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: 10186690
• 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/10186690
• 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-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; antimicrobial peptide; base; 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.

我们长期以来一直意识到HIV-1 Invelope糖蛋白（ENV）的惊人序列变化 基因，但是这种多样性的结构和功能含义才被掌握。结构 ENV的变化影响其与病毒健康和复制的所有关键驱动因素的互动，这不是 由序列变化捕获。这些差异是病毒表型特征（例如中和）的基础 敏感性，向流，感染力和传播性。虽然最近的研究提供了详细的结构 从一系列病毒分离株中的三聚体环境域的信息，结构代表静态的柏拉图式 Eng组装的理想。在天然条件下，HIV Env是一种高度动态的融合蛋白复合物 可以在抗原和功能上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指示，即ENV经历的倾向 大规模的动态运动本质上是高度分离的。这些变化直接影响给定的 Env与宿主中生物学因素的相互作用。在这里，我们将应用创新的结构分析 包括结构性质谱和冷冻EM在内的方法，以表征各种各样的多样性并确定 结构变化和动力学对抗体结合和抗原表现的后果 树突状细胞显示凝集素，DC-SIGN。我们已经开发了一种有效的方法来净化类似土著的方式 来自高度不同的HIV-1分离株的SOSIP三聚体，并在初步研究中证明了三聚体 即使来自中和抗性原发性分离株也显示出稳定性的显着差异，局部结构 BNAB表位，大规模构象呼吸和结构的动力学。我们将表征 从中和层频谱和全球面板中的Envs的结构动态曲线 耐药病毒的高度代​​表性的中和表型。 我们将测试产生有利免疫反应（广度和效力）的变体是否表现出来 结构动态特征。我们将评估动力学对抗体和直流反应性的影响 各种病毒分离株。了解Env结构和功能的变化将为 了解HIV致病性和病毒适应性的差异，并更好地理解如何 免疫系统与这种极度可变的抗原捕获。在这些领域的进展将告知我们 了解艾滋病毒的进化，并可能有助于指导基于ENV的疫苗和免疫原子的开发。