interleukin-8 and Hepatitis-C

The interactions that occur between viral proteins and host factors, such as cellular proteins and signal transduction machinery, have a significant influence on the replication, persistence, and pathogenesis of all viruses. This is exemplified by hepatitis C virus (HCV), which infects an estimated 3% of the world's population and is a significant cause of liver disease. HCV-host interactions also affect the outcome of interferon (IFN) antiviral therapy, which is effective only in certain patients. In this review, we focus on the HCV nonstructural 5A (NS5A) protein, a model for diverse virus-host interactions, and highlight the interaction of viruses, including HCV, with the chemokine system.

Topics: Antiviral Agents; Chemokines; eIF-2 Kinase; Hepacivirus; Hepatitis C; Host-Parasite Interactions; Humans; Interferons; Interleukin-8; Liver Diseases; Signal Transduction; Viral Nonstructural Proteins; Virus Replication

TNF-α is a proinflammatory cytokine, dramatically elevated during pathogenic infection and often responsible for inflammation-induced disease pathology. SOCS proteins are inhibitors of cytokine signaling and regulators of inflammation. In this study, we found that both SOCS1 and SOCS3 were transiently induced by TNF-α and negatively regulate its NF-κB-mediated signal transduction. We discovered that PBMCs from HCV-infected patients have elevated endogenous SOCS3 expression but less TNF-α-mediated IκB degradation and proinflammatory cytokine production than healthy controls. HCV protein expression in Huh7 hepatocytes also induced SOCS3 and directly inhibited TNF-α-mediated IL-8 production. Furthermore, we found that SOCS3 associates with TRAF2 and inhibits TRAF2-mediated NF-κB promoter activity, suggesting a mechanism by which SOCS3 inhibits TNF-α-mediated signaling. These results demonstrate a role for SOCS3 in regulating proinflammatory TNF-α signal transduction and reveal a novel immune-modulatory mechanism by which HCV suppresses inflammatory responses in primary immune cells and hepatocytes, perhaps explaining mild pathology often associated with acute HCV infection.

Topics: Cell Line; Female; Gene Expression Regulation; Hepacivirus; Hepatitis C; Hepatocytes; Humans; Inflammation; Interleukin-8; Male; NF-kappa B; Signal Transduction; Suppressor of Cytokine Signaling 3 Protein; Suppressor of Cytokine Signaling Proteins; TNF Receptor-Associated Factor 2; Tumor Necrosis Factor-alpha

This study aimed to determine the association of rs2430561 and rs4073 polymorphisms in the Interferon gamma (IFN-ɤ) and Interleukin 8 (IL-8) genes, respectively, with hepatitis C virus-related oral lichen planus and disease severity.. This is a case-control study. 60 subjects were equally divided into patients with and without oral lichen planus. They were further subdivided into hepatitis C virus seropositive and seronegative patients. All patients were genotyped for IFN-γ rs2430561 thymine to adenine (T > A) and IL-8 rs4073 adenine to thymine (A > T) polymorphisms. All patients with oral lichen planus had their lesions measured and documented using the Escudier scoring system.. Disease activity was significantly higher in the "oral lichen planus/hepatitis C virus-positive" patients than in the "oral lichen planus/hepatitis C virus-negative" patients (P = 0.003). IFN-γ rs2430561 T > A and IL-8 rs4073 A > T genotypes and allele frequencies were not associated with the oral lichen planus group or the normal group. Stratification of the two groups into HCV and non-HCV-infected patients or into erosive and non-erosive types revealed no significant associations. The "A-allele-containing" genotypes of IL-8 rs4073 A > T were significantly more prevalent in the patients with oral lichen planus than in those without.. Hepatitis C virus infection is associated with the development of erosive oral lichen planus. The A-allele of IL-8 rs4073 A > T may have a role in the development and progression of oral lichen planus.

Topics: Case-Control Studies; Disease Progression; Female; Genotype; Hepatitis C; Humans; Interferon-gamma; Interleukin-8; Lichen Planus, Oral; Male; Middle Aged; Polymorphism, Single Nucleotide; Real-Time Polymerase Chain Reaction

Injection drug use is a growing major public health concern. Injection drug users (IDUs) have a higher incidence of co-morbidities including HIV, Hepatitis, and other infections. An effective humoral response is critical for optimal homeostasis and protection from infection; however, the impact of injection heroin use on humoral immunity is poorly understood. We hypothesized that IDUs have altered B cell and antibody profiles.. A comprehensive systems biology-based cross-sectional assessment of 130 peripheral blood B cell flow cytometry- and plasma- based features was performed on HIV-/Hepatitis C-, active heroin IDUs who participated in a syringe exchange program (n = 19) and healthy control subjects (n = 19). The IDU group had substantial polydrug use, with 89% reporting cocaine injection within the preceding month. IDUs exhibited a significant, 2-fold increase in total B cells compared to healthy subjects, which was associated with increased activated B cell subsets. Although plasma total IgG titers were similar between groups, IDUs had significantly higher IgG3 and IgG4, suggestive of chronic B cell activation. Total IgM was also increased in IDUs, as well as HIV Envelope-specific IgM, suggestive of increased HIV exposure. IDUs exhibited numerous features suggestive of systemic inflammation, including significantly increased plasma sCD40L, TNF-α, TGF-α, IL-8, and ceramide metabolites. Machine learning multivariate analysis distilled a set of 10 features that classified samples based on group with absolute accuracy.. These results demonstrate broad alterations in the steady-state humoral profile of IDUs that are associated with increased systemic inflammation. Such dysregulation may impact the ability of IDUs to generate optimal responses to vaccination and infection, or lead to increased risk for inflammation-related co-morbidities, and should be considered when developing immune-based interventions for this growing population.

