Generation of a Single-Cycle Virus to Study the Pathogenesis of Nipah Virus

描述（由申请人提供）：1999年，马来亚猪爆发的呼吸系统疾病传播给人类，引起热性脑炎，死亡率很高。这次暴发的原因被确定为尼帕病毒，这是最近在澳大利亚的马和人身上出现的亨德拉病毒的亲戚。这两种病毒的宿主似乎都是果蝠，但传播给人类和动物的途径尚不清楚。自1999年以来，孟加拉国发生了三次NiV疫情。NiV感染对人类的死亡率很高，在40%至75%之间，并可在许多重要的农业动物中引起严重疾病。因此，NiV是用于生物恐怖的候选物。我们对NiV感染的理解一直受到其致死性和需要使用BSL-4条件进行研究的阻碍。这个应用程序的目标是使NiV研究更容易执行。根据从研究另一种副粘病毒呼吸道合胞病毒（RSV）中获得的知识，将构建含有NiV基因组拷贝的质粒，其中两个糖蛋白基因将被绿色荧光蛋白基因和抗生素抗性基因取代。由此产生的NiV“复制子”应该能够自主复制，就像现有的RSV复制子一样。糖蛋白将以反式形式提供，以生成“单循环病毒”（SCV）， RSV复制子已经成功地产生了。由于Ni-SCV仅在一轮内具有传染性，因此在较低水平的BSL条件下进行研究应该是可能的。一旦Ni-SCV产生，将Ni-SCV感染与NiV感染进行比较。这种新型的Ni-SCV系统将用于研究NiV的发病机制，包括NiV干扰先天免疫反应的能力。最初的研究将集中在P/C/V/W基因上，以剖析这些蛋白质在NiV感染背景下诱导干扰素的作用。在未来，Ni-SCV的研究可以扩展到探索趋化因子和细胞因子表达的诱导或抑制以及其他致病机制，并为筛选候选抗病毒药物提供一种手段。

DESCRIPTION (provided by applicant): In 1999, an outbreak of respiratory disease in Maylasian pigs spread to humans causing febrile encephalitis with a high mortality rate. The cause of this outbreak was identified as Nipah virus (NiV), a relative of Hendra virus that had recently emerged in horses and humans in Australia. The reservoir of both viruses appears to be fruit bats, though the route of transmission to humans and animals is not clear. Since 1999, there have been three outbreaks of NiV in Bangladesh. NiV infection has a high mortality rate in humans, between 40% and 75%, and can cause severe disease in a number of animals of agricultural importance. Therefore, NiV is a candidate for use in bioterror. Our understanding of NiV infection has been hampered by its lethal nature and the need to use BSL-4 conditions to study it. The goal of this application is to make NiV studies easier to perform. Based on knowledge gained from studying another paramyxovirus, respiratory syncytial virus (RSV), a plasmid will be constructed containing a copy of the NiV genome in which the two glycoprotein genes will be replaced with genes for the green fluorescent protein and for an antibiotic resistance gene. The resulting NiV "replicon" should be able to replicate autonomously, as does the existing RSV replicon. Glycoproteins will be provided in trans to generate "single-cycle virus" (SCV), as has been successful for the RSV replicon. Because the Ni-SCV will be infectious for one round only, studies under lower level BSL conditions should be possible. Once the Ni-SCV has been generated, Ni-SCV infection will be compared to infection with NiV. This novel Ni-SCV system will be used to study the pathogenesis of NiV, including the ability of NiV to interfere with the innate immune response. Initial studies will focus on the P/C/V/W genes to dissect the role of each of these proteins in the induction of interferon, in the context of NiV infection. In the future, work with the Ni-SCV could be expanded to explore the induction or suppression of chemokine and cytokine expression, and other pathogenic mechanisms, as well as to provide a means to screen candidate antiviral agents.