EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS--INHIBITION OF C

本研究的目的是探讨补体在多发性硬化（MS）动物模型实验性变态反应性脑脊髓炎（EAE）的细胞浸润、炎症和脱髓鞘中的作用。 有大量证据表明，在MS和EAE中，补体被激活，并可能导致与这些脱髓鞘疾病相关的炎症和细胞破坏。 我们实验室和其他实验室也有证据表明，补体蛋白和受体在中枢神经系统（CNS）中合成和表达，这表明局部补体产生可能有助于早期疾病的发展。确定补体是否参与脱髓鞘疾病的发病机制的一种方法是抑制其活化。 先前在EAE中抑制补体的尝试已证明该策略是可行的，但存在技术缺陷。 为了克服这些局限性，我们已经开发了一种转基因小鼠模型，其中可溶性鼠特异性抑制剂sCrry由星形胶质细胞使用胶质细胞酸性蛋白（GFAP）启动子构建体产生。 我们建议，使用这种新的转基因动物，在中枢神经系统特异性阻断补体激活，提供了一个独特的和更好的模型系统，以确定补体介导的各种作用，在EAE的炎症反应的发展。 我们假设使用该模型抑制补体激活将减少细胞浸润和脱髓鞘，这是EAE的特征。在本申请中，我们建议1）完成我们的sCrry表达的创始细胞系的分析，2）进行动力学分析，并比较sCrry和对照小鼠在EAE中的临床评分和组织病理学（浸润细胞类型的程度和鉴定、脱髓鞘和轴突横断的程度），和3）评估EAE中sCrry和对照小鼠中改变的基因表达（转基因表达与疾病的临床评分和组织病理学特征的相关性以及补体基因表达的检查）体内与疾病动力学和严重性相关）。 我们相信，本申请中提出的研究将产生关于补体在脱髓鞘疾病中的作用的重要新信息。 这些研究也将为将来研究补体抑制生物试剂作为MS治疗佐剂的潜力提供基础。

Our goal in this study is to determine the role(s) complement plays in the cellular infiltration, inflammation and demyelination in experimental allergic encephalomyelitis (EAE), the animal model for multiple sclerosis (MS). There is substantial evidence, in both MS and EAE, that complement is activated and may contribute to the inflammation and cellular destruction associated with these demyelinating diseases. There is also evidence, from our laboratory and others, that complement proteins and receptors are synthesized and expressed in the central nervous system (CNS), suggesting that local complement production may contribute to early disease development. One way to determine if complement contributes to the pathogenesis of demyelinating disease is to inhibit it's activation. Previous attempts to inhibit complement in EAE have demonstrated that strategy is feasible, but suffered from technical drawbacks. To overcome these limitations, we have developed a transgenic mouse model in which a soluble murine-specific inhibitor, sCrry, is produced by astrocytes using a glial fibrillary acidic protein (GFAP) promoter construct. We propose that blocking complement activation specifically in the CNS, using this new transgenic animal, provides a unique and better model system to determine the various roles that complement mediates in the development of the inflammatory response in EAE. We hypothesize that inhibiting complement activation using this model will reduce the cellular infiltration and demyelination, characteristic of EAE. In this application, we propose to 1) complete the analysis of our sCrry-expressing founder lines, 2) perform a kinetic analysis and comparison of clinical scores and histopathology of sCrry and control mice in EAE (extent and identification of infiltrating cell types, extent of demyelination and axonal transection), and 3) and assess altered gene expression in sCrry and control mice in EAE (correlation of transgene expression with clinical scores and histopathological features of the disease and examination of complement gene expression in vivo in relation to disease kinetics and severity). We believe the studies proposed in this application will generate significant new information on the role of complement in demyelinating disease. These studies will also provide the basis for future work examining the potential for complement-inhibiting biological reagents as therapeutic adjuvants in MS.