Defining parameters to induce breadth in knockin mice expressing HIV bnAb precursors

定义参数以诱导表达 HIV bnAb 前体的敲入小鼠的广度

基本信息

批准号：
10504569
负责人：
Laurent Karl Verkoczy
金额：
$ 27.85万
依托单位：
APPLIED BIOMEDICAL SCIENCE INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10504569
关键词：
Address Affinity Animal Model Antibodies Antibody Response Antigens B-Lymphocytes Binding Binding Sites Cell Survival Clinical Trials Consensus Crossbreeding Data Development Emu species Engineering Epitopes Exhibits Frequencies Funding Gene Targeting Goals HIV HIV Antibodies HIV-1 Hepatitis C Human Immunization Immunoglobulin Somatic Hypermutation Individual Infection Influenza Knock-in Knock-in Mouse Learning Modeling Mus Mutate Mutation Observational Study Pathway interactions Patients Peripheral Physiological Polymerase Polysaccharides Preventive vaccine Process Production Regimen Resolution Role Somatic Mutation Structure of germinal center of lymph node System Testing Time Vaccinee Vaccines Viral Virus Diseases Work base design experimental study in vivo insight mimicry mouse model mutant neutralizing antibody novel novel strategies optimism overexpression pandemic disease peripheral tolerance prevent recruit response synergism trait transmission process vaccine candidate vaccine development vaccine strategy

项目摘要

PROJECT SUMMARY / ABSTRACT This is a renewal R01 application to characterize an array of novel knockin (KI) mouse lines, we have created using a previously-described gene-targeting approach, to express unmutated precursors of four well- characterized, representative HIV-1 broadly neutralizing antibody (bnAb) lineages. Developing a preventative vaccine remains a critical priority to end the HIV pandemic, and it is widely held that such a vaccine will need to induce bnAb responses. While the eventual elicitation of bnAb responses in some HIV-1+ patients shows such responses are possible, no vaccine yet can elicit them. In this regard, animal models that can systematically identify impediments in bnAb precursor activation and maturation and iteratively test novel strategies to overcome such hurdles, would be highly beneficial for developing a bnAb-based vaccine. Three key roadblocks that merit further testing in such models are: i) B-cell tolerance controls, which can delete, inactivate, or modify bnAb lineage-expressing B-cells, before and during immunization, ii) high somatic mutation levels that accumulate in bnAbs over prolonged periods of infection, amounts prohibitive for vaccines to elicit, and whose exact level required for function is not known, and iii) no or minimal bnAb precursor affinity for standard Env immunogens. The overall objective of this proposal is to use novel bnAb precursor-directed immunogens to define minimal requirements to activate, and then induce breadth in, naïve precursor KI B-cells from CH103, CH31, CH01, and CH58 lineages (and determine if/what host controls limit both processes). The matured Abs of these lineages are all relatively less mutated, making them more attractive vaccine candidates. Furthermore, they target distinct Env regions, and express a different set of functional traits/neutralization profiles, thus embody the bnAb spectrum. We hypothesize that this unique set of immunogens and models will allow us to learn how to induce individual, immunization-directed bnAb pathways with moderate breadth, manageable mutation levels and no (or only cryptic) self-reactivity. To test this central hypothesis, we will use a multilayered approach where in Aim 1, we will first define to what extent naïve KI B-cells expressing CH103, CH31, CH01, or CH58 lineage precursors are under host controls. Then, in Aim 2, we will learn how best to activate these lineages with precursor-targeting immunogens, so they can be recruited into maturation pathways. Finally, in Aim 3, using immunogens that can initiate maturation, we will define minimal evolutionary trajectories to desired bnAb responses, under conditions where affinity maturation will be enhanced via cross-breeding KI mice to those overexpressing polymerase-ζ (with elevated Ig somatic mutation rates) and/or Eµ-bcl2 (having increased B-cell survival). Learning how to induce, and what limits bnAbs in these model settings will help define more tractable vaccine targets and strategies to elicit in clinical trials. More generally, the host-HIV interaction issues addressed here (poor precursor binding, self-reactivity/mimicry, and excess mutation) may be informative for viral infections like hepatitis C or influenza, also requiring bnAb responses and problematic as vaccine targets.

