人类乙肝病毒体外感染树嗣肝原代细胞的研究  

Study on the infection of primary tupaia hepatocytes with hepatitis B virus in vitro

作　　者：张晶晶[1,2] 杨光[1] 杨春[1] 蒿艳蓉[1] 曹骥[1] 欧超[1] 李瑗[1] 班克臣[1] Hubert E.Blum 苏建家[1] 

机构地区：[1]广西医科大学附属肿瘤医院,南宁530021 [2]德国弗莱堡大学附属医院

出　　处：《中华微生物学和免疫学杂志》2008年第11期984-988,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目（30560168）

摘　　要：目的为进一步研究人类乙肝病毒（HBV）提供接近自然感染状态的理想细胞模型。方法体外二步灌流法分离树嗣原代肝细胞，纯化后的乙肝患者血清感染上述肝细胞，Southern blot和Northern blot检测感染后细胞内的DNA和RNA，ELISA方法检测细胞上清的HBsAg，免疫组化检测细胞内HBsAg的表达。结果可检测出肝细胞内cccDNA（共价闭合环状DNA）、pgRNA（前基因组RNA）和sgRNA（亚基因组mRNA），感染后第7天，信号开始增强，持续到实验结束的第14天。细胞上清中HBsAg自第1天到第5天S／CO值逐渐下降，随后S／CO值逐渐升高。结论HBV可在原代树嗣肝细胞中复制和表达。Objective To provide a better cell model of closely nature infectious state for further research of hepatitis B virus （HBV）. Methods Primary tupaia hepatocytes were isolated by the two-step perfusion method. The hepatocytes were then infected with purified serum from patients with hepatitis B. DNA and RNA isolated from the hepatocytes were detected with Southern blot and Northern blot. HBsAg in supernatant was tested by immunohistochemical method. Results cccDNA, pgRNA and sgRNA could be detected by Southern blot and Northern blot, and strong signals could be seen from day 7 to day 14 post-infection. The S/CO value of HBsAg in supernatant decreased from day 1 to day 5 and then increased after 5 day. Conclusion Primary tupaia hepatocytes are competent for infection with HBV. HBV can stably replicate and express in HBV-infected tupaia hepatocytes.

关 键 词：乙肝病毒 感染 树嗣 原代肝细胞 

分 类 号：R51[医药卫生—内科学]