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Hepatitis C virus infection of human hepatoma cell line 7721 in vitro

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World Journal of Gastroenterology 2001年第5期7卷 685-689页

作者： Zhi-Qiang Song~1 Fei Hao~1 Feng Min~2 Qiao-Yu Ma~2 Guo-Dong Liu~2 Department of Dermatology~1Department of Infectious Diseases~2,Southwest Hospital,Third Military Medical University,Chongqing 400038,China Department of Dermatology Southwest Hospital Third Military Medical University Chongqing 400038 China. songzq@***

AIM: To establish a cell culture system with long-term replication of hepatitis C virus in vitro. METHODS: Human hepatoma cell line 7721 was tested for its susceptibility to HCV by incubating with a serum from a patient with chronic hepatitis C. Cells and supernatant were harvested at various phases during the culturing periods. The presence of HCV RNA, the expression of HCV antigens in cells and/or supernatant were examined by RTPCR, in situ hybridization and immunohistochemistry respectively. RESULTS: The intracellular HCV RNA was first detected on d2 after infection and then could be intermittently detected in both cells and supernatant over a period of at least three months. The expression of HCV NS3,CP10 antigens could be observed in cells. The fresh cells could be infected by supernatant from cultured infected cells and the transmission of viral genome from HCV-infected 7721 cells to PBMCs was also observed. CONCLUSION: The hepatoma line 7721 is not only susceptible to HCV but also supports its long-term replication in vitro.

关键词： Carcinoma, Hepatocellular Liver Neoplasms Antigens, Viral Cell Division hepacivirus development Hepatitis C Humans In Situ Hybridization In Vitro Phenotype RNA, Viral Research Support, Non-U.S. Gov't Tumor Cells, Cultured Virus Replication

Effect of in vitro interferon-beta administration on hepatitis C virus in peripheral blood mononuclear cells as a predictive marker of clinical response to interferon treatment for chronic hepatitis C

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World Journal of Gastroenterology 2004年第5期10卷 733-736页

作者： Kaori Mochizuki Tatehiro Kagawa Shinji Takashimizu Kazuya Kawazoe Sei-Ichiro Kojima Naruhiko Nagata Atsushi Nakano Yasuhiro Nishizaki Koichi Shiraishi Masaru Itakura Norihito Watanabe Tetsuya Mine Shohei Matsuzaki Department of Internal Medicine Division of GastroenterologyTokai University School of MedicineBohseidaiIsehara 259-1193Japan

AIM:To test whether in vitro incubation of peripheral blood mononuclear cells (PBMC) with interferon (IFN) could efficiently decrease hepatitis C virus-RNA (HCV-RNA) amount and to analyze whether this effect was associated with clinical response to ***:Twenty-seven patients with histologically proven chronic hepatitis C were given intravenous administration of 6 million units (MU) IFN-β daily for 6 weeks followed by three times weekly for 20 weeks. PBMC collected before IFN therapy were incubated with IFN-β and HCV-RNA in PMBC was semi-quantitatively ***: Twenty-five patients completed IFN *** patients (32%) had sustained loss of serum HCV-RNA with normal serum ALT levels after IFN therapy (complete responders).HCV-RNA in PBMC was detected in all patients,whereas it was not detected in PBMC from healthy *** vitro administration of IFN-β decreased the amount of HCV-RNA in PMBC in 18 patients (72%). Eight of these patients obtained complete response. On the other hand,none of the patients whose HCV-RNA in PBMC did not decrease by IFN-β was complete responders. Multiple logistic regression analysis revealed that the decrease of HCV-RNA amount in PBMC by IFN-β was the only independent predictor for complete response (P<0.05).CONCLUSION:The effect of in vitro IFN-β on HCV in PBMC reflects clinical response and would be taken into account as a predictive marker of IFN therapy for chronic hepatitis C.

