CD8 T cell mediated disruption of Blood Brain Barrier Tight Junctions

CD8 T 细胞介导的血脑屏障紧密连接破坏

基本信息

批准号：
9293869
负责人：
Aaron J Johnson
金额：
$ 39.75万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9293869
关键词：
AIDS Dementia Complex Acute Acute Hemorrhagic Leukoencephalitis Affect Alzheimer&apos s Disease Antigens Area Biological Models Blood - brain barrier anatomy Blood Vessels Brain C57BL/6 Mouse CD8B1 gene Cells Cerebral Malaria Chemicals Confocal Microscopy Cre-LoxP Data Disease Epilepsy Experimental Models Flow Cytometry Glioblastoma Goals HIV Hematopoietic Human Immune Immunohistochemistry Inflammation Inflammatory Investigation KDR gene Laboratories Magnetic Resonance Imaging Mammals Mediating Modeling Monitor Morbidity - disease rate Multiple Sclerosis Mus Neurologic Neurons Neuropilin-1 Pathology Peptides Permeability Plasmodium berghei Play Process Protein Biochemistry Research Resolution Role Signal Transduction Solid Stroke Syndrome T-Lymphocyte TMEV Testing Therapeutic Therapeutic Intervention Tight Junctions Transgenic Mice Traumatic Brain Injury Up-Regulation Variant Vascular Endothelial Cell Vascular Endothelial Growth Factors Vascular Permeabilities Viral Antigens Viral Hemorrhagic Fevers Work abstracting animal model development base behavioral study brain endothelial cell cell type in vivo in vivo imaging innovation insight intravital imaging intravital microscopy mouse model nervous system disorder neuroinflammation neurovascular unit novel perforin programs receptor two-photon

项目摘要

Abstract Blood brain barrier (BBB) disruption is an integral feature of neurological diseases as diverse as multiple sclerosis, acute hemorrhagic leukoencephalitis (AHLE), traumatic brain injury, stroke, cerebral malaria, viral hemorrhagic fevers, epilepsy, glioblastoma, alzheimer’s disease, and HIV dementia. A fundamental question in these diseases is the extent inflammatory immune cells contribute to CNS vascular permeability. This lack of understanding currently undermines therapeutic approaches to treat neurological disease in which uncontrolled BBB disruption contributes to pathology. My research program was the first to demonstrate that CD8 T cells have the capacity to disrupt BBB tight junctions using a novel murine model. Using this model, we have developed a tractable approach to dissect immune-mediated mechanisms of vascular endothelial growth factor (VEGF) mediated BBB disruption using a variation of Theiler’s Murine Encephalomyelitis Virus (TMEV) and the Plasmodium berghei ANKA (PbA) model systems. Our central hypothesis is CD8 T cells promote neuronal expression of VEGF which results in disruption of cerebral endothelial cell tight junctions and vascular permeability. We will test our central hypothesis through the following aims: Specific Aim #1 – Determine the extent direct engagement of antigen specific CD8 T cells with neurons promotes VEGF expression and ensuing vascular changes. Specific Aim #2 – Determine the extent neuronal expression of VEGF and its receptors contribute to CD8 T cell-initiated BBB disruption. Specific Aim #3 – Evaluate the contribution of neuronal VEGF to CD8 T cell-mediated BBB disruption in the Plasmodium berghei ANKA (PbA) model of experimental cerebral malaria. To our knowledge, no other laboratories have a similar mouse as PIFS that capitalizes on readily inducible acute CNS vascular permeability mediated by a well-defined immune cell type. Confirming the existence of homologous inflammatory mechanisms in humans would be the first step toward therapeutic intervention of neurological diseases in which neuroinflammation induced CNS vascular permeability plays a significant part. To accomplish these aims, we will employ: (a) flow cytometry, (b) behavioral studies, (c) high resolution confocal microscopy and immunohistochemistry, (d) 2-photon intravital microscopy (e) protein biochemistry, and (f) small mammal MRI.

抽象的血脑屏障 (BBB) 破坏是多种神经系统疾病的一个重要特征多发性硬化症、急性出血性白质脑炎 (AHLE)、创伤性脑损伤、中风、脑型疟疾、病毒性出血热、癫痫、胶质母细胞瘤、阿尔茨海默病和艾滋病毒这些疾病的一个基本问题是炎症免疫细胞的范围。目前对中枢神经系统血管通透性的缺乏了解会破坏这一点。治疗神经系统疾病的治疗方法，其中不受控制的血脑屏障破坏我的研究项目是第一个证明 CD8 T 细胞对病理学的贡献。具有使用新型小鼠模型破坏 BBB 紧密连接的能力，我们开发了一种易于处理的方法来剖析血管的免疫介导机制使用 Theiler 鼠的变种，内皮生长因子 (VEGF) 介导 BBB 破坏脑脊髓炎病毒 (TMEV) 和伯氏疟原虫 ANKA (PbA) 模型系统。我们的中心假设是 CD8 T 细胞促进 VEGF 的神经表达，导致脑内皮细胞紧密连接和血管通透性破坏。我们将通过以下目标来检验我们的中心假设：具体目标 #1 – 确定抗原特异性 CD8 T 细胞直接参与的程度神经元促进 VEGF 表达和随后的血管变化。具体目标 #2 – 确定 VEGF 及其受体的神经表达程度有助于 CD8 T 细胞引发的 BBB 破坏。具体目标 #3 – 评估神经元 VEGF 对 CD8 T 细胞介导的 BBB 的贡献实验性脑型疟疾的伯氏疟原虫 ANKA (PbA) 模型的破坏。据我们所知，没有其他实验室拥有与 PIFS 类似的鼠标，它利用了由明确的免疫细胞类型介导的易于诱导的急性中枢神经系统血管通透性。确认人类中同源炎症机制的存在将是第一个向神经系统疾病的治疗干预迈出一步，其中神经炎症诱导的中枢神经系统血管通透性发挥着重要作用，为了实现这些目标，我们将采用：（a）流式细胞术，（b）行为研究，（c）高分辨率共聚焦显微镜和免疫组织化学，(d) 2 光子活体显微镜检查，(e) 蛋白质生物化学，以及 (f) 小哺乳动物核磁共振成像。