HEPATITIS-B VIRUS-INFECTION OF ADULT HUMAN HEPATOCYTES CULTURED IN THE PRESENCE OF DIMETHYL-SULFOXIDE
HEPATITIS-B VIRUS-INFECTION OF ADULT HUMAN HEPATOCYTES CULTURED IN THE PRESENCE OF DIMETHYL-SULFOXIDE
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DOI：
10.1128/jvi.62.11.4136-4143.1988
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发表时间：
1988-11-01
影响因子：
5.4
通讯作者：
GUGUENGUILLOUZO, C
中科院分区：
文献类型：
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作者：
GRIPON, P;DIOT, C;GUGUENGUILLOUZO, C
我们研究了在纯培养或与乙型肝炎病毒（HBV）共培养中感染正常成人肝细胞的可能性。对来自不同供体的不同感染性血清和肝细胞群进行的几项检测仅发现了有限的HBV复制，不同细胞制备之间存在显著差异。在培养基中添加1.5%的二甲亚砜，显著促进了侵染过程。事实上，乙型肝炎e抗原的分泌，HBV DNA复制形式和主要HBV转录物的出现，以及完全HBV颗粒释放到培养基中都得到了证实。在二甲亚砜处理的细胞中，细胞内HBV DNA的显著增加可能与病毒吸附增强有关。当转染HepG2细胞系产生的病毒颗粒感染正常肝细胞时，获得了相同的结果。此外，对来自三个不同供体的肝细胞的比较分析表明，尽管在所有病例中都发现了大量的细胞内病毒DNA，但仅在一个病例中发现了病毒复制中间体。这些发现表明，这种产生HBV的细胞系可以作为传染性病毒的可再生来源，并且人肝细胞的原代培养是分析细胞内调节因子的独特工具，这些因子除了渗透步骤外，还可以调节HBV复制。
We investigated the possibility of infecting normal adult human hepatocytes maintained in pure cultures or in cocultures with hepatitis B virus (HBV). Several assays with different infectious sera and hepatocyte populations from various donors identified only limited HBV replication, with significant variations from one cell preparation to another. The addition of 1.5% dimethyl sulfoxide to the culture medium markedly enhanced the infection process. Indeed, hepatitis B e antigen secretion, the appearance of both HBV DNA replicative forms and major HBV transcripts, and the release of complete HBV particles into the medium were demonstrated. It is possible that the significant increase in intracellular HBV DNA in dimethyl sulfoxide-treated cells was related to enhanced adsorption of the virus. When viral particles produced by a transfected HepG2 cell line were used to infect normal hepatocytes, the same results were obtained. In addition, comparative assays with hepatocytes from three different donors showed that although high amounts of intracellular viral DNA were found in all cases, viral replicative intermediates were visualized in only one case. These findings suggest that this HBV-producing cell line could serve as a reproducible source of infectious virus and that primary culturing of human hepatocytes represents a unique tool for analyzing intracellular regulating factors which, in addition to the penetration step, modulate HBV replication.