Gene expression profiles in primary duodenal chick cells following transfection with avian influenza virus H5 DNA plasmid encapsulated in silver nanoparticles

In order to develop a systemically administered safe and effective nonviral gene delivery system against avian influenza virus (AIV) that induced cytokine expression, the hemagglutinin (H5) gene of AIV, A/Ck/Malaysia/5858/04 (H5N1) and green fluorescent protein were cloned into a coexpression vector pIRES (pIREGFP-H5) and formulated using green synthesis of silver nanoparticles (AgNPs) with poly(ethylene glycol) and transfected into primary duodenal cells taken from 18-day-old specific-pathogen-free chick embryos. The AgNPs were prepared using moderated temperature and characterized for particle size, surface charge, ultraviolet-visible spectra, DNA loading, and stability. AgNPs and AgNP-pIREGFP-H5 were prepared in the size range of 13.9 nm and 25 nm with a positive charge of +78 ± 0.6 mV and +40 ± 6.2 mV, respectively. AgNPs with a positive surface charge could encapsulate pIREGFP-H5 efficiently. The ultraviolet-visible spectra for AgNP-pIREGFP-H5 treated with DNase I showed that the AgNPs were able to encapsulate pIREGFP-H5 efficiently. Polymerase chain reaction showed that AgNP-pIREGFP-H5 entered into primary duodenal cells rapidly, as early as one hour after transfection. Green fluorescent protein expression was observed after 36 hours, peaked at 48 hours, and remained stable for up to 60 hours. In addition, green fluorescent protein expression generally increased with increasing DNA concentration and time. Cells were transfected using Lipocurax in vitro transfection reagent as a positive control. A multiplex quantitative mRNA gene expression assay in the transfected primary duodenal cells via the transfection reagent and AgNPs with pIREGFP-H5 revealed expression of interleukin (IL)-18, IL-15, and IL-12â.

[1]  A. Demirok,et al.  Comparison of three methods of tonometry in normal subjects: Goldmann applanation tonometer, non-contact airpuff tonometer, and Tono-Pen XL , 2014, Clinical ophthalmology.

[2]  A. Omar,et al.  Improved immune responses against avian influenza virus following oral vaccination of chickens with HA DNA vaccine using attenuated Salmonella typhimurium as carrier. , 2012, Comparative immunology, microbiology and infectious diseases.

[3]  A. Omar,et al.  Cytotoxicity and immunological responses following oral vaccination of nanoencapsulated avian influenza virus H5 DNA vaccine with green synthesis silver nanoparticles. , 2012, Journal of controlled release : official journal of the Controlled Release Society.

[4]  C. M. Drain,et al.  Fabrication of Size-Tunable Metallic Nanoparticles Using Plasmid DNA as a Biomolecular Reactor , 2011, Nanomaterials.

[5]  S. Singh,et al.  Functionalized Gold Nanoparticles and Their Biomedical Applications , 2011, Nanomaterials.

[6]  Jian Chen,et al.  Evaluation of Oral Immunization with Recombinant Avian Influenza Virus HA1 Displayed on the Lactococcus lactis Surface and Combined with the Mucosal Adjuvant Cholera Toxin Subunit B , 2011, Clinical and Vaccine Immunology.

[7]  K. Sandvig,et al.  New metal-based nanoparticles for intravenous use: requirements for clinical success with focus on medical imaging. , 2010, Nanomedicine : nanotechnology, biology, and medicine.

[8]  Chin-fen Yang,et al.  The hemagglutinin protein of influenza A/Vietnam/1203/2004 (H5N1) contributes to hyperinduction of proinflammatory cytokines in human epithelial cells. , 2010, Virology.

[9]  Moonjung Choi,et al.  Cellular uptake, cytotoxicity, and innate immune response of silica-titania hollow nanoparticles based on size and surface functionality. , 2010, ACS nano.

[10]  Fernando Silva,et al.  Long time effect on the stability of silver nanoparticles in aqueous medium: Effect of the synthesis and storage conditions , 2010 .

