Pathogenesis of Jeilongvirus

项目摘要 J副粘病毒（JPV）于20世纪70年代初在澳大利亚首次从啮齿动物中分离出来。它的基因组结构是 2005年确定。JPV基因组有8个基因，顺序为3 ′-N-P/V/C-M-F-SH-TM-G-L-5 ′。JPV 编码TM（跨膜）蛋白，其与任何已知蛋白质没有同源性，并且不存在于任何已知蛋白质中。 其他分类的副粘病毒。2006年，从人肾系膜组织中分离到北龙病毒（BeiPV 细胞（HMCs）作为来自大鼠细胞系的污染物。BeiPV与JPV具有相同的基因组结构。研究 表明BeiPV是啮齿类病毒。由于其独特的基因组结构，一个新的属，吉隆病毒， 将JPV和BeiPV划分为副粘病毒科。泰兰病毒（TlmPV），分离自 2011年在香港的一只锡金大鼠的肾脏中发现，其基因组结构与JPV和BeiPV相同， 表明它是Jeilongvirus属的成员。2014年，一个可能的吉隆病毒属成员是 从英国灰松鼠肾细胞的原代培养物中鉴定。此外， 已经在非洲、欧洲和中国的啮齿动物和蝙蝠中发现了类似JPV的病毒（个人 自2012年以来，Jelong病毒（Jelong communication）的研究表明，该病毒分布广泛，感染多种哺乳动物。 目前，人们对这类新出现的病毒知之甚少。抗JPV的抗体 在啮齿动物、猪和人类中检测到，表明JPV具有广泛的宿主范围和人畜共患性。 潜力在蝙蝠中发现吉隆病毒的事实说明了它们的人畜共患病潜力，因为蝙蝠 被认为是许多新出现的人畜共患病毒的天然宿主，如SARS-CoV，Hendra和 尼帕病毒和埃博拉病毒。在哺乳动物副粘病毒的每一个属中，都有重要的人类副粘病毒。 病原体因此，可以合理地预期，其中一种病毒在Jeilong病毒属中是致病性的。 人类JPV的研究具有重要意义，主要有以下几点：（1）如果出现致病性的人Jeilong病毒， （2）JPV可以作为研究JPV的模型，但JPV的致病机理尚不清楚。 副粘病毒的小疏水蛋白（SH）的功能;（3）JPV的TM是独特的， 副粘病毒家族中唯一在细胞间融合中起关键作用的病毒蛋白，但它不是 病毒与细胞融合所必需的。 我们选择JPV作为Jeilong病毒的原型，因为我们已经确定了JPV的一种毒株， 在实验室小鼠中致病。在本提案中，我们计划对JPV进行全面分析，重点是 了解SH和TM的功能及其在动物发病机制中的作用。为了实现这些目标， 我们已经建立了一种用于体内发病机制研究的动物模型和一种用于 操纵JPV的RNA基因组。此外，我们已经产生了多克隆和单克隆抗体， 所有的JPV蛋白质。在这一建议中，我们将重点关注以下具体目标：（1）阐明 （2）了解TM在体内外的功能。 JPV代表了一类尚未被研究的新病毒。我们建议的工作将指导我们 开发潜在的对策，以防其中一种在人类中致病，并提供新的 关于病毒蛋白质功能和进入过程的知识。

PROJECT SUMMARY J paramyxovirus (JPV) was first isolated from rodents in the early 1970s in Australia. Its genome structure was determined In 2005. The JPV genome has eight genes in the order of 3'-N-P/V/C-M-F-SH-TM-G-L-5'. JPV encodes a TM (transmembrane) protein that has no homology to any known proteins and does not exist in any other classified paramyxoviruses. In 2006, Beilong virus (BeiPV) was isolated from human kidney mesangial cells (HMCs) as a contaminant from a rat cell line. BeiPV has the same genome structure as JPV. Studies indicate that BeiPV is a rodent virus. Because of their unique genome structure, a new genus, Jeilongvirus, was proposed to classify JPV and BeiPV within the paramyxovirus family. Tailam virus (TlmPV), isolated from the kidney of a Sikkim rat in Hong Kong in 2011, has an identical genome structure as JPV and BeiPV, indicating that it is a member of Jeilongvirus genus. In 2014, a likely member of Jeilongvirus genus was identified from the primary culture of grey squirrel kidney cells from the UK. In addition, RNA sequences of JPV-like viruses have been identified in rodents and bats in Africa, Europe, and China (personal communication) since 2012, indicating that Jeilongvirus is widely distributed and infects a variety of mammals. At present, very little is known about this new and emerging class of viruses. Antibodies against JPV have been detected in rodents, pigs, and humans, suggesting that JPV has a broad host range and zoonotic potential. The fact that Jeilongviruses have been identified in bats illustrates their zoonotic potential, since bats are thought to be the natural reservoirs for many emerging zoonotic viruses such as SARS-CoV, Hendra and Nipah viruses and Ebola virus. In every genus of mammalian paramyxoviruses, there are important human pathogens. Thus, it is reasonable to expect that one of the viruses in the Jeilongvirus genus is pathogenic in humans. It is important to study JPV for following reasons: (1) in case a pathogenic human Jeilongvirus emerges, we will have knowledge about this class of viruses; (2) JPV can be used as a model for the study of the functions of the small hydrophobic (SH) protein of paramyxoviruses; and (3) TM of JPV is unique in that it is the only viral protein in the paramyxovirus family that plays a critical role in cell-to-cell fusion, but it is not essential for virus-to-cell fusion. We have chosen JPV as a prototype of Jeilongvirus, because we have identified a strain of JPV that is pathogenic in laboratory mice. In this proposal, we plan to carry out a comprehensive analysis of JPV, focusing on understanding the functions of SH and TM and their roles in pathogenesis in animals. Towards these goals, we have established an animal model for in vivo pathogenesis studies and a reverse genetics system for manipulating the RNA genome of JPV. In addition, we have generated polyclonal and monoclonal antibodies for all JPV proteins. In this proposal, we will focus on following specific aims: (1) Elucidating the functions of SH and the mechanisms of its functions and (2) Understanding the functions of TM in vitro and in vivo. JPV represents a new class of viruses that have not been studied. Our proposed work will guide us in developing potential countermeasures in case one of them is pathogenic in humans and provide new knowledge regarding viral protein functions and entry processes.