Project 3 - Ex vivo immune profiling of dengue viruses and vaccines

登革热是最常见的由蚊子传播的病毒性疾病，导致人类患病。登革热疾病可能 表现为一种非特定的发热性疾病，登革热(DF)，或一种更严重的感染，其特征是 出血或循环衰竭或休克、登革出血热(DHF)或登革休克综合征(DSS)。 目前还没有获得许可的登革热病毒(DENV)疫苗或疗法。我们的实验室有 优化和利用了人类体外病毒感染模型，这些模型对阐明病毒非常有用 DENV在特定细胞类型中逃避先天免疫的机制。了解病毒与宿主的相互作用 在DENV感染期间，参与产生免疫反应的分子机制是 将导致开发有效和安全的疫苗和疗法的关键因素。 这个项目将使用两个强大的人类免疫系统来分析体外对DENV感染的免疫反应。 PBMC和DC，它们是人类DENV感染的目标。这个受控制的DENV感染系统 捕获病毒与宿主交互的早期事件。我们将描述对DENV初级病毒的先天免疫反应 尼加拉瓜流行的病毒分离株(项目1)和DENV疫苗株(武田疫苗公司，项目2)。 然后，选定的菌株将用于深入表征先天免疫反应，即细胞 转录组、表观基因组和蛋白质组与基因组学(核心B)、蛋白质组学(核心C)合作 和免疫监测核心(核心D)在PBMC或DC中的病毒暴露。我们将鉴定宿主蛋白 在相同的DENV分离株中以不同的方式介导先天免疫反应并影响病毒复制 在DC中通过siRNA筛选疫苗菌株。数据将由数据分析和 建模核心(核心E)和数据管理和传播核心(核心F)以定义参数和 建立对DENV天然免疫至关重要的细胞和分子网络。网络将随后 与项目1(自然感染)和项目2(疫苗试验)体内研究产生的结果相结合。 此外，我们的体外系统将通过全球分析来识别多组基因，以设计有针对性的分析 用于项目1和2的体内研究，并验证对先天免疫重要的特定基因的作用 在体内发现了DENV感染。

Dengue is the most prevalent mosquito-borne viral disease causing disease in humans. Dengue disease may present as a non-specific febrile illness, dengue fever (DF), or as a more severe infection marked by hemorrhage or circulatory failure or shock, dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS). There are currently no licensed vaccines or therapeutics for dengue virus (DENV). Our laboratory has optimized and utilized human ex vivo models for virus infections, which have been very useful for elucidating mechanisms of innate immune evasion by DENV in specific cell types. Understanding virus-host interactions during DENV infections and the molecular mechanisms involved in the generation of immune responses is one of the crucial factors that will lead to the development of effective and safe vaccines and therapeutics. This project will analyze immune responses to DENV infections ex vivo using two robust systems of human PBMCs and DCs, which are targets for DENV infection in humans. This controlled system of DENV infection captures early events of the virus-host interaction. We will profile innate immune responses to DENV primary isolates circulating in Nicaragua (Project 1) and DENV vaccine strains (Takeda Vaccines Inc., Project 2). Selected strains will be then used for the in depth characterization of the innate immune response, cellular transcriptome, epigenome and proteome in collaboration with the Genomics (Core B), Proteomics (Core C) and immune monitoring Core (Core D) in PBMCs or DCs upon viral exposure. We will identify host proteins that differentially mediate innate immune responses and impact viral replication with the same DENV isolates and vaccines strains by siRNA screens in DCs. Data will be managed and analyzed by the Data Analysis and Modeling Core (Core E) and the Data Management and Dissemination Core (Core F) to define parameters and build cellular and molecular networks important for innate immunity to DENV. Networks will be subsequently integrated with those generated from in vivo studies in Projects 1 (natural infections) and 2 (vaccine trials). Also, our ex vivo systems will serve to identify sets of genes through global analysis to design targeted assays for in vivo studies in Projects 1 and 2 and to validate the role of specific genes important for innate immunity to DENV infection identified in vivo.