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首页> 外文期刊>World Journal of Gastroenterology >HepG2 cells support viral replication and gene expression of hepatitis C virus genotype 4 in vitro.
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HepG2 cells support viral replication and gene expression of hepatitis C virus genotype 4 in vitro.

机译：HepG2细胞在体外支持丙型肝炎病毒基因型4的病毒复制和基因表达。

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AIM: To establish a cell culture system with long-term replication of hepatitis C virus (HCV) genome and expression of viral antigens in vitro. METHODS: HepG2 cell line was tested for its susceptibility to HCV by incubation with a serum from a patient with chronic hepatitis C. Cells and supernatant were harvested at various time points during the culture. Culture supernatant was tested for its ability to infect naive cells. The presence of minus (antisense) RNA strand, and the detection of core and E1 antigens in cells were examined by RT-PCR and immunological techniques (flow cytometry and Western blot) respectively. RESULTS: The intracellular HCV RNA was first detected on d 3 after infection and then could be consistently detected in both cells and supernatant over a period of at least three months. The fresh cells could be infected with supernatant from cultured infected cells. Flow cytometric analysis showed surface and intracellular HCV antigen expression using in house made polyclonal antibodies (anti-core, and anti-E1). Western blot analysis showed the expression of a cluster of immunogenic peptides at molecular weights extended between 31 and 45 kDa in an one month old culture of infected cells whereas this cluster was undetectable in uninfected HepG2 cells. CONCLUSION: HepG2 cell line is not only susceptible to HCV infection but also supports its replication in vitro. Expression of HCV structural proteins can be detected in infected HepG2 cells. These cells are also capable of shedding viral particles into culture media which in turn become infectious to uninfected cells.
机译：目的：建立能够长期复制丙型肝炎病毒（HCV）基因组并在体外表达病毒抗原的细胞培养系统。方法：通过与慢性丙型肝炎患者血清一起温育，测试HepG2细胞系对HCV的敏感性。在培养过程中的不同时间点收集细胞和上清液。测试培养物上清液感染幼稚细胞的能力。分别通过RT-PCR和免疫技术（流式细胞仪和蛋白质印迹）检查了负（反义）RNA链的存在以及细胞中核心和E1抗原的检测。结果：感染后第3天首先检测到细胞内HCV RNA，然后在至少三个月的时间内可在细胞和上清液中均检测到。可以用培养的感染细胞的上清液感染新鲜细胞。流式细胞仪分析显示，使用自制的多克隆抗体（抗核心抗体和抗E1抗体）可表达表面和细胞内HCV抗原。 Western印迹分析显示在一个月大的感染细胞培养物中，一种免疫原性肽簇的表达分子量在31至45 kDa之间，而在未感染的HepG2细胞中则无法检测到。结论：HepG2细胞系不仅易感染HCV，还支持其在体外的复制。 HCV结构蛋白的表达可以在感染的HepG2细胞中检测到。这些细胞还能够将病毒颗粒脱落到培养基中，而培养基又对未感染的细胞具有传染性。

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