Van Tendeloo V F, Snoeck H W, Lardon F, Vanham G L, Nijs G, Lenjou M, Hendriks L, Van Broeckhoven C, Moulijn A, Rodrigus I, Verdonk P, Van Bockstaele D R, Berneman Z N
Laboratory of Experimental Hematology, University of Antwerp (UIA/UZA), Belgium.
Gene Ther. 1998 May;5(5):700-7. doi: 10.1038/sj.gt.3300626.
Human dendritic cells (DC) are highly professional antigen presenting cells for the priming of naive cytotoxic T cells. Gene transfer in DC would be a useful strategy to load DC with relevant de novo synthesized antigens for immunotherapeutical purposes. As a first step towards a DC-based gene therapy, we examined the efficiency of nonviral transfection in different types of cultured human dendritic cells with a humanized red-shifted green fluorescent protein reporter gene. Plasmid DNA transfection by electroporation or lipofection was used to transfect CD34+ progenitor cell-derived DC (PC-DC) and Langerhans' cells (PC-LC), as well as monocyte-derived DC (Mo-DC). While lipofection was unsuccessful in all types of DC, we obtained high-efficiency gene transfer by electroporation in PC-LC (16%) and PC-DC (12%). In contrast, electroporation was strikingly less efficient in Mo-DC (< or = 2%). The potent allostimulatory capacity of DC was still retained in electroporated PC-DC and PC-LC. In conclusion, electroporation of antigen expressing plasmid DNA is an efficient tool for nonviral gene transfer in PC-DC and PC-LC, but not in Mo-DC and could be useful for the development of DC-based tumor immunotherapy.
人树突状细胞（DC）是用于启动幼稚细胞毒性T细胞的高度专业的抗原呈递细胞。将基因导入DC可能是一种有用的策略，可用于为免疫治疗目的加载新合成的相关抗原。作为基于DC的基因治疗的第一步，我们用一种人源化的红移绿色荧光蛋白报告基因检测了不同类型培养的人树突状细胞中非病毒转染的效率。通过电穿孔或脂质体转染质粒DNA来转染CD34 +祖细胞衍生的DC（PC-DC）和朗格汉斯细胞（PC-LC）以及单核细胞衍生的DC（Mo-DC）。虽然脂质体转染在所有类型的DC中均未成功，但我们通过电穿孔在PC-LC（16%）和PC-DC（12%）中获得了高效的基因转移。相比之下，电穿孔在Mo-DC中的效率极低（≤2%）。电穿孔后的PC-DC和PC-LC仍保留了DC强大的共刺激能力。总之，表达抗原的质粒DNA的电穿孔是PC-DC和PC-LC中非病毒基因转移的有效工具，但不适用于Mo-DC，并且可能对基于DC的肿瘤免疫治疗的发展有用。