关键词： Adult Antiviral Agents dosage Drug Resistance, Viral Female hepacivirus Hepatitis C, Chronic Humans In Vitro Interferon-beta Leukocytes, Mononuclear Male Middle Aged Predictive Value of Tests RNA, Viral

Distribution and changes in hepatitis C virus genotype in China from 2010 to 2020

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World Journal of Clinical Cases 2022年第14期10卷 4480-4493页

作者： Jia Yang Hui-Xin Liu Ying-Ying Su Zhi-Sheng Liang Hui-Ying Rao Peking University Hepatology Institute Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver DiseasesPeking University People’s HospitalBeijing 100044China Department of Clinical Epidemiology and Biostatistics Peking University People’s HospitalBeijing 100044China National Institute of Diagnostics and Vaccine Development in Infectious Diseases School of Public HealthXiamen UniversityXiamen 361000Fujian ProvinceChina Department of Global Health School of Public HealthPeking UniversityBeijing 100044China

BACKGROUND Hepatitis C virus(HCV)causes a large number of infections *** infections seem to be increasing according to a report of the World Health Organization in *** direct-acting antivirals are quite effective for most genotypes of the HCV,some genotypes fail to ***,the trend of genotype distribution is vital to better control the development of this *** To analyze the distribution and trends of the HCV genotype before and after the emergence of direct-acting antivirals in *** We searched all literature published in five electronic databases-China National Knowledge Infrastructure,Wan Fang Data,VIP Chinese Journal Database,Chinese Biomedical Literature Service System,and PubMed-from January 1,2010 to December 31,*** search strategy combined medical subject headings and free-text terms,including“hepatitis C virus”or“HCV”and“genotype”or“subtype”and“China”or“Chinese”.Additional relevant articles were searched by manual *** were extracted to build a *** of the data were totaled according to regions,periods,routes of transmission,and *** percentages in various stratifications were *** There were 76110 samples from 30 provinces included in the *** 1(G1)accounted for 58.2%of cases nationwide,followed by G2,G6,G3b,G3a,unclassified and mixed infections(17.5%,7.8%,6.4%,4.9%,1.8%,and 1.2%,respectively).The constitution of genotype varied among different regions,with G6 and G3b being more common in the south and southwest,respectively(28.1%,15.4%).The past ten years have witnessed a decrease in G1 and G2 and an increase in G3 and G6 in almost all *** drug-use population had the most abundant genotypes,with G6 ranking first(33.3%),followed by G1 and G3b(23.4%,18.5%).CONCLUSION G3 and G6 pose a new challenge for HCV *** study revealed the distribution of HCV genotypes in China over the past 10 years,providing information for HCV management strategie

关键词： hepacivirus Genotype China Drug users Direct-acting antiviral Hepatitis C virus

Tricistronic hepatitis C virus subgenomic replicon expressing double transgenes

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World Journal of Gastroenterology 2014年第48期20卷 18284-18295页

作者： Xin Cheng Xiang-Cui Gao Jun-Ping Wang Xin-Ying Yang Yan Wang Bao-Sheng Li Fu-Biao Kang Hai-Jun Li Yue-Min Nan Dian-Xing Sun The Liver Disease Diagnosis and Treatment Center of PLA Bethune International Peace Hospital Department of Traditional and Western MedicalHepatology Third Hospital of Hebei Medical University

AIM: To construct a tricistronic hepatitis C virus (HCV) replicon with double internal ribosome entry sites (IRESes) of only 22 nucleotides for each, substituting the encephalomyocarditis virus (EMCV) IRESes, which ar... 详细信息

关键词： hepacivirus Replicon Internal ribosome entry site Tricistronic expression

Is laparoscopy an advantage in the diagnosis of cirrhosis in chronic hepatitis C virus infection?

