Guilhot S, Miller T, Cornman G, Isom H C
Department of Microbiology and Immunology Penn State College of Medicine Hershey 17033, USA.
Am J Pathol. 1996 Mar;148(3):801-14.
Three well differentiated SV40-immortalized rat hepatocyte cell lines, CWSV1, CWSV2, and CWSV14, and Hepatitis B Virus (HBV)-producing cell lines derived from them were examined for sensitivity to tumor necrosis factor (TNF)-alpha. CWSV1, CWSV2, and CWSV14 cells were co-transfected with a DNA construct containing a dimer of the HBV genome and the neo gene and selected in G418 to generate stable cell lines. Characterization of these cell lines indicated that they contain integrated HBV DNA, contain low molecular weight HBV DNA compatible with the presence of HBV replication intermediates, express HBV transcripts, and produce HBV proteins. The viability of CWSV1, CWSV2, and CWSV2 cells was not significantly altered when they were treated with TNF-alpha at concentrations as high as 20,000 U/ml. The HBV-expressing CWSV1 cell line, SV1di36, and the HBV-expressing CWSV14 cell line, SV14di208, were also not killed when treated with TNF-alpha. However, the HBV-expressing CWSV2 cell line, SV2di366, was extensively killed when treated with TNF-alpha at concentrations ranging from 200 to 20,000 U/ml. Analysis of several different HBV-producing CWSV2 cell lines indicated that TNF-alpha killing depended upon the level of HBV expression. The TNF-alpha-induced cell killing in high HBV-producing CWSV2 cell lines was accompanied by the presence of an oligonucleosomal DNA ladder characteristic of apoptosis.
对三种分化良好的经SV40永生化的大鼠肝细胞系CWSV1、CWSV2和CWSV14，以及由它们衍生出的乙型肝炎病毒（HBV）产生细胞系进行了肿瘤坏死因子（TNF）-α敏感性检测。将CWSV1、CWSV2和CWSV14细胞与包含HBV基因组二聚体和新霉素基因的DNA构建体共转染，并在G418中进行筛选以产生稳定细胞系。对这些细胞系的特性分析表明，它们含有整合的HBV DNA，含有与HBV复制中间体存在相符的低分子量HBV DNA，表达HBV转录本，并产生HBV蛋白。当用浓度高达20,000 U/ml的TNF-α处理时，CWSV1、CWSV2和CWSV2细胞的活力没有显著改变。表达HBV的CWSV1细胞系SV1di36和表达HBV的CWSV14细胞系SV14di208在用TNF-α处理时也未被杀死。然而，当用浓度为200至20,000 U/ml的TNF-α处理时，表达HBV的CWSV2细胞系SV2di366被大量杀死。对几种不同的产生HBV的CWSV2细胞系的分析表明，TNF-α诱导的杀伤取决于HBV的表达水平。在高HBV产生的CWSV2细胞系中，TNF-α诱导的细胞杀伤伴随着凋亡特有的寡核小体DNA梯带的出现。
