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

基本信息

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
项目类别：
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

项目摘要

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.

总体目标是开发将启动并选择HIV-1广泛中和抗体的免疫原子（BNAB）针对HIV Env GP41的远端膜近端外部区域（MPER）的谱系。远端 GP41 MPer抗体类型（例如10E8和DH511）是可取的，因为它们是最广泛的和潜在的bnabs分离。 BNAB免疫原设计谱系有两种策略。（1）定义BNAB 克隆谱系家谱，推断bnab未分离的普通祖先（UCA），然后选择自体 ENV结合 - 结束的B细胞谱系免疫原设计。（2）基于结构的设计，使用对设计与BNAB谱系成员结合的设计ENV的顺序谱系ABS。在这里我们建议结合两种策略设计免疫原的优势，可以启动和影响远端MPER bnabs。我们将使用新隔离的DH511谱系UCA，中间抗体（IAS）和BNAB作为设计顺序免疫原的试剂将选择类似DH511的前体并导致 BNAB开发（项目1，William Schief，PI），并在物理相关的情况下测试这些免疫原子 BNAB开发的敲入小鼠模型（项目2（Munir Alam，Pi，小动物模型核心，明天，pi，弗雷德·阿特（Fred Alt），co-i）。计算程序，抗原受体突变分析仪检测可能发生的低似然发生（项目2），该（项目2）允许定义关键抗体要诱导的体细胞突变将用于确定成功的疫苗需要的关键。目标。总体特定目的1。定义成功疫苗的关键IAS和抗体体细胞突变需要选择导致MPER BNAB诱导。（项目1和2）总体特定目的2。种系靶（GT）素数和增强免疫原的设计 DH511前体以及最佳亲和力的关键IAS和成熟的DH511 BNABS，可以选择正确/所需的IAS和BNAB。（项目1）总体特定目的3。解决DH511和DH511样谱系的共晶和冷冻结构带有ENV免疫原的抗体，沿BNAB成熟途径移动谱系并启用设计额外的免疫原子以完成诱导的MPER BNABS B细胞谱系的诱导。（项目1）总体特定目的4。从DH511 UCA和IA的免疫申请中选择最佳ENV免疫原子 VH和VL敲入小鼠。这将通过使用新颖的DH511 UCA VHDJH-RERANGING来实现 Ming Tian和Fred Alt最近在哈佛大学开发的小鼠。（小动物核心；项目1和2）。三个领先的HIV疫苗免疫原设计学术团队的合作将带来在基于结构和基于谱系的设计方面的专业知识，将是一种强大的方法通常，尤其是MPER BNAB的疫苗诱导疫苗诱导的问题。