HSV-1 Amplicon Vectors for HIV Vaccine Delivery

用于 HIV 疫苗递送的 HSV-1 扩增子载体

• 批准号: 6682052
• 负责人: MICHAEL C KEEFER
• 金额: $ 138.04万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-15 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6682052
• 关键词: AIDS vaccines; biotechnology; herpes simplex virus 1; transfection /expression vector; vaccine development; vector vaccine

DESCRIPTION (provided by applicant): The overall goal of this Program Project is to develop an HIV vaccine strategy based on the HSV -1 amplicon vector, and to test this approach in small animal and large animal models. We will test the HSV amplicon vector alone and in combination with a DNA vaccine provided by the NIAID Vaccine Research Center (VRC). We propose 3 Projects. In Project by Dewhurst, "Immunogenicity of HSV amplicon vectors in small animals", we will carry out studies in mice and guinea pigs to examine: 1) amplicons delivered mucosally, 2) their interaction with dendritic cells, 3) multigene SIV amplicon constructs, 4) a prime-boost experiment with a matching DNA vaccine, and 5) molecular adjuvants. In Project by Lewis, "Immunogenicity of HSV amplicon vectors in rhesus macaques", we will verify the immunogenicity of the HSV amplicon with SIV gene inserts. Specifically, we will examine amplicon delivery route, dose, multigene construct characteristics and the effect of homologous DNA prime- amplicon boost. Next we will perform a live virus challenge experiment in macaques immunized with the optimal multigenic SIV amplicon preparation with DNA priming. The challenge virus will be a highly pathogenic, molecularly cloned, SIVmac239 for assessment of protection from infection or disease progression. Finally, we will conduct studies to determine the immunogenicity of amplicon vectors encoding multiple HIV-1 clade C antigens (Gag, Tat, Nef, Env), alone or in combination with the VRC DNA prime. This data will allow us to make key decisions concerning a possible future human clinical trial using amplicon-based HIV vaccines. In Project by Federoff, " Amplicon Development and Enhancement", we will optimize the amplicon technology by: 1) developing a scalable process for high-titer amplicon production using helper virus-based/helper virus-free methods and well-characterized recombinase systems, 2) developing an affinity-based amplicon purification method, and 3) enhancing knowledge of co-packaged and purified protein constituents by proteomic analysis, which may enable the development of improved methods for amplicon production/purification.

描述(由申请人提供)： 该项目的总体目标是开发一种基于HSV-1扩增载体的HIV疫苗策略，并在小动物和大动物模型中测试这种方法。我们将单独测试HSV扩增载体，并与NIAID疫苗研究中心(VRC)提供的DNA疫苗一起测试。我们提出了3个项目。在Dehuurst的“HSV扩增子载体在小动物中的免疫原性”项目中，我们将在小鼠和豚鼠身上进行研究，以检查：1)经粘膜递送的扩增子，2)它们与树突状细胞的相互作用，3)多基因SIV扩增子的构建，4)匹配DNA疫苗的Prime-Boost实验，以及5)分子佐剂。在Lewis的“HSV扩增子载体在恒河猴中的免疫原性”项目中，我们将验证带有SIV基因插入的HSV扩增子的免疫原性。具体地说，我们将研究扩增子的传递途径、剂量、多基因结构特征以及同源DNA启动子增强的效果。接下来，我们将在用DNA启动的最佳多基因SIV扩增制剂免疫的猕猴中进行活病毒攻击实验。这种挑战病毒将是一种高致病性的分子克隆病毒SIVmac239，用于评估对感染或疾病进展的保护。最后，我们将进行研究，以确定编码多个HIV-1分支C抗原(Gag、Tat、Nef、Env)的扩增载体单独或与VRC DNA素联用的免疫原性。这些数据将使我们能够就未来可能进行的使用基于扩增子的艾滋病毒疫苗的人类临床试验做出关键决定。在费德洛夫的“扩增子开发和增强”项目中，我们将通过以下几个方面优化扩增子技术：1)利用基于辅助病毒/辅助病毒的方法和具有良好特性的重组酶系统开发可扩展的高滴度扩增子生产工艺，2)开发基于亲和力的扩增子纯化方法，以及3)通过蛋白质组分析增强对共包装和纯化蛋白质组分的了解，这可能使改进的扩增子生产/纯化方法的开发成为可能。