Original Articles
Copyright ©The Author(s) 2001. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jun 15, 2001; 7(3): 370-375
Published online Jun 15, 2001. doi: 10.3748/wjg.v7.i3.370
Hepatitis C virus in human B lymphocytes transformed by Epstein-Barr virus in vitro by in situ reverse transcriptase-polymerase chain reaction
Ji-Lin Cheng, Bao-Ling Liu, Yi Zhang, Wen-Bin Tong, Zheng Yan, Bai-Fang Feng
Ji-Lin Cheng, Bao-Ling Liu, Yi Zhang, Wen-Bin Tong, Zheng Yan, Bai-Fang Feng, Institute of Hepatology, People's Hospital, Medical Center of Beijing University, Beijing 100044, China
Author contributions: All authors contributed equally to the work.
Correspondence to: Ji-Lin Cheng, Department of Gastroenterology, Xinhua Hospital of Shanghai Second Medical University, Shanghai 200092, China
Received: September 29, 2000
Revised: October 3, 2000
Accepted: October 26, 2000
Published online: June 15, 2001

AIM: To study persistence and replication of hepatitis C virus (HCV) in patients' peripheral blood mononuclear cells (PBMC) cultured in vitro.

METHODS: Epstein-Barr virus (EBV) was used to transform the hepatitis C virus from a HCV positive patient to permanent lymphoblastoid cell lines (LCL). Positive and negative HCV RNA strands of the cultured cells and growth media were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) each month. Core and NS5 proteins of HCV were further tested using immunohistochemical SP method and in situ RT-PCR.

RESULTS: HCV RNA positive strands were consistently detected the cultured cells for one year. The negative-strand RNA in LCL cells and the positive-strand RNA in supernatants were observed intermittently. Immunohistochemical results medicated expression of HCV NS3 and C proteins in LCL cytoplasm mostly. The positive signal of PCR product was dark blue and mainly localized to the LCL cytoplasm. The RT-PCR signal was eliminated by overnight RNase digestion but not DNase digestion.

CONCLUSION: HCV may exist and remain functional in a cultured cell line for a long period.

Keywords: hepatitis C-like viruses, herpes virus 4, human, B-lymphocytes, cells cultured