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：目前HIV候选疫苗的一个关键缺失部分是引发广泛中和抗体的能力。在拟议的研究中，我们结合联合收割机疫苗设计，下一代测序（NGS）生物信息学，结构生物学和机制研究，以克服一个重要的障碍，在HIV疫苗的开发，引发抗体，识别天然构象的gp 41。gp 41的膜近端外部区域（MPER）是特别期望的疫苗靶标，因为它被许多广泛中和抗体识别，其中最有效的是10 E8。然而，抗体是如何引发MPER的几乎是未知的。同时，初步数据表明，10 E8表位涉及三聚体包膜刺突亚基之间的相互作用。MPER在其天然构象中的结构和功能也不完全了解。我们建议用三种三聚体的MPER对小鼠进行免疫接种，所述三种三聚体的MPER在稳定的天然刺突上、在脂质体上和在纳米颗粒上。我们将从免疫小鼠和非洲供体中鉴定针对与10 E8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测小鼠和人的B细胞库，以鉴定MPR特异性抗体的生殖系V区。我们将尝试通过抗原修饰来操纵小鼠中的MPER特异性B细胞应答。在合作中，我们将研究与10 E8复合的稳定的包膜尖峰的EM结构，同时使用一系列测定来确定10 E8的中和机制。