TICK DETERMINANTS OF POWASSAN VIRUS TRANSMISSION

项目摘要： 蜱传疾病继续在美国出现。Powassan病毒（POWV），由 莱姆病的传播媒介肩胛硬蜱是蜱传脑炎病毒（TBEV）复合体的成员。它 引起脑炎，包括约10%病例中的致命性神经侵入性疾病， 据记载，50%的幸存者有神经系统后遗症。很明显， 确诊病例数量的增加和地理范围的扩大（USGS，2015; CDC）。POWV 通过受感染的蜱传播给人类，并且POWV的成功传播可以在感染后三小时内发生。 肩胛硬蜱附着。成功的蜱虫喂养是由一系列的寄生虫促进的。 蜱唾液中的因子/蛋白质，其分泌到蜱在脊椎动物上产生的摄食池中 主持人皮肤作为宿主-病毒-蜱相互作用的界面;因此，本提案的目的是 在检查蜱组分的同时，研究感染了POWV的蜱对皮肤宿主的免疫调节作用 唾液，导致更有利环境病毒传播，并确定唾液成分， 增强POWV感染。我们将通过探索以下具体目标来检验我们的假设：（1）调查 POWV感染的蜱唾液在调节叮咬部位的免疫应答以促进POWV 传播和传播;（2）鉴定增强POWV的特定蜱唾液蛋白 传输拟议的研究是创新和新颖的，因为我们将首次研究 蜱-宿主界面上蜱传病毒增强作用的分子机制。我们还将 使用系统生物学方法同时研究POWV感染的宿主和媒介属性。 这是以前从未尝试过的。拟议的项目还将在现场识别POWV感染细胞 用激光扫描细胞仪来检测蜱虫的喂养，这将为我们提供精确的类型识别， 与早期蜱虫进食时间点相关的免疫细胞的数量、时间和位置。我们还将确定 增强POWV感染和传播的唾液蛋白，并检查这些蛋白的抗体 蛋白质将减弱病毒感染。这项拟议的工作将对该领域作出重大贡献 因为它将采用系统生物学方法，尖端技术和创新理念， 了解机制，并确定唾液成分，导致更有利的环境， 蜱传病毒传播总的来说，这些研究将填补我们目前知识/理解的空白， 蜱传病毒感染，继续增加流行，并提供了一个新的调查线， 蜱-病原体-宿主界面这反过来又会导致制定反措施， 病毒感染。

Project Summary: Tick borne diseases continue to emerge in the United States. Powassan virus (POWV), transmitted by the Lyme disease vector Ixodes scapularis, is a member of the tick-borne encephalitis virus (TBEV) complex. It causes encephalitis, including fatal neuroinvasive disease in approximately 10% of cases, and long-lasting neurological sequelae have been documented in 50% of survivors. POWV has recently re-emerged, as evident by the increase in number and expanding geographic range of confirmed cases (USGS, 2015; CDC). POWV transmitted to humans by infected ticks, and successful transmission of POWV can occur within three hours of Ixodes scapularis attachment. Successful tick feeding is facilitated by a collection of pharmacologically active factors/proteins in tick saliva, which are secreted into the feeding pool that the tick creates on the vertebrate host. The skin serves as the interface of host-virus-tick interactions; therefore, the objective of this proposal is to investigate cutaneous host immunomodulation by POWV-infected ticks while examining components of tick saliva that lead to a more favorable environment for virus transmission, and identify salivary components that enhances POWV infection. We will test our hypothesis by exploring the following specific aims: (1) Investigate the role of POWV infected tick saliva in modulating immune responses at the bite site to facilitate POWV transmission and dissemination; (2) Identify the specific tick salivary protein/s that potentiate POWV transmission. The proposed research is innovative and novel because we, for the first time, will examine the molecular mechanisms involved in potentiation of a tick borne virus at the tick-host interface. We will also investigate concurrently both host and vector attributes of POWV infection using a systems biology approach. This has never been attempted before. The proposed project will also identify POWV infected cells at the site of tick feeding using Laser Scanning Cytometer that will provide us with precise identification of the type, number, time, and location of immune cells associated with early tick feeding time points. We will also identify the salivary protein/s that enhances POWV infection and dissemination, and also check if antibodies to these proteins will attenuate virus infection. This proposed work will make a significant contribution to the field because it will employ systems biology approach, cutting-edge technologies and innovative ideas to understand the mechanisms and identify salivary components that lead to a more favorable environment for tick-borne virus transmission. Overall, these studies will fill the gaps in our current knowledge/understanding of tick-borne viral infection that continues to increase in prevalence, and provide a new line of investigation of the tick-pathogen-host interface. This, in turn, will lead to development of countermeasures that will attenuate the virus infection.