Dengue Human Immunology Project Consortium (DHIPC)

 描述（申请人提供）：我们的提案，人类免疫学项目联盟（DHIPC），是基于一个创新的战略，令人信服的初步研究，以及广泛的多学科合作网络，并将研究人类免疫反应1）在登革热病毒（DENV）感染期间或之后，2）在减毒活DENV疫苗接种之前和之后，使用高通量系统生物学方法，结合在充分表征的人类队列中进行的详细临床表型分析。长期目标是开发定义免疫应答的分子特征和与感染和疫苗接种结果相关的类别/指纹/概况。我们将使用“组学”技术平台，包括基因组学（核心B），RNAi筛选（项目3）和蛋白质组学（核心C），以及免疫分析（核心D），以研究来自健康供体的人类细胞（项目3）和DENV感染儿童的良好表征的人类队列，包括登革热流行地区的症状与非症状感染（项目1）。我们还包括人类减毒活DENV疫苗接种的研究（项目2），以预测免疫原性，并在未来将疫苗接种前或接种后不久确定的免疫特征与保护效力以及疫苗接种引发的不良事件相关联。数据分析和建模核心（Core E）将是将组学数据转化为生物学洞察力的引擎，数据管理传播核心（Core F）将作为中心生成的所有数据和资源的中央存储库，并确保这些材料不仅可以由该计划中的其他科学家访问，而且可以由更广泛的科学界访问。我们提出的系统免疫学研究的结果将为未来的假设驱动的研究奠定基础，并可能有助于开发诊断工具和定义与感染和疫苗接种结果相关的免疫特征。项目和核心由一个行政核心（核心A）监督，该核心具有报告和问责制的组织结构，以保持科学和财政的灵活性和责任。登革热具有重要的公共卫生意义，我们的项目有独特的机会将自然感染与体内和体外临床开发的减毒活疫苗进行比较，为研究感染和疫苗接种环境中的其他人类传染病建立新的范式。 项目-001：项目1 -自然登革病毒感染的免疫特征分析 项目负责人（PL）：伊娃哈里斯 描述（由申请方提供）：四种登革热病毒血清型（DENV 1 -4）引起人类最重要的蚊媒病毒性疾病，每年约有1亿例病例，但目前还没有批准的疫苗或抗病毒药。决定DENV感染后疾病结果的因素尚未得到很好的理解，并且被认为部分由宿主免疫应答介导。病毒因素和宿主先天免疫相互作用可能影响适应性免疫应答的质量，其可以提供保护或引起后续DENV感染的增强。登革人类免疫学项目联盟（DHIPC）的项目1专注于表征与自然DENV感染中的感染结果和疾病严重程度相关的免疫特征。项目1的总体方法是利用尼加拉瓜登革热长期持续研究的独特样本集，以发现与症状性登革病毒感染、严重登革热疾病和特异性适应性免疫反应相关的免疫因素。项目1将继续进行巴布亚新几内亚儿童登革热队列研究（PDCS），从记录良好的自然重复DENV感染中收集样本。它还将利用尼加拉瓜正在进行的以医院为基础的登革热研究，以调查严重与轻度登革热疾病，并将进行指数聚类研究，以提供无症状病毒血症个体的样本。所有样本均附有详细的临床数据。结合世界级的DHIPC核心，系统生物学方法将用于构建天然DENV感染期间先天免疫反应的特征。互补的尖端基因组学，转录组学，蛋白质组学，血清学和免疫表型分析方法的星座将使人类对登革热的免疫反应能够以前所未有的细节和复杂性进行解剖。目的1将表征具有不同临床结果的DENV感染中的免疫特征：使用索引群集研究的无症状和症状性DENV感染以及PDCS和基于医院的研究中的轻度与重度疾病。目的2将以感染的特定血清型顺序表征原发性和继发性DENV感染的免疫谱。目的3探讨天然免疫信号与抗体和细胞免疫应答之间的关系。将评价抗体中和、同种型和库，以及项目1中B细胞应答的幅度和表型特异性或交叉反应性。将通过细胞内阿法尼染色评价T细胞应答的多功能性。总体而言，项目1将能够识别可预测感染和疾病结果的生物标志物和免疫特征，从而有助于改善临床管理，疫苗设计和开发针对登革热的新型抗病毒疗法。

 DESCRIPTION (provided by applicant): Our proposal, Human Immunology Project Consortium (DHIPC), is based on an innovative strategy, compelling preliminary studies, and a wide network of multidisciplinary collaborations and will study the human immune responses 1) during or following dengue virus (DENV) infection, 2) before and after live attenuated DENV vaccination, using high-throughput systems biology approaches coupled with detailed clinical phenotyping in well-characterized human cohorts. The long-term goal is to develop molecular signatures that define immune response and categories/fingerprints/profiles that correlate with the outcome of infection and vaccination. We will use "omics" technology platforms including genomics (Core B), RNAi screens (Project 3) and proteomics (Core C), and immune profiling (Core D) to study human cells from healthy donors (Project 3) and well characterized human cohorts of DENV infected children that include symptomatic versus non-symptomatic infections in dengue endemic areas (Project 1). We also include studies of live attenuated DENV vaccinations in humans (Project 2), to predict immunogenicity, and in the future correlate immune profiles determined before or soon after vaccination with protective efficacy, and also with adverse events triggered by vaccination. The data analysis and modeling core (Core E) will be the engine for translating omics data into biological insight and the data Management Dissemination Core (Core F) will act as a central repository for all data and resources generated by the Center, and ensure that these materials are readily accessible by not only other scientists in the program, but also the broader scientific community. The results of our proposed systems immunology studies will create a foundation for future hypothesis-driven research and may help develop diagnostic tools and define immune profiles correlated with outcomes of