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

项目 3 - 登革热病毒和疫苗的离体免疫分析

基本信息

批准号：
10153665
负责人：
Ana Fernandez-Sesma
金额：
$ 30万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10153665
关键词：
Biological Biological Assay Biological Markers Cells Clinical Research Clinical Trials Collaborations Data Data Analyses Death Rate Dendritic Cells Dengue Dengue Fever Dengue Hemorrhagic Fever Dengue Infection Dengue Shock Syndrome Dengue Vaccine Dengue Virus Development Disease Event Failure Fever Generations Genes Genetic Genetic Determinism Genomics Geography Hemorrhage Human Immune Immune Evasion Immune Targeting Immune response Immunologic Monitoring Immunology Incidence Infection Innate Immune Response Laboratories Langerhans cell Life Cycle Stages Mediating Modeling Molecular Natural Immunity Nicaragua Patients Peripheral Blood Mononuclear Cell Property Proteins Proteome Proteomics Public Health Research Restriction Mapping Role Sampling Serotyping Severities Shock Skin Small Interfering RNA Supportive care System Systems Biology Therapeutic Tropical Climate Vaccination Vaccines Validation Viral Virus Diseases Virus Replication adaptive immune response cell type cohort data dissemination data management data modeling design epigenome functional genomics immune activation in vivo innate immune mechanisms macrophage monocyte mosquito-borne prototype severe dengue tool transcriptome vaccine candidate vaccine trial virus host interaction

项目摘要

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.

登革热是引起人类疾病的最流行的蚊媒病毒性疾病。登革热病可能表现为非特异性发热性疾病、登革热 (DF)，或更严重的感染，其特征为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)。目前还没有针对登革热病毒（DENV）的疫苗或疗法获得许可。我们实验室有优化并利用人类离体病毒感染模型，这对于阐明病毒感染非常有用 DENV 在特定细胞类型中的先天免疫逃避机制。了解病毒与宿主的相互作用在 DENV 感染期间，参与免疫反应产生的分子机制是之一导致开发有效和安全的疫苗和疗法的关键因素。该项目将使用两个强大的人类系统来分析对 DENV 感染的体外免疫反应 PBMC 和 DC 是人类 DENV 感染的目标。 DENV 感染的控制系统捕获病毒与宿主相互作用的早期事件。我们将分析对 DENV 原发性的先天免疫反应尼加拉瓜流行的分离株（项目 1）和 DENV 疫苗株（武田疫苗公司，项目 2）。选定的菌株将用于深入表征先天免疫反应、细胞与基因组学（核心 B）、蛋白质组学（核心 C）合作的转录组、表观基因组和蛋白质组病毒暴露后 PBMC 或 DC 中的免疫监测核心（核心 D）。我们将鉴定宿主蛋白差异介导先天免疫反应并影响相同 DENV 分离株的病毒复制并通过 DC 中的 siRNA 筛选来生产疫苗株。数据将由数据分析和分析部门进行管理和分析建模核心（核心 E）和数据管理和传播核心（核心 F）定义参数和建立对登革热病毒先天免疫至关重要的细胞和分子网络。网络随后将与项目 1（自然感染）和 2（疫苗试验）中体内研究产生的结果相结合。此外，我们的离体系统将通过全局分析来识别基因组，以设计有针对性的检测方法用于项目 1 和 2 中的体内研究，并验证对先天免疫重要的特定基因的作用体内鉴定出 DENV 感染。