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World Journal of Gastroenterology 2003年第4期9卷 745-750页

作者： Perdita Wietzke-Braun Felix Braun Peter Schott Giuliano Ramadori Department of Internal Medicine Division of Gastroenterology and EndocrinologyUniversity of GoettingenGermany

AIM:To evaluate the potential of laparoscopy in the diagnosis of cirrhosis and outcome of interferon treatment in HCV-infected patients. METHODS:In this retrospective study,diagnostic laparoscopy with laparoscopic liver biopsy was performed in 72 consecutive patients with chronic HCV *** presence or absence of drrhosis was analyzed macroscopically by laparoscopy and microscopically by liver biopsy *** and laboratory data and outcome of interferon-alfa treatment were compared between cirrhotic and noncirrhotic patients. RESULTS:Laparoscopically,cirrhosis was seen in 29.2 % (21/72)and non-cirrhosis in 70.8 %(51/72)of patients. Cirrhotic patients were significantly older with a significant longer duration of HCV infection than noncirrhotic patients. Laboratory parameters(AST,y-GT,y-globulin fraction)were measured significantly higher as well as significantly lower (prothrombin index,platelet count)in cirrhotic patients than in non-cirrhotic ***,cirrhosis was confirmed in 11.1%(8/72)and non cirrhosis in 88.9 %(64/72).Patients with macroscopically confirmed cirrhosis(n=21)showed histologically cirrhosis in 38.2 %(8/21)and histologically non- cirrhosis in 61.9 %(13/21).In contrast,patients with macroscopically non-cirrhosis(n=51)showed histologically non cirrhosis in all cases(51/51).Thirty-nine of 72 patients were treated with interferon-alfa,resulting in 35.9 %(14/39) patients with sustained response and 64.1%(25/39)with non ***-responders showed significantly more macroscopically cirrhosis than sustained *** contrast,there were no significant histological differences between non-responders and sustained responders. CONCLUSION:Diagnostic laparoscopy is more accurate than liver biopsy in recognizing cirrhosis in patients with chronic HCV *** biopsy is the best way to assess inflammatory grade and fibrotic *** invasive marker for staging,prognosis and management,and treatment outcome of chro

关键词： Adult Biopsy Comparative Study Female Genotype hepacivirus purification Hepatitis C, Chronic Humans Interferon-alpha Laparoscopy Liver Cirrhosis Male Middle Aged RNA, Viral Reproducibility of Results Retrospective Studies Treatment Outcome

HCV-RNA positivity in peripheral blood mononuclear cells of patients with chronic HCV-infection: does it really mean viral replication?

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World Journal of Gastroenterology 2001年第2期7卷 228-234页

作者： Volker Meier Sabine Mihm Perdita Wietzke-Braun Guliano Ramadori Georg-August-Universitat gottingen Zentrum innere medizinAbteilung Für Gastroenterologie Und EndokrinologieRobert-Koch-Straβe 4037075 GottingenGermany

AIM: To analyze the association of HCV-RNA with peripheral blood mononuclear cells (PBMC) and to answer the question whether HCV-RNA positivity in PBMC is due to viral replication. METHODS: HCV-RNA was monitored in serum and PBMC preparations from 15 patients with chronic HCV infection before, during and after an IFN-alpha therapy using a nested RT/PCR technique. In a second approach, PBMC from healthy donors were incubated in HCV positive plasma. RESULTS: In the IFN-alpha responding patients,HCV-RNA disappeared first from total RNA preparations of PBMC and then from serum. In contrast, in relapsing patients, HCV-RNA reappeared first in serum and then in PBMC. A quantitative analysis of the HCV-RNA concentration in serum was performed before and after transition from detectable to non detectable HCV-RNA in PBMC-RNA and vice versa. When HCV-RNA was detectable in PBMC preparations, the HCV concentration in serum was significantly higher than the serum HCV-RNA concentration when HCV-RNA in PBMC was not detectable. Furthermore, at no time during the observation period was HCV specific RNA observed in PBMC, if HCV-RNA in serum was under the detection limit. Incubation of PBMC from healthy donors with several dilutions of HCV positive plasma for two hours showed a concentration dependent PCR positivity for HCV-RNA in reisolated PBMC. CONCLUSION: The detectability of HCV-RNA in total RNA from PBMC seems to depend on the HCV concentration in serum. Contamination or passive adsorption by circulating virus could be the reason for detection of HCV-RNA in PBMC preparations of chronically infected patients.

