Structural and dynamic traits underlying phenotypic variation in HIV-1 Env

HIV-1 Env表型变异的结构和动态特征

• 批准号: 10643989
• 负责人: Kelly Keisen Lee
• 金额: $ 53.11万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-02 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10643989
• 关键词: Address; Adopted; Anatomy; Animals; Antibodies; Antibody Response; Antigen Presentation; Antigen-Presenting Cells; Antigens; Architecture; Area; Awareness; Behavior; Binding; Biological; Biological Assay; Biological Factors; Blood; Breathing; C-Type Lectins; CD209 gene; CD4 Antigens; Cell surface; Cells; Chimeric Proteins; Cryoelectron Microscopy; Defensins; Dendritic Cells; Deuterium; Development; Dissection; Electron Microscopy; Epitopes; Equilibrium; Evolution; Exhibits; Foundations; Genes; Glycoproteins; Goals; HIV; HIV envelope protein; HIV-1; Hydrogen; Immune; Immune Targeting; Immune response; Immune system; Individual; Lectin; Length; Maps; Mass Spectrum Analysis; Mediating; Molecular Conformation; Movement; Mucous Membrane; Nature; Pathogenesis; Pathogenicity; Phenotype; Physiology; Play; Polysaccharides; Property; Proteins; Recombinant Proteins; Resistance; Resolution; Role; Scheme; Serum Albumin; Serum Proteins; Shapes; Source; Structure; Surface Antigens; Testing; Tropism; Vaccines; Variant; Viral; Virus; Virus Replication; antibody and antigen binding; antimicrobial peptide; biophysical analysis; biophysical techniques; chemokine; comparative; cross reactivity; glycoprotein structure; glycosylation; grasp; immunogenicity; innovation; neutralizing antibody; novel; protein complex; receptor; trait; transmission process; vaccine development; vaccine trial; viral fitness; virological synapse

We have long been aware of the astounding sequence variation in the HIV-1 envelope glycoprotein (Env) gene, but the structural and functional implications of this diversity are only beginning to be grasped. Structural variation in Env impacts its interactions with all key drivers of viral fitness and replication, and this is not captured by variation in sequence. These differences underlie viral phenotypic traits such as neutralization sensitivity, tropism, infectivity, and transmissibility. While recent studies have provided detailed structural information for trimeric Env ectodomain from a range of viral isolates, the structures represent a static, Platonic ideal of the Env assembly. Under native conditions, HIV Env is a highly dynamic fusion protein complex that can flicker between antigenically and functionally distinct conformational states. Biophysical studies from our group and others are providing the first structure-based indications that the propensity for Env to undergo large-scale dynamic movements is highly isolate-specific in nature. These changes directly impact a given Env's interactions with biological factors in the host. Here we will apply innovative structural analytical approaches, including structural mass spectrometry and cryo-EM, to characterize Env diversity and to identify the consequences of structural variation and dynamics on antibody binding and antigen presentation by the dendritic cell displayed lectin, DC-SIGN. We have developed an effective approach for purifying the native-like SOSIP trimers from highly divergent HIV-1 isolates and have demonstrated in preliminary studies that trimers even from neutralization resistant primary isolates exhibit significant differences in stability, local structural dynamics of bNAb epitopes, large-scale conformational breathing, and structure. We will characterize structural dynamic profiles of the Envs from across the neutralization Tier spectrum and from the Global Panel of resistant viruses, which was composed to be highly representative of circulating neutralization phenotypes. We will test whether variants that produced favorable immune responses (breadth and potency) exhibit structural dynamic traits. And we will assess the effect of dynamics on antibody and DC-SIGN reactivity for diverse viral isolates. An understanding of variation in Env structure and function will provide a foundation for understanding differences in HIV pathogenicity and viral fitness, and a better understanding of how the immune system grapples with this extremely variable antigen. Progress in these areas will inform our understanding of HIV evolution, and may help guide development of Env-based vaccines and immunogens.

我们早就知道HIV-1包膜糖蛋白（Env）中令人震惊的序列变异 基因，但这种多样性的结构和功能的影响才刚刚开始被抓住。结构 Env的变异会影响其与病毒适应性和复制的所有关键驱动因素的相互作用， 通过序列中的变化来捕获。这些差异是病毒表型特征的基础， 敏感性、向性、感染性和传染性。虽然最近的研究提供了详细的结构 从一系列病毒分离物的三聚体Env胞外域的信息中，结构代表了静态的柏拉图式结构域。 理想的Env组件。在天然条件下，HIV Env是一种高度动态的融合蛋白复合物， 可以在抗原和功能不同的构象状态之间闪烁。我们的生物物理研究 小组和其他人提供了第一个基于结构的迹象，表明Env的倾向是经历 大规模动态运动本质上是高度孤立的。这些变化直接影响给定的 Env与宿主体内生物因子的相互作用。在这里，我们将采用创新的结构分析 方法，包括结构质谱和冷冻EM，以表征Env多样性和识别 结构变化和动力学对抗体结合和抗原呈递的影响 树突状细胞显示凝集素，DC-SIGN。我们已经开发出一种有效的方法来净化类土著 SOSIP三聚体来自高度不同的HIV-1分离株，并在初步研究中证明， 即使来自抗中和的初级分离物，在稳定性、局部结构 bNAb表位的动力学、大规模构象呼吸和结构。我们将描述 来自整个中和层谱和来自全球小组的Env的结构动态剖面 耐药病毒，其组成高度代表循环中和表型。 我们将测试产生有利免疫应答（广度和效力）的变体是否表现出 结构动态特征我们将评估动态对抗体和DC-SIGN反应性的影响， 不同的病毒分离株。了解Env结构和功能的变化将为 了解艾滋病毒致病性和病毒适应性的差异，并更好地了解 免疫系统与这种极其多变的抗原搏斗。这些领域的进展将为我们的 了解艾滋病毒的演变，并可能有助于指导基于Env的疫苗和免疫原的开发。