Hepatitis C virus in human B lymphocytes transformed by Epstein-Barr virus in vitro by in situ reverse transcriptase-polymerase chain reaction  

作　　者：Ji Lin Cheng Bao Ling Liu Yi Zhang Wen Bin Tong Zheng Yan Bai Fang Feng Institute of Hepatology,Peoples Hospital,Medical Center of Beijing University,Beijing 10(X)44,China 

出　　处：《World Journal of Gastroenterology》2001年第3期370-375,共6页世界胃肠病学杂志（英文版）

基　　金：The paper was support by a grant from the Ministry Youth Research of China,No.98-1-269

摘　　要：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.AIM To study persistence and replication ofheltitis C virus (HCV) in patients' peripheralblood mononuclear cells (PBMC) cultured invitro.METHODS Epstein-Barr virus (EBV) was usedto transform the hepatitis C virus from a HCVpositive patient to permanent lymphoblastoidcell lines (LCL). Positive and negative HCV RNAstrands of the cultured cells and growth mediawere detected by reverse transcriptase-polymerase chain reaction ( RT-PCR ) eachmonth. Core and NS5 proteins of HCV werefurther tested using immunohistochemical SPmethod and in situ RT-PCR.RESULTS HCV RNA positive strands wereconsistently detected the cultured cells for oneyear. The negative-strand RNA in LCL cells andthe positive-strand RNA in supernatants wereobserved intermittently. Immunohistochemicalresults medicated expression of HCV NS3 and Cproteins in LCL cytoplasm mostly. The positivesignal of PCR product was dark blue and mainlylocalized to the LCL cytoplasm. The RT-PCRsignal was eliminated by overnight RNasedigestion but not DNase digestion.CONCLUSION HCV may exist and remainfunctional in a cultured cell line for a longperiod.

关 键 词：B-LYMPHOCYTES Cells  Cultured Female HEPACIVIRUS development purification Herpesvirus 4  Human Humans Immunohistochemistry In Vitro Polymerase Chain Reaction RNA  Viral Research Support  Non-U.S. Gov't Reverse Transcriptase Polymerase Chain Reaction Transformation  Genetic Viral Core Proteins Viral Nonstructural Proteins Virus Replication 

分 类 号：R373.21[医药卫生—病原生物学]