Nanoparticle HIV Protein Vaccines for Cellular Responses

用于细胞反应的纳米颗粒 HIV 蛋白疫苗

• 批准号: 7052033
• 负责人: Russell J Mumper
• 金额: $ 32.15万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-15 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7052033
• 关键词: AIDS vaccines; confocal scanning microscopy; cooperative study; dendritic cells; enzyme linked immunosorbent assay; flow cytometry; gag protein; human immunodeficiency virus 1; immunocytochemistry; laboratory mouse; nanotechnology; nickel; peptide chemical synthesis; protein purification; recombinant proteins; surface coating; tissue /cell culture; vaccine development; vaccinia virus; virus protein; virus replication

DESCRIPTION (provided by applicant): Currently, there is no effective vaccine for HIV infection. HIV vaccines can potentially be used for prevention of infection or therapeutically to control the level of HIV replication post-exposure. The overall goal of this 5-year research proposal is to develop nanoparticle-based HIV-1 vaccines to elicit enhanced Th1, cytotoxic T lymphocyte (CTL), and humoral immune responses to recombinant Tat (1-72) and Gag p24 proteins. The immunogenicity of the developed nanoparticle-based HIV-1 Tat (1-72) and Gag p24 vaccine will further be enhanced by, 1) the attachment of a dendritic cell targeting ligand, mannopentaose, to the nanoparticles, and 2) coating the nanoparticles with CpG to target the toll-like receptor-9 (TLR-9). Optimized nanoparticles vaccines will be compared to a heterologous prime boost strategy based on vaccinia expressing antigen and adjuvanted protein. Recombinant HIV Tat (1-72) and Gag p24 proteins and his-tagged proteins will be synthesized and purified to homogeneity by Dr. Nath's laboratory at Johns Hopkins and tested for bioactivity. In Dr. Mumper's laboratory at the University of Kentucky, the proteins will be either coated on anionic nanoparticles (Type 1) or attached to nanoparticles made with a small amount of accessible nickel at the surface (Type 2). Both types of solid nanoparticles (<100 nm) will be made from novel oil-in-water microemulsion precursors. Dr. Mumper's lab will characterize the nanoparticles, perform in-vitro uptake and activation studies in mouse and human dendritic cells, and perform animal experiments to determine the humoral immune responses of the nanoparticle-based formulations, and perform immunohistochemical analyses. Dr. Woodward's laboratory at the University of Kentucky will be responsible for ELISPOT, CTL, multi-probe ribonuclease, tetramer, cell proliferation assays, as well as confocal microscopy and flow cytometry experiments. Dr. Nath's laboratory will perform studies demonstrating that sera from immunized mice can suppress Tat-induced LTR-transactivation, and will assess the effect of Tat and Gag p24 antisera on HIV replication. In addition, Dr. Nath's lab will produce vaccinia expressing antigen.

描述（申请人提供）：目前，尚无针对HIV感染的有效疫苗。 HIV 疫苗可潜在地用于预防感染或在治疗上控制暴露后的 HIV 复制水平。 这项为期 5 年的研究计划的总体目标是开发基于纳米颗粒的 HIV-1 疫苗，以引发增强的 Th1、细胞毒性 T 淋巴细胞 (CTL) 以及对重组 Tat (1-72) 和 Gag p24 蛋白的体液免疫反应。所开发的基于纳米颗粒的 HIV-1 Tat (1-72) 和 Gag p24 疫苗的免疫原性将通过以下方式进一步增强：1) 将树突状细胞靶向配体甘露五糖附着到纳米颗粒上，2) 涂覆纳米颗粒与 CpG 一起靶向 Toll 样受体 9 (TLR-9)。 优化的纳米粒子疫苗将与基于牛痘表达抗原和佐剂蛋白的异源初免加强策略进行比较。 重组 HIV Tat (1-72) 和 Gag p24 蛋白以及 his 标记蛋白将由 Nath 博士位于约翰·霍普金斯大学的实验室合成并纯化至同质，并测试其生物活性。在肯塔基大学 Mumper 博士的实验室中，蛋白质将被涂覆在阴离子纳米粒子（1 型）上，或者附着在表面由少量可接触镍制成的纳米粒子（2 型）上。两种类型的固体纳米颗粒（<100 nm）均由新型水包油微乳液前体制成。 Mumper 博士的实验室将表征纳米颗粒，在小鼠和人类树突状细胞中进行体外摄取和激活研究，并进行动物实验以确定基于纳米颗粒的制剂的体液免疫反应，并进行免疫组织化学分析。 Woodward 博士位于肯塔基大学的实验室将负责 ELISPOT、CTL、多探针核糖核酸酶、四聚体、细胞增殖测定以及共聚焦显微镜和流式细胞术实验。 Nath 博士的实验室将进行研究，证明来自免疫小鼠的血清可以抑制 Tat 诱导的 LTR 反式激活，并将评估 Tat 和 Gag p24 抗血清对 HIV 复制的影响。此外，Nath博士的实验室将生产表达抗原的牛痘。