Novel HIV 1 Envelope immunogens derived from broadly neutralizing plasmas

源自广泛中和血浆的新型 HIV 1 包膜免疫原

• 批准号: 8531847
• 负责人: D. Noah Sather
• 金额: $ 53.29万
• 依托单位: SEATTLE BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-16 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8531847
• 关键词: Animals; Antibodies; Antibody Formation; Antigens; Autologous; B-Lymphocytes; Binding; Binding Sites; Bone Marrow; CD4 Antigens; Cloning; Communities; Development; Epitope Mapping; Epitopes; Evaluation; Exhibits; Generations; Goals; HIV; HIV-1; Immune response; Immunity; Immunoglobulin G; Infection; Libraries; Monitor; Oryctolagus cuniculus; Phage Display; Phase; Plasma; Process; Protocols documentation; Recombinants; Reporting; Research; Research Personnel; Resources; Specificity; Spleen; Structure; Testing; Tissues; Vaccination; Vaccines; Virus; base; combat; design; env Gene Products; immunogenic; immunogenicity; neutralizing antibody; neutralizing monoclonal antibodies; novel; pandemic disease; prophylactic; receptor binding; response; success; tool; vaccination strategy; vaccine candidate

ABSTRACT The overall aim of our studies is to design novel HIV Envelope immunogens capable of eliciting broadly reactive neutralizing antibodies (NAbs). Although attempts at eliciting broadly neutralizing antibody responses by vaccination have been unsuccessful, exceptionally potent broadly neutralizing antibody responses have been detected in some chronically infected HIV+ subjects. We propose to test new immunogens that are derived from Envelope sequences that were isolated from a well- characterized HIV+ subject whose exceptional broadly neutralizing activity targeted the CD4 receptor binding site (CD4-BS). We hypothesize that HIV-1 Envelope immunogens derived from HIV-1 infected subjects whose plasma exhibits potent, broad cross-neutralizing activity to the CD4-BS will elicit NAbs that are capable of broad cross-neutralization. This proposal is separated into two distinct phases. In the first phase, we propose the following: First, construct and characterize soluble trimeric gp140 Env proteins derived from autologous Env sequences isolated from well-characterized broadly neutralizing plasmas. Second, we will use these trimeric gp140 Envs as immunogens and monitor their ability to elicit broadly neutralizing antibody responses in animals. We will carefully dissect the NAb responses elicited by vaccination to determine both the quality and the epitope targets of the elicited NAb responses. In the second phase, we will isolate new monoclonal neutralizing antibodies from animals that develop broadly neutralizing antibodies in response to vaccinations. We will utilize both phage display libraries and antigen-specific single B cell cloning to isolate antibodies from spleen and bone marrow tissue of immunized animals. Novel antibodies and Fabs will be isolated and extensively characterized for binding and neutralizing activity. Promising Fabs will be made into full immunoglobulin G molecules and extensively characterized for cross-neutralizing potential and binding epitope. Novel anti-HIV neutralizing antibodies from this study will provide valuable new tools to the research community, and help to define new neutralization epitopes on the HIV-1 Envelope spike.

抽象的 我们研究的总体目的是设计新型的HIV信封免疫原 能够引起广泛反应性中和抗体（NABS）。虽然尝试 通过疫苗接种引起广泛中和抗体反应的过程已经 失败的，极有效的广泛中和抗体反应已经 在某些长期感染的HIV+受试者中检测到。我们建议测试新 从孔中分离出来的包膜序列的免疫原子 表征了HIV+主题，其特殊的广泛中和活性针对 CD4受体结合位点（CD4-BS）。我们假设HIV-1信封 来自HIV-1感染受试者的免疫原子，其血浆表现出有效的疫苗， CD4-BB的广泛的交叉中和活动将引起能够的NABS 广泛的交叉中和化。 该建议分为两个不同的阶段。在第一阶段，我们 提出以下内容：首先，构建和表征可溶性三聚体GP140 env 源自从特征良好的自体env序列中得出的蛋白质 广泛中和等离子体。其次，我们将使用这些三聚gp140 envs 免疫原生物并监测其在中和抗体反应中广泛中和的能力 动物。我们将仔细剖析疫苗接种引起的NAB反应 确定引起的NAB响应的质量和表位目标。 在第二阶段，我们将把新的单克隆中和抗体隔离 响应疫苗接种而形成广泛中和抗体的动物。我们 将同时利用噬菌体显示库和抗原特异性的单个B细胞克隆 来自免疫动物的脾脏和骨髓组织的分离抗体。 小说 抗体和晶圆厂将被隔离并广泛特征以结合和 中和活性。 有希望的晶圆厂将被制成完整的免疫球蛋白G 分子和广泛的特征，用于交叉中和势和结合 表位。这项研究的新型抗HIV中和抗体将提供有价值的新抗体 研究社区的工具，并有助于定义新的中和表位 HIV-1信封尖峰。