Project 1 - Immune profiling of natural dengue virus infections

总结(项目1) 登革热病毒的四种血清型(DENV1-4)导致最重要的蚊媒病毒疾病 人类每年有大约1亿例病例，但还没有批准的疫苗或抗病毒药物存在。决定因素 DENV感染后的疾病结局尚不清楚，并被认为部分是通过 宿主免疫反应。病毒因素和宿主先天免疫相互作用可能会影响病毒的质量 获得性免疫反应，可在随后的DENV中提供保护或导致增强 感染。登革热人类免疫学项目联盟(DHIPC)的项目1重点是 与自然DENV感染结局和疾病严重程度相关的免疫特征 感染。项目1的总体方法是利用来自长期的独特样本集 尼加拉瓜正在进行的登革热研究使发现与症状相关的免疫因素成为可能 DENV感染、严重登革热和特异性适应性免疫反应。项目1将继续 尼加拉瓜儿童登革热队列研究(PDCS)从有充分记录的自然重复病例中收集样本 DENV感染。它还将利用尼加拉瓜正在进行的基于医院的登革热研究，以使 对严重登革热和轻度登革热进行调查，并将进行指数分类研究以提供样本 来自无症状的病毒携带者。所有样本都将提供详细的临床数据。与 世界级的DHIPC核心，将使用系统生物学的方法来构建先天的签名 自然感染DENV期间的免疫反应。互补的尖端基因组的星座， 转录组、蛋白质组、血清学和免疫表型方法将使人类免疫 对登革热的反应将以前所未有的细节和复杂性进行剖析。目标1将 临床转归不同的DENV感染的免疫特征：无症状和 使用指数聚类研究的症状性DENV感染以及PDDC和 基于医院的研究。AIM 2将描述初次和继发性DENV感染的免疫特征， 感染的特定血清型顺序。目标3将调查先天免疫信号之间的联系 抗体和细胞介导的免疫反应。抗体中和、同种类型和谱系将是 评估，以及B细胞反应的大小和血清型特异性或交叉反应 项目1将通过细胞内细胞因子染色来评估T细胞反应的多功能性。总的来说， 项目1将能够识别预测感染和疾病的生物标志物和免疫特征 结果，从而有助于改善临床管理，疫苗设计和开发的新 针对登革热的抗病毒疗法。

SUMMARY (PROJECT 1) 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 cytokine 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.