Topics: Adult; B-Lymphocytes; CD40 Ligand; Comorbidity; Cross-Sectional Studies; Female; Hepatitis C; Heroin; HIV Antibodies; HIV Infections; Humans; Immunity, Humoral; Immunoglobulin G; Immunoglobulin M; Inflammation; Interleukin-8; Male; Narcotics; New York; Substance Abuse, Intravenous; Transforming Growth Factor alpha; Tumor Necrosis Factor-alpha; Young Adult

We aimed to investigate any association between hepatitis C virus (HCV) infection and non-Hodgkin’s\ lymphoma (NHL) in the view of cytokines that control inflammation/angiogenesis and their correlation with\ certain CD markers. NHL patients with or without HCV infection were studied. CD5, CD30, CD3, CD20 and\ CD45 were immunohistochemically evaluated. Plasma levels of vascular endothelial and platelet derived growth\ factors (VEGF, and PDGF), tumor necrosis factor (TNF-α), transforming growth factor (TGF-β), interleukin-6\ (IL-6), IL-8, IL-4, IL-12 and interferon gamma (IFN-γ) were detected by enzyme-linked immunosorbent assay\ (ELISA). HCV+ve NHL patients showed a significant reduction in VEGF, PDGF, IFN-γ, CD5 and CD45 and a\ significant increase in IL-12 and IL-8. In conclusion, there was a significant change in cytokine secretion and\ expression of CD markers in HCV+ve NHL patients. Based on our results, HCV infection in NHL patients\ requires more in-depth investigations to explore any role in lymphoma progression.

Topics: Adolescent; Adult; Aged; Antigens, CD; Biomarkers, Tumor; Female; Hepacivirus; Hepatitis C; Humans; Inflammation; Interferon-gamma; Interleukin-12; Interleukin-4; Interleukin-6; Interleukin-8; Lymphoma, Non-Hodgkin; Male; Middle Aged; Neovascularization, Pathologic; Platelet-Derived Growth Factor; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A; Young Adult

Hepatitis B virus/hepatitis C virus (HBV/HCV) dual infection is common among high-risk individuals. To characterize the virological and immunological features of patients with HBV/HCV dual infection, we enrolled 1,049 individuals who have been identified as injection drug users. Patients were divided into single and dual infection groups according to the serological markers. We found the average HCV RNA level was significantly lower; however, HBV viral load was significantly higher in HBV/HCV dual-infected patients (n = 42) comparing HCV single infection (n = 340) or HBV single infection (n = 136). The level of anti-HBs in patients who experienced spontaneous HBV clearance was higher than that in HCV single-infected patients with HBV spontaneous clearance. The level of anti-HCV E2 in HBV/HCV dual infection was lower than that detected in HCV single infection. Serum levels of IL-6, IL-8, and TNF-α were significantly lower in HBV/HCV dual-infected patients than in patients infected with HBV or HCV alone. Taken together, two viral replications are imbalanced in dual infected patients. The anti-HBs and anti-HCV E2 antibody production were impaired and proinflammatory IL-6, IL-8, and TNF-α also downregulated due to dual infection. These findings will help further understanding the pathogenesis of HBV/HCV dual infection.

Topics: Adult; Antibody Formation; Cytokines; Female; Hepacivirus; Hepatitis B; Hepatitis B virus; Hepatitis C; Humans; Interleukin-6; Interleukin-8; Male; RNA, Viral; Serologic Tests; Tumor Necrosis Factor-alpha; Viral Load; Virus Replication

Fibrosis progression in hepatitis C virus (HCV)-infected patients varies greatly between individuals. Chemokines recruit immune cells to the infected liver and may thus play a role in the fibrosis process.. To investigate plasma levels of a diverse chemokine panel in relation to liver fibrosis.. African-American and Caucasian HCV genotype 1 infected patients were treated with peginterferon (pegIFN) and ribavirin (RBV) for 48 weeks (VIRAHEP-C cohort). Plasma levels of 13 cytokines were studied at baseline (n = 386). Subsequently, GROα levels were assessed in a sub cohort (n = 99) at baseline, and at 4 and 12 weeks after start of pegIFN/RBV treatment.. Increased severity of fibrosis (Ishak fibrosis score 0-2 vs. 3-6) was associated with increased plasma IP-10 (CXCL10) and IL-8 (CXCL8) levels, and decreased plasma levels of the chemokine growth-related oncogene (GRO, CXCL1-3). Plasma GRO levels were also positively correlated with platelet counts, and were higher in African-American as compared to Caucasian patients. In response to pegIFN/RBV treatment, GROα levels increased in Caucasian but not African-American patients from week 4 onwards.. The association with severity of fibrosis and platelet count positions plasma GRO as a potential biomarker for liver fibrosis in HCV-infected patients. The secretion of GRO by platelets may explain the correlation between GRO plasma level and platelet count. The ethnic difference in GRO levels both pre-treatment and in response to pegIFN/RBV might be driven by a genetic polymorphism in GROα associated with higher plasma levels and more common in the African-American population.

