Expression of hepatitis C virus proteins in epithelial intestinal cells in vivo

Previous work on hepatitis C virus (HCV) led to the discovery of a new form of virus particle associating virus and lipoprotein elements. These hybrid particles (LVP for lipo-viro-particles) are enriched in triglycerides and contain at least apolipoprotein B (apoB), HCV RNA and core protein. These findings suggest that LVP synthesis could occur in liver and intestine, the two main organs specialized in the production of apoB-containing lipoprotein. To identify the site of LVP production, the genetic diversity and phylogenetic relationship of HCV quasispecies from purified LVP, whole serum and liver biopsies from chronically infected patients were studied. HCV quasispecies from LVP and liver differed significantly, suggesting that LVP were not predominantly synthesized in the liver but might also originate in the intestine. The authors therefore searched for the presence of HCV in the small intestine. Paraffin-embedded intestinal biopsies from 10 chronically HCV-infected patients and from 12 HCV RNA-negative controls (10 anti-HCV antibody-negative and two anti-HCV antibody-positive patients) were tested for HCV protein expression. HCV NS3 and NS5A proteins were stained in small intestine epithelial cells in four of the 10 chronically infected patients, and not in controls. Cells expressing HCV proteins were apoB-producing enterocytes but not mucus-secreting cells. These data indicate that the small intestine can be infected by HCV, and identify this organ as a potential reservoir and replication site. This further emphasizes the interaction between lipoprotein metabolism and HCV, and offers new insights into hepatitis C infection and pathophysiology.

Download Full-text

DETEKSI ANTIBODI MULTIPEL HEPATITIS C DALAM DARAH DONOR

INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY ◽

10.24293/ijcpml.v21i3.738 ◽

2016 ◽

Vol 21 (3) ◽

pp. 261

Author(s):

Ranti Permatasari ◽

Aryati Aryati ◽

Budi Arifah

Keyword(s):

Hepatitis C ◽

Predictive Value ◽

Core Protein ◽

Rapid Test ◽

Antibody Test ◽

Diagnostic Value ◽

Hcv Infection ◽

Antibody Detection ◽

Hcv Rna ◽

Hcv Antibody

Hepatitis C (HCV) infection could be spread by blood transfusion. Screening of HCV in donor blood could prevent HCV infection to the recipient. HCV antibody test using rapid test of multiple antibody detection by immunochromatography method is an easy and rapid test that could detect four HCV antibodies separately. The aim of this study was to evaluate the diagnostic value of antibody HCV using multiple antibody detection rapid test in diagnosing HCV infection. This was an analytical observational study with a cross sectional design. The samples consisted of 42 donors’ blood serum from the Surabaya Branch of the Indonesian Red Cross which underwent HCV infection test using ELISA method. The samples were then tested using PCR HCV RNA as the gold standard and antibody HCV multiple antibodydetection rapid test The diagnostic value of HCV antibody test using multiple antibody detection rapid test by immunochromatography method showed a diagnostic sensitivity of 100%, diagnostic specificity of 75%, positive predictive value of 66.7% and negative predictive value of 100%, a diagnostic efficiency of 83.3%, with a positive probability ratio of 4 times. The most often positive antibody pattern was four (4) positive antibodies (core protein, NS3, NS4 and NS5). Core protein (CP) and NS3 were the most often positive antibodies. Based on this study result, the HCV antibody test using multiple antibody detection rapid test by immunochromatography method has a good diagnostic value.

Download Full-text

Second-generation hepatitis C virus (HCV) antibody titers and serum HCV RNA levels in low-risk blood donors

Hepatology ◽

10.1016/0270-9139(94)90733-1 ◽

1994 ◽

Vol 19 (4) ◽

pp. I142

Author(s):

N Yuki

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Blood Donors ◽

Second Generation ◽

Low Risk ◽

Hcv Rna ◽

Antibody Titers ◽

Hcv Antibody ◽

Rna Levels

Download Full-text

932. Universal Hepatitis C Virus Screening in a Tennessee Tertiary Care Emergency Department

Open Forum Infectious Diseases ◽

10.1093/ofid/ofy209.072 ◽

2018 ◽

Vol 5 (suppl_1) ◽

pp. S31-S32 ◽

Cited By ~ 1

Author(s):

Cody A Chastain ◽

Jakea Johnson ◽

Karen Miller ◽

Katie Moore ◽

Amanda Lako ◽

...