项目概要/摘要这是一个更新的 R01 应用程序，用于表征我们创建的一系列新型敲入 (KI) 小鼠品系使用先前描述的基因靶向方法，表达四个良好的未突变前体具有代表性的 HIV-1 广泛中和抗体 (bnAb) 谱系的开发。疫苗仍然是结束艾滋病毒大流行的首要任务，人们普遍认为这种疫苗需要诱导 bnAb 反应，而最终在一些 HIV-1+ 患者中引发 bnAb 反应表明了这一点。反应是可能的，但尚无疫苗可以引起这些反应，在这方面，动物模型可以系统地引起这些反应。识别 bnAb 前体激活和成熟的障碍，并迭代测试新策略克服这些障碍，将非常有利于开发基于 bnAb 的疫苗的三个关键障碍。值得在此类模型中进一步测试的是：i) B 细胞耐受控制，可以删除、失活或修改表达 bnAb 谱系的 B 细胞，在免疫之前和期间，ii) 高体细胞突变水平在长时间的感染过程中，bnAb 中的积累量超过了疫苗所能引发的水平，其功能所需的确切水平尚不清楚，并且 iii) bnAb 前体对标准 Env 没有亲和力或亲和力极小该提案的总体目标是使用新型 bnAb 前体定向免疫原。定义激活来自 CH103 的幼稚前体 KI B 细胞的最低要求，然后诱导广度， CH31、CH01 和 CH58 谱系（并确定宿主控制是否/什么限制这两个过程）。这些谱系中的突变相对较少，使它们成为更具吸引力的候选疫苗。它们针对不同的 Env 区域，并表达一组不同的功能特征/中和特征，因此我们勇敢地说，这套独特的免疫原和模型将使我们能够了解如何诱导个体、免疫导向的 bnAb 途径，其宽度适中、易于管理为了检验这个中心假设，我们将使用多层的。在目标 1 中，我们将首先定义幼稚 KI B 细胞表达 CH103、CH31、CH01、或 CH58 谱系前体受宿主控制然后，在目标 2 中，我们将学习如何最好地激活这些。具有前体靶向免疫原的谱系，因此它们可以被招募到成熟途径中。目标 3，使用可以启动成熟的免疫原，我们将定义所需的最小进化轨迹 bnAb 反应，在亲和力成熟将通过杂交 KI 小鼠增强的条件下那些过度表达聚合酶-ζ（Ig 体细胞突变率升高）和/或 Eμ-bcl2（增加了解如何诱导 bnAb，以及在这些模型设置中限制 bnAb 的因素将有助于定义更多信息。在临床试验中引发的易处理的疫苗目标和策略更普遍的是，宿主与艾滋病毒的相互作用问题。这里讨论的（前体结合不良、自身反应性/模仿和过度突变）可能会为以下方面提供信息：丙型肝炎或流感等病毒感染也需要 bnAb 反应，并且作为疫苗靶点存在问题。

项目成果

期刊论文数量（15）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Induction of HIV-1 broad neutralizing antibodies in 2F5 knock-in mice: selection against membrane proximal external region-associated autoreactivity limits T-dependent responses.

DOI：
10.4049/jimmunol.1300971
发表时间：
2013-09-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
0
作者：
Verkoczy L;Chen Y;Zhang J;Bouton-Verville H;Newman A;Lockwood B;Scearce RM;Montefiori DC;Dennison SM;Xia SM;Hwang KK;Liao HX;Alam SM;Haynes BF
通讯作者：
Haynes BF

Rescue of HIV-1 broad neutralizing antibody-expressing B cells in 2F5 VH x VL knockin mice reveals multiple tolerance controls.

DOI：
10.4049/jimmunol.1101633
发表时间：
2011-10-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
0
作者：
Verkoczy L;Chen Y;Bouton-Verville H;Zhang J;Diaz M;Hutchinson J;Ouyang YB;Alam SM;Holl TM;Hwang KK;Kelsoe G;Haynes BF
通讯作者：
Haynes BF

Autoreactivity and broad neutralization of antibodies against HIV-1 are governed by distinct mutations: Implications for vaccine design strategies.

DOI：
10.3389/fimmu.2022.977630
发表时间：
2022
期刊：
FRONTIERS IN IMMUNOLOGY
影响因子：
7.3
作者：
Li, Xiaojun;Liao, Dongmei;Li, Zhengyang;Li, Jixi;Diaz, Marilyn;Verkoczy, Laurent;Gao, Feng
通讯作者：
Gao, Feng

Antibody polyspecificity and neutralization of HIV-1: a hypothesis.

HIV-1 的抗体多特异性和中和作用：一个假设。