Topics: Adult; Aged; Antiviral Agents; Biomarkers; Black or African American; Chemokine CXCL1; Chemokines; Drug Therapy, Combination; Female; Hepacivirus; Hepatitis C; Humans; Interferon alpha-2; Interferon-alpha; Interleukin-8; Interleukins; Liver Cirrhosis; Male; Middle Aged; Platelet Count; Polyethylene Glycols; Polymorphism, Genetic; Recombinant Proteins; Ribavirin; White People; Young Adult

The rate of hepatocellular carcinoma (HCC) is increasing in Egypt where the major risk factor is chronic infection with hepatitis C virus (HCV). The development of effective markers for the detection of HCC could have an impact on cancer mortality and significant public health implications worldwide. The objective of this study is to investigate the role of interleukin-8, alpha-fetoprotein (AFP), oxidative stress markers, and some trace elements in Egyptian patients with hepatocellular carcinoma infected with hepatitis C virus.. This study comprised 40 patients with HCC (20 with cirrhosis and 20 without cirrhosis) and 20 patients with hepatitis C virus. They were 39 males and 21 females with ages ranging from 22 to 71 years. Twenty apparently healthy volunteers with matched age and sex were taken as control group. Serum concentration levels of IL-8 and AFP were measured using an enzyme-linked immunosorbent assay (ELISA). Antioxidants were measured using spectrophotometric analysis and trace elements by using atomic absorption spectrophotometry.. A highly significant elevation was found in interleukin-8, alpha- fetoprotein, iron, and malondialdehyde in patients with HCC compared to control subjects. On the other hand, serum levels of reduced glutathione, catalase, superoxide dismutase, total antioxidant capacity, and zinc were significantly decreased in patients with HCC compared to control subjects. A positive correlation was found between serum level of IL-8 and each of GSH (r = -0.534 and p = 0.000), SOD (r = -0.295 and p = 0.021), CAT (r = -0.545 and p = 0.000), and Zn (r = 0.422 and p = 0.001) in all patient groups.. The ability to measure IL-8 in serum could be useful as a marker of HCC in patients. The levels of antioxidants such as CAT, SOD, and GSH in HCC patients when compared to control groups play a vital and important role in the prevention of liver cancer. Interleukin-8, some antioxidants (MDA, GSH, CAT and SOD), and some trace elements (Fe and Zn) might be simultaneously evaluated in order to enhance the detection of HCC.

Topics: Adult; Aged; alpha-Fetoproteins; Biomarkers, Tumor; Carcinoma, Hepatocellular; Case-Control Studies; Catalase; Enzyme-Linked Immunosorbent Assay; Female; Glutathione; Hepatitis C; Humans; Interleukin-8; Liver Neoplasms; Male; Middle Aged; Oxidative Stress; Superoxide Dismutase; Young Adult; Zinc

Autoimmune and non-autoimmune thyroiditis frequently occur in persons with hepatitis C virus (HCV) infection. Treatment with interferon alpha (IFNα) is also associated with significant risk for the development of thyroiditis. To explore HCV-thyroid interactions at a cellular level, we evaluated whether a human thyroid cell line (ML1) could be infected productively with HCV in vitro.. ML1 cells showed robust surface expression of the major HCV receptor CD81. Using a highly sensitive, strand-specific reverse transcription polymerase chain reaction assay, positive-sense and negative-sense HCV RNA were detected in ML1 cell lysates at days 3, 7, and 14 postinfection with HCV. HCV core protein was expressed at high levels in ML1 supernatants at days 1, 3, 5, 7, and 14 postinfection. The nonstructural protein NS5A was also detected in ML1 cell lysates by Western blotting. HCV entry into ML1 cells was shown to be dependent on the HCV entry factors CD81 and SR-B1/CLA1, while IFNα inhibited HCV replication in ML1 cells in a dose-dependent manner. Supernatants from HCV-infected ML1 cells were able to infect fresh ML1 cells productively, suggesting that infectious virions could be transferred from infected to naïve thyroid cells in vivo. Additionally, HCV infection of ML1 cells led to increased expression of the pro-inflammatory cytokine IL-8.. For the first time, we have demonstrated that HCV can infect human thyroid cells in vitro. These findings strongly suggest that HCV infection of thyrocytes may play a role in the association between chronic HCV infection and thyroid autoimmunity. Furthermore, the thyroid may serve as an extrahepatic reservoir for HCV viral replication, thus contributing to the persistence of viral infection and to the development of thyroid autoimmunity.

Topics: Autoimmunity; Cell Line; Hepacivirus; Hepatitis C; Humans; Interferon-alpha; Interleukin-8; Scavenger Receptors, Class B; Tetraspanin 28; Thyroid Gland; Thyroiditis; Viral Core Proteins; Viral Nonstructural Proteins; Virus Replication

The manner in which ribavirin (RBV) enhances the antiviral effects of interferon (IFN) against hepatitis C virus (HCV) remains unknown. We investigated whether RBV modifies IFN-stimulated genes (ISGs) in vivo and in vitro.. We measured the messenger RNA (mRNA) levels of ISGs in T lymphocytes from patients with HCV infection who were receiving IFN-α therapy with or without RBV. We added RBV and/or IFN-α to a plasmid-based HCV replication system containing a full-length HCV genotype 1a sequence in HepG2 and Huh7 cell lines and the JFH-1 HCV genotype 2a sequence in Huh7 cell lines and measured levels of ISGs and autocrine IFN-β.. The expression of protein kinase R and myxovirus resistance A mRNA was enhanced more with IFN-α and RBV than by IFN-α alone in assays in vivo and in vitro. Such enhancement depended on autocrine IFN-β being enhanced by RBV. RBV upregulated interleukin 8 (IL-8) in the absence of IFN-α. The IL-8 upregulation induced by RBV was responsible for the activation of activator protein 1 (AP-1).. Ribavirin augments the anti-HCV effects of IFN-α induced by ISGs through enhancing autocrine IFN-β. Moreover, RBV can enhance IL-8 through activating AP-1. Improved understanding of ISG modulation by RBV would help to establish a means of eliminating HCV.

