Immune Regualtion of Neuronal Injury and Repair

这是一个修订后的竞争性更新申请的一个项目的基础上，我们发现的abeneficial 外周免疫系统在小鼠面部运动神经元（FMN）修复过程中的作用。在 在最初的资助期间，确定了所涉及的免疫细胞和分子成分：CD 4 + Th 2效应细胞是面神经损伤后FMN存活所必需的，通过抗原依赖性过程 需要外周激活和中枢再激活。脑源性神经营养因子（BDNF）是一种 重要分子重要的是，从轴突切断小鼠中提取的淋巴结细胞的FACS分析表明， Th 1/Th 2细胞分泌型白细胞介素是对周围神经损伤的反应。功能 挤压损伤引起的面瘫的恢复在免疫缺陷小鼠中受损，但可以 通过免疫重建恢复至野生型（WT）。假设CD 4+效应T细胞 在运动神经元修复过程中发挥独特的作用，Tha细胞介导FMN 通过中枢BDNF依赖性过程和Thi细胞介导的功能性存活 通过参与病变部位的促炎反应而恢复。4个目标将测试这一点 假说.目的#1是确定CD 4+效应T细胞定位在细胞内的机制。 面神经损伤后的面运动核。实验将利用免疫缺陷小鼠重建 与表达GFP的CD 4 + T细胞，在与Thi/Th 2细胞因子核糖核酸探针的sifu杂交中，趋化因子RT- PCR和趋化因子中和抗体/敲除小鼠。目的#2.确定机制 面神经损伤后潜在的CD 4 + T细胞介导的FMN存活，并将使用WT和嵌合 BDNF阴性小鼠，以测试CD 4 +T细胞衍生的BDNF在FM拯救中的作用。目标#3是确定 免疫细胞介导颅运动神经元（FMN）免于轴突切断诱导的细胞死亡， 一般是脊柱MN。WT、免疫缺陷和重组免疫缺陷小鼠将用于 确定坐骨神经损伤后免疫系统对坐骨神经MN活力的影响。目标#4是 确定Thi效应细胞是否介导外周神经损伤诱导的功能恢复 瘫痪STAT 4、T-bet和STAT 6缺陷小鼠将用于选择性重建实验， 确定Thi/Th.2效应细胞在运动功能恢复中的作用，用行为测试评估， 在面部或坐骨神经挤压伤后免疫系统既有保护性，也有破坏性。 神经疾病（如肌萎缩性侧索硬化症，一种致命的MN疾病）和/或创伤（如 脊髓损伤），但这种相互矛盾的行为的监管性质尚未确定。 了解免疫系统如何使受损的神经系统受益， 制定有效的治疗策略，以抵消疾病或损伤的进展，并减少残疾。

This is a revised competitive renewal application of a project based on our discovery of abeneficial role for the peripheral immune system in mouse facial motoneuron (FMN)reparative processes. In the initial funding period, the immune cellular and molecular components involved were identified:the CD4+ effector Th2 cell is necessary for FMN survival after facial nerve injury, via an antigen-dependent process requiring peripheral activation and central re-activation. & brain-derived neurotrophic factor (BDNF) is an essential molecule. Importantly, FACS analysis of lymph node cells taken from axotomized mice indicates that Thi/Th2 cytokine-secreting cells are generated in response to peripheral nerve injury. Functional recovery from facial paralysis induced by crush injury is impaired in immunodeficient mice, but can be restored to wildtype (WT) with immune reconstitution. It is hypothesizedthat CD4+effector T cells play distinct roles in motoneuron reparative processes, with the Tha cell mediating FMN survival through a central BDNF-dependent process and the Thi cell mediating functional recovery byparticipation in the lesion site pro-inflammatory response. 4 aims will test this hypothesis. Aim #1 is to determine the mechanism underlying CD4+ effector T cell localization within the facial motor nucleus after facial nerve injury. Experiments will utilize immunodeficient mice reconstituted with GFP-expressing CD4+Tcells, in sifu hybridization with Thi/Th2 cytokine riboprobes, chemokine RT- PCR, and chemokine neutralization antibodies/knockout mice. Aim #2.is to determine the mechanism underlying CD4+ T cell-mediated FMNsurvival after facial nerve injury, and will use WT and chimeric BDNF-negative mice to test the role of CD4+T cell-derived BDNF in FMNrescue. Aim #3 is to determineif immune cell-mediated rescue of cranial motoneurons (FMN)from axotomy-induced cell death can be generalized to a spinal MN. WT,immunodeficient,and reconstituted immunodeficientmicewill be used to determine the impact ofthe immune system on sciatic MNviability after sciatic nerve injury. Aim #4 is to determine if the Thi effector cell mediates functionalrecoveryfrom peripheral nerveinjury-induced paralysis. STAT4,T-bet, and STAT6deficient mice will be used with selective reconstitution experiments to determine the role of Thi/Th.2 effector cells in recovery of motor function, assessed with behavioral tests, after a facial or sciatic nerve crush injury. The immune system can have both protective and destructive effects in neural disease (such as amyotrophic lateral sclerosis, a fatal MNdisease) and/or trauma (such as spinal cord injury), but the regulatory nature ofsuch contradictory actions has yet to be determined. Understanding how the immune system benefits the injured nervous system holds great promise in the development of effective treatment strategies to offset disease or injury progression, and reduce disability.