STABILITY AND STRUCTURE OF GP 120 ON INACTIVATED HIV

GP 120 对灭活 HIV 的稳定性和结构

• 批准号: 2752144
• 负责人: HAYNES W SHEPPARD
• 金额: $ 16.71万
• 依托单位: PUBLIC HEALTH FOUNDATION ENTERPRISES
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2000-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2752144
• 关键词: AIDS vaccines; HIV envelope protein gp120; HIV infections; active immunization; antibody titering; bioengineering /biomedical engineering; cell line; gene expression; genetic strain; human immunodeficiency virus 1; humoral immunity; laboratory rabbit; leukocytes; neutralizing antibody; phenotype; protein structure; vaccine development; virion; virus RNA

DESCRIPTION: (Adapted from applicant's abstract): A traditionally successful approach to viral vaccines, the whole inactivated virus (WIV), has received relatively little attention in the overall effort to develop an effective HIV/AIDS vaccine. Early successes with WIV in the SIV model were confounded by xenogeneic cellular antigens derived from the human cell lines used to propagate the SIV. Growing SIV in allogeneic or autologous monkey cells has shown partial protection in some experiments. In HIV, only one inactivated virus product (the Salk/IRC Immunogen) has been tested in humans and this material is not "whole" inactivated virus since the envelope glycoprotein has been completely removed. Furthermore, this product is being tested as a therapeutic rather than a prophylactic immunogen. Since published studies of WIV using HIV-1 are extremely limited, a great deal more needs to be done to evaluate this HIV/AIDS vaccine concept. One well known technical challenge to the development of WIV is the propensity of the envelope glycoprotein of HIV-1 8gp 120) to "fall off" the virion during the process of concentration , inactivation, and purification. A second potential problem is the loss of the "native" oligomeric structure of the gp 120 during inactivation steps that involve protein denaturation (e.g. formalin). Recent studies suggest that important antigenic epitopes may be lost when gp120 is converted from oligomeric to monomeric conformation. The proposed studies will be aimed at the identification and/or selection of HIV-1 variants, representing all major subtypes (A-F), which express a phenotype of high gp 120 stability that can be maintained through multiple passages in normal peripheral blood lymphocytes or cell lines. Starting with such HIV-1 variants should provide a significant advantage in the preparation of HIV that retains full antigenicity which may be critical for protection. The applicants will assess several inactivation procedures that preferentially target nucleic acid rather than protein (e.g. psoralen-UV, beta-propiolactone, gamma irradiation), oligomeric conformation of gp120. Finally, they will assess the immunogenicity of several WIV preparations in small animals. Initial studies will focus on subtype-C, one of the most prevalent subtypes in developing countries.

描述：（改编自申请人摘要）： 传统上成功的病毒疫苗方法， 灭活病毒（WIV），已经得到了相对较少的关注， 为研制有效的艾滋病毒/艾滋病疫苗作出全面努力。早期成功 在SIV模型中，WIV与异种细胞抗原混淆 来源于用于繁殖SIV的人类细胞系。SIV成长 在同种异体或自体猴细胞中， 一些实验。在HIV中，只有一种灭活病毒产物（Salk/IRC 免疫原）已经在人体中进行了测试，这种材料并不“完整” 灭活病毒，因为包膜糖蛋白已完全 删除.此外，该产品正在测试作为一种治疗，而 而不是预防性免疫原自从发表了使用HIV-1的WIV研究以来 非常有限，需要做更多的工作来评估这一点。 艾滋病毒/艾滋病疫苗概念。一个众所周知的技术挑战， WIV的发展是HIV-1包膜糖蛋白的倾向 8 gp 120）以在浓缩过程中“脱落”病毒体， 灭活和纯化。第二个潜在的问题是， GP 120的“天然”寡聚体结构在灭活步骤期间 涉及蛋白质变性（例如福尔马林）。最近的研究表明 当gp 120从 低聚物到单体构象。拟议的研究将针对 在鉴定和/或选择HIV-1变异体时，代表所有 主要亚型（A-F），表达高gp 120稳定性表型 可以通过正常外周血中的多个通道来维持 血液淋巴细胞或细胞系。从这种HIV-1变异开始， 在制备HIV中提供了显著的优势， 完全抗原性，这对于保护可能是关键的。申请人 将评估几种灭活程序， 核酸而不是蛋白质（例如，peptien-UV，β-丙内酯， γ辐射），GP 120的寡聚构象。最后，他们会 评估几种WIV制剂在小动物中的免疫原性。 最初的研究将集中在C亚型，这是最普遍的一种。 发展中国家的亚型。