Engineered Envelope Glycoprotein Trimers for HIV-1 Vaccine Immunogens

用于 HIV-1 疫苗免疫原的工程包膜糖蛋白三聚体

基本信息

批准号：
8743611
负责人：
Min Lu
金额：
$ 19.68万
依托单位：
RBHS-NEW JERSEY MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-01-23 至 2016-01-22
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8743611
关键词：
Address Administrative Supplement Affect Animals Antibodies Antibody Formation Antigens Area Award Binding C-terminal Cavia Cell fusion Cells Cleaved cell Clinical Trials Complex Consensus DNA Data Development Disease Engineering Epitopes Foundations Funding Genetic Variation Glycoproteins Goals Grant HIV Envelope Protein gp120 HIV-1 Human Hurricane Immune response Immunization Immunology Infection Knowledge Lead Measurement Mediating Methods Modeling Modification Molecular Conformation Natural Immunity Oryctolagus cuniculus Outcome Particulate Plasma Pleomorphism Preparation Preventive Process Protein Engineering Proteins Public Health Recombinants Request for Applications Research Science Serum Son of Sevenless Proteins Specificity Structure Surface Testing Vaccines Viral Load result Virion Virus base cell mediated immune response cost design dimorphism env Gene Products env Glycoproteins flexibility immunogenicity improved insight nanoscale neutralizing antibody nonhuman primate novel parent grant programs research study structural biology transmission process vector virus genetics

项目摘要

This application requests an Administrative Supplement to Grant R21 AI087472 “Engineered envelope glycoprotein trimers for HIV-1 vaccine immunogens”, a grant awarded in 2009, and now in a second no-cost extension. As a result of power outages during Hurricane Sandy, sera collected from rabbits immunized with two modified gp140 proteins were irretrievably lost. While preliminary measurements of anti-Env antibody titers in the rabbit serum suggest promising outcomes for this project, we are unable to complete our analysis to assess the extent and breadth of the serum neutralizing activity. The supplemental funds will enable us to repeat our rabbit immunization experiments. These immunogenicity studies will provide a platform for new R01 research, intended to design and develop HIV-1 Env immunogens that present key quaternary epitopes in a stable state capable of eliciting broadly neutralizing HIV-1 antibodies in humans. Experiment 1: To produce and characterize homogeneous preparations of biochemically stabilized HIV-1 gp140 trimers. Experiment 2: To conduct immunogenicity studies in rabbits to determine whether stable native gp140 trimers can improve neutralizing antibody responses. Our overall goal is to understand how specific gp41-gp41 interactions modulate the trimerization and stability of the prefusion Env complex and to apply this knowledge to design native Env-trimer mimics with enhanced immunogenicity of neutralizing epitopes. To do this, we will express and purify to homogenicity modified HIV-1 JR-FL gp140 trimers, and evaluate their immunogenicity in rabbits. These interlinked experiments are intended to generate proof-of-concept information in the rabbit model that can be used to address our overarching hypothesis that specific cooperative inter-subunit interactions in native Env trimer lead to its inherent instability and can be engineered to enhance immunogenicity. The with-cost extension of support would significantly overcome losses of materials incurred by Sandy that we need desperately in order to fulfill the central objective of the parent grant.

本申请请求行政补充，以授予 R21 AI087472“工程信封 HIV-1 疫苗免疫原的糖蛋白三聚体”，2009 年授予的一项资助，现在是第二次免费资助由于桑迪飓风期间停电，从接种疫苗的兔子身上采集了血清。在初步测量抗 Env 抗体滴度时，两种修饰的 gp140 蛋白不可挽回地丢失。兔血清中的结果表明该项目有希望的结果，但我们无法完成我们的分析评估血清中和活性的程度和广度将使我们能够重复我们的兔子免疫实验，这些免疫原性研究将为新的 R01 提供一个平台。研究，旨在设计和开发 HIV-1 Env 免疫原，该免疫原在能够在人类体内引发广泛中和 HIV-1 抗体的稳定状态。实验 1：生产和表征生化稳定的均质制剂 HIV-1 gp140 三聚体。实验2：在家兔中进行免疫原性研究，以确定天然gp140是否稳定三聚体可以改善中和抗体反应。我们的总体目标是了解特定的 gp41-gp41 相互作用如何调节 gp41-gp41 的三聚化和稳定性预融合环境复合物，并应用这些知识来设计具有增强的天然环境三聚体模拟物中和表位的免疫原性为此，我们将表达并纯化同源性修饰的 HIV-1。 JR-FL gp140 三聚体，并评估其在兔子中的免疫原性。在兔子模型中生成概念验证信息，可用于解决我们的总体问题假设天然 Env 三聚体中特定的亚基间协同相互作用导致其固有的不稳定性并且可以通过工程设计来增强免疫原性，并且有成本地扩展支持将显着增加。克服桑迪造成的材料损失，我们迫切需要这些材料来实现中心目标的父母补助金。