Env-Specific B Cell Repertoire Responses to NFL Trimer vaccines

对 NFL 三聚体疫苗的环境特异性 B 细胞库反应

• 批准号: 9111304
• 负责人: Jiang Zhu
• 金额: $ 55.32万
• 依托单位: INTERNATIONAL AIDS VACCINE INITIATIVE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9111304
• 关键词: Address; Affinity Chromatography; Animal Model; Animals; Antibodies; Antibody Response; Antigens; B cell repertoire; B-Lymphocytes; Binding; Blood; Cell Count; Cell Lineage; Cell Separation; Data; Development; Epitopes; Evaluation; Evolution; Gene Expression Profile; Gene Expression Profiling; Genes; HIV; HIV-1; Human; Immune response; Immunization; Inbred Mouse; Individual; Infection; Length; Light; Link; Macaca; Macaca mulatta; Messenger RNA; Microfluidics; Modeling; Monoclonal Antibodies; Mus; Oryctolagus cuniculus; Outcome; Pathway interactions; Pattern; Phase; Property; RNA; Reporting; Research Personnel; Series; Serum; Sorting - Cell Movement; Structure; System; Technology; Vaccination; Vaccine Design; Vaccines; Vesicular stomatitis Indiana virus; Virus; Virus-like particle; base; deep sequencing; design; env Glycoproteins; humanized mouse; in vivo; longitudinal analysis; mouse model; neutralizing antibody; neutralizing monoclonal antibodies; new technology; next generation sequencing; nonhuman primate; novel; programs; response; single cell analysis; vaccine candidate; vaccine delivery; vaccine development; vaccine trial; vector

Project Summary Broadly neutralizing antibodies (bnAbs) recognize conserved epitopes on the envelope glycoprotein (Env) of the human immunodeficiency virus type-1 (HIV-1). Atomic structures of the BG505 SOSIP.664 gp140 trimer have provided a rational framework for Env-based HIV-1 vaccine design. The Wyatt group recently developed a cleavage-independent, native flexibly linked (NFL) trimer, which offers a promising alternative for Env-based vaccine design. Vaccine delivery is a critical issue yet to be addressed in developing well-folded trimers toward vaccine products. In a recent review, Schiller et al. promoted a virus-like particle (VLP) approach for HIV-1 vaccine delivery. Other delivery systems include mRNA and virus vectors. For the NFL trimer, it remains unclear which delivery system will produce the most robust immune response to enable protection against HIV- 1 infection. We hypothesize that a quantitative readout of B-cell response will facilitate rational evaluation of vaccine candidates since a robust cross-neutralizing antibody response is expected for an effective HIV-1 vaccine. Next-generation sequencing (NGS) has been widely used to study the diversity and maturation of bnAbs. We have developed a series of novel NGS technologies for the analysis of dynamic B-cell responses in natural infection, animal immunization, and human vaccine trials. In Project 3 of this HIVRAD application, we will investigate the Env-specific B-cell response to four vaccines based on the NFL trimer and different delivery systems in various animal models – inbred mice, VelocImmune mice, rabbits and non-human primates (NHPs). In Aim 1, we will deep sequence the B-cell response of inbred mice, VelocImmune mice, and rabbits immunized with four vaccines. We will characterize the basic repertoire properties such as germline gene usage, degree of SHM, and CDR3 length and identify distinctive repertoire patterns associated with each vaccine. For each animal model, we will compare the B-cell responses to four vaccines. We will compare the B-cell responses in hu-mice and human vaccine trials. In Aim 2, we will deep sequence the B-cell response in NHPs immunized with four vaccines. Similarly, we will characterize the basic repertoire properties and identify distinctive patterns associated with four vaccines in the NHP model. We will compare the B-cell responses to four vaccines and compare the B-cell responses in NHPs and human vaccine trials. In Aim 3, we will study the developmental pathways of Env-specific B-cell lineages elicited by four vaccines. We will trace the lineages of monoclonal antibodies (mAbs) identified by single B-cell sorting (Project 2, Wyatt) and microfluidics-based single-cell analysis (Project 3, Zhu) in NGS repertoires. We will investigate whether the vaccine-elicited neutralizing mAbs resemble known bnAbs and their developmental pathways. Project 3, together with Project 1 (RNA and VSV vector), Project 2 (NFL trimer, VLP, and serum Ab analysis), and two Cores on in-vivo study and gene expression profiling, will constitute a comprehensive HIVRAD program towards an effective HIV-1 vaccine.

项目摘要 广泛中和抗体（BNAB）在包膜糖蛋白（ENV）上公认的保守表位 人类免疫缺陷病毒类型1（HIV-1）。 BG505 SOSIP的原子结构。664GP140 Trimer 已经为基于ENV的HIV-1疫苗设计提供了合理的框架。怀亚特集团最近开发了 独立于切割的本地链接（NFL）三聚机，为基于ENV的替代品提供了希望的替代方案 疫苗设计。疫苗输送是一个关键问题 疫苗产品。在最近的评论中，Schiller等人。促进了HIV-1的病毒样粒子（VLP）方法 疫苗输送。其他输送系统包括mRNA和病毒载体。对于NFL夹子，它仍然存在 尚不清楚哪种输送系统将产生最强大的免疫反应，以保护对HIV- 1感染。我们假设B细胞响应的定量读数将有助于合理评估 候选疫苗的候选物，因为有效的HIV-1预计可靠的跨中和抗体反应 疫苗。下一代测序（NGS）已被广泛用于研究多样性和成熟 bnabs。我们开发了一系列新型NGS技术，用于分析动态B细胞响应 自然感染，动物免疫抑制和人类疫苗试验。在此Hivrad申请的项目3中，我们 将根据NFL触发器和不同的输送研究ENV特定的B细胞对四种疫苗的反应 各种动物模型中的系统 - 近交小鼠，速度小鼠，兔子和非人类素数（NHP）。 在AIM 1中，我们将深入序列的近交小鼠的B细胞响应，速度免疫小鼠和兔子免疫。 有四种疫苗。我们将表征基本的曲目特性，例如种系基因使用，程度 SHM和CDR3长度，并确定与每种疫苗相关的独特曲目模式。每个 动物模型，我们将比较B细胞反应与四种疫苗。我们将比较B细胞的响应 HU-MICE和人类疫苗试验。在AIM 2中，我们将深入序列NHPS免疫中的B细胞响应 有四种疫苗。同样，我们将表征基本的曲目属性并确定独特的模式 与NHP模型中的四种疫苗相关。我们将比较B细胞的响应与四种疫苗和 比较NHP和人类疫苗试验中的B细胞反应。在AIM 3中，我们将研究发展 四种疫苗引起的ENV特异性B细胞谱系的途径。我们将追踪单克隆的谱系 通过单个B细胞排序（项目2，WYATT）和基于微流体的单细胞鉴定的抗体（mAb） NGS曲目中的分析（项目3，ZHU）。我们将调查疫苗引起的中和mAb是否 类似于已知的BNAB及其发育途径。项目3，以及项目1（RNA和VSV） 向量），项目2（NFL Trimer，VLP和血清AB分析），以及两个体内研究和基因的核心 表达谱分析将构成针对有效的HIV-1疫苗的全面Hivrad计划。