Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 95, Issue 5, Pages 1253-1263Publisher
SPRINGER
DOI: 10.1007/s00253-012-4042-z
Keywords
Upconversion nanoparticles; Gene delivery; DNA carrier; Foot-and-mouth disease virus; DNA vaccine
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Funding
- National Natural Science Foundation of China [31100688, 21101140]
- Gansu Provincial Sci. & Tech. Department [1102NKDA033, 1102NKDA034, 1002NKDA037]
- Ministry of Science and Technology of China [2008FY130100]
- National Modern Meat Caprine Industrial Technology System [nycytx-39]
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The potential of the upconversion nanoparticles NaYF4:Yb/Er@silica(UCPs)/plasmid DNA (pcDNA3.1/VP1-GFP) complex in inducing immune responses was evaluated using the UCPs as carriers of the foot-and-mouth disease virus (FMDV AsiaI/Jiangsu2005) DNA vaccine. The UCPs protection against DNaseI degradation was measured using an in vitro inhibition assay. The expression of the plasmid in vivo was determined via confocal microscopy. Its biocompatibility was evaluated through cytotoxicity assay. Based on the results, the aminosilane-modified UCPs can electrostatically bind, condense, and protect plasmid DNA. Cell viability assays demonstrated that the cytotoxicity of the UCPs/plasmid DNA complex is lower than that of the cationic lipid/plasmid DNA complex, and that the transfection efficiency of UCPs is the same as that of the cationic lipid. Furthermore, the UCP/plasmid DNA complex was intramuscularly administered to guinea pigs. Humoral and cellular immune responses were detected using indirect enzyme-linked immunosorbent assay (ELISA), micro-neutralization assay, and T-lymphocyte proliferation assay. Anti-FMDV specific antibodies, neutralizing antibodies, and T-lymphocyte proliferation responses were induced after vaccination. In the challenge test, all the guinea pigs vaccinated with the UCPs/plasmid DNA complex were fully protected from the FMDV challenge. The current study encourages the use of UCPs as an effective nanosystem for gene delivery to cells for in vitro and in vivo vaccination, and other therapeutic applications.
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