Detection of Antibody to Hepatitis C Virus in Prospectively Followed Transfusion Recipients with Acute and Chronic Non-A, Non-B Hepatitis

Alter, Harvey J.; Purcell, Robert H.; Shih, J. Wai-Kuo; Melpolder, Jacqueline C.; Houghton, Michael; Choo, Qui‐Lim; Kuo, George

doi:10.1056/nejm198911303212202

Cited by 1,577 publications

(398 citation statements)

References 13 publications

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“…2 is a major public health problem, with 170 million chronically infected people worldwide (1,2). The current treatment by combined interferon-ribavirin therapy fails to cure the infection in 30 -50% of cases (3,4), particularly those with HCV genotypes 1 and 2.…”

Section: Infection By the Hepatitis C Virus (Hcv)mentioning

confidence: 99%

Heat-shock Protein 90 Is Essential for Stabilization of the Hepatitis C Virus Nonstructural Protein NS3

Ujino

Yamaguchi

Shimotohno

et al. 2009

Journal of Biological Chemistry

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The hepatitis C virus (HCV) is a major cause of chronic liver disease. Here, we report a new and effective strategy for inhibiting HCV replication using 17-allylaminogeldanamycin (17-AAG), an inhibitor of heat-shock protein 90 (Hsp90). Hsp90 is a molecular chaperone with a key role in stabilizing the conformation of many oncogenic signaling proteins. We examined the inhibitory effects of 17-AAG on HCV replication in an HCV replicon cell culture system. In HCV replicon cells treated with 17-AAG, we found that HCV RNA replication was suppressed in a dose-dependent manner, and interestingly, the only HCV protein degraded in these cells was NS3 (nonstructural protein 3). Immunoprecipitation experiments showed that NS3 directly interacted with Hsp90, as did proteins expressed from ⌬NS3 protease expression vectors. These results suggest that the suppression of HCV RNA replication is due to the destabilization of NS3 in disruption of the Hsp90 chaperone complex by 17-AAG. Infection by the hepatitis C virus (HCV)2 is a major public health problem, with 170 million chronically infected people worldwide (1, 2). The current treatment by combined interferon-ribavirin therapy fails to cure the infection in 30 -50% of cases (3, 4), particularly those with HCV genotypes 1 and 2. Chronic infection with HCV results in liver cirrhosis and can lead to hepatocellular carcinoma (5, 6). Although an effective combined interferon-␣-ribavirin therapy is available for about 50% of the patients with HCV, better therapies are needed, and preventative vaccines have not yet been developed.HCV is a member of the Flaviviridae family and has a positive strand RNA genome (7, 8) that encodes a large precursor polyprotein, which is cleaved by host and viral proteases to generate at least 10 functional viral proteins: core, E1 (envelope 1), E2, p7, NS2 (nonstructural protein 2), NS3, NS4A, NS4B, NS5A, and NS5B (9, 10). NS2 and the amino terminus of NS3 comprise the NS2-3 protease responsible for cleavage between NS2 and NS3 (9, 11), whereas NS3 is a multifunctional protein consisting of an amino-terminal protease domain required for processing NS3 to NS5B (12, 13). NS4A is a cofactor that activates the NS3 protease function by forming a heterodimer (14 -17), and the hydrophobic protein NS4B induces the formation of a cytoplasmic vesicular structure, designated the membranous web, which is likely to contain the replication complex of HCV (18,19). NS5A is a phosphoprotein that appears to play an important role in viral replication (20 -23), and NS5B is the RNA-dependent RNA polymerase of HCV (24, 25). The 3Ј-untranslated region consists of a short variable sequence, a poly(U)-poly(UC) tract, and a highly conserved X region and is critical for HCV RNA replication and HCV infection (26 -29).Hsp90 (heat-shock protein 90) is a molecular chaperone that plays a key role in the conformational maturation of many cellular proteins. Hsp90 normally functions in association with other co-chaperone proteins, which together play an important role in folding newly ...