[11]  H. Autrup,et al.  PVP-coated silver nanoparticles and silver ions induce reactive oxygen species, apoptosis and necrosis in THP-1 monocytes. , 2009, Toxicology letters.

[12]  D. Kapczynski,et al.  Strategies and challenges for eliciting immunity against avian influenza virus in birds , 2008, Immunological reviews.

[13]  S. Meneghetti,et al.  Synthesis of colloids based on gold nanoparticles dispersed in castor oil , 2008 .

[14]  P. Bartlett,et al.  Layered double hydroxide nanoparticles as cellular delivery vectors of supercoiled plasmid DNA , 2007, International journal of nanomedicine.

[15]  Y. Lai,et al.  Avian influenza virus hemagglutinin display on baculovirus envelope: cytoplasmic domain affects virus properties and vaccine potential. , 2007, Molecular therapy : the journal of the American Society of Gene Therapy.

[16]  V. Apostolopoulos,et al.  Poly-L-lysine-coated nanoparticles: a potent delivery system to enhance DNA vaccine efficacy. , 2007, Vaccine.

[17]  A. Slita,et al.  DNA-polycation complexes: effect of polycation structure on physico-chemical and biological properties. , 2007, Journal of biotechnology.

[18]  H. B. Liu,et al.  Biosynthesis and characterization of Ti/Ni bimetallic nanoparticles , 2006 .

[19]  Y. Tabata,et al.  Self assembly of DNA nanoparticles with polycations for the delivery of genetic materials into cells. , 2006, Journal of nanoscience and nanotechnology.

[20]  Baokang Jin,et al.  One-step synthesis of monodisperse silver nanoparticles beneath vitamin E Langmuir monolayers. , 2006, The journal of physical chemistry. B.

[21]  Aibing Yu,et al.  Inorganic nanoparticles as carriers for efficient cellular delivery , 2006 .

[22]  K. Leong.  Polymeric Controlled Nucleic Acid Delivery , 2005 .

[23]  Gunnar P. H. Dietz,et al.  Delivery of bioactive molecules into the cell: the Trojan horse approach , 2004, Molecular and Cellular Neuroscience.

[24]  L. Anderson,et al.  Pulmonary delivery of respiratory syncytial virus DNA vaccines using macroaggregated albumin particles. , 2004, Vaccine.

[25]  Bing Xu,et al.  Presenting Vancomycin on Nanoparticles to Enhance Antimicrobial Activities , 2003 .

[26]  Kemin Wang,et al.  Bioconjugated nanoparticles for DNA protection from cleavage. , 2003, Journal of the American Chemical Society.

[27]  Thomas Kissel,et al.  In vitro cytotoxicity testing of polycations: influence of polymer structure on cell viability and hemolysis. , 2003, Biomaterials.

[28]  V. Labhasetwar,et al.  Size-dependency of nanoparticle-mediated gene transfection: studies with fractionated nanoparticles. , 2002, International journal of pharmaceutics.

[29]  T. Chakraborty,et al.  Transfer of eukaryotic expression plasmids to mammalian host cells by bacterial carriers. , 2001, Current opinion in biotechnology.

[30]  T. He,et al.  Adenoviral vector-mediated gene transfer for human gene therapy. , 2001, Current gene therapy.

[31]  W. Mark Saltzman,et al.  Enhancement of transfection by physical concentration of DNA at the cell surface , 2000, Nature Biotechnology.

[32]  G. Kearns,et al.  Substrates for direct imaging of chemically functionalized SiO2 surfaces by transmission electron microscopy. , 2006, Analytical chemistry.

[33]  Jin Woo Kim,et al.  Test Tube Selection of Large Independent Sets of DNA Oligonucleotides , 2006, Nanotechnology: Science and Computation.

[34]  P. Barrow,et al.  Invasion of Vero cells by Salmonella species. , 1989, Journal of medical microbiology.