Applying High-Performance Protein Engineering Tools to HIV Immunogen Design

将高性能蛋白质工程工具应用于 HIV 免疫原设计

• 批准号: 8686742
• 负责人: Margaret E Ackerman
• 金额: $ 75.24万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-18 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8686742
• 关键词: Address; Affinity; Antibodies; Antibody Formation; Antibody Repertoire; Antigens; Automobile Driving; B-Lymphocytes; Basic Science; Benchmarking; Clinic; Clinical; Development; Diagnosis; Disease; Engineering; Epidemic; Epitopes; Exhibits; Generations; HIV; HIV vaccine; HIV-1; Human; Immune; Immune response; Immune system; Immunity; Immunoglobulin Fragments; In Vitro; Individual; Infection; Lead; Libraries; Mediating; Methods; Molecular Conformation; Pathway interactions; Performance; Population Heterogeneity; Production; Protein Engineering; Proteins; Reagent; Receptors, Antigen, B-Cell; Sampling; Site; Solutions; Source; Structure; Surface; Surveys; Technology; Therapeutic; Therapeutic antibodies; Translating; Translations; Vaccination; Vaccine Design; Vaccines; Variant; Virus; Yeasts; base; combinatorial; design; flexibility; immunogenic; improved; innovation; insight; meetings; neutralizing antibody; novel; population based; pressure; response; screening; success; tool; vaccine development

DESCRIPTION (provided by applicant): Passively transferred neutralizing antibodies (nAbs) have provided definitive protection from HIV-1 infection, yet identification and mechanistic study of these nAbs has not resulted in the development of a protective vaccine. We hypothesize that previous immunogen design efforts have achieved limited success due to their limited scope and restricted focus. The lack of a high-performance screening platform places severe constraints on immunogen design and has impeded the translation of findings from basic science into vaccine development. When attempting to elicit protective antibodies against a highly diverse virus in the context of a disease in which naturally occurring protective immune responses are largely absent, we posit that the scale of technology and methods used must be adequately matched to the scale of the task. Combinatorial, high-throughput protein engineering platforms can achieve the landscape coverage that may be necessary for successful development of a preventative HIV vaccine, and allow us to translate our understanding of nAbs into the induction of protective antibody responses. We therefore propose to develop high-performance protein engineering tools to screen billions of HIV envelope trimer sequence variants according to defined criteria, allowing us to evolve envelope immunogens tailored to effectively present functionally relevant and immunogenic epitopes capable of driving the generation of protective antibodies. The multi-pronged strategy we describe both leverages the growing reagent toolkit for traditional immunogen design, including more than a dozen new broad nAbs, and improved means to functionally parse Abs present in diverse, polyclonal samples; and further explores a complementary strategy based on selective engagement of na¿ve and germline antibody repertoires which we believe addresses the fundamental limitation of previous immunogen design efforts and represents an innovative and unbiased forward engineering strategy to induce the generation of neutralizing antibodies. Ultimately, the tools developed by these studies represent adaptable platforms capable of rapidly selecting envelope variants from vast sequence diversity according to flexible design criteria, and may open a path to a fundamental breakthrough in immunogen design by bridging the translational gap that has come to be known as "the nAb problem".

描述（由适用提供）：被动转移的中和抗体（NABS）提供了免受HIV-1感染的明确保护，但是对这些NABS的识别和机械研究并未导致保护性疫苗的发展。我们假设以前的免疫原设计工作因其有限的范围和限制重点而取得了有限的成功。缺乏高性能筛选平台对免疫原具的严重限制，并阻碍了将基础科学发现转化为疫苗开发的结果。在试图在自然存在的受保护的免疫复杂的疾病的背景下，在很大程度上不存在疾病的情况下，在试图对高度潜水的病毒进行保护的抗体时，我们认为所使用的技术和方法的规模必须与任务规模充分匹配。组合，高通量蛋白工程平台可以实现成功开发预防性HIV疫苗所必需的景观覆盖范围，并允许我们将对NAB的理解转化为受保护抗体反应的诱导。因此，我们建议根据定义的标准开发高性能蛋白工程工具，以筛选数十亿的HIV围栏触发序列变体，从而使我们能够进化量身定制的包膜免疫原，以有效地存在功能相关和免疫原性的表位，能够促进产生的抗体抗体的产生。我们描述的是多管齐下的策略，这两种策略都利用了不断增长的试剂工具包用于传统的免疫原设计，包括十几个新的宽阔的NAB，以及改进的手段在潜水员，多克隆样品中的功能性ABS。并进一步探讨了一项基于NA¿VE和种系抗体曲目的选择性参与的完整策略，我们认为这涉及以前的免疫原设计工作的基本限制，并代表了一种创新且无偏向的远期工程策略，以引起中和抗体的产生。最终，这些研究开发的工具代表了能够根据柔性设计标准从巨大序列多样性中快速选择的包膜变体的适应性平台，并可能通过弥合被称为“ NAB问题”的翻译差距来打开免疫原设计基本突破的途径。