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

• 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
• 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

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）：该提案介绍了细胞和分子机制，通过这些机制，修饰的自我蛋白质特异性效应T细胞（TEFF）亚群调节帕金森氏病（PD）模型中的神经变性。越来越多的证据表明，神经毒性炎症活性会影响PD的发病机理和进展。神经炎症过程还产生了氧化和改良的自中心神经系统（CNS）蛋白，从而导致功能障碍，折叠，聚集和保留这些氧化产物。在PD中，硝化的？ - 突触核蛋白（N-？ - SYN）被发现在黑质中的多巴胺能神经元的细胞质和Lewy体内聚集，并通过垂死和受损的神经元释放到外交环境中。先前的研究表明，在1-甲基4-苯基1,2,3,6-四氢吡啶（MPTP）中毒后，TEFFS的过继转移加剧了小胶质细胞介导的神经炎症，并扩增多巴胺能神经化学，并具有加速和延长的神经疗法。此外，与分泌IFN-C（TH1类型）的TEFF类型相比，分泌IL-17（TH17类型）的TEFF将神经变性加剧的水平更大。因此，我们认为N - ？ - SYN特异性Th17效应T细胞通过小胶质细胞介导的神经毒性或神经元的直接细胞毒性的过度激活而间接地加剧神经变性。为了检验该假设，我们将首先确定N - ？ - SYN特异性效应T细胞是否通过加剧小胶质细胞或直接细胞毒性对多巴胺能神经元进行神经毒性介导了神经毒性，通过产生N - ？SYN特异性Th17特异性Th17效应T细胞系，并测试具有神经元和神经元的内部培养物中的这种线，以确定神经元和神经元的培养基。然后，我们将确定是否接触单元格 Th17介导的神经退行性加剧需要再次使用由Th17效应T细胞，小胶质细胞和神经元组成的神经退行性的加重，或者促炎细胞因子。这些共培养将在直接接触或transwell配置中与细胞配置，以确定T细胞直接通过T细胞接触细胞对细胞接触的必要性，以加剧其神经退行性效应。最后，我们将验证与神经炎症性和神经退行性的Th17介导的对测试因素的介导的反应有关的特定因素的机制，我们将针对PD动物模型中的这些因素，并利用抗体阻断测定法，以及抗体阻断，SIRNNA，SIRGENIC模型以及TH17介导的介导的Neuronemation neuroinemation neuroinemational of Neuroinematimation neuroinemations of Neuroinemant or Neuroinemant of Neuroinemfation neuroinemfation neuroInsmodeanm。这些实验将共同进行，将允许对N - ？ - SYN特异性效应T细胞如何诱导反映帕金森氏病的模型中的神经变性。