Evaluation of Antibody Responses Elicited by Zika Vaccination in Flavivirus-naïve and -experienced Individuals

项目标题：评估寨卡病毒疫苗在黄病毒中引起的抗体反应-幼稚和- 有经验的个人 项目摘要 寨卡病毒(ZIKV)是黄病毒属的成员，最近引发了广泛的神经系统病例 病理学和先天神经缺陷。作为回应，一个多边调查联盟设计并 开发了多种候选疫苗，可以诱导出强大的ZIKV中和抗体，这些抗体被证明是 在动物模型中与疾病保护相关。尽管取得了这些进展，但目前尚不清楚ZIKV如何 免疫抗体反应在有或没有接触过其他黄病毒的人中形成，如 如登革热病毒(DENV)、日本脑炎(JEV)或黄热病病毒(YFV)；尤其是这些病毒 所有这些都与寨卡病毒有显著的流行病学重叠。我们的长期目标是理解潜在的幽默 黄病毒疫苗接种产生的机制，可提供对黄病毒幼稚的长期保护 和/或经验丰富的人群。这些信息将指导黄病毒地方性疾病的疫苗接种策略 在一些地区或前往疫区旅行的幼稚的黄病毒患者中也会发生这种情况。这项提议的总体目标 本研究旨在评价ZIKV疫苗诱导的黄病毒B细胞库的特异性和功能。 天真和有经验的人。为了实现这些目标，这项研究将利用尖端技术 使用RNAseq和下一代对黄病毒特异性单个B细胞的B细胞受体(BCR)进行测序 测序(NGS)。BCR将在接种寨卡病毒疫苗的个人之间进行比较，并与之前公布的结果进行比较 单抗，以确定B细胞谱系的患病率、基因分配、体细胞分化程度 高度突变(SHM)和重链互补决定区3的长度(HCDR3)。公共B 细胞谱系将以单抗的形式表达，并对其特异性、功能、结构、 以及在小鼠模型中防御黄病毒挑战的能力。共有40个样本将从以下方面进行评估 在3个不同的寨卡病毒I期临床试验中代表5个群体的个人：a)黄病毒-幼稚的个人 使用寨卡病毒纯化灭活全病毒疫苗(ZPIV)b)黄病毒幼稚的人接种疫苗 接种了腺载体寨卡病毒M-E(Ad26.ZIKV.M-Env)c)居住在波多黎各的人 有登革热感染经历的人接种了ZPIV疫苗d)JEV(IXIARO®)接种了ZPIV的个人 接种和e)YFV(YV-VAX®)疫苗接种ZPIV疫苗的个体。黄病毒-幼稚，寨卡病毒 将在目标1和有黄病毒经验的个人中探索接种疫苗的个人(a-b组)，方法是 将在目标2中探讨先前感染(c组)或先前接种疫苗(d-e组)的情况。目标3将检查晚期 最后一次接种ZPIV疫苗后的时间点(6个月)以确定循环B细胞的寿命 AIMS 1和AIMS 2中描述的谱系特征。评估对寨卡病毒疫苗反应的流行B细胞谱系将 揭示居住在不同地区的个体之间体液反应的特异性、功能和持久性。 这些研究将为ZIKV疫苗在两种黄病毒中的效力和持久性提供见解。 幼稚和有经验的人，并可能转化为产生长期保护的疫苗战略。

Project Title: Evaluation of Antibody Responses Elicited by Zika Vaccination in Flavivirus-naïve and - experienced Individuals Project Summary Zika virus (ZIKV) is a member of the flavivirus genus that recently precipitated widespread cases of neurological pathology and congenital neurologic defects. In response, a multilateral coalition of investigators designed and developed multiple vaccine candidates that elicited potent ZIKV-neutralizing antibodies, which were shown to correlate with disease protection in animal models. Despite these advances, it remains unclear how the ZIKV immunization antibody response is shaped in humans with and without prior exposure to other flaviviruses, such as dengue virus (DENV), Japanese encephalitis (JEV) or yellow fever virus (YFV); particularly as these viruses all have significant epidemiologic overlap with ZIKV. Our long-term goal is to understand the underlying humoral mechanisms generated by flavivirus vaccination, which can provide long-term protection in flavivirus-naïve and/or -experienced populations. Such information would guide vaccination strategies in flavivirus-endemic areas or among flavivirus-naïve individuals traveling to endemic areas. The overall objective of this proposed research is to evaluate the specificity and function of the B cell repertoire elicited by ZIKV vaccination in flavivirus- naïve and -experienced individuals. To achieve these goals, this research will utilize leading-edge technologies to sequence B cell receptors (BCRs) from flavivirus-specific, single B cells using RNAseq and Next Generation Sequencing (NGS). BCRs will be compared between Zika vaccinated individuals, and to previously published monoclonal antibodies, to determine the prevalence of B cell lineages, gene assignment, degree of somatic hypermutation (SHM), and lengths of heavy chain complementary-determining region 3 (HCDR3). Common B cell lineages will be expressed as monoclonal antibodies and evaluated for their specificity, function, structural, and ability to protect against flavivirus challenges in mouse models. A total of 40 samples will be evaluated from individuals representing 5 groups in 3 different Phase I Zika vaccine clinical trials: a) Flavivirus-naïve individuals who were vaccinated using a Zika purified inactivated whole virus vaccine (ZPIV) b) Flavivirus-naïve individuals who were vaccinated with an adeno-vectored Zika M-E (Ad26.ZIKV.M-Env) c) Individuals living in Puerto Rico with prior dengue infection who were ZPIV vaccinated d) JEV (IXIARO®) vaccinated individuals who were ZPIV vaccinated and e) YFV (YV-VAX®) vaccinated individuals who were ZPIV vaccinated. Flavivirus-naïve, Zika vaccinated individuals (Groups a-b) will be explored in Aim 1 and Flavivirus-experienced individuals, either by prior infection (Group c) or by prior vaccination (Groups d-e), will be explored in Aim 2. Aim 3 will examine a late timepoint (6 months) following the last ZPIV vaccination to determine the longevity of the circulating B cell lineages characterized in Aims 1 and 2. Evaluating prevalent B cell lineages responding to Zika vaccination will reveal the specificity, function and durability of the humoral response among individuals living in different regions. These studies will provide insights into the potency and durability of ZIKV vaccine responses in both flavivirus- naïve and -experienced individuals and may translate into vaccine strategies that yield long-lived protection.