CD8 T cell mediated disruption of Blood Brain Barrier Tight Junctions

摘要 血脑屏障(BBB)破坏是各种神经系统疾病的一个不可或缺的特征 多发性硬化症，急性出血性白质脑炎，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 痴呆症。这些疾病的一个基本问题是炎性免疫细胞的程度 有助于中枢神经系统的血管通透性。这种缺乏理解的态度目前破坏了 治疗失控血脑屏障中断的神经系统疾病的治疗方法 对病理学有贡献。我的研究项目是第一个证明CD8 T细胞 有能力利用一种新的小鼠模型破坏血脑屏障紧密连接。使用这个模型， 我们开发了一种简单易用的方法来剖析免疫介导的血管机制。 内皮生长因子(VEGF)介导的血脑屏障破坏作用 脑脊髓炎病毒(TMEV)和伯氏疟原虫(PBA)模型系统。 我们的中心假设是CD8 T细胞促进神经元表达血管内皮生长因子 导致脑血管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： 特定目标#1-确定抗原特异性CD8 T细胞与 神经元促进血管内皮生长因子的表达和随后的血管变化。 特定目标#2-确定血管内皮生长因子及其受体在神经元中的表达程度 导致CD8T细胞启动的血脑屏障破坏。 特定目标#3-评估神经元血管内皮生长因子在CD8 T细胞介导的血脑屏障中的作用 实验性脑型疟疾伯氏疟原虫(PBA)模型的破坏。 据我们所知，没有其他实验室有像PIF这样的老鼠 易于诱导的急性中枢神经系统血管通透性由明确定义的免疫细胞类型介导。 证实人类体内存在同源炎症机制将是第一个 迈向神经炎症性神经系统疾病的治疗干预 诱导的中枢神经系统血管通透性起着重要作用。为了实现这些目标，我们将 使用：(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)双光子活体显微镜，(E)蛋白质生化，和(F)小 哺乳动物核磁共振。

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.