Dissecting Polyclonal Sera to Reveal Correlates of Productive Immune Responses to HIV

剖析多克隆血清以揭示 HIV 生产性免疫反应的相关性

• 批准号: 10716061
• 负责人: Lars Oliver Hangartner
• 金额: $ 87.06万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-16 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10716061
• 关键词: Amino Acids; Antibodies; Antibody Binding Sites; Antibody Response; Antigens; B cell repertoire; B-Lymphocytes; B-cell receptor repertoire sequencing; Bar Codes; Blood Circulation; Cells; Clinical; Clinical Research; Complementarity Determining Regions; Cryoelectron Microscopy; Data; Electron Microscopy; Engineering; Epitope Mapping; Epitopes; Evaluation; Funding; Future; Genes; Glycoproteins; HIV; HIV vaccine; Human; Image; Immune response; Immunogenetics; Immunoglobulin A; Immunoglobulin Somatic Hypermutation; Immunoglobulins; Knock-in Mouse; Maps; Methodology; Methods; Miniaturization; Molecular; Monoclonal Antibodies; Mucous Membrane; Negative Staining; Polishes; Polysaccharides; Preparation; Production; Productivity; Property; Publications; Resolution; Sampling; Site; Specificity; Structure; Surface; Technology; Testing; Vaccine Design; Vaccines; Viral; data format; data repository; database query; design; improved; miniaturize; mouse model; neutralizing antibody; nonhuman primate; pathogen; phase I trial; polyclonal antibody; pre-clinical; single-cell RNA sequencing; vaccination strategy; vaccine development

Project Summary/Abstract Rational, structure-based immunogen design for HIV vaccine development has began to deliver promising results in both, pre-clinical non-human primate (NHP) and clinical studies such as the IAVI G001 phase I trial [NCT03547245]. Moreover, advances in single cell RNAseq and electron microscopy-based polyclonal antibody mapping (EMPEM) have enabled analysis of both vaccine and pathogen-induced antibody responses at unprecedented resolution, further enabling rational vaccine design. EMPEM and single cell B cell analysis deliver fundamentally different data formats that if properly integrated, can illuminate immune responses at unprecedented detail. We recently extended the original negative stain EMPEM method, capable of distinguishing polyclonal antibody epitope specificities and angle of approach, to high resolution cryoEM-based EMPEM to yield molecular details of epitope-paratope interfaces. In the best cases we can derive some sequence information for the complementarity determining regions (CDRs) from structural data, identify Vh/Vl gene usage, and query databases of antibody sequences to directly generate monoclonal antibodies. As such, we can short cut the laborious efforts to generate epitope specific monoclonal antibodies using conventional methods. These molecular details are critical for analyzing shepherding or polishing immunization strategies that are now being attempted for HIV vaccine development, wherein immunogens are designed to elicit and mature antibodies with specific features in CDRs that resemble known broadly neutralizing antibodies (bnAbs). In this renewal application we propose to extend the resolution, throughput, and utility of our EMPEM approach. Moreover, since most pathogens will enter the body through mucosal surfaces, analysis of (vaccine-induced) mucosal antibodies will be explored. The data generated will provide new correlates of productive immune responses to the portfolio of exciting HIV immunogens with epitope focusing or germline targeting properties that will help down select and prioritize amongst the various approaches being pursued.

项目总结/摘要 用于HIV疫苗开发的基于结构的合理免疫原设计已经开始交付 在临床前非人灵长类动物（NHP）和临床研究（如IAVI）中均获得了有希望的结果 G 001 I期试验[NCT 03547245]。此外，单细胞RNA测序和电子技术的进展 基于显微镜的多克隆抗体作图（EMPEM）已经能够分析疫苗和 病原体诱导的抗体反应在前所未有的分辨率，进一步使合理的疫苗 设计EMPEM和单细胞B细胞分析提供完全不同的数据格式， 正确整合，可以在前所未有的细节上阐明免疫反应。我们最近延长了 独创负染EMPEM法，能区分多克隆抗体表位 特异性和角度的方法，以高分辨率cryoEM为基础的EMPEM产生分子细节 表位-互补位界面。在最好的情况下，我们可以得到一些序列信息， 从结构数据中扩增互补决定区（CDR），鉴定Vh/Vl基因使用，并查询 抗体序列数据库，以直接产生单克隆抗体。因此，我们可以做空 减少了使用常规方法产生表位特异性单克隆抗体的费力工作， 方法.这些分子细节对于分析牧羊或抛光免疫至关重要 目前正在尝试的HIV疫苗开发策略，其中免疫原是 设计用于引发和成熟抗体，所述抗体在CDR中具有类似于广泛已知的特异性特征， 中和抗体（bnAb）。在这次更新申请中，我们建议延长决议， 我们的EMPEM方法的吞吐量和实用性。此外，由于大多数病原体会进入人体， 通过粘膜表面，将探索（疫苗诱导的）粘膜抗体的分析。数据 所产生的将提供新的相关性生产性免疫反应的投资组合，令人兴奋的艾滋病毒 具有表位聚焦或种系靶向特性的免疫原， 优先考虑正在采取的各种办法。