐明il - 10的功能作用在WT面部核FNA)后,2)确定CD4 + T细胞救FMN FNA-induced通过激活细胞死亡RAG-2 KO小鼠il - 10的面部核内级联,3)确定后的CD4 + T细胞介导神经保护机制防止FNA的pre-symptomatic SOD1鼠标,和4)测试的能力在vitro-generated,神经保护SOD1 CD4 + T细胞改变疾病发病和/或发展SOD1老鼠。将使用IL-10报告基因小鼠和条件IL-10敲除小鼠，以及WT、ID和SOD1小鼠的过继转移实验，以及激光捕获显微解剖。基于本次修订产生的新数据表明SOD1 T细胞是正常的，但抗原呈递细胞（APC）是有缺陷的，SOD1 T细胞和APC将在体外使用相关和不相关抗原以及在体内使用过继转移来检测表型和功能。最后，在SOD1小鼠发病前使用神经保护性的na特异性T细胞或APC进行过继免疫治疗，并评估对疾病发病、进展和寿命的影响。就预期结果而言，这项工作有望确定FNA后CD4+ T介导的神经保护的关键机制因素，揭示FNA后SOD1 ALS小鼠模型中阻止CD4+ T细胞有效活化/分化的外周免疫系统缺陷，并确定FNA抗原体外产生的神经保护性SOD1 T细胞过继免疫治疗的治疗潜力。ALS每年侵袭3万美国人（每天15例），通常发生在他们生命中最富有成效的时期，给患者、他们的家庭和医疗保健系统带来了巨大的成本。神经免疫相互作用缺陷的识别将产生重要的积极影响，因为它们可以成为ALS治疗干预的目标。

DESCRIPTION (provided by applicant): We have established a physiologically relevant mechanism of CD4+ T cell-mediated neuroprotection involving axotomized wildtype (WT) mouse facial motoneurons (FMN) that may have significance in the treatment of amyotrophic lateral sclerosis (ALS), a fatal motoneuron disease. Superimposition of facial nerve axotomy (FNA) on the transgenic mouse model of ALS (SOD1G93A) during pre-symptomatic stages indicates that SOD1 mice behave like immunodeficient (ID) mice in terms of increased FMN loss and decreased functional recovery, through a mechanism that, paradoxically, is not inherent within the motoneuron itself, but, instead, involves a defect in peripheral immune: CNS glial cell interactions. The long-term goal of this project is to utilize our WT mouse model of immune-mediated neuroprotection after FNA as a template to elucidate how a malfunctioning peripheral immune system contributes to motoneuron cell loss in the SOD1 mouse model of ALS. Our central hypothesis is that CD4+ effector T cells provide neuroprotection to motoneurons through regulation of the central microglial and astrocytic response to peripheral nerve damage. There are 4 specific aims designed to: 1) elucidate the functional role of IL-10 in the WT facial nucleus following FNA, 2) determine if CD4+ T cells rescue FMN from FNA-induced cell death in RAG-2 KO mice through activation of the IL-10 cascade within the facial nucleus, 3) determine the mechanism that prevents CD4+T cell-mediated neuroprotection following FNA in the pre-symptomatic SOD1 mouse, and 4) test the ability of in vitro-generated, neuroprotective SOD1 CD4+ T cells to alter disease onset and/or progression in SOD1 mice. IL-10 reporter and conditional IL-10 knockout mice, along with adoptive transfer experiments with WT, ID and SOD1 mice, and laser capture microdissection will be used. Based on new data generated for this revision suggesting that SOD1 T cells are normal, but antigen-presenting cells (APC) are defective, SOD1 T cells and APC will be examined phenotypically and functionally both in vitro using relevant and irrelevant antigens and in vivo using adoptive transfer. Finally, adoptive immunotherapy with either neuroprotective FNA-specific T cells or APC into SOD1 mice will be done prior to disease onset, and the effects on disease onset, progression, and lifespan assessed. With respect to expected outcomes, the work is expected to identify key mechanistic elements in CD4+ T-mediated neuroprotection following FNA, reveal peripheral immune system defects following FNA in the SOD1 mouse model of ALS that prevent effective CD4+ T cell activation/differentiation and determine the therapeutic potential of adoptive immunotherapy with neuroprotective SOD1 T cells generated in vitro by FNA antigens. ALS strikes 30,000 Americans each year (15 cases a day), often at peak productive points in their lives and with enormous cost to affected individuals, their families, and the health care system. Identification of defects in neural:immune interactions will have an important positive impact because they can then be the target of therapeutic intervention in ALS treatment.