Stat 1 modification for antiviral defense

病毒是急性和慢性疾病最常见的原因之一，新发现的病毒 继续导致对现有疫苗产生抵抗力的突发疾病。尽管这个问题的范围很广， 大多数病毒感染，特别是新型感染的诊断准确性和治疗效果， 效果非常有限。为了解决这个问题，我们制定了一项替代战略， 来定义和改善抗病毒宿主反应。我们特别关注呼吸道感染 因为这是一种常见自然感染途径，也可能是生物恐怖主义威胁的传播途径。在这 在这方面，我们已经开发了一种用于人气道上皮细胞的原代培养和感染的系统， 表现出对在体内发现的行为的高度保真度。我们对这种上皮细胞模型的分析与 相应的小鼠模型表明，对呼吸道病毒感染的抗病毒防御依赖于 关键在于干扰素（IFN）激活气道上皮细胞中的Stall信号分子。基于 根据这些发现，我们认为改善气道上皮细胞的Stall功能可以增强抗病毒作用， 防御因此，我们设计了一种具有战略性双半胱氨酸突变体的改良Stall（设计为 Slal 1-CC），其对内源性干扰素水平高度反应。因此，Stall-CC的表达应 增强干扰素信号传导功能并提供对病毒复制的更好控制。的确，我们的 初步研究表明，Stall-CC表达显著降低病毒复制水平 在体外（使用转导的细胞）和体内（使用转基因或基因转移技术）。比如说， 携带Stall-CC转基因或用Stall-CC基因转移载体处理的小鼠被完全保护 对抗迄今为止所研究的三种病毒中的每一种造成的致命感染。此外，我们有 观察到Stall-CC表达没有毒性，与直接给药的情况相反， 干扰素本身的过度表达。在这个建议中，我们的目标是将我们的方法扩展到紧急的研究。 具有流行能力的病原体在人群中传播。为此，我们的目标是 建立这些新病毒的感染能力和诊断特征，这是一种新的治疗策略， 以及对这些病原体的先天免疫的更好理解。

Viruses are among the most frequent causes of acute and chronic illness, and newly discovered viruses continue to cause emergent diseases that resist established vaccines. Despite the scope of this problem, the accuracy of diagnosis and efficacy of treatment for most viral infections, especially new types of infections, is very limited in effectiveness. To address this issue, we have developed an alternative strategy that is aimed at defining and then improving the antiviral host response. We focus particularly on respiratory infection since this is a common route of natural infection and a likely route for delivery of bioterrorist threats. In that regard, we have developed a system for primary-culture and infection of human airway epithelial cells that exhibits high fidelity to behavior found in vivo. Our analysis of this epithelial cell model in concert with a corresponding mouse model indicates that antiviral defense against respiratory viral infection depends critically on interferon (IFN) activation of the Stall signaling molecule in airway epithelial cells. Based on these findings, we proposed that improving Stall function in airway epithelial cells would enhance antiviral defense. Accordingly, we engineered a modified Stall with strategic double-cysteine mutalions (designaled Slal1-CC) that is hyperresponsive to endogenous interferon levels. Expression of Stall-CC should thereby enhance interferon-signaling function and provide for better control of viral replication. Indeed, our Preliminary Studies demonstrate that Stall-CC expression markedly decreases Ihe level of viral replication both in vitro (using transduced cells) and in vivo (using transgenic or gene transfer technology). For example, mice carrying the Stall-CC transgene or treated with a Stall-CC gene-transfer vector are fully protected against otherwise lethal infections due to each of the three viruses studied thus far. Furthermore, we have observed no toxicities of Stall-CC expression, in contrast to the situation for direct administralion or overexpression of interferon itself. In this proposal, we aim to extend our approach to the study of emergent pathogens with the capability for epidemic spread through the human population. In doing so, we aim to establish infection capabilities and diagnostic signatures for these new viruses, a new therapeutic strategy for these pathogens, and a better understanding of innate immunity to these agents.