Topics: Adult; Aged; Antiviral Agents; Female; Gene Expression Profiling; Hepacivirus; Hepatitis C; Humans; Interferon-alpha; Interferon-beta; Interleukin-8; Male; Middle Aged; Ribavirin; T-Lymphocytes; Transcriptional Activation; Virus Replication

Hepatitis C virus (HCV), a major cause of liver disease worldwide, is frequently resistant to the antiviral alpha interferon (IFN). Previous studies have shown that HCV NS5A protein induces expression of the proinflammatory interleukin-8 (IL-8) and this may affect the therapeutic outcome. The aim of the present study was to investigate the relationship among levels of IL-8 in serum of subjects with past exposure to HCV indicated by positive IgG to HCV and negative PCR, patients with chronic HCV infections and in responder to combined alfa IFN and ribavirin therapy. The study included 48 Egyptian subjects with evidence of HCV infection. They were classified according to viremia to patients with chronic hepatitis C with positive viremia and immune subjects with positive HCV IgG alone. Chronic HCV patients were followed after therapy and 16 healthy adults as control group. It was found that viral load was dependent factor for the level of IL 8 (P = 0.003) and there was significant correlation between levels of IL-8 before treatment and after treatment. The present study highlights the kinetic of serum levels of interleukin-8 in patients with chronic hepatitis C genotype 4 before and after therapy with combined alfa interferon and ribavirin. It was demonstrated that HCV infection was associated with higher levels of interleukin-8 in pretreatment and posttreatment period. Moreover, immune subjects also had higher level of interleukin-8, indicating its role in immunity to HCV infection.

Topics: Adult; Antibodies, Viral; Antiviral Agents; Drug Therapy, Combination; Enzyme-Linked Immunosorbent Assay; Female; Hepacivirus; Hepatitis C; Humans; Immunoglobulin G; Interferon-alpha; Interleukin-8; Male; Middle Aged; Polymerase Chain Reaction; Ribavirin; Treatment Outcome

The aim of this study was to explore a possible association between the pattern of serum cytokines with the virological and biochemical status of hepatitis C virus (HCV)-seropositive blood donors.. 23 non-viremic and 33 viremic HCV-seropositive blood donors based on HCV-RNA tests, and 29 healthy individuals were included. Cytometric bead array assays were performed to detect cytokines.. The subjects were classified as low, medium or high cytokine producers based on the tertile distribution. The absence of detectable viremia was associated with high IL-1β and IL-8 producers. Conversely, elevated levels of IL-6, IL-10 and IL-12 were associated with detectable viremia. An increased frequency of high IL-1β producers was observed frequently in the non-viremic recombinant immunoblot assay (RIBA)-indeterminate subjects, while the high IL-4, IL-6, IL-8, IL-10 and IL-12 producers were more frequent in the non-viremic RIBA-positive subjects. Furthermore, the levels of IL-1β and IL-8 were higher in viremic subjects with a low level of alanine-aminotransferase (ALT), whereas the level of IFN-γ was increased among viremic subjects with a high ALT level.. IL-1β and IL-8 were more likely to be associated with a non-viremic or less severe HCV infection, whereas IL-2 and IFN-γ levels correlated with a high ALT level.

Topics: Adult; Blood Donors; Female; Hepacivirus; Hepatitis C; Humans; Interferon-gamma; Interleukin-8; Male; Middle Aged; RNA, Viral; Viremia

In vitro study has shown that mechanisms for inhibiting interferon (IFN)-α antiviral action by non-structural 5A protein include interaction with IFN-induced RNA-dependent protein kinase and induction of interleukin (IL)-8 expression. Mutations in the non-structural 5A IFN sensitivity-determining region (ISDR) were reported to correlate with sustained virological response (SVR). IL-8 is associated with the inhibition of IFN-α action. We investigated whether pretreatment ISDR mutations and hepatic IL-8 messenger RNA (mRNA) expression had an effect on the SVR rate under combination therapy.. A total of 53 HCV-1b patients who completed 24 weeks of pegylated-IFN-α2b plus ribavirin, a 24-week follow-up and had enough tissue specimens were enrolled. Liver biopsy was performed within 6 months before antiviral therapy. Hepatic IL-8 mRNA expression was measured by real-time reverse transcriptase PCR.. Of 53 patients, 30 exhibited SVR. Multivariate analysis revealed that hepatic IL-8 mRNA expression <1.5×10(-4) (OR 6.66, 95% CI 1.77-25.05) and ISDR mutations ≥4 (OR 12.20, 95% CI 1.23-125.00) were independent predictors of SVR. Fibrosis scores and alanine aminotransferase levels were predictive of hepatic IL-8 mRNA expression by multiple linear regression analysis (r(2)=0.204).. SVR to combination therapy in hepatitis C 1b patients was associated with down-regulated hepatic IL-8 mRNA expression and ISDR mutations. Fibrosis scores and alanine aminotransferase levels were predictive of hepatic IL-8 mRNA expression.

