HBV cccDNA在2.2.15细胞表达的动态观察

目的研究2.2.15细胞中是否存在HBVcccDNA,探讨2.2.15细胞内HBV产物的动态表达规律。方法采用PCR方法检测2.2.15细胞内cccDNA,Taqman定量PCR技术检测2.2.15细胞内及培养上清中HBVDNA含量,EIA方法检测培养上清中HBsAg、HBeAg的动态表达,并进行定量资料相关性统计学分析。结果2.2.15细胞及培养上清中存在cccDNA,2.2.15细胞培养上清中HBVDNA与HBsAg、HBeAg之间存在相关性(r=0.833,P<0.05和r=0.939,P<0.01),而细胞内HBVDNA与培养上清HBVDNA及HBsAg、HBeAg之间无相关性(r=0.024,P>0.05和r=0.177,P>0.05)。结论为阐明2.2.15细胞内HBV的复制规律提供一定依据。

Dynamic expression of hepatitis B virus covalently closed circular DNA in 2.2.15 cell

Objective To determine the presence of covalently closed circular DNA (cccDNA), and to investigate the expression kinetics of HBV DNA, HBsAg and HBeAg in 2.2.15 cell. Methods HBV cccDNA was assessed by polymerase chain reaction, HBV DNA was measured by Taqman quantitative PCR,and HBsAg and HBeAg was measured by EIA. Results HBV cccDNA was found in both intracellular and extracelluar space. There was a good correlation between HBsAg, HBeAg and HBV DNA in the supernatant of 2.2.15 cell (r= 0.833,P<0.05 and r= 0.939, P< 0.01 for HBsAg and HBeAg, respectively), whereas there was no significant correlation between intracellular HBV DNA levels and virus antigen levels (r = 0.024, P= 0.955 and r = 0.177; P = 0.625 for HBsAg and HBeAg, respectively). Conclusion HBV cccDNA was detectable in the culture medium and intracellularly in 2.2.15 cells, and these data provided an indication of HBV replication in 2.2.15 cell.