Development of Fiber-Mutant Adenovirus Vectors for targeting gene therapy

我们已经证明了腺病毒载体（Ad）基因修饰的树突状细胞（dc）在免疫治疗中的有效性，而高剂量Ad需要充分的基因转导到树突状细胞中。RT-PCR分析显示，dc对ad介导的基因转移的相对抗性是由于缺乏柯萨奇-腺病毒受体的表达，并且dc表达了足够的α (v)-整合素。因此，我们研究了纤维突变体Ad在纤维旋涡中含有Arg-Gly-Asp （RGD）序列是否能有效地将LacZ基因高水平转导并表达到dc中。在小鼠DC系和正常人DC （NHDC）中，纤维突变Ad的基因传递效率高于常规Ad。此外，在8000个载体颗粒/细胞下，用纤维突变Ad和常规Ad转导的NHDC导致β -半乳糖苷酶活性相差70倍。我们提出，以α (v)整合素为目标的Ad是一种非常强大的工具，可用于实施基于dc的疫苗接种策略。

We have demonstrated the usefulness of dendritic cells (DCs) genetically modified by adenovirus vectors (Ad) to immunotherapy, while sufficient gene transduction into DCs is required for high doses of Ad. The RT-PCR analysis revealed that the relative resistance of DCs to Ad-mediated gene transfer is due to the absence of Coxsackie-adenovirus receptor expression, and that DCs expressed adequate alpha(v)-integrins. Therefore, we investigated whether fiber-mutant Ad containing the Arg-Gly-Asp (RGD) sequence in the fiber knob can efficiently transduce and express high levels of the LacZ gene into DCs. The gene delivery by fiber-mutant Ad was more efficient than that by conventional Ad in both murine DC lines and normal human DCs (NHDC). Furthermore, NHDC transduced with fiber-mutant Ad and conventional Ad at 8000-vector particles/cell resulted in a 70-fold difference in beta-galactosidase activity. We propose that alpha(v)-integrin-targeted Ad is a very powerful tool with which to implement DC-based vaccination strategies.