Keyword(s):

Emergency Department ◽

Hepatitis C Virus ◽

Hepatitis C ◽

Tertiary Care ◽

Linkage To Care ◽

Hcv Rna ◽

Grant Recipient ◽

Age Cohort ◽

Hcv Antibody ◽

Hcv Screening

Abstract Background Despite hepatitis C virus (HCV) age cohort and risk factor screening recommendations, many at-risk individuals remain undiagnosed. Current screening practices may not adequately capture those at high risk for infection, especially in regions with increasing injection drug use (IDU). Universal HCV screening in a Tennessee tertiary care emergency department (ED) was introduced to help define regional epidemiology and to improve diagnosis and linkage to care. Methods This screening program was implemented in the Vanderbilt University Medical Center ED. Adult patients who underwent phlebotomy for clinical purposes were offered HCV screening. Samples were initially tested for HCV antibodies; if positive, samples were reflexed for HCV RNA testing. Patients with positive HCV RNA tests (i.e., active HCV infection) were notified, counseled, and offered linkage to care. Results A total of 11,637 screening tests were performed between April 1, 2017 and March 31, 2018, with 1,008 (8.7%) HCV antibody positive and 488 (4.2%) RNA positive. Of note, 81 (0.7%) were HCV antibody positive but RNA testing could not be performed due to insufficient sample volume. Several notable populations had high rates of HCV (Table 1). Importantly, 3.9% of people not born between 1945 and 1965 were HCV RNA positive, and they were the majority (63.5%) of patients with active HCV (Table 2). A minority (31.6%) of those with active HCV had a known history of IDU (Table 2). Conclusion HCV is common among patients presenting for emergency care at a Tennessee tertiary care ED. Universal screening identified many infections that would have been missed using age cohort and risk factors alone. ED HCV screening may be a useful method to augment guideline-based testing and intervene among populations not consistently screened. Disclosures C. A. Chastain, Gilead Sciences, Inc.: Grant Investigator and Research Contractor, Grant recipient and Research support. J. Johnson, Gilead Sciences, Inc.: Grant Investigator, Grant recipient. K. Miller, Gilead Sciences, Inc.: Grant Investigator, Grant recipient. J. H. Han, Gilead Sciences, Inc.: Grant Investigator, Grant recipient. W. H. Self, Gilead Sciences, Inc.: Grant Investigator, Grant recipient.

Download Full-text

The presence of hepatitis C virus (HCV) antibody in human immunodeficiency virus-positive hemophilic men undergoing HCV "seroreversion"

Blood ◽

10.1182/blood.v82.3.1010.1010 ◽

1993 ◽

Vol 82 (3) ◽

pp. 1010-1015 ◽

Cited By ~ 36

Author(s):

MV Ragni ◽

OK Ndimbie ◽

EO Rice ◽

FA Bontempo ◽

S Nedjar

Keyword(s):

Human Immunodeficiency Virus ◽

Hepatitis C Virus ◽

Hepatitis C ◽

Second Generation ◽

Hcv Rna ◽

Negative State ◽

Immunodeficiency Virus ◽

Before And After ◽

Hcv Antibody ◽

Cd4 Lymphocyte

Abstract Hepatitis C virus (HCV) is a major cause of transfusion-induced chronic liver disease in hemophiliacs, with 70% to 90% being anti-HCV positive. Seroreversion or loss of antibody response to HCV has been observed in a small proportion of human immunodeficiency virus-positive [HIV(+)] anti-HCV(+) hemophilic men. Despite the seroreversion to an anti-HCV- negative state, such patients continue to show serum alanine aminotransferase (ALT) elevations and biopsy evidence of cirrhosis and/or chronic active hepatitis. To determine the cause for the loss of anti-HCV antibody, we compared first- and second-generation anti-HCV enzyme immunosorbent assay (EIA 1.0 and 2.0), second-generation recombinant immunoblot (RIBA 2.0), and HCV-RNA amplification using polymerase chain reaction (PCR) in 19 “seroreverters” before and after seroreversion. There was no difference between 19 seroreverters and 59 persistently anti-HCV-positive hemophiliacs in mean ALT (1.1 +/- 0.1 XUL v 2.0 +/- 0.2 XUL; chi 2 = 1.80, P > .05), in mean CD4 (188 +/- 36/microL v 232 +/- 28/microL; t = 0.965, P > .05), or in the rate of progression to acquired immunodeficiency syndrome (13 of 19 [68.4%] v 30 of 59 [50.9%]; chi 2 = .987, P > .05, respectively). Before seroreversion, all 19 seroreverters (100%) were positive for EIA 1.0 and 2.0 and PCR, and all but 2 of 19 (89.5%) were RIBA 2.0 positive, whereas, after seroreversion, none were positive for EIA 1.0, 15 of 19 (78.9%) were positive for EIA 2.0, 8 of 18 (44.4%) were positive for RIBA 2.0, and 18 of 19 (94.7%) were positive for PCR. There was a lower CD4 lymphocyte number after seroreversion in those who were RIBA 2.0 negative as compared with those who were RIBA 2.0 positive (32 +/- 10/microL v 171 +/- 52/microL; t = 2.638, P > .05). These results indicate that HIV(+) anti-HCV(+) hemophilic men who undergo “HCV seroreversion” are truly infectious and anti-HCV positive by second- generation tests. Anti-HCV detection in immunosuppressed hosts is significantly improved by second-generation EIA and RIBA assays.

