HIV ANTIGEN DISPLAY USING RECOMBINANT FLOATING VESICLES

使用重组浮动囊泡显示 HIV 抗原

• 批准号: 2544531
• 负责人: SHILADITYA DASSARMA
• 金额: $ 7.7万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-02-01 至 2001-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2544531
• 关键词: AIDS vaccines; HIV envelope protein; antigen presentation; laboratory mouse; laboratory rabbit; microorganism immunology; peptide library; plasmids; recombinant proteins; vaccine development; vesicle /vacuole

DESCRIPTION (adapted from applicant's abstract): The AIDS pandemic and associated generation of multiple antibiotic resistance among pathogens, in addition to recalcitrant endemic diseases such as malaria, all indicate that innovative approaches to vaccine development are needed. As a step toward this goal, we are developing an antigen delivery system for HIV-1 and HIV-2 clad viruses using a novel, particulate virus-like organelle called the gas vesicle, which is naturally used for flotation by aquatic prokaryotes. We have extensively studied the gas vesicles of salt-loving halophilic archaebacteria both genetically and biochemically, and found that these extremely stable proteinaceous structures are encoded by a cluster of 13 or 14 genes, including two genes, gvpA and gvpC, encoding major structural proteins. When we cross-linked TNP to gas vesicles and injected mice, we found high-level and long-term immune response in the absence of any adjuvant. When a peptide-encoding sequence was genetically engineered into gvpC, it was effectively presented to the mouse immune system. In order to facilitate the production of recombinant gas vesicles displaying HIV antigenic epitopes, we incorporated four unique restriction sites in gvpC and one site in the C-terminal coding region of gvpA. During the period of this proposal, we plan to (1) evaluate the best sites for genetic engineering of gas vesicles using selected SIV and HIV sequences, (2) produce epitope libraries of SIV and HIV Env and Gag proteins in gas vesicles and possible other sequences, and (3) evaluate the magnitude, duration, and memory of systemic response, and stimulation of mucosae and of a cell mediated response in mice. The use of SIV will permit subsequent testing of antigen delivery in a primate model. We hope to establish the feasibility of antigen presentation using halobacterial gas vesicles and lay the groundwork for the development of an HIV vaccine delivery system. If successful, this system would permit extremely large-scale production of a sale, inexpensive, and stable vaccine.

描述（改编自申请人的摘要）：艾滋病的流行和 病原体之间的多重抗生素耐药性的相关产生， 除了疟疾等难以控制的地方病外，所有这些都表明， 需要对疫苗开发采取创新办法。 作为迈向 为此，我们正在开发一种针对HIV-1和HIV-2的抗原递送系统， 使用一种新的，颗粒状的病毒样细胞器称为气体 囊泡，这是天然用于漂浮水生原核生物。 我们 广泛研究了嗜盐嗜盐菌的气泡 古细菌的遗传和生物化学，并发现这些 非常稳定的蛋白质结构由13个或更多个的簇编码。 14个基因，包括两个基因，gvpA和gvpC，编码主要结构蛋白， proteins. 当我们将TNP与气泡交联并注射小鼠时， 发现高水平和长期的免疫反应在没有任何 佐剂。 当一个肽编码序列被基因工程改造成 gvpC，它被有效地呈递给小鼠免疫系统。 为了 促进展示HIV的重组气体囊泡的产生 抗原表位，我们将四个独特的限制性位点，在gvpC 和gvpA的C-末端编码区中的一个位点。 期内 在这项建议中，我们计划（1）评估遗传学的最佳位点， 使用选定的SIV和HIV序列的气囊泡工程化，（2） 在气体中产生SIV和HIV Env和Gag蛋白的表位库 囊泡和可能的其他序列，和（3）评估大小， 持续时间和记忆的全身反应，和刺激粘膜和 一种细胞介导的反应。 SIV的使用将允许随后的 在灵长类动物模型中测试抗原递送。 我们希望建立 利用嗜盐菌气囊泡和层进行抗原呈递的可行性 为开发艾滋病毒疫苗输送系统奠定基础。 如果 如果成功，该系统将允许超大规模生产 销售、廉价和稳定的疫苗。