IMPROVED ADENOVIRAL VECTORS FOR HEPATIC GENE THERAPY

描述：重组腺病毒载体提供了人类基因的潜力 因为它们能够在高浓度下刺激许多组织， 体内效率 使用这些载体的热情已经 通过针对载体的强大免疫反应进行调节 由于载体衍生物的低水平合成， 抗原 事实上，腺病毒介导的基因转移是持久的， 缺乏抗原依赖性免疫力的动物需要产生一种 抗原性较低的载体。 凯博士和他的同事最近开发了一种 制备不含载体的高滴度腺病毒载体的方法 负责产生免疫反应的基因组序列。 出乎意料的是，这些缺失的载体，虽然在基因转移方面有效， 具有最小毒性或无毒性，在免疫活性或 免疫缺陷动物 这是因为这些被删除的载体可以 在体外和体内转导的细胞中不复制它们的基因组。 因此 假设腺病毒DNA的持续存在与 它的复制能力。 这项建议的主要目标是：1） 确定稳定性所需的腺病毒基因的最小数量， 把这些放回向量中，以形成一个可以持久存在的最小向量; 2)确定允许载体基因组持久性的机制; 3） 确定急性毒性和抗原依赖性免疫反应 针对新的载体;和4）开发缺失的载体系统， 产生整合载体系统。 这些研究的结果将 对腺病毒基因的发展具有重要意义 疗法

DESCRIPTION: Recombinant adenovirus vectors offer potential for human gene therapy because of their ability to transduce many tissues at high efficiency in vivo. The enthusiasm for use of these vectors has been tempered by a powerful immunologic response directed against vector containing cells because of the low level synthesis of vector derived antigens. The fact that adenovirus-mediated gene transfer is persistent in animals lacking antigen-dependent immunity establishes the need to produce a less antigenic vector. Dr. Kay and his colleagues recently developed a method for creating high titer adenovirus vectors that were devoid of vector genomic sequences responsible for producing the immune response. Unexpectedly, these deleted vectors, although efficient at gene transfer, with minimal or no toxicity, do not persist in vivo in immunocompetent or immunodeficient animals. This occurred because these deleted vectors could not replicate their genome in transduced cells in vitro and in vivo. Thus it is hypothesized that persistence of adenoviral DNA is inherently related to its ability to replicate. The major goals of this proposal are: 1) to determine the minimal number of adenovirus genes needed for stability and place these back into the vector to make a minimal vector that can persist; 2) determine the mechanism(s) allowing for vector genome persistence; 3) determine the acute toxicity and antigen-dependent immunological responses directed against the new vector; and 4) develop the deleted vector system to produce an integrating vector system. The results of these studies will have important implications for the development of adenoviruses for gene therapy.