Pathogenesis of Jeilongvirus

项目概要 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'。联合PV 编码一种 TM（跨膜）蛋白，该蛋白与任何已知蛋白没有同源性，并且不存在于任何已知蛋白中。 其他分类的副粘病毒。 2006年从人肾系膜中分离到北龙病毒（BeiPV） 细胞（HMC）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 是一种啮齿动物病毒。由于其独特的基因组结构，一个新属——杰龙病毒， 建议将 JPV 和 BeiPV 分类为副粘病毒科。 Tailam 病毒 (TlmPV)，分离自 2011年香港锡金大鼠的肾脏，与JPV和BeiPV具有相同的基因组结构， 表明它是节龙病毒属的成员。 2014年，杰龙病毒属的一个可能成员是 从英国灰松鼠肾细胞的原代培养物中鉴定出。此外，RNA 序列 JPV 样病毒已在非洲、欧洲和中国的啮齿动物和蝙蝠中发现（个人资料） 自2012年以来的通讯）表明，杰龙病毒分布广泛，感染多种哺乳动物。 目前，人们对这一类新出现的病毒知之甚少。 JPV 抗体有 在啮齿类动物、猪和人类中均已检测到，表明 JPV 具有广泛的宿主范围和人畜共患性 潜力。在蝙蝠体内发现节龙病毒的事实说明了它们的人畜共患潜力，因为蝙蝠 被认为是许多新兴人畜共患病毒（例如 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，可以合理地预期节龙病毒属中的一种病毒在体内具有致病性。 人类。研究JPV很重要，原因如下：（1）以防致病性人类杰龙病毒 一旦出现，我们就会对这一类病毒有了了解； (2) JPV可以作为研究的模型 副粘病毒小疏水（SH）蛋白的功能； (3) JPV 的 TM 的独特之处在于 副粘病毒家族中唯一在细胞间融合中发挥关键作用的病毒蛋白，但它不是 对于病毒与细胞的融合至关重要。 我们选择 JPV 作为杰龙病毒的原型，因为我们已经鉴定出一种 JPV 毒株，它是 对实验室小鼠具有致病性。在本提案中，我们计划对JPV进行全面分析，重点关注 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， 我们建立了体内发病机制研究的动物模型和反向遗传学系统 操纵 JPV 的 RNA 基因组。此外，我们还生成了多克隆和单克隆抗体 对于所有 JPV 蛋白。在本提案中，我们将重点关注以下具体目标：（1）阐明 SH及其功能机制；（2）了解TM的体外和体内功能。 JPV 代表了一类尚未被研究的新病毒。我们提出的工作将指导我们 制定潜在的对策，以防其中一种对人类致病，并提供新的 有关病毒蛋白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.