PROJECT 2:Structural studies of SOSIP trimers

项目2：SOSIP三聚体的结构研究

基本信息

批准号：
10427133
负责人：
IAN A WILSON
金额：
$ 119.94万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-06-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10427133
关键词：
Adjuvant Affinity Animals Antibodies Antibody Response Antigen Presentation Antigens Autologous B-Cell Antigen Receptor B-Lymphocytes Binding Biochemical Biological Assay Biophysics Calorimetry Chemicals Collaborations Complex Coronavirus Cryoelectron Microscopy Data Differential Scanning Calorimetry Ebola virus Electron Microscopy Engineering Ensure Epitopes Exercise Feedback Ferritin Formulation Foundations Funding Generations Glycoproteins Goals HIV-1 Hepatitis C virus Human Hydrogen Immunization Immunize Immunologics Infection Letters Macaca Mass Spectrum Analysis Methods Molecular Sieve Chromatography Negative Staining Outcome Particulate Pathway interactions Procedures Property Research Institute Resolution Roentgen Rays Series Sorting - Cell Movement Stress Tests Structural Biologist Structure Surface Surface Plasmon Resonance Techniques Testing Titrations Vaccine Design Vaccines Validation Viral Viral Envelope Proteins Work X-Ray Crystallography base biophysical analysis biophysical techniques biophysical tools crosslink design experience immunogenicity improved in vivo influenzavirus iron oxide nanoparticle light scattering nanoparticle neutralizing antibody next generation particle preservation rational design response simian human immunodeficiency virus trafficking vaccine trial ward

项目摘要

Project 2 Abstract Project 2 will generate structural information to validate and improve the design of new generations of SOSIP trimers in close interaction with Project 1 and Core B. Project 2 will take a highly integrative approach involving cryo- and negative-stain-electron microscopy (NS-EM), X-ray crystallography and a range of other biophysical tools (SEC-MALS, ITC, DSC and SPR) to provide a comprehensive biophysical understanding of trimers and trimer-bnAb complexes. The data will be shared with Project 1 to improve the stability, antigenicity and immunogenicity of new SOSIP trimer designs. Dr. Ian A. Wilson will direct Project 2 at The Scripps Research Institute and Dr. Andrew B. Ward will be co-Leader. The Specific Aims are: Aim 1: To biophysically and structurally characterize new SOSIP trimers, including germline targeting SOSIP trimers. We will generate high-resolution structures by cryo-EM or X-ray crystallography to enable further engineering SOSIP trimers, and low-resolution structures by NS-EM to provide rapid feedback about the quality of SOSIP trimers. We will also use NS-EM to assess the quality of trimers after stress tests that mimic vaccine storage and formulation conditions such as the effect of adjuvant mixing. These assays are critical prior to animal immunization studies, to ensure only high-quality trimers are evaluated. Aim 2: To determine high-resolution structures of SHIV infection-elicited, vaccine-induced NAbs, or naïve precursor antibodies isolated with GT-SOSIP trimers. We will solve structures of neutralizing Abs (NAbs) isolated from trimer-immunized and SHIV-infected animals to assess their similarity to known bNAbs and their potential for developing breadth. The goals are to further optimize the presentation of bNAb epitopes on SOSIP trimers and facilitate the design of sets or series of trimers for steering responses toward bnAbs in prime-boost-boost strategies that mimic infection. We will solve structures of SOSIP trimer complexes with germline precursor antibodies to compare with affinity-matured bnAbs. The goal is to design SOSIP trimer boosts for guiding precursor antibodies toward breadth. Aim 3: To design, optimize and determine structures of SOSIP trimers displayed on nanoparticle platforms. We will incorporate SOSIP trimers into various proteinaceous nanoparticles (NPs), such as the two-component self-assembling NPs, via an iterative cycle of structure-based design and testing. The goal is to preserve the desirable antigenic properties of SOSIP trimers, while gaining the immunological benefits of particulate antigen presentation. Working with Project 1 and Core B, we will evaluate a range of designs and thereby generate an arsenal of SOSIP-NP immunogens that will be tested in animals. The outcomes of those studies will further guide the design of new SOSIP-NP constructs.

项目2摘要项目 2 将生成结构信息以验证和改进新一代 SOSIP 的设计三聚体与项目 1 和核心 B 密切相互作用。项目 2 将采取高度综合的方法，涉及冷冻电子显微镜和负染色电子显微镜 (NS-EM)、X 射线晶体学和一系列其他生物物理学工具（SEC-MALS、ITC、DSC 和 SPR）提供对三聚体和三聚体-bnAb 复合物的数据将与项目 1 共享，以提高稳定性、抗原性和稳定性。 Ian A. Wilson 博士将指导斯克里普斯研究中心的项目 2。研究所和 Andrew B. Ward 博士将担任联合领导者，具体目标是：目标 1：从生物物理和结构上表征新的 SOSIP 三聚体，包括种系我们将通过冷冻电镜或 X 射线晶体学生成高分辨率结构。能够通过 NS-EM 进一步设计 SOSIP 三聚体和低分辨率结构，以提供快速反馈关于 SOSIP 三聚体的质量，我们还将在压力测试后使用 NS-EM 来评估三聚体的质量。模拟疫苗储存和配制条件，例如佐剂混合的影响，这些测定至关重要。在动物免疫研究之前，确保仅评估高质量的三聚体。目标 2：确定 SHIV 感染引起、疫苗诱导的高分辨率结构 NAb，或用 GT-SOSIP 三聚体分离的天然前体抗体，我们将解析其结构。从三聚体免疫和 SHIV 感染的动物中分离出中和抗体 (NAb)，以评估其与已知的 bNAb 及其发展广度的潜力，目标是进一步优化 bNAb 的展示。 SOSIP 三聚体上的 bNAb 表位，有助于设计三聚体组或系列，以引导反应模拟感染的初免-加强-加强策略中的 bnAb 我们将解析 SOSIP 三聚体复合物的结构。与种系前体抗体与亲和力成熟的 bnAb 进行比较，目标是设计 SOSIP 三聚体。引导前体抗体走向广度的增强。目标 3：设计、优化和确定 SOSIP 三聚体的结构我们将把 SOSIP 三聚体整合到各种蛋白质纳米颗粒 (NP) 中，例如通过基于结构的设计和测试的迭代循环的两组分自组装纳米颗粒。目标是保留 SOSIP 三聚体所需的抗原特性，同时获得免疫学益处与项目 1 和核心 B 合作，我们将评估一系列设计和产生一系列 SOSIP-NP 免疫原，并在动物中进行测试。研究将进一步指导新 SOSIP-NP 结构的设计。