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

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World Journal of Gastroenterology 2001年第3期7卷 370-375页

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 Institute of Hepatology People's Hospital Medical Center of Beijing University of Beijing China.

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.

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

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Induction of systemic lupus erythematosus syndrome in bALb/c mice by immunization with active chromatin

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Acta Pharmacologica Sinica 2004年第6期25卷 105-109页

作者： Hong LI, Yun-yi ZHANG, Ya-nan SUN, Xi-yi HUANG, Yong-feng JIA, Duan LI2 Department of Pharmacology, Schoolof Pharmacy, Fudan University, Shanghai 200032, China Department of Pharmacology School of Pharmacy Fudan University 上海 200032 Department of Pharmacology School of Pharmacy Fudan University Shanghai 200032

AIM: To establish an animal model for systemic lupus erythematosus (SLE)-like syndrome in mice. METHODS: bALb/c mice were immunized with active chromatin isolated from ConA-actived syngeneic spleno-lymphocytes. Plasma samples of mice were tested by enzyme-linked immunosorbent assays (ELISA) for the presence of IgG anti-dsDNA, -ssDNA, and anti-histone antibodies. Tumor necrosis factor-α (TNF-α) in serum was measured by ELISA. Spleno-lymphocyte proliferation assays and the levels of interferon-γ (IFN-γ) in supernatants were tested respectively. Proteinuria was measured. Kidneys were examined by direct immunohistochemical method and light microscopy. RESULTS: Anti-ds DNA, ssDNA, and histone antibodies were induced in active chromatin-immu- nized mice, the proliferation response of splenocytes to ConA and LPS were reduced, levels of interferon-γ in supernatants and TNF-α in serum were lowered. Lupus nephritis was assessed by the presence of Ig deposits, glomerular pathology and proteinuria. CONCLUSION: The active chromatin-induced SLE-like mouse model was similar to idiopathic SLE in human.

关键词： systemic lupus erythematosus chromatin autoantibody tumor necrosis factor interferon type II b-lymphocytes T-lymphocytes female enzyme-linked immunosorbent assay

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b-cell clonality in the liver of hepatitis C virus-infected patients

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World Journal of Gastroenterology 2009年第13期15卷 1636-1640页

作者： He-bin Fan You-Fu Zhu An-Shen Chen Mu-Xiu Zhou Fu-Ming Yan Xiao-Ju Ma Hao Zhou Department of Infectious Disease The People'sLiberation Army 161 Hospital Wuhan 430010 Hubei ProvinceChina Department of Infectious Disease NanfangHospital Southern Medical University Guangzhou 510515Guangdong Province China Department of Pathology The People'sLiberation Army 161 Hospital Wuhan 430010 Hubei ProvinceChina

AIM:The association of hepatitis C virus(HCV) infection with typeⅡmixed cryoglobulinemia is well established,but the role of HCV in b-cell lymphoma remains *** patients with HCV infection,b-cell clonal expansions have been detected in peripheral blood and bone marrow,and a high prevalence of b-cell non-Hodgkin's lymphomas has been *** biopsies in chronic HCV infection frequently show portal lymphoid infiltrates with features of b follicles,whose clonality has not yet been *** object of this study was to determine the frequency of liver-infiltrating monoclonal b-cells in 40 patients with HCV ***:Eight hundred and forty-eight patients were studied prospectively,including 40 HCV-positive patients and 808 patients with chronic hepatitis b virus(HbV)*** study for b-and T-cell markers was performed on the paraffin-embedded liver tissue *** clonality of lymphoid b-cells was tested using a polymerase chain reaction(PCR)approach designed to identify immunoglobulin heavy chain gene(IgH) ***:Liver-infiltrating monoclonal b-cells were detected in the liver for 4(10%)of 40 HCV-positive patients but were present in only 3(0.37%)of 808 liver biopsy specimens with chronic HbV ***-square testing showed that the monoclonal b-cells infiltration in the liver was more frequent in the HCV-infected patients(P=0.000).A clonal IgH rearrangement was detected in 5(71.4%)of 7 liver biopsy specimens with monoclonal b-cells *** 2 of 5 patients with both a clonal b-cell expansion and monoclonal b-cells infiltration in the liver,a definite b-cell malignancy was finally ***:Liver-infiltrating monoclonal b-cells are detected in the liver of patients with chronic HCV and HbV infection.A high percentage of patients with monoclonal b-cells infiltration and b-cell clonality in the liver were finally diagnosed as having a definite b-cell malignancy.

关键词： Hepatitis Hepatitis C virus b-lymphocytes Polymerase chain reaction Gene rearrangement CIonality

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Identification of the epitopes on HCV core protein recognized by HLA-A2 restricted cytotoxic T lymphocytes

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World Journal of Gastroenterology 2001年第4期7卷 583-586页

作者： Hong-Chao Zhou De-Zhong Xu Xue-Ping Wang Jing-Xia Zhang Ying-Huang Yong-Ping Yan Yong Zhu bo-Quan Jin Department of Epidemiology,the Fourth Military Medical University,Xi’an 710033,Shaanxi Province,ChinaDepartment of Immunology,the Fourth Military Medical University,Xi’an 710033,Shaanxi Province,China Department of Immunology the Fourth Military Medical University Xi'an 710033 Shaanxi Province China. juliu@***

AIM: To identify hepatitis C virus(HCV) core protein epitopes recognized by HLA-A2 restricted cytotoxic T lymphocyte (CTL). METHODS: Utilizing the method of computer prediction followed by a 4h(51)Cr release assay confirmation. RESULTS: The results showed that peripheral blood mononuclear cells (PbMC) obtained from two HLA-A2 positive donors who were infected with HCV could lyse autologous target cells labeled with peptide "ALAHGVRAL (core 150-158)". The rates of specific lysis of the cells from the two donors were 37.5% and 15.8%, respectively. blocking of the CTL response with anti-CD4 mAb caused no significant decrease of the specific lysis. but blocking of CTL response with anti-CD8 mAb could abolish the lysis. CONCLUSION: The peptide (core 150-158) is the candidate epitope recognized by HLAA2 restricted CTL.

关键词： Amino Acid Sequence Antibodies, Viral b-lymphocytes Cell Line Epitope Mapping HLA-A2 Antigen Hepacivirus Hepatitis C Humans Peptide Fragments Predictive Value of Tests Research Support, Non-U.S. Gov't T-lymphocytes, Cytotoxic Viral Core Proteins

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