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Section: Infection By the Hepatitis C Virus (Hcv)mentioning

confidence: 99%

Heat-shock Protein 90 Is Essential for Stabilization of the Hepatitis C Virus Nonstructural Protein NS3

Ujino

Yamaguchi

Shimotohno

et al. 2009

Journal of Biological Chemistry

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“…Hepatitis C virus (HCV), 1 a member of the Flaviviridae virus family (1), is the major agent responsible for parenterally transmitted non-A non-B hepatitis (2). The viral genome is represented by a single-stranded positive-sense RNA that is translated into a large polyprotein subsequently cleaved into the different viral proteins, structural and nonstructural (3).…”

Section: Introductionmentioning

confidence: 99%

Specific detection of hepatitis C virus minus strand RNA in hematopoietic cells.

Lerat¹,

Berby²,

Trabaud³

et al. 1996

J. Clin. Invest.

260

188

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The presence of hepatitis C virus (HCV) negative strand RNA in extrahepatic compartments based on PCR detection assays has been suggested in many reports with a very heterologous detection rate (from 0 to 100%). In this study, we have analyzed the presence of HCV negative strand in hepatic (liver biopsies, n ϭ 20) and extrahepatic (sera, n ϭ 32; PBMC, n ϭ 26 and fresh bone marrow cells, n ϭ 8) compartments from infected patients with three different reverse transcriptase (RT)-PCR-based assays using primers located in the 5 Ј noncoding region, with or without a tag sequence, or in the nucleocapsid (CAP). Samples were selected to display different viral loads (10 5 -3 ϫ 10 7 genomic equivalent/ml or gram) and viral genotypes ( n ϭ 5). Using synthetic as well as biological templates, we could document extensive artifactual detection of negative strand RNA, due to self priming and mispriming events, when either 5 Ј noncoding region primer pair was used, whereas both artifacts were dramatically reduced (

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“…Hepatitis C virus (HCV) is the major etiological agent responsible for parenterally transmitted non-A, non-B hepatitis [1,2]. HCV is a small enveloped, positive-strand RNA virus, which appears to have a similar genetic organization to the pesti-or flaviviruses [3].…”

Section: Introductionmentioning

confidence: 99%

Negative-strand HCV RNA Was Not Detected in Bone Marrow Cells of Patients with HCV Infection.

et al. 1998

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Summary:To determine whether hepatitis C virus (HCV) replicates in bone marrow, we investigated positive-and negative-strand HCV RNA in bone marrow cells and fluids, and sera from patients with HCV infection. The study population consisted of 15 patients positive for antibodies to HCV (anti-HCV). Positive-and negative-strands HCV RNA were detected using highly strandspecific rTth reverse transcription-polymerase chain reaction (rTth RT-PCR) followed by Southern blotting analysis. Positive-strand HCV RNA was detected in 12 (80%) serum samples, in 13 (86.7%) bone marrow fluid specimens, and in 6 (40.0%) bone marrow cell samples. Negativestrand HCV RNA was detected in 9 (60.0%) serum samples, 11 (91.7%) fluid specimens, while it was not detected in bone marrow cells. The absence of negative-strand HCV RNA in bone marrow cells suggested that HCV does not replicate in these cells. Negative-strand HCV RNA detected in serum and bone marrow fluid samples may have been due to contamination with circulating HCV RNA from hepatocytes.

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Detection of Antibody to Hepatitis C Virus in Prospectively Followed Transfusion Recipients with Acute and Chronic Non-A, Non-B Hepatitis

Cited by 1,577 publications

References 13 publications

Heat-shock Protein 90 Is Essential for Stabilization of the Hepatitis C Virus Nonstructural Protein NS3

Heat-shock Protein 90 Is Essential for Stabilization of the Hepatitis C Virus Nonstructural Protein NS3

Specific detection of hepatitis C virus minus strand RNA in hematopoietic cells.

Negative-strand HCV RNA Was Not Detected in Bone Marrow Cells of Patients with HCV Infection.

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