Improved Mimotope Structures and Scaffolds for Prime-Boost Strategies

改进的 Mimotope 结构和支架用于 Prime-Boost 策略

• 批准号: 7290750
• 负责人: Ralf Wagner
• 金额: $ 33.37万
• 依托单位: NOVARTIS VACCINES AND DIAGNOSTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-20 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7290750
• 关键词: AIDS vaccines; B lymphocyte; HIV envelope protein; Macaca; antigen antibody reaction; antigens; bioengineering /biomedical engineering; biological models; biomimetics; clinical research; epitope mapping; human immunodeficiency virus 1; laboratory rabbit; neutralizing antibody; simian immunodeficiency virus; transfection /expression vector; vaccine development; vaccine evaluation; vector vaccine; viruslike particle

Efforts to generate vaccines that elicit broadly neutralizing antibodies have been thwarted by features of the HIV-1 envelope glycoprotein (Env) that limit access to conserved, neutralizing epitopes and difficulties in mimicking neutralizing structures in vaccine constructs. These challenges apply especially to the pretransmembrane region (PTM) of gp41, which is the target of approximately half of the broadly neutralizing monoclonals that have been identified (2F5, 4E10, Z13), and the coiled-coil domain of the gp41 prehairpin intermediate, which is the target of the broadly-active fusion inhibitor T20 (Enfuvirtide) and a recently described neutralizing antibody D5. Here we propose to create and evaluate rationally-designed gp41 immunogens that present PTM, coiled-coil, and other HIV mimotopes in stable, highly exposed, oligomeric structures for eliciting broadly neutralizing antibodies. These studies will further investigate the influence of the virion membrane and virus-like particle (VLP) structure on antibody induction, especially for PTM determinants, by comparing gp41 oligomers in the context of VLPs or soluble immunogens. The immunogens utilize a novel, oligomeric gp41 scaffold, in which the N- and C-heptad repeats (HR) of SIV serve as a trimerization domain and the transmembrane domain (TM) and cytoplasmic tail of HIV serve as targeting signals for incorporationg oligomers into VLPs. We have shown that this gp41 scaffold allows insertion of large regions of Env between the trimerizaton and TM domains, without impairing oligomer assembly or incorporation of the gp41 immunogen into VLPs. The gp41 scaffold also contains an alternative insertion site, between the N- and C-HR, which may also be used to provide a highly exposed, loop context for evaluating some epitopes. Specific aims include creating and evaluating the following Env determinants presented in the oligomeric gp41 scaffold in VLP and soluble formats: 1) PTM region containing the 2F5 and 4E10 epitopes; 2) N-HR coiled-coil region 3) combination inserts involving PTM, coiled coil, and other HIV neutralizing mimotopes. Immunogens will be extensively characterized, evaluated as single immunogens, and directly compared with other immunogens in the program. Promising constructs will be further assessed in prime-boost combinations involving other vectors and immunogens in the program, with different formulations and adjuvants.

产生引起广泛中和抗体的疫苗的努力已受到特征的挫败 HIV-1信封糖蛋白（ENV）限制了访问保守的，中和表位和困难 模仿疫苗构建体中中和结构。这些挑战特别适用于 GP41的前跨膜区域（PTM），这是大约一半中和的目标 已鉴定的单克隆（2f5，4e10，Z13）和gp41 prehairpin的盘绕螺旋域 中间体，这是广泛活跃融合抑制剂T20（Enfuvirtide）的靶标 描述了中和抗体D5。在这里，我们建议创建和评估合理设计的GP41 稳定，高度暴露的低聚物中存在PTM，盘绕螺旋和其他HIV模仿的免疫原子 引发广泛中和抗体的结构。这些研究将进一步研究 抗体诱导上的病毒膜膜和病毒样颗粒（VLP）结构，尤其是PTM 决定因素，通过在VLP或可溶性免疫原的背景下比较GP41低聚物。这 免疫原利用一种新颖的寡聚GP41支架，其中N-和C-Heptad重复（HR）SIV 用作艾滋病毒的修剪域和跨膜结构域（TM）和细胞质尾巴 将掺入VLP的靶向信号。我们已经表明，此GP41脚手架允许 插入trimerizaton和TM域之间的大型ENV区域，而不会损害低聚物 将GP41免疫原的组装或掺入VLP中。 GP41支架还包含替代方案 n-和c-HR之间的插入站点，也可用于提供高度暴露的循环环境 用于评估一些表位。具体目的包括创建和评估以下ENV决定因素 以VLP和可溶性格式的寡聚GP41支架呈现：1）含有2f5和的PTM区域 4e10 eptopes; 2）N-HR盘绕线圈区域3）涉及PTM，盘绕线圈和其他HIV的组合插件 中和模型。免疫原子将被广泛特征，评估为单个免疫原， 并直接与该程序中的其他免疫原比较进行比较。有希望的结构将进一步评估 在涉及其他媒介和免疫原的原始组合中，有不同 配方和佐剂。