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Efficient delivery of vector DNA to dendritic cells using silica‐nanoparticles
Author(s) -
Messmer Davorka,
Huang ChienTze,
Seible Daniel,
Yang Jian,
Saenz Rebecca,
Esener Sadik,
Trogler William
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1077.22
Subject(s) - gene delivery , chemistry , flow cytometry , electroporation , adsorption , nanoparticle , dna , microbiology and biotechnology , biophysics , in vivo , transfection , materials science , nanotechnology , biochemistry , gene , biology , organic chemistry
Objective: Optimize plasmid DNA delivery to DCs using silica‐nanoparticles (NPs) for tumor immunotherapy. Methods: Human monocyte‐derived DCs were generated from PBMCs by culturing adherent cells in GM‐CSF and IL‐4 for 5 to 7 days. SiO 2 nanoparticles were generated using polymer beads as template. The surface was functionalized with amino silane or amino phosphoric acid and the polymer core was removed by calcination or organic solvents. Results: We sought to determine the optimal parameters in terms of size and surface modification of silica‐NPs with regard to DNA adsorption. DNA could be adsorbed to silica‐NPs of different sizes and surface modifications including amine groups and poly‐L‐lysine (pLL) lead to increased DNA adsorption to the NPs. NPs of 45, 100, and 200 nm diameter were effectively taken up by DCs and no obvious preference was observed. Interestingly pLL‐modified NPs were not taken up by DCs. Confocal microscopy indicated that NPs were also present in the nucleus of the DC which showed high viability for at least 7 days. To determine whether the NPs can be used to deliver DNA into DCs and lead to gene expression of the transgene, immature DCs were exposed to NP‐DNA complexes containing gfp‐plasmid and analyzed for Gfp expression by flow cytometry. 45% of the DCs expressed GFP at 48h. Conclusion: SiO 2 ‐nanoparticles of 80 and 120 nm are more efficient than electroporation to introduce a transgene into DCs in vitro and possibly in vivo for DNA vaccines.