CD 8 T Cell Mediated Disruption of Blood Brain Tight Junction

描述（由申请人提供）：血脑屏障（BBB）破坏是许多严重神经系统疾病的常见病理特征。尽管由于这些与CNS血管通透性相关的神经系统疾病而导致的发病率和死亡率的巨大负担，但对炎症细胞如何促进BBB破坏的潜在分子机制仍然知之甚少。我们已经开发了一种新的中枢神经系统血管通透性的小鼠模型，提供了一个易于处理的方法来定义特定的炎症介质，破坏血脑屏障在体内使用泰勒病毒感染。这种BBB破坏的小鼠模型是由CNS浸润性CD 8 T细胞的肽特异性刺激引发的，并且不受TNF-α、IFN-β、LT R和IL- 1。然而，穿孔素对于CNS血管通透性和BBB紧密连接的保存都是至关重要的。值得注意的是，尽管具有相同的CD 8 + T细胞应答和CTL活性，但MHC相同的C57 BL/6和129 Svlm小鼠对PIFS的易感性显著不同。该提议的目的是验证我们的基本假设，即CD 8 T细胞利用穿孔素启动脑内皮细胞BBB紧密连接的破坏。C57 BL/6和129 SvIm小鼠背景上的特异性修饰基因对于该过程的发生也至关重要。我们将使用这个模型来确定：1。BBB破坏所必需的穿孔素的细胞来源，2.）造血细胞在促进BBB破坏中的作用，以及3.）控制血脑屏障破坏的遗传因子的染色体定位。我们将使用传统的遗传分析，蛋白质生物化学，成像，细胞免疫学测定，分子生物学和逆转录病毒表达，以确定介导破坏血脑屏障的炎症因子。鉴定导致致命性CNS血管通透性的强有力因素将确定BBB治疗性修饰的靶点。这些疾病的特征在于破坏血脑屏障，包括中风、病毒性出血热、脑型疟疾、HIV痴呆、多发性硬化症（MS）和休克。增加对血脑屏障通透性的了解也有利于化疗。公共卫生相关性：炎症介导的血脑屏障（BBB）破坏是一个知之甚少，但相对常见的许多神经系统疾病的特征。我们已经开发了一种新的中枢神经系统血管通透性的小鼠模型，可用于定义特定的细胞相互作用和炎症介质，导致体内血脑屏障的破坏。定义这些因子是对BBB通透性进行新的治疗性修饰的第一步。

DESCRIPTION (provided by applicant): Disruption of the Blood Brain Barrier (BBB) is common pathologic feature of numerous serious neurological diseases. Despite the enormous burden of morbidity and mortality due to these neurological diseases associated with CNS vascular permeability, the underlying molecular mechanisms of how inflammatory cells promote BBB disruption remain poorly understood. We have developed a novel murine model of CNS vascular permeability that provides a tractable approach to defining specific inflammatory mediators that disrupt the BBB in vivo using Theiler's virus infection. This murine model of BBB disruption is initiated by peptide-specific stimulation of CNS infiltrating CD8 T cells and is not mediated by TNF-a, IFN-?, LT¿R and IL- 1. However, perforin is critical for both CNS vascular permeability and preservation of BBB tight junctions. Notably, MHC-identical C57BL/6 and 129 Svlm mice differ dramatically in susceptibility to PIFS, despite having identical CD8+ T cell responses and CTL activity. The goal of this proposal is to test our underlying hypothesis that CD8 T cells utilize perforin to initiate the disruption of cerebral endothelial cell BBB tight junctions. Specific modifier genes on the C57BL/6 and 129 SvIm mouse background are also critical for this process to occur. We will determine using this model: 1.) the cellular source of perforin necessary for BBB disruption, 2.) the role of hematopoetic cells in promoting disruption of the BBB, and 3.) the chromosome location of genetic factors that govern BBB disruption. We will use conventional genetic analysis, protein biochemistry, imaging, cellular immunology assays, molecular biology and retrovirus expression to determine the inflammatory factors that mediate disruption of the BBB. Identification of powerful factors that contribute to fatal CNS vascular permeability would define targets for therapeutic modification of the BBB. Such diseases characterized by disruption of the BBB including stroke, viral hemorrhagic fevers, cerebral malaria, HIV dementia, multiple sclerosis (MS), and shock. Increased understanding of BBB permeability could also benefit chemotherapy. PUBLIC HEALTH RELEVANCE: Inflammation mediated blood brain barrier (BBB) disruption is a poorly understood, yet relatively common feature of numerous neurologic diseases. We have developed a novel murine model of CNS vascular permeability that can be utilized to define specific cellular interactions and inflammatory mediators that result in disruption of the BBB in vivo. Defining such factors is the first step towards novel therapeutic modification of BBB permeability.