Engineered Envelope Glycoprotein Trimers for HIV-1 Vaccine Immunogens

此应用程序请求对Grant R21 AI087472“工程信封”的管理补充 用于HIV-1疫苗免疫原的糖蛋白三聚体“，这是2009年授予的一项赠款，现在是第二次免费 分机。由于飓风桑迪期间停电，从免疫的兔子身上收集的血清 两个修饰过的gp140蛋白已经无可挽回地丢失了。而抗环境病毒抗体效价的初步测定 在兔血清中显示这个项目有希望的结果，我们无法完成我们的分析 评估血清中和活性的范围和广度。补充资金将使我们能够 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环境免疫原，在 能够在人类体内引起广泛中和HIV-1抗体的稳定状态。 实验一：生物化学稳定的均一制剂的制备和表征 HIV-1 gp140三聚体。 实验二：在兔体内进行免疫原性研究，以确定稳定的天然gp140 三聚体可以提高中和抗体反应。 我们的总体目标是了解特定的gp41-gp41相互作用如何调节三聚化和稳定性。 并将这一知识应用于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，我们将表达和纯化到同源性的修饰的HIV-1 JR-FL gp140三聚体，并评价其在兔体内的免疫原性。这些相互关联的实验旨在 为了在兔子模型中生成概念验证信息，这些信息可以用来解决我们的主要问题 假设天然环境三聚体中特定的协作亚基间相互作用导致其固有的不稳定性 并且可以被改造成增强免疫原性。在有成本的情况下延长支助将大大 克服我们迫切需要的桑迪造成的物资损失，以实现中心目标 父母的资助。

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.