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.

项目摘要 /摘要这是一个续订R01应用程序，用于表征一系列新型敲击蛋白（Ki）鼠标线，我们创建了使用以前描述的基因靶向方法，表达四个孔的未分离的前体表征，代表性的HIV-1广泛中和抗体（BNAB）谱系。开发预防措施疫苗仍然是结束艾滋病毒大流行的关键优先事项，人们普遍认为这种疫苗需要诱导BNAB响应。虽然事件在某些HIV-1+患者中引起BNAB反应的启发表明反应是可能的，没有疫苗可以引起它们。在这方面，可以系统地的动物模型确定BNAB前体激活和成熟的障碍，并迭代地测试新策略克服这种障碍，将对开发基于BNAB的疫苗非常有益。三个关键的障碍在此类模型中的进一步测试是：i）B细胞公差控制，可以删除，灭活或修改在免疫之前和期间，表达BNAB谱系的B细胞，ii）高细胞突变水平在长时间的感染期内积聚在BNAB中，禁止疫苗引起疫苗，并且其功能所需的确切级别尚不清楚，iii）否或最小的BNAB前体亲和力该提案的总体目的是将新型BNAB前体指导的免疫原对象定义最小的要求，以激活，然后诱导广度，从CH103中幼稚的前体Ki B细胞， CH31，CH01和CH58谱系（并确定/哪种主机控制是否限制了这两个过程）。成熟的腹肌这些谱系的突变都相对较少，使它们更具吸引力的候选疫苗。此外，他们针对不同的环境区域，并表达不同的功能性状/中和概况，因此体现BNAB谱。我们假设这套独特的免疫原子和模型将使我们能够了解如何以中等广度，易于管理的方式诱导个人免疫化指导的BNAB途径突变水平和无（或仅是加密）自我反应性。为了检验此中心假设，我们将使用多层在AIM 1中，我们将首先定义在何种程度上表达CH103，CH31，CH01，或CH58谱系前体处于宿主对照状态。然后，在AIM 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，因此可以募集到成熟途径中。最后，在 AIM 3，使用可以启动成熟的免疫原子，我们将定义最小的进化轨迹 BNAB响应，在通过交叉繁殖的Ki小鼠提高亲和力成熟的条件下那些过表达聚合酶ζ（Ig体细胞突变率升高）和/或Eµ-BCl2（增加 B细胞生存）。学习如何影响，以及这些模型设置中的bnabs的限制将有助于定义更多在临床试验中引起的可导致疫苗目标和策略。更一般地，宿主-HIV互动问题在此处解决（前体的结合差，自我反应性/模仿和过量突变）可能是有用的诸如乙型肝炎或影响的病毒感染，还需要BNAB反应和问题作为疫苗靶标。

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。