A novel ex vivo model of West Nile virus infection in human lymphoid tissue

描述（由申请人提供）：西尼罗病毒（WNV）是中枢神经系统（CNS）的高毒力人类病原体，是美国流行性脑炎的最常见原因。由于没有针对西尼罗河病毒的疫苗或特异性抗病毒治疗，了解其发病机制是当务之急。感染后，西尼罗河病毒被运送到局部引流淋巴结，在那里病毒的初级扩增和关键的宿主防御反应被触发；然而，对于人类在这个交汇处发生的事件知之甚少。为了研究这一点，我们建立了西尼罗河病毒感染的离体人淋巴组织模型。在初步研究中，我们发现西尼罗河病毒在所有供体中都能强劲复制，并且可以检测到西尼罗河病毒感染的细胞。在本研究中，我们将使用两种不同致病性的西尼罗河病毒株来表征细胞对西尼罗河病毒感染的反应，并识别淋巴组织内的允许靶细胞（目的1），测量感染期间的炎症反应（目的2），并确定淋巴结内病毒复制优先发生的解剖环境（目的3）。了解人类系统中的这些病毒和细胞方面，以及这些方面在神经侵入性和非神经侵入性西尼罗河病毒株之间的差异，不仅可以初步了解早期感染事件，还可以为人类体内易感性提供新的假设。

DESCRIPTION (provided by applicant): West Nile virus (WNV) is a highly virulent human pathogen of the central nervous system (CNS) and the most common cause of epidemic encephalitis in the United States. Since there are no vaccines or specific antiviral treatments available for WNV, understanding its pathogenesis is a high priority. Following infection, WNV is transported to the local draining lymph nodes, where primary amplification of the virus and key host defense responses are triggered; however, there is very little known about the events that take place at this junction in humans. To study this, we developed an ex vivo human lymphoid tissue model of WNV infection. In preliminary studies, we show that WNV replicates robustly in all donors tested, and WNV-infected cells can be detected. In this application, we will use two strains of WNV that differ in their pathogenic potential in humans to characterize the cellular response to WNV infection and identify the permissive target cells within the lymphoid tissue (Aim 1), measure the inflammatory response during the infection (Aim 2), and determine the anatomical environment within the lymph node where virus replication preferentially occurs (Aim 3). Understanding these viral and cellular aspects in a human system and how these differ between a neuroinvasive and non-neuroinvasive strain of WNV will not only provide the first glimpse into early infection events, but also lead to new hypotheses for in vivo susceptibility in humans.