Oral efficacy and intestinal mucosal immune responses induced by the complex of DNA encoding Green Fluorescent Protein disulfide-crosslinked low molecular linear polyetheleneimine, and sodium hyaluronate

• Main Authors: Kuan-Long Wu, 吳冠龍
• Other Authors: Yi-Ning Chen
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/53270759868027462758

碩士 === 中原大學 === 生物科技研究所 === 103 === A gene delivery system carrying the gene segment of the antigen protein into the host body to express in non-viral way to stimulate mucosal and systemic immune responses has been developed. The current study used the ternary complex（DNA-CLPEI-HA）composed of plasmid DNA, disulfide-crosslinked low molecular linear polyetheleneimine(CLPEI) ,and hyaluronic acid (HA), which exhibited high gene transfection efficiency and low toxicity in cell culture systems. The examination by electron microscopes showed the physical properties of DNA-CLPEI-HA complex. The stability of DNA in the complex after oral delivery was determined by DNA electrophoresis and the digestions of restriction enzyme. The transfection efficiency of green fluorescence protein (GFP) gene delivered by DNA-CLPEI-HA complex in the intestinal epithelial cells in cell culture systems and the cell viability of the complex were examined in vitro. The expression of GFP and the immune responses induced in mice was evaluated by the levels of anti-GFP IgG antibody. The DNA-CLPEI-HA complex had a particle size of about 300 ~ 400nm. The DNA binding with CLPEI and HA at the mixing ratio N / P ratio 100 can be effectively protected from the digestion of restriction enzymes. Good transfection efficiency and low cytotoxicity were observed in cell culture system. The complex caused mild to no damage to the intestine in mice ingested orally the complex. Thus, the oral gene delivery system developed in this study had potentials to become a platform for oral vaccines used to fight intestinal pathogens.