Download Full-text

Poly(I:C) and Lipopolysaccharide Innate Sensing Functions of Circulating Human Myeloid Dendritic Cells Are Affected In Vivo in Hepatitis C Virus-Infected Patients

Journal of Virology ◽

10.1128/jvi.01741-06 ◽

2007 ◽

Vol 81 (11) ◽

pp. 5537-5546 ◽

Cited By ~ 31

Author(s):

Ian Gaël Rodrigue-Gervais ◽

Loubna Jouan ◽

Geneviève Beaulé ◽

Dominike Sauvé ◽

Julie Bruneau ◽

...

Keyword(s):

Dendritic Cells ◽

Hepatitis C Virus ◽

Hepatitis C ◽

Poly I:C ◽

Hcv Rna ◽

Genomic Rna ◽

Tnf Α ◽

Wide Range ◽

Rna Levels

ABSTRACT The role of peripheral dendritic cells (DCs) in hepatitis C virus (HCV) infection is unclear. To determine if persistent infection exerts an inhibitory pressure on HCV-specific innate responses, we analyzed DC function in blood through quantification of cell-associated HCV RNA levels in conjunction with multiparametric flow cytometry analysis of pathogen recognition receptor-induced cytokine expression. Independently of the serum viral load, fluorescence-activated cell sorter-purified total DCs had a wide range of cell-associated HCV genomic RNA copy numbers (mean log10, 5.0 per 106 cells; range, 4.3 to 5.8). Here we report that for viremic patients with high viral loads in their total DCs, the myeloid DC (MDC) subset displayed impaired expression of interleukin-12 (IL-12) and tumor necrosis factor alpha (TNF-α) but normal IL-6 or chemokine CCL3 expression in response to poly(I:C) and lipopolysaccharide (LPS). IL-6-expressing cells from this subgroup of viremic patients demonstrated a significant increase (sixfold more) in TNF-α− IL-12− cell frequency compared to healthy donors (mean, 38.8% versus 6.5%; P < 0.0001), indicating a functional defect in a subpopulation of cytokine-producing MDCs (∼6% of MDCs). Attenuation of poly(I:C) and LPS innate sensing was HCV RNA density dependent and did not correlate with viremia or deficits in circulating MDC frequencies in HCV-infected patients. Monocytes from these patients were functionally intact, responding normally on a per-cell basis following stimulation, independent of cell-associated HCV RNA levels. Taken together, these data indicate that detection of HCV genomic RNA in DCs and loss of function in the danger signal responsiveness of a small proportion of DCs in vivo are interrelated rather than independent phenomena.

Download Full-text

Cytokine-Modulated Natural Killer Cells Differentially Regulate the Activity of the Hepatitis C Virus

International Journal of Molecular Sciences ◽

10.3390/ijms19092771 ◽

2018 ◽

Vol 19 (9) ◽

pp. 2771 ◽

Cited By ~ 1

Author(s):

Yoo Cho ◽

Hwan Lee ◽

Hyojeung Kang ◽

Hyosun Cho

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Nk Cells ◽

Natural Killer ◽

Core Protein ◽

Lymphoid Cells ◽

Hcv Rna ◽

Cell Infection ◽

Hcv Core Protein

HCV genotype 2a strain JFH-1 replicates and produces viral particles efficiently in human hepatocellular carcinoma (huh) 7.5 cells, which provide a stable in vitro cell infection system for the hepatitis C virus (HCVcc system). Natural killer (NK) cells are large lymphoid cells that recognize and kill virus-infected cells. In this study, we investigated the interaction between NK cells and the HCVcc system. IL-10 is a typical immune regulatory cytokine that is produced mostly by NK cells and macrophages. IL-21 is one of the main cytokines that stimulate the activation of NK cells. First, we used anti-IL-10 to neutralize IL-10 in a coculture of NK cells and HCVcc. Anti-IL-10 treatment increased the maturation of NK cells by enhancing the frequency of the CD56+dim population in NK-92 cells. However, with anti-IL-10 treatment of NK cells in coculture with J6/JFH-1-huh 7.5 cells, there was a significant decrease in the expression of STAT1 and STAT5 proteins in NK-92 cells and an increase in the HCV Core and NS3 proteins. In addition, rIL-21 treatment increased the frequency of the CD56+dim population in NK-92 cells, Also, there was a dramatic increase in the expression of STAT1 and STAT5 proteins in rIL-21 pre-stimulated NK cells and a decrease in the expression of HCV Core protein in coculture with J6/JFH-1-huh 7.5 cells. In summary, we found that the functional activation of NK cells can be modulated by anti-IL-10 or rIL-21, which controls the expression of HCV proteins as well as HCV RNA replication.

