Eliciting antibodies to probe native trimeric gp41 for HIV vaccine design

引发抗体来探测天然三聚体 gp41，用于 HIV 疫苗设计

• 批准号: 8790381
• 负责人: MICHAEL B ZWICK
• 金额: $ 56.51万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-15 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8790381
• 关键词: African; Antibodies; Antibody Formation; Antigens; B cell repertoire; B-Lymphocyte Epitopes; B-Lymphocytes; Binding; Bioinformatics; Biological Assay; Cell Lineage; Cell Separation; Cloning; Collaborations; Combined Vaccines; Complex; Cytoplasmic Tail; Data; Development; Elements; Epitopes; HIV Envelope Protein gp41; HIV vaccine; Human; Immune Sera; Immunization; Lead; Length; Liposomes; Membrane; Membrane Structure and Function; Mining; Modification; Molecular; Molecular Conformation; Mus; Peptides; Phylogeny; Population; Property; Protocols documentation; Receptor Cell; Regimen; Resistance; Resolution; Roentgen Rays; Sampling; Series; Serum; Site; Structure; Techniques; Testing; Time; Vaccine Design; Vaccines; Virion; Virus; cohort; design; env Glycoproteins; insight; nanoparticle; neutralizing antibody; next generation sequencing; novel; polyclonal antibody; prospective; public health relevance; response; scaffold; stoichiometry; structural biology; translational study; vaccine candidate; vaccine development; ward

DESCRIPTION: A crucial missing component of current HIV vaccine candidates is an ability to elicit broadly neutralizing antibodies. In the proposed studies we combine vaccine design, next generation sequencing (NGS) bioinformatics, structural biology and mechanistic studies to overcome an important roadblock in HIV vaccine development, that of eliciting antibodies that recognize native conformations of gp41. The membrane proximal external region (MPER) of gp41 is a particularly desirable vaccine target as it is recognized by numerous broadly neutralizing antibodies, the most potent of which is 10E8. However, how antibodies are elicited to the MPER is virtually unknown. Meanwhile, preliminary data suggest that the 10E8 epitope involves interactions between subunits of the trimeric envelope spike. The structure and function of the MPER in its native conformations is also incompletely understood. We propose to immunize mice with three trimeric presentations of the MPER on stabilized native spikes, on liposomes and on nanoparticles. We will identify novel MPER antibodies to epitopes overlapping with 10E8 from immunized mice and from African donors. We will use NGS/bioinformatics analyses to deeply probe the B cell repertoires of the mice and humans to identify germline V-regions of MPER-specific antibodies. We will attempt to manipulate MPER specific B cell responses in mice through antigen modification. In collaboration we will study the EM structure of stabilized envelope spikes in complex with 10E8, while using a battery of assays to define the mechanism of neutralization by 10E8.

描述：当前候选艾滋病毒疫苗的一个重要缺失部分是引发广泛中和抗体的能力。在拟议的研究中，我们结合了疫苗设计、下一代测序（NGS）生物信息学、结构生物学和机制研究，以克服 HIV 疫苗开发中的一个重要障碍，即引发识别 gp41 天然构象的抗体。 gp41 的近膜外部区域 (MPER) 是特别理想的疫苗靶点，因为它可以被许多广泛中和抗体识别，其中最有效的是 10E8。然而，如何引发针对 MPER 的抗体实际上尚不清楚。同时，初步数据表明 10E8 表位涉及三聚体包膜刺突亚基之间的相互作用。 MPER 的天然构象的结构和功能也尚未完全了解。我们建议用稳定的天然尖峰、脂质体和纳米粒子上的三种 MPER 三聚体表达来免疫小鼠。我们将从免疫小鼠和非洲供体中鉴定出针对与 10E8 重叠的表位的新型 MPER 抗体。我们将使用 NGS/生物信息学分析来深入探究小鼠和人类的 B 细胞库，以确定 MPER 特异性抗体的种系 V 区。我们将尝试通过抗原修饰来操纵小鼠中 MPER 特异性 B 细胞反应。在合作中，我们将研究与 10E8 复合的稳定包膜尖峰的 EM 结构，同时使用一系列测定来定义 10E8 的中和机制。