Ladner S K, Otto M J, Barker C S, Zaifert K, Wang G H, Guo J T, Seeger C, King R W
Avid Therapeutics, Inc., Philadelphia, Pennsylvania 19104, USA.
Antimicrob Agents Chemother. 1997 Aug;41(8):1715-20. doi: 10.1128/AAC.41.8.1715.
We report the development and isolation of a cell line, termed HepAD38, that replicates human hepatitis B virus (HBV) under conditions that can be regulated with tetracycline. In the presence of the antibiotic, this cell line is free of virus due to the repression of pregenomic (pg) RNA synthesis. Upon removal of tetracycline from the culture medium, the cells express viral pg RNA, accumulate subviral particles in the cytoplasm that contain DNA intermediates characteristic of viral replication, and secrete virus-like particles into the supernatant. Since the HepAD38 cell line can produce high levels of HBV DNA, it should be useful for analyses of the viral replication cycle that depend upon viral DNA synthesis in a synchronized fashion. In addition, this cell line has been formatted into a high-throughput, cell-based assay that permits the large-scale screening of diverse compound libraries for new classes of inhibitors of HBV replication.
我们报告了一种细胞系HepAD38的建立和分离，该细胞系在可通过四环素调控的条件下复制人乙型肝炎病毒（HBV）。在抗生素存在的情况下，由于前基因组（pg）RNA合成受到抑制，该细胞系无病毒。从培养基中去除四环素后，细胞表达病毒pg RNA，在细胞质中积累含有病毒复制特征性DNA中间体的亚病毒颗粒，并将病毒样颗粒分泌到上清液中。由于HepAD38细胞系可产生高水平的HBV DNA，它对于以同步方式依赖病毒DNA合成的病毒复制周期分析应是有用的。此外，该细胞系已被构建成一种基于细胞的高通量检测方法，可用于大规模筛选各种化合物文库以寻找新型HBV复制抑制剂。
