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

• 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
• 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

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病毒蛋白上存在一簇重叠的四级中和表位（QNE） 包络尖峰它们由gp 120的V2和V3可变区的部分组成， 通过GP 120三聚体中的分子间相互作用来稳定。该艾滋病毒防治和发展方案项目2的目标是， 产生可以引发有效中和抗QNE抗体的免疫原。抗QNE单克隆抗体 单克隆抗体（mAb）是非常有效的，比以前描述的HIV mAb有效1,000倍。我们的团队是 在追求和实现这一目标的独特地位，因为：（一）我们已经产生和特点>100 人抗HIV单克隆抗体，（ii）我们描述了第一个人QNE单克隆抗体，2909，（iii）我们已经成功地 开发并使用了一种基于结构的合理免疫原设计策略，用于生产一组 V3-支架免疫原，和（iv）我们已经证明这些合理设计的免疫原能够 在动物中诱导交叉进化枝中和抗体。这是我们工作的一个自然和合乎逻辑的延伸， 为QNE支架免疫原的开发提供了合理的设计策略。在确定了 两种QNE mAb，人mAb 2909和恒河猴mAb 2.5B的晶体结构，并创建了3D 在gpl 20的背景下，QNE的结构模型，我们现在建议设计，工程，合成和 测试QNE-支架蛋白的免疫原性。在目标2.1中，我们会这样做。通过以下方式改进我们的QNE模型： 使用生物物理和生物化学方法研究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来设计和创建QNE支架蛋白质， 将QNE的成分嫁接到其中。用适当的mAb和HIV+检测抗原性后 血清中，我们将在目标2.3中使用重组QNE支架蛋白来选择和表征新的QNE 单克隆抗体在目标2.4中，我们将在非人灵长类动物中测试重组QNE抗原的免疫原性， 确定它们诱导具有保护性抗病毒功能的有效Ab的能力。这些研究将有可能 为艾滋病毒/艾滋病疫苗提供了一种宝贵的试剂。