infection and vaccination. The Projects and Cores are overseen by an Administrative Core (Core A), which has an organizational structure for reporting and accountability to maintain scientific and fiscal flexibility and responsibility. Dengue disease is of great public health importance and our program has the unique opportunity to compare natural infections with live attenuated vaccines under clinical development both in vivo and ex vivo, building a new paradigm for studying other human infectious diseases in the settings of infection and vaccination. Project-001: Project 1 - Immune Profiling of Natural Dengue Virus Infections Project Leader (PL): Eva Harris DESCRIPTION (as provided by applicant): The four dengue virus serotypes (DENV1-4) cause the most important mosquito-borne viral disease of humans, with ~100 million cases annually, yet no approved vaccines or antivirals exist. Factors that determine disease outcome following DENV infection are not well understood and are thought to be mediated in part by host immune responses. Viral factors and host innate immune interactions likely influence the quality of the adaptive immune response, which can either protection provide or cause enhancement in a subsequent DENV infection. Project 1 of the Dengue Human Immunology Project Consortium (DHIPC) focuses on characterizing immune signatures associated with infection outcomes and disease severity in natural DENV infections. The overall approach of Project 1 is to take advantage of unique sample sets from long-term ongoing studies of dengue in Nicaragua to enable discovery of immune factors associated with symptomatic DENV infection, severe dengue disease, and specific adaptive immune responses. Project 1 will continue the Nicaraguan Pediatric Dengue Cohort Study (PDCS) to collect samples from well-documented natural repeat DENV infections. It will also leverage an on-going hospital-based study of dengue in Nicaragua to enable investigation of severe vs. mild dengue disease and will conduct an index cluster study to provide samples from asymptomatic viremic individuals. Detailed clinical data will accompany all samples. In conjunction with the world-class DHIPC Cores, a systems biology approach will be used to construct a signature of innate immune responses during natural DENV infection. The constellation of complementary cutting-edge genomic, transcriptomic, proteomic, seromic, and immunophenotyping methods will enable the human immune response to dengue to be dissected with unprecedented detail and sophistication. Aim 1 will characterize immune profiles in DENV infections with different clinical outcomes: asymptomatic and symptomatic DENV infection using the index cluster study and mild vs. severe disease in the PDCS and the hospital-based study. Aim 2 will characterize immune profiles in primary and secondary DENV infections, in specific serotype order of infection. Aim 3 will investigate the association between innate immune signatures and antibody and cell-mediated immune responses. Antibody neutralization, isotype, and repertoire will be evaluated, as well as the magnitude and serotype-specificity or cross-reactivity of the B cell response within Project 1. T cell responses will be evaluated for multi-functionality by intracellular cytokne staining. Overall, Project 1 will enable identification of biomarkers and immune signatures predictive of infection and disease outcome and thus contribute to improving clinical management, vaccine design and development of novel antiviral therapies against dengue.