Iacovacci S, Bertolini L, Manzin A, Valli M B, Battaglia M, Ponzetto A, Clementi M, Carloni G
Institute of Experimental Medicine, C.N.R., Rome, Italy.
Res Virol. 1997 Mar-Apr;148(2):147-51. doi: 10.1016/s0923-2516(97)89901-3.
The ability of hepatitis C virus (HCV) to replicate in two B-cell lines, CE and TOFE, derived from bone marrow of healthy subjects was compared using qualitative and quantitative molecular methods. The presence of intracellular negative-stranded HCV RNA (replicative intermediate) was investigated by nested polymerase chain reaction (PCR) in the infected cultures at different times after infection. The amounts of positive-stranded HCV RNA (genomic RNA copies) synthesized and released from cells one week after in vitro infection were determined by competitive PCR after reverse transcription of viral RNA for the 5' viral untranslated region. In both cell lines, HCV RNA replication took place, but the TOFE cell line appeared to be a more efficient virus producer than the CE cell line. The TOFE cell line could be a valuable and reliable tool for basic and clinical HCV studies.
使用定性和定量分子方法比较了丙型肝炎病毒(HCV)在源自健康受试者骨髓的两种B细胞系CE和TOFE中复制的能力。通过巢式聚合酶链反应(PCR)在感染后不同时间的感染培养物中研究细胞内负链HCV RNA(复制中间体)的存在。在病毒RNA逆转录为5'病毒非翻译区后,通过竞争性PCR测定体外感染一周后从细胞合成并释放的正链HCV RNA(基因组RNA拷贝)的量。在两种细胞系中均发生了HCV RNA复制,但TOFE细胞系似乎比CE细胞系更有效地产生病毒。TOFE细胞系可能是用于基础和临床HCV研究的有价值且可靠的工具。