Article

Enhancement of gene transfection into human dendritic cells using cationic PLGA nanospheres with a synthesized nuclear localization signal.

Laboratory of Pharmaceutics and Drug Delivery, Department of Pharmaceutical Science, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan.
International journal of pharmaceutics (impact factor: 2.96). 07/2009; 379(1):187-95. DOI:10.1016/j.ijpharm.2009.06.015 pp.187-95
Source: PubMed

ABSTRACT Effective delivery of DNA encoding antigen into the dendritic cells (DCs), which are non-dividing cells, is very important for the development of DNA vaccines. In a previous study, we developed the PLGA nanospheres that contained a cationic nanomaterial and showed high transfection efficiency in COS7 cells, which divide. In the present study, to produce an effective vector for the DNA vaccines, the gene expression and intracellular trafficking of pDNA complexed with PLGA/PEI nanospheres, in combination with an NF-kappaB analog as a nuclear localization signal (NLS) and electroporation were evaluated in human monocyte-derived DCs (hMoDCs). Cellular uptake of pDNA both in COS7 cells and hMoDCs was enhanced using the PLGA/PEI nanospheres. On the other hand, the PLGA/PEI nanospheres significantly promoted the transfection in COS7 cells, but had almost no effect on transfection in hMoDCs. The intranuclear transport of pDNA by PLGA/PEI nanospheres in COS7 cells was significantly higher than that in hMoDCs. These results indicate that pDNA complexed with PLGA/PEI nanospheres cannot enter into the nuclei of non-dividing cells. However, PLGA/PEI nanospheres combinated with NLS and electroporation (experimental permeation enhancer) greatly elevated the transfection efficiency by improvement of not only intracellular uptake but also intranuclear transport of pDNA in the hMoDCs. Thus, this delivery system using nanospheres combined with synthesized NLS might be applicable to DC-based gene vaccines when much non-invasive application such as needle-free injector should be required.

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Keywords

Cellular uptake
 
COS7 cells
 
DC-based gene vaccines
 
dendritic cells
 
DNA encoding antigen
 
Effective delivery
 
effective vector
 
experimental permeation enhancer
 
hMoDCs
 
human monocyte-derived DCs
 
NF-kappaB analog
 
non-dividing cells
 
non-invasive application
 
nuclear localization signal
 
pDNA
 
pDNA complexed
 
PLGA nanospheres
 
PLGA/PEI nanospheres
 
PLGA/PEI nanospheres combinated
 
synthesized NLS