[Ultrasound-mediated microbubble destruction enhances LMP-1 gene transfection into dendritic cells in vivo].
OBJECTIVE
To investigate the transfection efficiency and the optimal conditions of delivering latent membrane protein-1 (LMP-1) gene to dendritic cells (DCs) by ultrasound exposure combined with contrast agent.


METHODS
Human DCs were cultured in vivo and transfected with the recombinant plasmid pEGFP-C3-LMP1 under varying conditions including ultrasound intensities, exposure time and microbubble contrast agent concentration. The transfection efficiency was assessed by fluorescent microscopy and flow cytometry, and the cell viability by trypan blue exclusion test.


RESULTS
An exposure time of 60 s at MI 1.0 with a microbubble contrast agent concentration of 20% resulted in the optimal effect of delivering the recombinant plasmid pEGFP-C3-LMP1 into the DCs, with a transfection efficiency of (14.37∓2.12)%. Over 90% of the transfected cells were viable after the transfection.


CONCLUSIONS
Microbubble contrast agent combined with ultrasound exposure can enhance the delivery of recombinant plasmid pEGFP-C3-LMP1 into the DCs.
Xu
Rui-feng
RF
Department of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. xuruifeng2003@163.com
Shi
Min
M
You
Chang-xuan
CX
Lü
Cheng-wei
CW
Luo
Rong-cheng
RC
Liao
Wang-jun
WJ
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics*; D002478:Cells, Cultured; D003287:Contrast Media / Q000008:administration & dosage / Q000494:pharmacology*; D003598:Cytoskeletal Proteins / Q000235:genetics*; D003713:Dendritic Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D060588:LIM Domain Proteins / Q000235:genetics*; D045423:Microbubbles; D010957:Plasmids; D014162:Transfection; D014465:Ultrasonics
D048868:Adaptor Proteins, Signal Transducing; D003287:Contrast Media; D003598:Cytoskeletal Proteins; D060588:LIM Domain Proteins; C484380:PDLIM7 protein, human