To evaluate the clinical feasibility of the antibody titer against a chimeric polypeptide (named Core 518), in which a domain of Core and NS3 of hepatitis C virus (HCV) was fused, ELISA was performed in a total of 76 serum samples. Each serum was serially diluted using two-fold dilution method with distilled water into 10 concentrations. They were all positive for second generation anti-HCV assay (HCV EIA II; Abbott Laboratories). Genotyping RT-PCR, quantitative competitive RT-PCR, and RIBA (Lucky Confirm; LG Biotech) were also assayed. Anti-Core 518 antibody was detected in x 12800 or higher dilutions of sera from 35 of 43 chronic hepatitis C (81.4%) and nine of 16 hepatocellular carcinoma sera (56.3%), one of four cirrhosis (25%), 0 of four acute hepatitis C, and one of nine healthy isolated anti-HCV-positive subjects (p=0.0000). The anti-Core 518 antibody titers were well correlated with the presence of HCV RNA in serum (p=0.002). The anti-Core 518 antibody titers decreased significantly in nine of ten responders to IFN-alpha treatment. Monitoring anti-Core 518 titers may be helpful not only for differentiating the status of HCV infection among patients with various type C viral liver diseases, but also for predicting responses to IFN-alpha treatment.
Adult ; Aged ; Female ; Genotype ; Hepatitis C/immunology* ; Hepatitis C/drug therapy* ; Hepatitis C/diagnosis ; Hepatitis C/blood ; Hepatitis C Antibodies/immunology* ; Hepatitis C Antibodies/blood ; Hepatitis C Antigens/immunology* ; Hepatitis C-Like Viruses/immunology* ; Hepatitis C-Like Viruses/genetics ; Human ; Immunoblotting ; Interferon Alfa-2a/therapeutic use* ; Male ; Middle Age ; RNA, Viral/blood ; Recombinant Fusion Proteins/immunology ; Viral Core Proteins/immunology* ; Viral Nonstructural Proteins/immunology*

Genetic changes between codons 2209 and 2248 of NS5A of genotype 1b hepatitis C virus (HCV-1b) have been reported to be associated with the sensitivity to interferon-alpha (IFN-alpha). The present study was performed to analyze such relationship in Korean patients with chronic hepatitis C and HCV-1b (n=19), including 12 chronic hepatitis C patients treated with IFN-alpha, 3 chronic hepatitis C patients without treatment as controls, and 4 patients with hepatocellular carcinoma (HCC). Two serum samples, before and after the treatment, were analyzed for the mutations by reverse transcription-polymerase chain reaction, cloning and sequencing. The mutations were identified in 32% (6/19), including five intermediate type (1-3 mutations) and one mutant type (4 or more). In 12 patients treated with IFN-alpha, the number of amino acid substitutions in NS5A2209-2248 was not associated with outcome of the treatment.
Adult ; Aged ; Amino Acid Sequence ; Antiviral Agents/therapeutic use* ; Base Sequence ; Carcinoma, Hepatocellular/virology* ; Carcinoma, Hepatocellular/blood ; Codon ; Female ; Genotype ; Hepatitis C, Chronic/virology* ; Hepatitis C, Chronic/drug therapy* ; Hepatitis C, Chronic/blood ; Hepatitis C-Like Viruses/isolation & purification ; Hepatitis C-Like Viruses/genetics* ; Hepatitis C-Like Viruses/classification ; Human ; Interferon-alpha/therapeutic use* ; Liver Neoplasms/virology* ; Liver Neoplasms/blood ; Male ; Middle Age ; Molecular Sequence Data ; Mutation* ; Prognosis ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUND: Although the polyproteins of hepatitis C virus(HCV) are processed and formed in nearly equimolar amounts, individual functional proteins have a discrepancy in their time of appearance following HCV infection and eliciting immune response. This study was conducted to compare the reactivity toward regional specific HCV protein in relation to virological characteristics, including HCV genotype and HCV replication. METHODS: Sera from forty-five patients with chronic HCV infection were analyzed through the experiments of the recombinant immunoblot assay(RIBA-2), HCV genotyping and HCV RNA quantitation. RESULTS: The frequencies of seropositivity to C22-3, C33C, C100-3 and 5-1-1 proteins were 91.1+ACU-, 91.1+ACU-, 64.4+ACU- and 53.3+ACU-, respectively, of all the patients, and thus the antibodies to C22-3 and C33C proteins were found more frequently (p +ADw- 0.05). The antibody responses between core or NS3 proteins and NS4 proteins showed more discrepancy in the HCC group than that in the CH group, implying a possibility of oncogenic potential of core or NS3 gene in hepatocarcinogenesis. The detection rate of antibodies to C22-3 and C33C, in accordance with serum HCV RNA levels, was significantly higher in highly viremic patients than that in low viremic patients (p +ADw- 0.05). Antibodies to C22-3, C33C, C100-3 and 5-1-1 were also found more frequently in patients with HCV genotype 1b, compared to those with HCV genotype 2a (p +ADw- 0.05). CONCLUSION: These results suggest that antibody detection of HCV may depend on the virological characteristics of HCV, the levels of HCV replication and HCV genotype and, therefore, HCV RNA detection using RT-PCR technique is essential for confirmatory diagnosis for HCV infection. Furthermore, the HCV core or NS3 Protein may play important role in hepatocarcinogenesis.
Adult ; Aged ; Female ; Genotype ; Hepatitis C Antibodies/blood+ACo- ; Hepatitis C, Chronic/virology ; Hepatitis C, Chronic/immunology+ACo- ; Hepatitis C-Like Viruses/physiology ; Hepatitis C-Like Viruses/genetics ; Human ; Male ; Middle Age ; RNA, Viral/blood ; Viral Core Proteins/immunology+ACo- ; Viral Nonstructural Proteins/immunology+ACo- ; Virus Replication

