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.

项目概要 广泛中和抗体 (bnAb) 可识别包膜糖蛋白 (Env) 上的保守表位 人类免疫缺陷病毒 1 型 (HIV-1) BG505 SOSIP.664 gp140 三聚体的原子结构。 为 Wyatt 小组最近开发的基于 Env 的 HIV-1 疫苗设计提供了合理的框架。 一种独立于切割的天然柔性连接 (NFL) 三聚体，为基于 Env 的三聚体提供了一种有前景的替代方案 疫苗设计是开发折叠良好的三聚体过程中尚未解决的一个关键问题。 在最近的一篇评论中，Schiller 等人推广了一种针对 HIV-1 的病毒样颗粒 (VLP) 方法。 疫苗递送系统包括 mRNA 和病毒载体，但仍保留 NFL 三聚体。 尚不清楚哪种传递系统将产生最强大的免疫反应，从而能够预防艾滋病毒- 我们发现 B 细胞反应的定量读数将有助于合理评估。 候选疫苗，因为预计有效的 HIV-1 会产生强大的交叉中和抗体反应 下一代测序（NGS）已被广泛用于研究疫苗的多样性和成熟度。 我们开发了一系列新颖的 NGS 技术，用于分析动态 B 细胞反应。 在此 HIVRAD 应用的项目 3 中，我们进行了自然感染、动物免疫和人类疫苗试验。 将研究基于 NFL 三聚体和不同递送方式的四种疫苗的 Env 特异性 B 细胞反应 各种动物模型中的系统 - 近交系小鼠、VeloImmune 小鼠、兔子和非人类灵长类动物 (NHP)。 在目标 1 中，我们将对近交系小鼠、VeloImmune 小鼠和免疫兔子的 B 细胞反应进行深度测序 我们将描述四种疫苗的基本特性，例如种系基因的使用、程度。 SHM 和 CDR3 长度并确定与每种疫苗相关的独特的库模式。 动物模型中，我们将比较 B 细胞对四种疫苗的反应。 在目标 2 中，我们将深入测序 NHP 免疫的 B 细胞反应。 同样，我们将描述四种疫苗的基本特性并确定独特的模式。 我们将比较 B 细胞对四种疫苗的反应，并与 NHP 模型中的四种疫苗相关。 比较 NHP 和人类疫苗试验中的 B 细胞反应。在目标 3 中，我们将研究发育过程。 四种疫苗引发的 Env 特异性 B 细胞谱系的途径 我们将追踪单克隆疫苗的谱系。 通过单 B 细胞分选（项目 2，Wyatt）和基于微流体的单细胞鉴定抗体 (mAb) NGS 库中的分析（项目 3，朱） 我们将研究疫苗是否引发中和单克隆抗体。 类似于已知的 bnAb 及其发育途径，以及项目 1（RNA 和 VSV）。 载体）、项目 2（NFL 三聚体、VLP 和血清抗体分析）以及体内研究和基因的两个核心 表达谱分析将构成一个全面的 HIVRAD 计划，旨在开发有效的 HIV-1 疫苗。