Topics: Aged; Antiviral Agents; Female; Gene Expression Regulation; Hepacivirus; Hepatitis C; Humans; Interferons; Interleukin-8; Liver; Male; Middle Aged; Mutation; RNA, Messenger; Treatment Outcome; Viral Nonstructural Proteins

Progressive deposition of liver fibrosis is a common feature of chronic hepatitis associated with hepatitis C virus (HCV) infection, and it may eventually lead to cirrhosis and liver failure. Although this fibrogenic process appears to be linked to HCV protein expression and replication via indirect mechanisms, i.e., to be mediated by virally-driven inflammation, a direct role of HCV in inducing fibrosis deposition has never been entirely excluded.. We established an in vitro system in which the human hepatic stellate cell line LX-2 was cultured in the presence of conditioned medium from human hepatoma Huh-7 cells transduced with a lentiviral vector expressing HCV core proteins of different genotypes.. Treatment of LX-2 cells, with conditioned medium from Huh-7 cells expressing HCV core protein, led to the activation of alpha-smooth muscle actin expression. Among the chemokines secreted by cells transduced with HCV core, interleukin-8 was identified as the strongest inducer of alpha-smooth muscle actin expression in LX-2 and primary hepatic stellate cells. This effect was accompanied by a decrease in cell migration and increased focal contact organisation.. The expression of the HCV core in hepatocytes may contribute to the establishment of a profibrogenic microenvironment.

Topics: Carcinoma, Hepatocellular; Cell Movement; Cloning, Molecular; Developing Countries; DNA Primers; DNA, Complementary; Hepacivirus; Hepatitis C; Humans; Incidence; Interleukin-8; Liver Neoplasms; Phenylurea Compounds; Polymerase Chain Reaction; Receptors, Interleukin-8A; Receptors, Interleukin-8B; RNA, Messenger; RNA, Viral; Triazoles; Tumor Cells, Cultured; Viral Core Proteins

Interferon (IFN) response rate in hepatitis C virus (HCV) patients has been varied with genotypes. In this study, we investigated the effects of HCV NS5A protein on IFN resistance and compared the genotypic differences of NS5A. We showed that IFN-α-, poly I:C-, and Sendai virus-induced ISRE transcriptional activities were inhibited by both genotype 1b and 2a NS5A protein. We demonstrated that not only genotype 1b but also genotype 2a NS5A exerted the similar extent of IFN-α-induced antiviral activity. We showed that NS5A derived from both genotype 1b and 2a showed no significant differential IFN responses as seen in HCV patients. These data imply that some other host factor may be involved in genotypic differences of IFN antagonism in HCV patients.

Topics: Active Transport, Cell Nucleus; Antiviral Agents; Cell Line; Cell Nucleus; Drug Resistance, Viral; Gene Silencing; Hepacivirus; Hepatitis C; Humans; Interferon-alpha; Interleukin-8; Phosphorylation; Poly I-C; Sendai virus; STAT1 Transcription Factor; Transcription, Genetic; Viral Nonstructural Proteins

Topics: Adipokines; Hepatitis C; Humans; Insulin Resistance; Interleukin-8; Leptin; Liver

In the majority of cases, infection with hepatitis C virus (HCV) becomes chronic and is often associated with impaired innate and adaptive immune responses. The mechanisms underlying viral persistence and lack of protective immunity are poorly understood. Considering that dendritic cells (DCs) play critical roles in initiating and modulating immune responses, we explored the effect of HCV proteins on DC gene and protein expression, phenotype, and function. Human DCs were generated following plastic adherence of monocytes and culture with granulocyte-macrophage colony-stimulating factor and interleukin-4 (IL-4) from normal subjects. Autologous nonadherent peripheral blood mononuclear cells were infected with vaccinia constructs expressing various HCV proteins (core-E1, NS5A, NS5B) or an irrelevant protein beta-galactosidase (beta-gal) as the control, induced to undergo apoptosis, then co-cultured with DCs. Between 2% and 10% of the genes probed in a cDNA nylon array were differentially regulated within DCs that had engulfed HCV proteins. In particular, the presence of intracellular NS5A led to increased transcriptional and protein expression of IL-8 (CXCL-8), a chemokine with proinflammatory and anti-interferon properties, and impaired interferon induction of signal transducers and activators of transcription 1 (STAT1) serine and tyrosine and STAT2 tyrosine phosphorylation.. These data provide novel mechanisms by which HCV subverts antiviral host immunity.

Topics: Antigen-Presenting Cells; Apoptosis; Cells, Cultured; Coculture Techniques; Dendritic Cells; Genetic Vectors; Hepatitis C; Hepatitis C Antigens; Humans; Immunophenotyping; Interferon-alpha; Interleukin-8; Monocytes; Oligonucleotide Array Sequence Analysis; Phagocytosis; Phosphorylation; Recombinant Proteins; Reverse Transcriptase Polymerase Chain Reaction; Serine; Signal Transduction; STAT1 Transcription Factor; STAT2 Transcription Factor; Tyrosine; Vaccinia virus

