Transgenic Mouse Model for Echovirus Pathogenesis

描述(申请人提供)：动物病毒启动其复制能力 通过附着到细胞膜受体进行循环，然后进入 手机。对脊髓灰质炎病毒受体PVR的鉴定对于 脊髓灰质炎转基因小鼠模型的建立。虽然这是一个很棒的 Deal对脊髓灰质炎病毒感染的发病机制知之甚少 了解艾柯病毒是如何致病的。ECHO病毒很重要 人类病原体，导致无菌性脑膜炎、新生儿败血症、 心肌炎、红疹、呼吸系统疾病和胃肠道疾病。 因为大多数埃科病毒株只在人类身上复制，所以小鼠模型不能。 可用于研究ECHO病毒的致病机制。这项工作的目的是 建议分离出表达人类细胞受体的转基因小鼠 为研究ECHO病毒的致病机制建立新的动物模型。 我们建议分离表达异二聚体VLA-2的转基因小鼠 整合素由α2和β1亚基组成，已知为细胞 Echo病毒1型和8型的受体。计划有三个具体目标。第一, 编码VLA-2的α2和β1亚基的人cDNA将被放置在 人β-肌动蛋白启动子的转录调控，并用于 建立同时表达这两种蛋白的转基因小鼠。第二，纸巾和 将对VLA-2转基因小鼠的器官进行Northern和Western印迹检测 VLA-2RNA和蛋白表达的免疫组织化学分析。 第三，VLA-2转基因小鼠对ECHO病毒1型的易感性 不同途径的感染以及病毒复制的位置 接种后，将被测定。此外，Echo病毒能够 通过轴突从肌肉和肠道传播到中枢神经系统 运输将被确定。第四，将进行实验以确定是否 转基因VLA-2在ECHO病毒1型感染过程中的免疫应答 老鼠。基因敲除小鼠将被用来研究免疫反应在 限制或加重艾柯病毒的发病机制。转基因动物模型将 使对ECHO病毒复制部位、病毒机制的研究成为可能 传播，并以艾柯病毒的组织嗜性为基础。成立一家小型企业 动物模型可能会提供有关ECHO病毒发病机制的信息， 刺激Echo病毒疫苗或抗病毒药物的开发。

DESCRIPTION (provided by applicant): Animal viruses initiate their replicative cycles by attaching to a cellular membrane receptor, followed by entry into the cell. Identification of the poliovirus receptor, Pvr, was essential for the establishment of a transgenic mouse model for poliomyelitis. Although a great deal is known about the pathogenesis of poliovirus infection, relatively little is understood about how echoviruses cause disease. Echoviruses are important human pathogens and are responsible for aseptic meningitis, neonatal sepsis, myocarditis, exanthemas, respiratory illness and gastrointestinal disease. Because most echovirus strains only replicate in humans, a mouse model has not been available for studying echovirus pathogenesis. The object of work in this proposal is to isolate transgenic mice that express human cell receptors for echovirus, to establish a new animal model for studying echovirus pathogenesis. We propose to isolate transgenic mice that express VLA-2, a heterodimeric integrin consisting of alpha2 and beta1 subunits, which is known to be the cell receptor for echovirus types 1 and 8. Three specific aims are planned. First, human cDNA encoding the alpha2 and beta1 subunits of VLA-2 will be placed under the transcriptional control of the human beta-actin promoter, and used to establish transgenic mice that express both proteins. Second, tissues and organs of VLA-2 transgenic mice will be examined by Northern and Western blot analysis and immunohistochemistry for expression of VLA-2 RNA and protein. Third, the susceptibility of VLA-2 transgenic mice to echovirus type 1 infection by different routes, as well as the location of virus replication after inoculation, will be determined. In addition, the ability of echovirus to spread from muscle and intestine to the central nervous system by axonal transport will be determined. Fourth, experiments will be done to determine if an immune response is mounted during echovirus 1 infection of VLA-2 transgenic mice. Knock-out mice will be used to study the role of the immune response in limiting or exacerbating echovirus pathogenesis. Transgenic animal models would enable studies on the sites of echovirus replication, the mechanisms of viral spread, and the basis of echovirus tissue tropism. Establishment of a small animal model may yield information about echovirus pathogenesis that could stimulate development of echovirus vaccines or antiviral drugs.