Rational Design of Immunogens Targeting HIV-1 Quaternary Neutrlizing Epitopes

针对 HIV-1 四级中和表位的免疫原的合理设计

基本信息

批准号：
8789433
负责人：
XIANGPENG KONG
金额：
$ 53.39万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-01-23 至 2016-01-22
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8789433
关键词：
AIDS Vaccines AIDS/HIV problem Animals Antibodies Antigens Biochemical Biological Assay Bos taurus structural-GP protein Chimeric Proteins DNA Data Databases Development Docking Engineering Engraftment Epitopes Eukaryotic Cell Generations Goals HIV HIV-1 Human Imagery Immune Sera Immunization Immunoglobulin Variable Region In Vitro Infection Instruction Macaca Macaca mulatta Maps Membrane Proteins Methods Modeling Molecular Monoclonal Antibodies Oryctolagus cuniculus Positioning Attribute Prevalence Prevention Process Production Reagent Recombinants Regimen Scaffolding Protein Serum Structural Models Structure Testing V3 Loop Vaccines Viral Viral Physiology Viral Proteins Virion Work base biophysical techniques design engineering design human monoclonal antibodies immunogenicity in vivo intermolecular interaction monomer mutant nonhuman primate protein structure scaffold success three-dimensional modeling

项目摘要

A cluster of overlapping quaternary neutralizing epitopes (QNEs) is present on the HIV-1 virus protein envelope spikes. They are composed of portions of the V2 and V3 variable regions of gpl 20 and appear to be stabilized by the intermolecular interactions in the gpl 20 trimer. The goal of Project 2 of this HIVRAD is to create immunogens that can elicit potent neutralizing anti-QNE Abs. Anti-QNE monoclonal antibodies (mAbs) are extremely potent, being 1,000-fold more potent than previously described HIV mAbs. Our team is in a unique position to pursue and achieve this goal since: (i) we have generated and characterized >100 human anti-HIV mAbs, (ii) we described the first human QNE mAb, 2909, (iii) we have successfully developed and used a structure-based rational immunogen design strategy for the production of a panel of V3-scaffold immunogens, and (iv) we have shown these rationally designed immunogens to be capable of inducing cross-clade neutralizing Abs in animals. It is a natural and logical extension of our work to apply our rational design strategy to the development of QNE-scaffold immunogens. Having already determined the crystal structures of two QNE mAbs, human mAb 2909 and rhesus mAb 2.5B, and having created a 3D structural model of the QNE in the context of gpl 20, we now propose to design, engineer, synthesize and test the immunogenicity of QNE-scaffold proteins. We will, in Aim 2.1. refine our model of the QNE by investigating the antigen-antibody interactions of QNE mAbs using biophysical and biochemical methods. In Aim 2.2. we will design and create QNE-scaffold proteins by identifying suitable protein scaffolds and grafting the components of QNE into them. After testing for antigenicity with appropriate mAbs and HIV+ sera, we will, in Aim 2.3, use the recombinant QNE-scaffold proteins to select and characterize new QNE mAbs. In Aim 2.4, we will test the recombinant QNE antigens for immunogenicity in non-human primates to determine their ability to induce potent Abs with protective anti-viral functions. These studies will potentially provide an invaluable reagent for inclusion in an HIV/AIDS vaccine.

HIV-1病毒蛋白上存在一群重叠的四季中和表位（QNE）信封尖峰。它们由GPL 20的V2和V3变量区域的部分组成，似乎通过GPL 20夹子中的分子间相互作用稳定。 Hivrad项目2的目标是产生可以引起有效中和抗QNE ABS的免疫原子。抗QNE单克隆抗体（mAb）非常有效，比以前描述的HIV mAB高1,000倍。我们的团队是自从以下独特的位置以追求和实现这一目标，因为：（i）我们已经生成和特征> 100 人类抗HIV mabs，（ii）我们描述了第一个人类QNE mab，2909，（iii）我们已经成功开发并使用了基于结构的理性免疫原设计策略来生产一批面板 V3-SCADGOLD免疫原子，（iv）我们已经证明了这些合理设计的免疫原能够在动物中诱导中和ABS的跨层。应用我们的工作是我们工作的自然而合乎逻辑的扩展 QNE-SCADGOLD免疫剂的开发的合理设计策略。已经确定了两个Qne mabs的晶体结构，人MAB 2909和恒河mab 2.5b，并创建了3D QNE的结构模型在GPL 20的背景下，我们现在建议设计，工程师，合成和测试QNE-SCADGOLD蛋白的免疫原性。我们将在AIM 2.1中。通过使用生物物理和生化方法研究QNE mAb的抗原抗体相互作用。在目标2.2。我们将通过识别合适的蛋白质支架和将Qne的组件嫁接到其中。测试适当mAb和HIV+的抗原性后 Sera，我们将在AIM 2.3中使用重组QNE-SCADGOLD蛋白来选择和表征新的QNE mabs。在AIM 2.4中，我们将测试重组QNE抗原在非人类灵长类动物中的免疫原性确定它们通过保护性抗病毒功能诱导有效ABS的能力。这些研究可能会提供一项宝贵的试剂，以纳入HIV/AIDS疫苗。