The chemokine CXCL-8 (interleukin-8) is induced by many viruses, including hepatitis C virus (HCV). In the current study, we examined CXCL-8 levels in the context of acute and chronic HCV replication in vitro. Two different small interfering RNAs were used to silence CXCL-8 mRNA and protein expression in Huh7 and BB7 replicon cells. HCV RNA synthesis in BB7 cells was inhibited by CXCL-8 knockdown. Furthermore, antibody neutralization of endogenous CXCL-8 activity inhibited HCV replication, while addition of recombinant human CXCL-8 stimulated NS5A protein expression. Moreover, CXCL-8 protein levels correlated positively with HCV RNA levels in four independent subgenomic and genomic replicon lines (R = 0.41, P = 0.0013). However, CXCL-8 mRNA levels correlated inversely with CXCL-8 protein and HCV RNA levels in all replicon lines and in Huh7 cells. Transient replication assays with strongly permissive and weakly permissive Huh7 cells and three independent subgenomic replicons with various replicative capacities revealed that CXCL-8 protein levels were higher in weakly than in strongly permissive cells. The JFH-1 subgenomic replicon, which replicated to high levels in both strongly and weakly permissive Huh7 cells, induced CXCL-8 protein to high levels in both cell types. The data indicate that in the replicon system, CXCL-8 protein levels are positively associated with chronic HCV replication and that CXCL-8 removal inhibits HCV replication. During acute HCV replication, CXCL-8 production may be inhibitory to viruses with low replicative capacity. The data underscore the complex regulation of CXCL-8 mRNA and protein expression and further suggest that in addition to contributing to HCV pathology via proinflammatory actions, CXCL-8 may have opposing antiviral and proviral effects depending on the level of HCV replication, the cellular context, and whether the infection is acute or chronic.

Topics: Cell Line, Tumor; Hepacivirus; Hepatitis C; Humans; Interleukin-8; Liver; Replicon; RNA, Small Interfering; RNA, Viral; Virus Replication

Chronic hepatitis C carries the risk to develop mixed cryoglobulinemia (MC) and B-cell non-Hodgkin's lymphoma (B-NHL), possibly because viral antigens stimulate the host's inflammatory response via extracellular pattern recognition receptors (PRR). To clarify this issue, we studied whether recognition of hepatitis C virus (HCV) proteins by PRR is involved in the pathogenesis of HCV-associated MC or B-NHL.. Peripheral blood mononuclear cells of patients with HCV-associated B-NHL (n = 12), MC (n = 14), uncomplicated hepatitis C (n = 12), and healthy volunteers (n = 12) were incubated with the recombinant HCV proteins E2, core, and NS3 to study induction of cytokine production, stimulation of B-cell proliferation, and immunoglobulin secretion. In addition, serum levels of interleukin-6 (IL-6) were measured by ELISA.. HCV core was the only studied protein, which induced production of IL-6 and IL-8 in CD14(+) cells. IL-6 induction was mediated via Toll-like receptor 2 (TLR2) and lead to increased B-cell proliferation in vitro. TLR2 expression on monocytes and IL-6 serum concentrations were increased in all groups of HCV-infected patients compared with healthy controls and were highest in MC (P < 0.05).. Increased secretion of IL-6 via stimulation of TLR2 by HCV core protein may play a role in the pathogenesis of hepatitis C-associated MC and B-NHL.

Topics: Adult; Aged; Aged, 80 and over; Cell Proliferation; Cryoglobulinemia; Female; Hepacivirus; Hepatitis C; Humans; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Lymphoma, B-Cell; Lymphoma, Non-Hodgkin; Male; Middle Aged; Recombinant Proteins; Toll-Like Receptor 2; Up-Regulation; Viral Core Proteins

We evaluated the association between variations in hepatitis C virus (HCV) core protein and hepatitis severity in patients with chronic HCV infection who achieved remission without viral eradication and had a biochemical response to interferon (IFN) therapy, to evaluate the effect of HCV core sequence in the absence of the influence of host factors. METHODS. Using serum from 10 patients with a biochemical response and 10 patients with no response, we measured serum levels of interleukin (IL)-1 beta , IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IFN- gamma , and tumor necrosis factor- alpha before and after IFN therapy. Expression vectors with the core region were transfected into Huh7 cells, and cytokine induction was evaluated by reporter assay.. In biochemical responders, only IL-8 levels decreased after IFN therapy (P=.04). Changes in the C-terminal hydrophobic region were observed more frequently in biochemical responders. Activation of the IL-8 promoter by HCV core protein was significantly decreased in biochemical responders after IFN therapy (P=.04). When 69 C-terminal amino acids from before IFN therapy were replaced with those from after IFN therapy in 3 biochemical responders, their ability to transactivate IL-8 decreased.. Differences in amino acids in the HCV core protein correlates with hepatitis activity through the modulation of IL-8 induction in HCV-infected patients.

Topics: Adult; Amino Acid Sequence; Female; Genes, Reporter; Genetic Variation; Hepacivirus; Hepatitis C; Hepatitis C Antigens; Humans; Interferons; Interleukin-8; Male; Middle Aged; Molecular Sequence Data; Promoter Regions, Genetic; Transcriptional Activation; Treatment Outcome; Viral Core Proteins

In order to examine the immunopathogenesis of hepatitis C virus (HCV)-related liver injury in renal-transplant patients, intra-hepatic cytokine profiles were examined in 38 liver biopsies from 38 patients by measuring messenger RNA (mRNA) concentrations by a real-time PCR method of a Th1 cytokine (i.e., interferon (IFN)-gamma), a Th2 cytokines (i.e., interleukine (IL)-10), a proinflammatory cytokine (i.e., IL-8), and a potent fibrogenic factor (transforming growth factor [TGF]-beta). There was no significant difference in TGF-beta, IFN-gamma, IL-10, or IL-8 levels of expression according to liver-activity grade, liver-fibrosis stage, the concentration of HCV RNA at liver biopsies, or the HCV genotype. However, IFN-gamma/beta-actin mRNA concentration was higher than the IL-10/beta-actin mRNA concentration in patients with F3 Metavir score. Median IFN-gamma/beta-actin mRNA concentration tended to be higher in patients with A3 and A4 Metavir activity grades compared with those with A0 and A1 activity grades. There was a significant correlation between the duration of HCV infection and both TGF-beta/beta-actin (r(2)=0.19, P=0.04) and IL-8/beta-actin mRNA concentrations (r(2)=0.19, P=0.03). IFN-gamma/beta-actin mRNA concentration also increased according to the duration of HCV (r(2)=0.19, P=0.07). Finally, there was a significant correlation between the duration of HCV infection and liver fibrosis stage (r(2)=0.17, P=0.045). Intrahepatic Th1 cytokine profile seems to be predominant in patients with extensive fibrosis and activity scores, suggesting that it might be responsible for liver injury in renal transplant patients.

