MULTI-TIER AIDS VACCINE--MANUFACTURE AND TESTING

该项目的长期目标是开发一种疫苗，以保护人类免受自然界中发现的不同抗原形式的HIV-1感染。这种保护需要引发针对不同HIV- 1包膜蛋白的免疫应答。我们认为，必须在疫苗中呈现多种HIV-1包膜蛋白，以诱导针对HIV-1变体范围的免疫记忆。我们进一步提出，可以通过利用一种以上的疫苗递送系统来优化不同应答的幅度。我们的临床前研究评价了三重疫苗递送系统，其具有（i）重组DNA疫苗，随后是（ii）重组牛痘病毒疫苗，随后是（iii）纯化蛋白疫苗（D-V-P），证明了小鼠中高水平的HIV-1中和抗体。该项目的目标是将这一疫苗战略转化为人类应用。一种多包膜重组牛痘病毒疫苗（PolyEnv 1）目前正在进行临床试验（项目4）。因此，在本项目中，我们将制备用于人类的DNA和纯化的蛋白质。该项目的前两个目标描述了临床级材料的设计和生产。虽然技术驱动，这两个目标是至关重要的小规模人体试验（项目4）的进行。第三个目标是检验这些目标对于进行小规模人体试验至关重要的假设（项目）。第三个目的是检验我们的3层多包膜疫苗策略可以保护非人灵长类动物免受异源病毒感染的假设。这一目标的实现对于疫苗方法的临床进展至关重要。本项目的具体目标是：目标1：制备安全、免疫原性好且适用于人类的多包膜DNA疫苗。目的2：制备一种安全、免疫原性好、适用于人的多囊膜蛋白疫苗。目的3：确定三层多包膜疫苗是否保护非人灵长类动物免受异源病毒攻击。

The long term objective of this project is to develop a vaccine to protect humans from antigenically different forms of HIV-1 found in nature. Such protection will require elicitation of immune responses against diverse HIV- 1 envelop proteins. We submit that multiple HIV-1 envelope proteins must be presented in a vaccine to induce immunological memory against the range of HIV-1 variants. We further proposed that the magnitude of diverse responses may be optimized by utilization of more than one vaccine delivery system. Our pre-clinical studies evaluation a three-tired vaccine delivery system with (i) recombinant DNA vaccine followed by (ii) recombinant vaccinia virus vaccine followed by (iii) purified protein vaccine (D-V-P) demonstrate high levels of HIV-1 neutralizing antibodies in mice. The goal of this project is to translate this vaccine strategy to human application. A multi-envelope recombinant vaccinia virus vaccine (PolyEnv1) is currently in clinical trial (Project 4). Accordingly, in this project we will prepare DNA and purified proteins fro use in humans. The first two aims of this project describe the design and production of clinical grade material. While technically driven these two aims are critical for the conduct of small scale human trials (Project 4). The third aim tests the hypothesis that these aims are critical for the conduct of small scale human trials (Project). The third aim tests the hypothesis that our 3-tiered, multi- envelope vaccine strategy can protect non-human primates from heterologous virus. The implementation of this aim is critical for clinical advanced of the vaccine approach. Specific aims developed in this project are: Aim 1: To prepare a multi-envelope DNA vaccine that is safe, immunogenic and suitable for use in humans. Aim 2: To prepare a multi-envelope protein vaccine that is safe, immunogenic and suitable for use in humans. Aim 3: To determine whether a three-tiered multi-envelope vaccine protects non-human primates from heterologous virus challenge.