Download Full-text

Novel Robust Hepatitis C Virus Mouse Efficacy Model

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00413-06 ◽

2006 ◽

Vol 50 (10) ◽

pp. 3260-3268 ◽

Cited By ~ 22

Author(s):

Qing Zhu ◽

Yoko Oei ◽

Dirk B. Mendel ◽

Evelyn N. Garrett ◽

Montesa B. Patawaran ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Protease Inhibitor ◽

Small Animal ◽

Rna Replication ◽

Treatment Withdrawal ◽

Whole Body ◽

Hcv Rna ◽

Whole Body Imaging

ABSTRACT The lack of a robust small-animal model for hepatitis C virus (HCV) has hindered the discovery and development of novel drug treatments for HCV infections. We developed a reproducible and easily accessible xenograft mouse efficacy model in which HCV RNA replication is accurately monitored in vivo by real-time, noninvasive whole-body imaging of gamma-irradiated SCID mice implanted with a mouse-adapted luciferase replicon-containing Huh-7 cell line (T7-11). The model was validated by demonstrating that both a small-molecule NS3/4A protease inhibitor (BILN 2061) and human alpha interferon (IFN-α) decreased HCV RNA replication and that treatment withdrawal resulted in a rebound in replication, which paralleled clinical outcomes in humans. We further showed that protease inhibitor and IFN-α combination therapy was more effective in reducing HCV RNA replication than treatment with each compound alone and supports testing in humans. This robust mouse efficacy model provides a powerful tool for rapid evaluation of potential anti-HCV compounds in vivo as part of aggressive drug discovery efforts.

Download Full-text

Identification of in vivo interaction between Hepatitis C Virus core protein and 5′ and 3′ UTR RNA

Virus Research ◽

10.1016/j.virusres.2009.07.023 ◽

2009 ◽

Vol 145 (2) ◽

pp. 285-292 ◽

Cited By ~ 16

Author(s):

Kyung Lee Yu ◽

Soo In Jang ◽

Ji Chang You

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Core Protein ◽

Virus Core

Download Full-text

Characterization of Low- and Very-Low-Density Hepatitis C Virus RNA-Containing Particles

Journal of Virology ◽

10.1128/jvi.76.14.6919-6928.2002 ◽

2002 ◽

Vol 76 (14) ◽

pp. 6919-6928 ◽

Cited By ~ 461

Author(s):

P. André ◽

F. Komurian-Pradel ◽

S. Deforges ◽

M. Perret ◽

J. L. Berland ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Apolipoprotein B ◽

Core Protein ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Hcv Rna ◽

Low Density ◽

Density Fractions

ABSTRACT The presence of hepatitis C virus (HCV) RNA-containing particles in the low-density fractions of plasma has been associated with high infectivity. However, the nature of circulating HCV particles and their association with immunoglobulins or lipoproteins as well as the characterization of cell entry have all been subject to conflicting reports. For a better analysis of HCV RNA-containing particles, we quantified HCV RNA in the low-density fractions of plasma corresponding to the very-low-density lipoprotein (VLDL), intermediate-density lipoprotein, and low-density lipoprotein (LDL) fractions from untreated chronically HCV-infected patients. HCV RNA was always found in at least one of these fractions and represented 8 to 95% of the total plasma HCV RNA. Surprisingly, immunoglobulins G and M were also found in the low-density fractions and could be used to purify the HCV RNA-containing particles (lipo-viro-particles [LVP]). Purified LVP were rich in triglycerides; contained at least apolipoprotein B, HCV RNA, and core protein; and appeared as large spherical particles with a diameter of more than 100 nm and with internal structures. Delipidation of these particles resulted in capsid-like structures recognized by anti-HCV core protein antibody. Purified LVP efficiently bind and enter hepatocyte cell lines, while serum or whole-density fractions do not. Binding of these particles was competed out by VLDL and LDL from noninfected donors and was blocked by anti-apolipoprotein B and E antibodies, whereas upregulation of the LDL receptor increased their internalization. These results suggest that the infectivity of LVP is mediated by endogenous proteins rather than by viral components providing a mechanism of escape from the humoral immune response.

Download Full-text