Topics: Adult; Aged; Cytokines; Female; Hepacivirus; Hepatitis C; Humans; Interferon-gamma; Interleukin-10; Interleukin-8; Kidney Transplantation; Liver; Liver Cirrhosis; Male; Middle Aged; RNA, Messenger; RNA, Viral; Time Factors; Transforming Growth Factor beta

Hepatitis C virus (HCV) has been reported to replicate in monocytes/macrophages in infected patients. However, it is unclear whether macrophages are susceptible to infection in vitro and whether such an infection is consequential. Sera from 26 HCV-infected patients were incubated with primary human macrophages collected from healthy donors. Virus negative strand was detected by a Tth enzyme-based strand-specific assay and virus sequences were analysed by single strand conformation polymorphism (SSCP) and sequencing. Concentrations of the cytokines tumour necrosis factor-alpha (TNF-alpha) and interleukin (IL)-1beta, IL-6, IL-8, IL-10 and IL-12p70 were measured in culture supernatants and respective mRNAs were analysed in cell extracts by quantitative RT-PCR. For 15 sera, HCV RNA was detectable in 2- and 3-week cultures from at least one donor. Virus negative strand was detected in 29 % of macrophage samples in this group. In four cases, HCV RNA sequences amplified from macrophages differed from those amplified from sera suggesting evolution during infection. Concentrations of TNF-alpha and IL-8 were found to be significantly higher in supernatants from HCV-infected cultures. In conclusion, these preliminary data suggest that primary human macrophages are susceptible to HCV infection in vitro and this infection is associated with the induction of cytokines TNF-alpha and IL-8.

Topics: Cells, Cultured; Culture Media, Conditioned; Cytokines; Hepacivirus; Hepatitis C; Humans; Interleukin-8; Macrophages; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Viral; Tumor Necrosis Factor-alpha

Topics: Hepatitis B; Hepatitis C; Humans; Interleukin-1; Interleukin-2; Interleukin-8; Lymphocytes; Military Personnel; Russia

Hepatitis C virus (HCV), a major cause of liver disease worldwide, is frequently resistant to the antiviral alpha interferon (IFN). We have recently found that the HCV NS5A protein induces expression of the proinflammatory chemokine IL-8 to partially inhibit the antiviral actions of IFN in vitro. To extend these observations, in the present study we examined the relationship between levels of IL-8 in serum, HCV infection, and biochemical response to IFN therapy. Levels of IL-8 were significantly elevated in 132 HCV-infected patients compared to levels in 32 normal healthy subjects and were also significantly higher in patients who did not respond to IFN therapy than in patients who did respond to therapy. This study suggests that HCV-induced changes in levels of chemokine and cytokine expression may be involved in HCV antiviral resistance, persistence, and pathogenesis.

Topics: Drug Resistance, Microbial; Hepatitis C; Humans; Interferon-alpha; Interleukin-8; Tumor Necrosis Factor-alpha

To investigate the cellular immunological changes in children with viral hepatitis, interleukin 6 (IL-6), interleukin 8 (IL-8), interferon alpha (IFN alpha), and tumor necrosis factor alpha (TNF alpha) in supernatant of cultured peripheral blood mononuclear cells (PBMCs) of 49 children with hepatitis A, B or C were measured. The levels of IL-6, IL-8, TNF alpha in PBMCs of the 3 viral hepatitis groups were increased and the level of IFN alpha decreased as compared with those of normal control group. But there were no significant differences among the 3 viral hepatitis groups. It was concluded that cellular immunological disorders were related to the onset and the induced damage of the viral hepatitis in children.

Topics: Adolescent; Child; Child, Preschool; Female; Hepatitis A; Hepatitis B, Chronic; Hepatitis C; Hepatitis, Viral, Human; Humans; Infant; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Male; Tumor Necrosis Factor-alpha

The increased frequency of autoantibodies and B cell non-Hodgkins lymphoma (B-NHL) in hepatitis C virus (HCV) infection suggests dysregulated humoral immunity. Soluble CD23 (sCD23) is involved in B cell activation and proliferation and the serum levels are raised in autoimmune diseases and B cell lymphoproliferative disease. We compared the serum levels of sCD23 in patients with HCV infection with those in patients with alcoholic cirrhosis (AC) and in healthy controls. Serum levels of interleukin (IL) 8, IL10, granulocyte macrophage-colony stimulating factor, and interferon-gamma were assessed simultaneously to check for generalized nonspecific immune stimulation. In contrast to the essentially normal serum levels of these latter cytokines, the levels of sCD23 were raised in the patients with HCV compared to those with AC and the normal controls (medians 34.0, 10.1, and 11.1 arbitrary units, respectively; HCV vs AC P < 0.0004, HCV vs controls P < 0.0001, AC vs controls P > 0.8). These results confirm HCV-induced humoral immune dysregulation and invite comparison with primary Sjögrens syndrome and Epstein-Barr virus infection, both of which are also associated with raised levels of serum sCD23, autoantibodies, and B-NHL.

