腺病毒载体介导gfp基因在树突细胞的转导及其影响因素的分析

目的：研究腺病毒载体介导外源基因在人树突细胞转染的有效方法。方法：绿色荧光蛋白（gfp）报告基因重组腺病毒的构建采用直接连接法。人树突细胞的制备通过分离人外周血单核细胞，然后在体外经过诱导过程再生。结果：经腺病毒介导实现了gfp基因在树突细胞的转导和表达。病毒滴度对转导效率影响较大，只有使用高滴度（MOI〉100）的重组腺病毒才能获得较高的转导效率（40％以上）；脂质体和多聚赖氨酸可以明显提高转导效率（提高50％左右）。转导效率最高可达65％左右。结论：由腺病毒介导进行树突细胞的转基因需要较高的病毒滴度；脂质体和多聚赖氨酸可以提高基因的转导效率。

Adenoviris-mediated gfp gene transduction in dendritic cells and analysis of some factors involved

Objective:To obtain an efficient method for transfecting the foreign gene into human dendritic cells.Methods:The recombinant adenovirus containing green fluorescence protein(GFP) gene was constructed using the ligation method. The dendritic cells were generated from the monocytes isolated from human peripheral blood.Results:The gfp gene transduction and expression in dendritic cells were achieved by adenovirus mediation. The viral titer had a great affect on the transduction. Only the high titer virus used(MOI>100) results in the reasonable transduction(above 40%). The transduction was significantly increased by using liposome and polylysine(increasing about 50%), the maximum transduction efficiency could reach 65%.Conclusion:The high viral titer was needed for transduction into human dendritic cells. The liposome and polylysine increase the transduction.