关键词： Adult Aged Antiviral Agents Female hepacivirus Hepatitis C, Chronic Humans Interferon-alpha Leukocytes, Mononuclear Male Middle Aged RNA, Viral Research Support, Non-U.S. Gov't Reverse Transcriptase Polymerase Chain Reaction Viral Load Virus Replication

Full-length core sequence dependent complex-type glycosylation of hepatitis C virus E2 glycoprotein

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World Journal of Gastroenterology 2002年第3期8卷 499-504页

作者： CarolineStaib GerdSutter GSF-Institut

AIM: To study HCV polyprotein processing is important for the understanding of the natural history of HCV and the design of vaccines against HCV. The purpose of this study is to investigate the affection of context sequences on hepatitis C virus (HCV) E2 processing. METHODS: HCV genes of different lengths were expressed and compared in vaccinia virus/T7 system with homologous patient serum S94 and mouse anti-serum M( E2116) raised against *** -derived E2 peptide, *** analysis and GNA ( Galanthus nivalus ) lectin binding assay were performed to study the post-translational processing of the expressed products. RESULTS: E2 glycoproteins with different molecular weights (-75 kDa and -60 kDa) were detected using S94 and M( E2116), respectively. Deglycosylation analysis showed that this difference was mainly due to different glycosylation. Endo H resistance and its failure to bind to GNA lectin demonstrated that the higher molecular weight form (75 kDa) of E2 was complex-type glycosylated, which was readily recognized by homologous patient serum S94. Expression of complex-type glycosylated E2 could not be detected in all of the core-truncated constructs tested, but readily detected in constructs encoding full-length core sequences. CONCLUSION: The upstream conserved full-length core coding sequence was required for the production of E2 glycoproteins carrying complex-type N-glycans which reacted strongly with homologous patient serum and therefore possibly represented more mature forms of E2. As complex-type N-glycans indicated modification by Golgi enzymes, the results suggest that the presence of full-length core might be critical for E1/E2 complex to leave ER. Our data may contribute to a better understanding of the processing of HCV structural proteins as well as HCV morphogenesis.

关键词： Animals Cell Line Glycosylation Hela Cells hepacivirus Hepatitis C Antibodies Humans Molecular Weight Protein Processing, Post-Translational Research Support, Non-U.S. Gov't Viral Envelope Proteins

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

SEN virus does not affect treatment response in hepatitis C virus coinfected patients but SEN virus response depends on SEN virus DNA concentration

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World Journal of Gastroenterology 2004年第13期10卷 1893-1897页

作者： Abdurrahman Sagin Ortwin Adams OliverKirschberg AndreasErhardt TobiasHeintges Dieter Hussinger Klinik fr Gastroenterologie Hepatologie und InfektiologieUniversittsklinikum DsseldorfMoorenstr 540225 DsseldorfGermany Institut fr Virologie der Heinrich-Heine-Universitt Dsseldorf Moorenstr 540225 DsseldorfGermany

AIM: To clarify the effect of SEN virus (SENV) infection on a combination therapy including interferon alfa (IFN-α) or pegylated-IFN with ribavirin in patients with chronic hepatitis and the effect of a combination therapy on ***: SENV DNA was determined by polymerase chain reaction in serum samples from 95 patients with chronic hepatitis C. Quantitative analysis was done for SENV H ***: Twenty-one (22%) of 95 patients were positive for SENV DNA. There was no difference in clinical and biochemical parameters between patients with HCV infection alone and coinfected patients. The sustained response rate for HCV clearance after combination therapy did not differ between patients with SENV (52%) and without SENV(50%, n.s.). SENV DNA was undetectable in 76% of the initially SENV positive patients at the end of follow-up. SENV H response to combination therapy was significantly correlated with SENV DNA level (P=-0.05).CONCLUSION: SENV infection had no influence on the HCV sustained response rate to the combination *** rate of SENV to the combination therapy depends on SENV DNA level.

关键词： Adult Antiviral Agents DNA Virus Infections DNA Viruses purification DNA, Viral Drug Therapy, Combination Female hepacivirus Hepatitis C, Chronic Humans Interferon Alfa-2a Interferon-alpha Male Middle Aged Polyethylene Glycols Prevalence RNA, Viral Ribavirin