NOVEL VIRAL VECTORS FOR AIDS VACCINES

项目2中提议的研究的长期目标是开发 并优化预防免疫的新策略 使用腺相关病毒(AAV)和 痘病毒作为疫苗载体。我们要利用猿猴 以免疫缺陷病毒(SIV)猕猴艾滋病模型为开发工具。 就概念而言，项目0005可分为两部分：(A) 开发和检测AAV载体；及(B)检测痘病毒 载体(在Therion Biologics开发)。 (A)AAV是一种复制缺陷的细小病毒，具有独特的特征 使其作为稳定传递外源DNA的载体具有吸引力(in 在这种情况下，艾滋病毒基因)传递到细胞，总体上，作为疫苗载体。 首先，AAV不会在受感染的宿主中引起任何疾病(安全)。 其次，AAV在19号染色体上以特定的方式整合(长- 术语表达)。第三，AAV感染大多数(如果不是全部)哺乳动物细胞 (广义取向)。第四，AAV非常稳定和耐热 (不需要冷链)。最后，克隆的AAV DNA前病毒是 转基因后具有传染性，在体外可以很容易地操纵 (表达系统可以优化)。这一行动的具体目标 研究的目的是：(I)推导和表征AAV载体 SIV/HIV基因的表达；(Ii)rAAV的制备和鉴定 RAAV/SIV的免疫原性和免疫效力 在小鼠和猕猴身上；(Iv)开发新的战略，以有效地 包装rAAV；以及，(V)产生rAAV/HIV-1用于在非人类身上进行测试 HIV-1感染的灵长类动物模型。 (B)项目0005第二部分的目标是在工作的基础上 之前在本NCVDG的主持下演出。具体来说，我们 将继续研究使用活重组疫苗的策略 病毒载体(RVV)和假病毒粒子(非传染性SIV颗粒 从细胞培养中的rVV产生)。此外，我们还将测试新的rvv 表达辅助的SIV/HIV基因(nef、vif、vpx)和新的 减毒痘病毒载体。最后，我们将探索顺序 用痘病毒载体和rAAV进行免疫。 通过结合一些独特的艾滋病毒免疫接种方法，我们将 获得有关最佳方法和组合的宝贵知识 一种安全有效的艾滋病毒疫苗。

The long term goal of the proposed research in Project 2 is to develop and optimize novel strategies for prophylactic immunization against the human immunodeficiency virus (HIV) using adeno-associated virus (AAV) and poxviruses as vaccine vectors. We will exploit the simian immunodeficiency virus (SIV) macaque model of AIDS as development tool. For conceptual purposes, Project 0005 can be considered in 2 parts: (A) development and testing of AAV vectors; and, (B) testing of poxvirus vectors (developed at Therion Biologics). (A) AAV is a replication defective parvovirus with unique features that make it attractive as a vector for the stable delivery of foreign DNA (in this case, HIV genes) to cells, and overall, as a vaccine vehicle. First, AAV does not cause any disease in the infected host (safety). Second, AAV integrates in a site-specific fashion on chromosome 19 (long- term expression). Third, AAV infects most (if not all) mammalian cells (broad tropism). Fourth, AAV is extremely stable and heat resistant (cold chain not required). Finally, cloned AAV DNA proviruses are infectious after transfection and can be easily manipulated in vitro (expression systems can be optimized). The specific objectives of this research are to: (i) derive and characterize AAV vectors that direct expression of SIV/HIV genes; (ii) generate and characterize rAAV particles in vitro; (iii) test immunogenicity and efficacy of rAAV/SIV in mice and macaques; (iv) develop new strategies for the efficient packaging of rAAV; and, (v) generate rAAV/HIV-1 for testing in non-human primate models of HIV-1 infection. (B) The goal of the second part of Project 0005 is to build on work previously performed under the auspices of this NCVDG. Specifically, we will continue to investigate strategies using live recombinant vaccinia virus vectors (rVV) and pseudovirions (non-infectious SIV particles generated from rVV in cell culture). In addition, we will test new rVV which express auxiliary SIV/HIV genes (nef, vif, vpx) and novel attenuated poxvirus vectors. Lastly, we will explore sequential immunizations with poxvirus vectors and rAAV. By combining a number of unique approaches to HIV immunization, we will gain invaluable knowledge concerning the optimal approach and composition for a safe and effective HIV vaccine.