新型埃博拉病毒样颗粒构建及与宿主细胞互作效应分析

Ebola hemorrhagic fever is caused by Ebola viruses, which is a severe contact disease in humans and non-human primates with case fatality rates of up to 90% in human outbreaks, and also currently one of the most serious diseases to human. Work with infectious Ebola viruses has to be performed in a maximum containment laboratory under biosafety level (BSL) 4 conditions, which restricts the work. In order to study the viruses or to develop novel therapeutics under BSL2 conditions, researchers rely either on recombinant over-expression of Ebola virus proteins, or on lifecycle modeling systems, both of which can be worked with in negative infectious Ebola virus. Our study expects to determine the key genes and amino acids which impact virulence of Ebola virus through construction multiple life cycles of Ebola virus-like particles. Detection type I interferon response after Ebola virus-like particles infection decide the virus gene that participate in avoiding host innate immune response. To determine the different expressed proteins after Ebola virus infection, we can use proteomic technology to acquire the key regulatory proteins data and intend to clarify the mechanism of Ebola infection and host immune protection.

埃博拉出血热是一种由埃博拉病毒引起，出现于人和灵长动物身上一种高度接触性烈性传染病，病死率可高达90％，是目前对人类危害最严重的疾病之一。与感染埃博拉病毒的工作仅限于世界范围内生物安全等级最高的封闭性实验室内（BSL4）进行而严重制约着这项工作的开展。为了在BSL2条件下研究埃博拉病毒，开发新的治疗手段，通过对埃博拉病毒蛋白重组表达或对其生命周期建模系统，这两种方法都可以使不具有感染性的埃博拉病毒颗粒被加工。本研究拟通过构建多生命周期重组埃博拉病毒样颗粒，确定决定埃博拉病毒毒力的关键基因和氨基酸位点。同时检测埃博拉病毒感染后宿主I型干扰素反应，确定参与埃博拉病毒逃避宿主天然免疫反应的病毒基因。通过蛋白质组学技术对埃博拉病毒引起宿主差异表达蛋白进行分析建谱，获得机体在抵御埃博拉病毒侵袭过程中的关键调节蛋白相关数据，为阐明埃博拉感染致病机制及机体的免疫防护作用提供理论依据。

埃博拉出血热是一种由埃博拉病毒引起，出现于人和灵长动物身上一种高度接触性烈性传染病，病死率可高达90％，是目前对人类危害最严重的疾病之一。与感染埃博拉病毒的工作仅限于世界范围内生物安全等级最高的封闭性实验室内（BSL4）进行而严重制约着这项工作的开展。为了在BSL2条件下研究埃博拉病毒，本项目成功构建了具有多个生命周期的埃博拉病毒样颗粒，并通过对埃博拉感染后宿主RNA表达谱分析，发现circRNA-chr19 通过与miR-30b-3p互作调控宿主CLDN18的表达，miR-150-3p可以靶向GP和VP40，抑制埃博拉病毒复制；同时还发现埃博拉sGP蛋白抑制宿主干扰素表达，在埃博拉病毒免疫逃逸中发挥了重要作用。为我们下一步埃博拉病毒致病机理及机体的免疫防护作用提供理论依据。