HSV-1 Amplicon Vectors for HIV Vaccine Delivery

描述（由申请人提供）： 该计划项目的总体目标是开发基于HSV-1扩增子载体的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扩增子载体，并与NIAID疫苗研究中心（VRC）提供的DNA疫苗联合测试。我们提出三个项目。在Dewhurst的项目“Immunogenicity of HSV amplicon vectors in small animals”中，我们将在小鼠和豚鼠中进行研究，以检查：1）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扩增子构建体，4）使用匹配的DNA疫苗的初免-加强实验，以及5）分子佐剂。在刘易斯的项目“恒河猴中HSV扩增子载体的免疫原性”中，我们将验证具有SIV基因插入物的HSV扩增子的免疫原性。具体而言，我们将检查扩增子递送途径、剂量、多基因构建体特征和同源DNA初免-扩增子加强的效果。接下来，我们将在用DNA引发的最佳多基因SIV扩增子制备物免疫的猕猴中进行活病毒攻击实验。攻毒病毒将是高致病性的分子克隆SIVmac 239，用于评估对感染或疾病进展的保护作用。最后，我们将进行研究以确定编码多种HIV-1进化枝C抗原（Gag、达特、Nef、Env）的扩增子载体单独或与VRC DNA引物组合的免疫原性。这些数据将使我们能够就未来可能使用基于扩增子的HIV疫苗进行人体临床试验做出关键决定。在Federoff的项目“扩增子开发和增强”中，我们将通过以下方式优化扩增子技术：1）使用基于辅助病毒/无辅助病毒的方法和充分表征的重组酶系统开发用于高滴度扩增子生产的可扩展方法，2）开发基于亲和力的扩增子纯化方法，和3）通过蛋白质组学分析增强共包装和纯化的蛋白质组分的知识，这可以使得能够开发用于扩增子生产/纯化的改进方法。

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.