T cell-mediated mechanisms of neurodegeneration in models of Parkinson's disease

帕金森病模型中 T 细胞介导的神经变性机制

基本信息

批准号：
9208175
负责人：
Rebecca Wilshusen
金额：
$ 2.71万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-03-01 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9208175
关键词：
1-Methyl-4-phenylpyridinium Adaptive Immune System Address Adoptive Transfer Affect Animal Model Antigen-Presenting Cells Apoptotic Attenuated Biological Assay Blocking Antibodies Cell Culture Techniques Cell Line Cell Survival Cell-Mediated Cytolysis Cells Cervical lymph node group Coculture Techniques Cytoplasm Development Disease Disease Progression Environment Eragrostis Functional disorder Hour Immunization In Vitro Individual Infectious Agent Inflammatory Interferons Interleukin-17 Intoxication Laboratories Lead Lewy Bodies Link Lymphoid Tissue Macrophage Activation Measures Mediating Microglia Midbrain structure Modeling Molecular Mus Natural Immunity Nerve Degeneration Neuraxis Neurons Nitrates Oxidative Stress Oxides Parkinson Disease Pathogenesis Phenotype Process Proteins Small Interfering RNA Substantia nigra structure T-Lymphocyte Testing Toxic effect Transgenic Model Tyrosine 3-Monooxygenase Validation Western Blotting adaptive immunity alpha synuclein cytokine cytotoxicity dopaminergic neuron experimental study improved insight mouse model neuroinflammation neuronal survival neuropathology neurotoxic neurotoxicity neutralizing antibody novel palliative public health relevance response symptom treatment synuclein

项目摘要

DESCRIPTION (provided by applicant): This proposal addresses the cellular and molecular mechanisms by which modified self-protein specific effector T cell (Teff) subsets modulate neurodegeneration in models of Parkinson's disease (PD). Increasing evidence suggests that neurotoxic inflammatory activities affect pathogenesis and progression of PD. Neuroinflammatory processes also produce oxidized and modified self-central nervous system (CNS) proteins which lead to dysfunction, mis-folding, aggregation, and retention of those oxidized products. In PD, nitrated ?-synuclein (N-?-syn) is found aggregated within the cytoplasm and Lewy bodies of dopaminergic neurons within the substantia nigra and is release to the extraneuronal environment by dying and damaged neurons. Previous studies have shown that after 1-methyl 4-phenyl 1,2,3,6-tetrahydropyridine (MPTP) intoxication, adoptive transfer of Teffs exacerbates microglial-mediated neuroinflammation and amplifies dopaminergic neurodegeneration with accelerated and prolonged neuropathology. Additionally, Teffs that secrete IL-17 (Th17 type) exacerbate neurodegeneration to greater levels than those Teff types that secrete IFN-c (Th1 type). Thus, we posit that N-?-syn specific Th17 effector T cells exacerbate neurodegeneration indirectly via hyperactivation of microglial-mediated neurotoxicity or alternatively by direct cytotoxicity of neurons. In order to test this hypothesis we will first determine whether N-?-syn specific effector T cells mediate neurotoxicity via exacerbation of microglia or by direct cytotoxicity to dopaminergic neurons by generating a N-?-syn specific Th17 effector T cell line and testing this line in in vitro cultures with both neurons and microgli to determine neuron cell survival and toxicity. We will then determine whether cell-to-cell contact or proinflammatory cytokines are required for Th17-mediated exacerbation of neurodegeneration with the use of co-cultures composed again of Th17 effector T cells, microglia, and neurons. These co-cultures will be configured with the cells in either direct contact or transwell configurations in order to determine the necessity of direct cell-to-cell contact by T cells to exacerbate their neurodegenerative effects. Lastly we will validate the mechanisms of specific factors linked to the exacerbation of neuroinflammatory and neurodegenerative Th17- mediated responses to test said factors we will target those factors in animal models of PD as well as utilize antibody blocking assays, siRNA, and transgenic models to block Th17-mediated exacerbation of neuroinflammation or neurodegeneration. These experiments, taken together, will permit novel insight into how N-?-syn specific effector T cells induce neurodegeneration in models reflective of Parkinson's disease.

描述（由申请人提供）：该提案阐述了修饰的自身蛋白特异性效应T细胞（Teff）子集在帕金森病（PD）模型中调节神经变性的细胞和分子机制。越来越多的证据表明神经毒性炎症活动影响帕金森病的发病机制和进展。神经炎症过程还会产生氧化和修饰的自身中枢神经系统（CNS）蛋白，导致这些氧化产物功能障碍、错误折叠、聚集和保留。在PD中，硝化α-突触核蛋白（N-α-syn）被发现聚集在黑质内多巴胺能神经元的细胞质和路易体内，并通过死亡和受损的神经元释放到神经元外环境。先前的研究表明，1-甲基4-苯基1,2,3,6-四氢吡啶（MPTP）中毒后，Teffs的过继转移会加剧小胶质细胞介导的神经炎症，并放大多巴胺能神经变性，从而加速和延长神经病理学。此外，分泌 IL-17（Th17 型）的 Teff 比分泌 IFN-c（Th1 型）的 Teff 类型更严重地加剧神经变性。因此，我们假设 N-α-syn 特异性 Th17 效应 T 细胞通过小胶质细胞介导的神经毒性的过度激活或通过神经元的直接细胞毒性间接加剧神经变性。为了检验这一假设，我们首先通过生成 N-α-syn 特异性 Th17 效应 T 细胞系并测试该细胞系，确定 N-α-syn 特异性效应 T 细胞是否通过小胶质细胞恶化或通过对多巴胺能神经元的直接细胞毒性介导神经毒性。与神经元和小胶质细胞进行体外培养，以确定神经元细胞的存活和毒性。然后我们将确定细胞与细胞之间是否接触使用再次由 Th17 效应 T 细胞、小胶质细胞和神经元组成的共培养物时，Th17 介导的神经变性恶化需要促炎细胞因子或促炎细胞因子。这些共培养物将与细胞直接接触或跨孔配置，以确定 T 细胞直接细胞与细胞接触以加剧其神经退行性影响的必要性。最后，我们将验证与神经炎症和神经退行性 Th17 介导反应加剧相关的特定因素的机制，以测试所述因素，我们将在 PD 动物模型中针对这些因素，并利用抗体阻断测定、siRNA 和转基因模型来阻断Th17 介导的神经炎症或神经变性的恶化。这些实验结合在一起，将能够对 N-α-syn 特异性效应 T 细胞如何在反映帕金森病的模型中诱导神经变性提供新的见解。