摘要
目的研究乙型肝炎病毒C蛋白在NK细胞中的表达及其对NK细胞功能的影响。方法用脂质体转染法将HBVC基因真核表达载体pHBI-CMV-HBC转入NK-92细胞,通过Western Blot检测C基因在NK-92细胞中表达.用ELISA法检测NK细胞分泌IFN-γ水平,MTT法检测NK细胞对HepG2细胞的细胞毒作用。结果Western Blot证实转染有pHBI-CMV-HBC的NK-92细胞能表达HBVC蛋白。转染了重组真核表达质粒的NK-92细胞IFN-γ水平均高于空质粒对照组和空白对照组(P〈0.01)。与两对照组相比,重组真核表达质粒转染组NK-92细胞对于HepG2细胞的细胞毒活性明显提高,差异具有统计学意义(P〈0.01)。结论HBc基因瞬时高表达可影响NK-92细胞分泌IFN-γ和细胞毒功能。
Objective To investigate the influence of hepatitis B virus C protein on the function of natural killer cell. Methods Recombinant eukaryotic expression plasmid pHBI-CMV-HBC was constructed and confirmed by double restrictive enzyme digestion and DNA sequencing analysis. Then the recombinant plasmid was tmnsfected into NK-92 cells with lipofectamine encapsuled. The transfected NK-92 cells containing expressive HBV C protein was confirmed by Western Blot analysis. ELISA was employed to determine the IFN-γ level secreted by NK-92 cells. And finally the cytotoxicities of NK cells were analysed by MTT colorimetry, with the hepatoblastoma cell line (HepG2) as target cell. Results Western blotting confirmed the expression of HBV C protein in the NK-92 cells transfected with pHBI-CMV-HBC. NK cytotoxicities and IFN-7 secretion level of NK-92 cells transfected with recombinant plasmid significantly increased compared to control NK-92 cells transfected with blank plasmid ( P 〈 0.01 ) and untransfected NK-92 cells( P 〈 0.01 ). Conclussion Transient expression of HBC can increase IFN-γ secretion and cytotoxicities of NK-92 cells.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
北大核心
2008年第4期275-277,共3页
Chinese Journal of Experimental and Clinical Virology
基金
国家教育部留学回国人员基金(编号2006KY12.1.4.2)