Mizutani T, Kato N, Ikeda M, Sugiyama K, Shimotohno K
Virology Division, National Cancer Center Research Institute, Tokyo, Japan.
Biochem Biophys Res Commun. 1996 Oct 23;227(3):822-6. doi: 10.1006/bbrc.1996.1591.
We recently found that human T-cell leukemia virus type I-infected cloned MT-2C cells could support the replication of hepatitis C virus (HCV) up to 30 days postinoculation (p.i.). To obtain a more persistent HCV replication system using MT-2C cells, we examined the effect of various culture conditions after virus inoculation. We found that persistent HCV infection (at least 80 days) could be reproducibly achieved by reducing the temperature from 37 degrees C to 32 degrees C. Under this culture condition, HCV RNA could be detected in the cells up to 198 days p.i. Sequence analysis of HCV hypervariable region 1 revealed that certain HCV species became predominant during the culture period. We also demonstrated that virus transmission by the cell-free mode could be successfully repeated at least 4 times at 32 degrees C as opposed to only twice at 37 degrees C. This HCV-infected culture system will be useful for the various biological studies, including investigations into the mechanisms of HCV replication and multiplication.
我们最近发现，人类I型嗜T细胞病毒感染的克隆MT - 2C细胞在接种后（p.i.）长达30天能够支持丙型肝炎病毒（HCV）的复制。为了使用MT - 2C细胞获得更持久的HCV复制系统，我们检测了病毒接种后各种培养条件的影响。我们发现，通过将温度从37摄氏度降低到32摄氏度，可以重复性地实现HCV的持续感染（至少80天）。在这种培养条件下，在接种后长达198天的细胞中都能检测到HCV RNA。对HCV高变区1的序列分析表明，在培养期间某些HCV毒株变得占主导地位。我们还证明，与在37摄氏度仅能成功重复两次相比，在32摄氏度无细胞模式的病毒传播至少可以成功重复4次。这种HCV感染的培养系统将有助于进行各种生物学研究，包括对HCV复制和增殖机制的研究。
