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

• 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
• 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

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.

登革热是最流行的蚊子传播的病毒性疾病，引起人类疾病。登革热可能 表现为非特异性发热性疾病，登革热（DF），或表现为更严重的感染， 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）疫苗或治疗剂。本实验室 优化并利用了用于病毒感染的人离体模型，这对于阐明 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 在登革病毒感染和参与免疫反应的产生的分子机制是一个 这些关键因素将导致开发有效和安全的疫苗和疗法。 该项目将使用两个强大的人类免疫系统分析对DENV感染的免疫应答。 PBMC和DC是人类DENV感染的靶点。这种控制登革病毒感染的系统 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 尼加拉瓜流行的分离株（项目1）和DENV疫苗株（Takeda Vaccines Inc.，项目2）。 然后将选择的菌株用于深入表征先天免疫应答、细胞免疫应答和细胞免疫应答。 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质 差异介导先天免疫应答并影响相同DENV分离株的病毒复制 并通过在DC中的siRNA筛选疫苗株。数据将由数据分析部管理和分析， 建模核心（核心E）和数据管理和传播核心（核心F）来定义参数和 构建对DENV先天免疫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）中体内研究产生的结果相结合。 此外，我们的离体系统将用于通过全局分析来识别基因组，以设计靶向测定法 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， 体内鉴定的DENV感染。