CD8 T cell mediated disruption of Blood Brain Barrier Tight Junctions

摘要 血脑屏障（BBB）破坏是神经系统疾病的一个组成特征， 多发性硬化、急性出血性脑白质炎（阿勒）、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HIV 痴呆这些疾病的一个基本问题是炎症免疫细胞 有助于CNS血管渗透性。这种缺乏了解的情况目前正在破坏 治疗其中不受控制的BBB破坏的神经疾病的治疗方法 有助于病理学。我的研究项目首次证明了CD 8 T细胞 具有使用新的鼠模型破坏BBB紧密连接的能力。利用这个模型， 我们已经开发了一种易于处理的方法来剖析免疫介导的血管炎机制， 内皮生长因子（VEGF）介导的血脑屏障破坏，使用Theiler小鼠的变体 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CD 8 T细胞促进VEGF的神经元表达， 导致脑内皮细胞紧密连接和血管通透性的破坏。 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CD 8 T细胞与抗原特异性CD 8 T细胞直接接合的程度。 神经元促进VEGF表达和随后的血管变化。 具体目标#2 -确定VEGF及其受体的神经元表达程度 有助于CD 8 T细胞引发的BBB破坏。 具体目标#3 -评价神经元VEGF对CD 8 T细胞介导的BBB的贡献 在实验性脑型疟疾的伯氏疟原虫ANKA（PbA）模型中的破坏。 据我们所知，没有其他实验室拥有像PIFS这样的小鼠， 由明确定义的免疫细胞类型介导的易诱导急性CNS血管通透性。 证实人类存在同源炎症机制将是第一个 向神经系统疾病治疗性干预迈进，其中神经炎症 诱导的CNS血管渗透性起重要作用。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，（e）蛋白质生物化学，和（f）小 哺乳动物MRI。

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.