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  

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

作　　者：仲飞[1] 仲振宇[2] 梁爽[2] 李秀锦[3] 

机构地区：[1]河北农业大学动物科技学院基础兽医学部,保定071001 [2]河北大学生命科学学院,保定071001 [3]燕山大学环境与化学工程学院生物工程系

出　　处：《中国免疫学杂志》2006年第11期991-996,共6页Chinese Journal of Immunology

摘　　要：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。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.

关 键 词：树突细胞 基因转导 腺病毒载体 

分 类 号：R392.11[医药卫生—免疫学]