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重组腺相关病毒2型转导人外周血单核细胞来源树突状细胞的研究

Infection Efficiency of Recombinant Adeno-associated Virus 2-mediated Exogenous Genes Transfer to Human Peripheral Blood Monocyte-derived Dendritic Cells

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【作者】 胡贵方康禹俞守义彭建强马鑫吴小兵

【Author】 HU Gui-fang~1,KANG Yu~2,YU Shou-yi~1,PE NG Jian-qiang~2,MA Xin~3,WU Xiao-bing~3(1Department of Tropical Army Hygiene of First Military College of Medici ne, China; 2.Benyuan Zhengyang Company of Genetic Technology, Ltd. China;,National Institute for Viral Dise ase Control and Prevention,China C DC,Beijing 100052,China)

【机构】 第一军医大学热带军队卫生学系本元正阳基因技术股份有限公司中国疾病预防控制中心病毒病预防控制所病毒基因工程国家重点实验室中国疾病预防控制中心病毒病预防控制所病毒基因工程国家重点实验室 北京 100052北京 100052

【Abstract】 To find out the infection efficiency of recombinant adeno-as sociated virus 2-mediated exogenou s genes in human peripheral blood monocyte-derived dendritic cells(D Cs),the process of transfection wa s investigated by FITC-labeled rAA V2,and observed under confocal mic roscope.Newly separated dendritic cells were tranfected by rAAV2-luc and rAAV2-GFP at different MOI,and transfection efficiency were detec ted by luminometer for rAAV2-luc a nd flow cytometry for rAAV2-GFP.Th e results were elucidated in four different assay systems:(1)60min w as needed for rAAV2 to bind on den dritic cells,and got into cells in the following 10min;(2)the express ion of luc could be detected at th e MOI as low as 1×10~5v.g/cell,and the expression plateau was reached by the MOI of 10~6~10~7v.g/cell,furt her increase of MOI had no functio n on expression level;(3)transgene expression was detected after 48h, and maintained a higher expression level from 96h to 240h after infec tion;(4)7days postinfection of rAA V2-GFP,5%~18% dendritic cells were GFP positive. These data suggest th at rAAV2 vector can efficiently in fect monocyte-derived dendritic ce lls and mediate exogenous gene exp ression,and that the application o f rAAV2 as vector may be useful fo r gene transfer to dendritic cells in ex vivo immunotherapy.

【基金】 国家863高科技发展计划基因治疗重大关键技术资助项目(编号:863-BH03-05-02)
  • 【文献出处】 病毒学报 ,Chinese Journal of Virology , 编辑部邮箱 ,2003年04期
  • 【分类号】R392
  • 【被引频次】1
  • 【下载频次】93
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