Immunogen Design for Induction of HIV distal gp41 broadly neutralizing antibodies

用于诱导 HIV 远端 gp41 广泛中和抗体的免疫原设计

• 批准号: 10132973
• 负责人: S. Munir ALAM
• 金额: $ 155.24万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-09 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10132973
• 关键词: Affinity; Animal Model; Animals; Antibodies; Antibody Affinity; Antibody Response; Antigen Receptors; Antigens; Autologous; B-Lymphocytes; Binding; Cell Lineage; Cellular biology; Clone Cells; Collaborations; Complex; Coupled; Cryoelectron Microscopy; Crystallization; Detection; Development; Directed Molecular Evolution; Distal; Genealogy; Goals; HIV; HIV vaccine; HIV-1; Hydrophobicity; Immune Tolerance; Immunization; Knock-in Mouse; Lead; Lipids; Macaca mulatta; Membrane; Membrane Lipids; Monoclonal Antibodies; Mus; Mutation; Pathway interactions; Physiological; Process; Proteins; Reagent; Regimen; Site-Directed Mutagenesis; Somatic Mutation; Structure; Surface; Testing; Vaccines; Virion; X-Ray Crystallography; Yeasts; base; computer program; design; expectation; information model; iterative design; member; mouse model; neutralizing antibody; novel; novel strategies; programs; vaccine development; ward

The overall objective is to develop immunogens that will initiate and select HIV-1 broad neutralizing antibody (bnAb) lineages directed to the distal membrane proximal external region (MPER) of HIV Env gp41. Distal gp41 MPER antibody types such as 10E8 and DH511 are desirable because they are among the most broad and potent bnAbs isolated. There are two strategies for bnAb immunogen design lineages. (1) Define bnAb clonal lineage genealogies, infer the bnAb unmutated common ancestor (UCA) and select autologous Envs that bind—termed B cell lineage immunogen design. (2) Structural-based design, using structures of sequential lineage Abs to design Envs that bind to bnAb lineage members. Here we propose to combine the strengths of both strategies to design immunogens that can initiate and induce distal MPER bnAbs. We will use the newly isolated DH511 lineage UCA, intermediate antibodies (IAs) and bnAbs as reagents upon which to design sequential immunogens that will select DH511-like precursors and lead to bnab development (Project 1, William Schief, PI), and to test these immunogens in physiologically relevant knock-in mouse model of bnAb development (Project 2 (Munir Alam, PI, Small Animal Models Core, Ming Tian, PI, Fred Alt, Co-I). A computational program, Antigen Receptor Mutation Analyzer for Detection of Low Likelihood Occurrences (ARMADiLLO) (Project 2) that allows for definition of the critical antibody somatic mutations to be induced will be used to determine key IAs that a successful vaccine will need to target. Overall Specific Aim 1. Define the key IAs and antibody somatic mutations that a successful vaccine will need to select to lead to distal MPER bnAb induction. (Projects 1 and 2) Overall Specific Aim 2. Design of germline targeting (GT) prime and boost immunogens that bind to the DH511 precursors and to key IAs and mature DH511 bnAbs in optimal affinities and can select the correct/desired IAs and bnAbs. (Project 1) Overall Specific Aim 3. Solve co-crystal and cryoEM structures of DH511 and DH511-like lineage antibodies with Env immunogens that move the lineage along the bnAb maturation pathway and enable design of additional immunogens to complete the induction of distal MPER bnAbs B cell lineages. (Project 1) Overall Specific Aim 4. Selection of optimal Env immunogens from immunizations of DH511 UCA and IA VH and VL knock-in mice. This will be accomplished by use of the novel DH511 UCA VHDJH-rearranging mouse recently developed by Ming Tian and Fred Alt at Harvard. (Small Animal Core; Projects 1 and 2). This collaboration of three leading academic teams in HIV vaccine immunogen design will bring together expertise in structure-based and lineage-based design, and will be a powerful approach to the problem of vaccine induction of disfavored antibody lineages in general and distal MPER bnAbs in particular.

总体目标是开发能够启动和选择HIV-1广泛中和抗体的免疫原 针对HIV Env gp 41的远端膜近端外部区域（MPER）的bnAb谱系。远侧 gp 41 MPER抗体类型如10 E8和DH 511是理想的，因为它们是最广泛的 并分离出有效的bnAb。bnAb免疫原设计谱系有两种策略。(1)定义bnAb 克隆谱系谱系，推断bnAb未突变的共同祖先（UCA），并选择自体 结合称为B细胞谱系免疫原的包膜设计。(2)基于结构的设计，使用 序列谱系Ab以设计结合bnAb谱系成员的Env。这里我们建议将联合收割机 这两种策略的优势，设计免疫原，可以启动和诱导远端MPER bnAbs。我们将使用新分离的DH 511谱系UCA、中间抗体（IA）和bnAb作为 设计顺序免疫原的试剂，这些免疫原将选择DH 511样前体并导致 bnab开发（项目1，William Schief，PI），并在生理学相关环境中测试这些免疫原。 bnAb发育的敲入小鼠模型（项目2（MunirAlam，PI，小动物模型核心， Ming Tian，PI，Fred Alt，Co-I）。抗原受体突变检测分析仪 低发生可能性（ARMADiLLO）（项目2），允许定义关键抗体 诱导的体细胞突变将用于确定成功疫苗所需的关键IA， 目标 具体目标1。定义成功的疫苗所需的关键IA和抗体体细胞突变 需要选择导致远端MPER bnAb诱导。（项目1和2） 具体目标2。设计结合至免疫原的生殖系靶向（GT）初免和加强免疫原。 DH 511前体和关键IA和成熟DH 511 bnAb的最佳亲和力，并可以选择 正确/所需的IA和bnAb。（项目1） 具体目标3。解析DH 511和类DH 511谱系的共晶和cryoEM结构 具有Env免疫原的抗体，所述Env免疫原使谱系沿着bnAb成熟途径移动， 设计额外的免疫原以完成远端MPER bnAb B细胞谱系的诱导。（项目1） 具体目标4。从DH 511 UCA和IA免疫中筛选最佳Env免疫原 VH和VL基因敲入小鼠。这将通过使用新的DH 511 UCA VHDJH重排来实现 最近由哈佛的Ming Tian和Fred Alt开发的老鼠。（小动物核心;项目1和2）。 三个领先的艾滋病毒疫苗免疫原设计学术团队的合作将带来 结合了基于结构和基于血统的设计方面的专业知识，这将是一种强大的方法， 疫苗诱导一般不利的抗体谱系和特别是远端MPER bnAb的问题。