Modified Envelope Glycoproteins for HIV-1 Vaccine

用于 HIV-1 疫苗的修饰包膜糖蛋白

• 批准号: 6746563
• 负责人: JOHN P MOORE
• 金额: $ 32.71万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-15 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6746563
• 关键词: AIDS vaccines; CD4 molecule; HIV envelope protein; HIV envelope protein gp120; HIV envelope protein gp41; antibody formation; biotechnology; chimeric proteins; clinical research; conformation; disulfide bond; enzyme linked immunosorbent assay; guinea pigs; human immunodeficiency virus 1; human tissue; humoral immunity; laboratory rabbit; monoclonal antibody; neutralizing antibody; protein engineering; protein sequence; protein structure function; receptor binding; recombinant proteins; vaccine development

DESCRIPTION (provided by applicant): This is an application for the first competitive renewal of RO1 AI45463 "Modified envelope glycoproteins for HIV- 1 vaccine". Our overall goals are to gain additional knowledge of what forms of the HIV- 1 envelope glycoproteins might make effective immunogens for the induction of broadly neutralizing antibodies, and to learn about the properties of these proteins. Towards these goals, we propose three Specific Aims. 1: To study and compare the properties of cleaved, stabilized and uncleaved forms of oligomeric I-IIV-1 envelope glycoproteins. We seek to study purified monomeric, dimeric and trimeric forms of both cleaved and uncleaved HIV-1 JR-FL gpl40 proteins. An emphasis will be to better define the differences between cleaved and uncleaved trimers. We will t study the receptor-binding properties of the different Env forms; their reactivities with neutralizing and non-neutralizing MAbs; their stabilities to heat and detergent, and the pathways by which they dissociate under these conditions; their performance in ultra-centrifugation assays of molecular size; their appearance in electron micrographs both alone and as antibody complexes. 2:To evaluate the immunological responses elicited by cleaved, stabilized and uncleaved forms of oligomeric HIV-1 envelope glycoproteins in small animals. We will conduct immunogenicity studies in rabbits and guinea-pigs to determine whether cleaved, uncleaved and further modified forms of the HIV-1 Env complex can elicit neutralizing antibodies. These studies will involve immunization of the animals in a DNA-prime, protein-boost format, using in the priming phase both membrane-bound and soluble forms of HIV- 1 Env as a comparison. We will also evaluate the immunogenicity of Env complexes captured onto nanometer-sized beads to form particulate antigens. 3: To design, express and evaluate antigenic variants of the I-IIV-1 Env complex which contain additional modifications that may improve their immunogenicity. Unmodified forms of the HIV-1 Env complex may fail to induce broadly neutralizing antibodies, however they are presented. In anticipation of this outcome, we will make oligomeric forms of Env that contain additional modifications designed to enhance their immunogenicity. We will remove specific glycans and/or variable loops, create complexes containing CD4-mimetics, and attempt to induce Env configurations that resemble those that exist subsequent to CD4 binding.

描述（由申请人提供）：这是RO1 AI45463“修改的HIV-1疫苗的修饰信封糖蛋白”的首次竞争更新的应用程序。我们的总体目标是获得更多了解HIV-1包膜糖蛋白的形式的知识，可能使有效的免疫原诱导诱导广泛中和抗体，并了解这些蛋白质的特性。达到这些目标，我们提出了三个具体目标。 1：研究和比较寡聚I-I-I-I-I-1包膜糖蛋白的分裂，稳定和未裂解形式的特性。我们寻求研究裂解和未溶解的HIV-1 JR-FL GPL40蛋白的纯化的单体，二聚体和三聚体形式。重点是更好地定义切割和未裂解的三聚体之间的差异。我们将研究不同ENV形式的受体结合特性；它们具有中和和非中和mab的重新激活；它们的加热和洗涤剂的稳定性，以及它们在这些条件下分离的途径；它们在分子大小的超中心分析测定法中的性能；它们在电子显微照片中的出现单独和作为抗体复合物。 2：评估小动物中的寡聚HIV-1包膜糖蛋白的裂解，稳定和未溶解形式引起的免疫反应。我们将在兔子和豚鼠中进行免疫原性研究，以确定HIV-1 Env复合物的裂解，未裂解和进一步修饰的形式是否可以引起中和中和抗体。这些研究将涉及以DNA-PRIME，蛋白质增强格式对动物进行免疫，并在启动阶段使用膜结合和可溶性的HIV-1 Env进行了比较。我们还将评估捕获在纳米尺寸珠上的ENV复合物的免疫原性，形成颗粒抗原。 3：设计，表达和评估I-IIV-1 Env复合物的抗原变异，其中包含可能改善其免疫原性的其他修饰。 HIV-1 Env复合物的未修饰形式可能无法诱导广泛中和抗体，但是出现了。为了预期这种结果，我们将制作旨在增强其免疫原性的其他修饰的寡聚形式的ENV形式。我们将删除特定的聚糖和/或可变环，创建包含CD4摄影的复合物，并尝试诱导类似于CD4结合后存在的ENV配置。