Nucleofection of a DNA vaccine into human monocyte-derived dendritic cells.
An efficient method for delivering DNA vaccines into dendritic cells is considered to be of paramount importance. Electroporation-based technology (nucleofection) has gained increasingly popularity, but few reports focused on the possible functional consequences related to this method. In this study, the nucleofection technique was used to transfer the recombinant plasmid into hMoDCs for phenotype expression analysis and immunopotency detection. The results showed that the nucleofection of increasing concentrations of plasmid DNA decreased the viability of the hMoDCs. The welfare of nucleofected hMoDCs depended on the dosage of the plasmid and the plasmid's retention time within the cells. Accompanied by the process of nucleofection, it would bring some non-specific changes. The methodology reported here is suggestive of a feasible system for DNA vaccine transfer into hMoDCs with the caution of certain undesired effect.
Department of Biotechnology, Capital Institute of Pediatrics, Beijing 100020, China.
Qiu
Q
Xie
Jia
J
Luo
Zhiqiang
Z
Zhu
Guoliang
G
Wang
Jianhua
J
Wang
Bo
B
Niu
D002460:Cell Line; D003713:Dendritic Cells / Q000276:immunology*; D018274:Electroporation / Q000379:methods*; D015870:Gene Expression; D006801:Humans; D016130:Immunophenotyping; D016753:Interleukin-10 / Q000096:biosynthesis / Q000276:immunology; D018664:Interleukin-12 / Q000096:biosynthesis / Q000276:immunology; D009000:Monocytes / Q000276:immunology*; D012333:RNA, Messenger / Q000378:metabolism; D019444:Vaccines, DNA / Q000235:genetics* / Q000276:immunology
C508609:IL10 protein, human; D012333:RNA, Messenger; D019444:Vaccines, DNA; D016753:Interleukin-10; D018664:Interleukin-12