EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS--INHIBITION OF C

我们在这项研究中的目标是确定(S)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)的细胞渗透、炎症和脱髓鞘中所起的作用。在MS和EAE中都有大量证据表明，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。来自我们实验室和其他实验室的证据也表明，补体蛋白和受体在中枢神经系统(CNS)中合成和表达，表明局部补体的产生可能有助于早期疾病的发展。确定补体是否与脱髓鞘疾病的发病机制有关的一种方法是抑制其激活。以前抑制EAE补体的尝试已经证明，该策略是可行的，但存在技术缺陷。为了克服这些局限性，我们开发了一种转基因小鼠模型，在该模型中，星形胶质细胞利用胶质纤维酸性蛋白(GFAP)启动子构建产生一种可溶的小鼠特异性抑制物sCrry。我们认为，利用这种新的转基因动物，在中枢神经系统特异性地阻断补体激活，为确定补体在EAE炎症反应发展中的各种中介作用提供了一个独特的、更好的模型系统。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。在这一应用中，我们建议1)完成我们的sCrry表达创立者系的分析，2)在EAE中对sCrry和对照小鼠的临床评分和组织病理学进行动力学分析和比较(浸润性细胞类型的范围和鉴定、脱髓鞘程度和轴突横切)，以及3)在EAE中评估sCrry和对照小鼠中基因表达的变化(转基因表达与疾病临床评分和组织病理学特征的相关性，以及体内补体基因表达与疾病动力学和严重程度的关系)。我们相信，在这项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。这些研究还将为今后研究补体抑制生物试剂作为MS的治疗佐剂的可能性提供基础。

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.