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: 8409958
• 负责人: Margaret E Ackerman
• 金额: $ 47.83万
• 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/8409958
• 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; Screening procedure; 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; 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". PUBLIC HEALTH RELEVANCE: Our proposed studies aim to improve our understanding of the interaction between the HIV envelope and the immune system, providing new insight into envelope-based vaccine design, facilitating development of a protective vaccine to contain the HIV epidemic.

描述（由申请人提供）：被动转移的中和抗体（nAb）已经提供了针对 HIV-1 感染的明确保护，但这些 nAb 的鉴定和机制研究尚未导致保护性疫苗的开发。我们与之前的免疫原设计工作进行了斗争。由于范围有限和关注点有限，缺乏高性能的筛选平台严重限制了免疫原的设计，并阻碍了将基础科学的发现转化为疫苗的开发。在自然发生的保护性免疫反应基本上不存在的疾病背景下，高度多样化的病毒，我们认为所使用的技术和方法的规模必须与任务的规模充分匹配。实现成功开发预防性艾滋病毒疫苗所需的景观覆盖，并使我们能够将对单克隆抗体的理解转化为诱导保护性抗体反应，因此，我们建议开发高性能蛋白质工程工具来筛查数十亿艾滋病毒。根据定义的标准，包膜三聚体序列变体，允许我们开发出适合有效呈现功能相关和免疫原性表位的包膜免疫原，这些表位能够驱动保护性抗体的产生。我们描述的多管齐下的策略都利用了用于传统免疫原设计的不断增长的试剂工具包，包括十多种新的广泛的nAb，以及改进的功能性解析不同多克隆样品中存在的 Ab 的方法，并进一步探索基于 na¿ 的选择性参与的补充策略；我们相信这些研究解决了以前免疫原设计工作的根本局限性，并代表了一种创新且公正的正向工程策略来诱导中和抗体的产生，最终，这些研究开发的工具代表了能够快速选择包膜的适应性平台。根据灵活的设计标准，从巨大的序列多样性中提取变体，并可能通过弥合被称为“nAb 问题”的翻译缺口，为免疫原设计的根本性突破开辟道路。 公共健康相关性：我们提出的研究旨在提高我们对 HIV 包膜和免疫系统之间相互作用的理解，为基于包膜的疫苗设计提供新的见解，促进开发遏制 HIV 流行的保护性疫苗。