[Persistence of hepatitis C virus type II in patient's peripheral blood B lymphocytes transformed by Epstein-Barr virus]

Abstract

Objective: To study the persistence and replication of hepatitis C virus (HCV)-RNA in human peripheral blood B cells transformed by Epstein-Barr virus (EBV) and cultured in vitro.

Methods: EBV was used for infecting B lymphocytes from one hepatitis C patient with HCV positive in the peripheral blood mononuclear cells (PBMC) and transforming them into lymphoblasts capable of being propagated indefinitely. Then, HCV RNA of the cultured cells and supernatants was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) every one month. HCV gene was typed by enzyme separating method. Electron microscopy and immunoelectron microscopy were employed to locate HCV in the cells and observe the character of its configuration.

Results: HCV positive-strand RNA was detected in the cultured cells for 1 year. Interestingly, the HCV positive-strand RNA was identified in supernatants and the negative-strand RNA was also observed in the cultured cells intermittently. HCV gene was type II. Electron microscopy observed HCV spherical virus-like particles with a diameter of approximately 45 nm to 70 nm, individual particles 110 nm, in the LCL cytoplasmic vesicles.

Conclusion: HCV may exist in the cultured cell line for a longer period and reproduce in and secrete out of them. HCV locates mainly in the cytoplasm. The cell line might be useful in analyzing the mechanisms of HCV persistence in PBMC.