Topics: Adult; Alanine Transaminase; Aspartate Aminotransferases; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Hepatitis C; Humans; Interferon-gamma; Interleukin-10; Interleukin-8; Liver Cirrhosis, Alcoholic; Male; Middle Aged; Receptors, IgE

Topics: Actuarial Analysis; Cyclosporine; Cytokines; Drug Therapy, Combination; Extracellular Matrix Proteins; Fibronectins; Gene Expression Regulation; Glutathione Transferase; Graft Rejection; Hepatitis C; Humans; Hyaluronic Acid; Immunosuppressive Agents; Interferon-gamma; Interleukin-6; Interleukin-8; Laminin; Liver Transplantation; Prednisolone; Recurrence; Retrospective Studies; Survival Rate; Tacrolimus; Time Factors

The present study investigated plasma levels of interleukin-8 (IL-8) in patients with post-hepatitic cirrhosis and correlated it with the severity of liver diseases and haemodynamic parameters. Plasma IL-8 levels were significantly higher in 57 post-hepatitic cirrhotic patients (7.5 +/- 1.8 pg/mL; P < 0.005) than those in 41 healthy subjects (2.0 +/- 0.2 pg/mL). Elevated (> 5 pg/mL) plasma IL-8 levels were found in up to 30% of cirrhotic patients. In cirrhotic patients, plasma IL-8 levels progressively increased in relation to the severity of liver dysfunction (4.5 +/- 1.0, 4.9 +/- 1.4 and 20.5 +/- 8.3 pg/mL for Pugh's class A, B and C, respectively; P < 0.005). A significant correlation was observed between plasma IL-8 levels and serum bilirubin levels (r = 0.72; P < 0.001). There were no differences in the hepatic venous pressure gradient (15.4 +/- 1.1 vs 15.1 +/- 0.9 mmHg; P > 0.05) and systemic vascular resistance (1119 +/- 118 vs 1199 +/- 54 dyn.s/cm5; P > 0.05) between cirrhotic patients with and without elevated plasma IL-8 levels. In addition, plasma IL-8 levels did not correlate with hepatic venous pressure gradient (r = 0.26; P > 0.05) and systemic vascular resistance (r = -0.24; P > 0.05). These results demonstrate that plasma IL-8 levels are increased in patients with post-hepatitic cirrhosis. The severity of liver cirrhosis is an important factor for the occurrence of enhanced IL-8 levels. IL-8 does not play a role in the hyperdynamic circulation observed in patients with post-hepatitic cirrhosis.

Topics: Bilirubin; Female; Hepatitis B; Hepatitis C; Humans; Hypertension, Portal; Interleukin-8; Liver; Liver Circulation; Liver Cirrhosis; Male; Middle Aged; Vascular Resistance; Venous Pressure

Cytotoxic T lymphocytes (CTL) are important to the control of viral replication and their presence may be important to disease outcome. An understanding of the spectrum of proteins recognized by hepatitis C virus (HCV)-specific CTL and the functional properties of these cells is an important step in understanding the disease process and the mechanisms of persistent infection, which occurs in the majority of HCV-infected individuals. In this report we identify HCV-specific CTL responses restricted by the HLA class I molecules A2, A3, A11, A23, B7, B8, and B53. The epitopes recognized by these intrahepatic CTL conform to published motifs for binding to HLA class I molecules, although in some cases we have identified CTL epitopes for which no published motif exists. The use of vectors expressing two different strains of HCV, HCV-1 and HCV-H, revealed both strain-specific and cross-reactive CTL. These HCV-specific CTL were shown to produce cytokines including IFN-gamma, TNF-alpha, GM-CSF, IL-8, and IL-10 in an antigen- and HLA class I-specific manner. These studies indicate that the CTL response to HCV is broadly directed and that as many as five different epitopes may be targeted in a single individual. The identification of minimal epitopes may facilitate peptide-specific immunization strategies. In addition, the release of proinflammatory cytokines by these cells may contribute to the pathogenesis of HCV-induced liver damage.

Topics: Amino Acid Sequence; Cytokines; Epitope Mapping; Granulocyte-Macrophage Colony-Stimulating Factor; Hepacivirus; Hepatitis C; Histocompatibility Antigens Class I; Humans; Interleukin-10; Interleukin-8; Molecular Sequence Data; T-Lymphocytes, Cytotoxic; Viral Vaccines

Interleukin-8 (IL-8) and granulocyte-macrophage colony-stimulating factor (GM-CSF) are important mediators of inflammation and immune response in human disease. To demonstrate their importance in pathophysiological processes in liver disease, we measured the circulating levels of IL-8 and GM-CSF in patients with hepatocellular carcinoma (HCC) and chronic active hepatitis (CAH). IL-8 and GM-CSF levels in serum samples were determined with highly specific and sensitive enzyme-linked immunosorbent assays. IL-8 levels were more elevated in serum samples of patients with HCC and CAH associated with hepatitis C virus infection than HCC and CAH associated with hepatitis B virus infection. However, in all patients with autoimmune CAH and in some patients with HCC and CAH, GM-CSF levels were elevated over the baseline levels measured in all of the normals, but this difference was not statistically significant for any group. We conclude that IL-8 and GM-CSF are increased in some patients with liver diseases, and as such they may play a significant role in host defense and disease.

Topics: Carcinoma, Hepatocellular; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Hepatitis B; Hepatitis C; Hepatitis, Chronic; Humans; Interleukin-8; Liver Neoplasms; Male; Middle Aged; Reference Values