The role of T cells in exercise-induced recovery after stroke

T 细胞在中风后运动恢复中的作用

• 批准号: 428668629
• 负责人: Professorin Dr. Wiebke Hansen
• 金额: --
• 依托单位: Institut für Medizinische Mikrobiologie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/428668629?language=en
• 关键词: role; cells; exercise; induced; recovery

Background and preliminary dataExercise therapy improves the neurological recovery following stroke. In addition, exercise modulates the immune system in healthy subjects and attenuates the inflammatory response in various disease models. In our preliminary experiments, we demonstrated that T cells, especially regulatory T cells (Tregs), are a prerequisite for exercise-enhanced stroke recovery after photothrombotic stroke in mice. Our preliminary findings also showed that exercise influences the occurrence of T cells, astrocytes and microglia. In particular, we found an increase of Tregs in cervical lymph nodes and brain parenchyma in mice subjected to exercise therapy after stroke. Further experiments showed a strong impact of exercise on the activity of cortical neurons. Most importantly, we found a normalization of stroke-induced hyperexcitability of periinfarct neurons in response to physical exercise.ObjectivesOur proposed project shall illuminate the interplay between exercise, T cells and neuronal plasticity following stroke. We will analyze effects of exercise therapy on T cell localization in the ischemic brain and the molecular and functional analysis of T cell-dependent effects of exercise. Finally, we will determine T cell-mediated exercise effects on neuronal plasticity, i.e. axonal sprouting and neuronal excitability.Work programThe temporal and spatial distribution of T cells and their subsets in the ischemic brain and meninges in exercise and control animals will be compared using immunohistochemistry, flow cytometry and whole brain 3-D imaging. Gene expression profiles of T cell subsets and detailed functional analysis including activation, migration, metabolism and signaling will be performed. To further clarify the role of Tregs for exercise-enhanced stroke recovery, we will deplete them at different time points in DEREG mice. In parallel, non-functional Tregs from scurfy mice will be compared to wild type Tregs regarding their ability to mediate exercise effects in Rag1-/- mice after stroke. Antigenic-specificity will be investigated by using 2D2/Rag1 mice. Axonal plasticity of corticospinal tract neurons will be analyzed following stereotactic injection of axonal tracers. To complement axonal plasticity studies, the effect of exercise therapy on the translatome of cortical layer 5 pyramidal neurons will be analyzed in Glt25d2 bacTRAP mice. Effects on neuronal activity and connectivity will be determined by electrophysiological experiments, including single-cell patch-clamp recordings, voltage-sensitive dye experiments, neuronal Ca2+-imaging, and in vivo electrophysiology.ConclusionsOur proposed experiments shall clarify the role of T cells for exercise-mediated long-term recovery following ischemic stroke, thus providing the basis for future recovery-enhancing therapies that modulate identified molecular targets – so called exercise mimetics.

背景和初步资料运动疗法可改善脑卒中后神经功能的恢复.此外，运动调节健康受试者的免疫系统，并减弱各种疾病模型中的炎症反应。在我们的初步实验中，我们证明了T细胞，特别是调节性T细胞（TCFs），是小鼠光血栓性中风后运动增强中风恢复的先决条件。我们的初步研究结果还表明，运动影响T细胞，星形胶质细胞和小胶质细胞的发生。特别是，我们发现中风后接受运动治疗的小鼠颈部淋巴结和脑实质中的TdR增加。进一步的实验表明，运动对皮层神经元的活动有很大的影响。最重要的是，我们发现了一个正常化的中风引起的hyperexcitability的梗死周围神经元在响应体育exercise.ObjectivesOur提出的项目应阐明运动，T细胞和神经元可塑性中风后之间的相互作用。我们将分析运动疗法对缺血性脑中T细胞定位的影响，以及运动对T细胞依赖性影响的分子和功能分析。最后，我们将确定T细胞介导的运动对神经元可塑性的影响，即轴突发芽和neuronalexcitability.Work programTheT细胞及其亚群在缺血性脑和脑膜运动和对照动物的时间和空间分布将使用免疫组织化学，流式细胞术和全脑3-D成像进行比较。将进行T细胞亚群的基因表达谱和详细的功能分析，包括活化、迁移、代谢和信号传导。为了进一步阐明TdR在运动增强中风恢复中的作用，我们将在DEREG小鼠的不同时间点耗尽它们。平行地，将来自皮屑小鼠的非功能性TdR与野生型TdR关于它们在中风后介导Rag 1-/-小鼠中的运动效应的能力进行比较。将使用2D 2/Rag 1小鼠研究抗原特异性。皮质脊髓束神经元的轴突可塑性将在立体定向注射轴突示踪剂后进行分析。为了补充轴突可塑性研究，将在Glt 25 d2 bacTRAP小鼠中分析运动疗法对皮质第5层锥体神经元的translatome的影响。对神经元活性和连通性的影响将通过电生理学实验来确定，包括单细胞膜片钳记录、电压敏感染料实验、神经元Ca 2+成像和体内电生理学实验。从而为将来调节已鉴定的分子靶点的增强恢复的疗法--所谓的运动模拟物--提供了基础。