BACKGROUND: Transforming growth factor beta-1 (TGF beta 1) has been suggested to play a role in the development, growth or progression of hepatocellular carcinoma (HCC). Genotype and serum titer of HCV also affect the occurrence of HCC in chronic hepatitis C. In this study, we were to evaluate the effects of genotype or serum titer of HCV on the expression of TGF beta 1. We also intended to examine the correlation between the up-regulation of TGF beta 1 and the association with HCC in patients with chronic hepatitis C. METHODS: We studied 19 patients with chronic hepatitis C and 18 with HCC associated with HCV infection. HCV genotype was determined by line probe reverse hybridization assay and the amount of HCV-RNA was quantitated by branched DNA signal amplification assay. Serum TGF beta 1 level was measured by enzyme linked immunosorbent assay. RESULTS: HCV genotypes of patients with HCC were similar to those without it. Serum HCV-RNA titer was higher in genotype 1b than in non-1b (p < 0.05). Serum TGF beta 1 levels were higher in HCC than in chronic hepatitis (p < 0.05). However, there was no significant difference in the serum TGF beta 1 level between genotype 1b and non-1b. Also, it was not correlated with the serum HCV-RNA titer or alanine aminotransferase levels. CONCLUSION: TGF beta 1 seems to be overexpressed in HCC compared to that of chronic hepatitis C: it was not affected by serum ALT levels, genotype or serum HCV titer. It is suggested that TGF beta 1 may be associated with the malignant transformation of hepatocyte or the progression of HCV-associated HCC.
Adult ; Aged ; Alanine Transaminase/blood ; Carcinoma, Hepatocellular/virology ; Carcinoma, Hepatocellular/metabolism* ; Female ; Genotype ; Hepatitis C, Chronic/virology ; Hepatitis C, Chronic/metabolism* ; Hepatitis C-Like Viruses/genetics ; Hepatitis C-Like Viruses/classification ; Human ; Liver Neoplasms/virology ; Liver Neoplasms/metabolism* ; Male ; Middle Age ; RNA, Viral/blood ; Transforming Growth Factor beta/blood*

To determine the prevalence and clinical relevance of HGV infection in dialysis patients, we performed a cross-sectional study of 61 HD patients and 79 Continuous Ambulatory Peritoneal Dialysis (CAPD) patients. HGV-RNA was identified by reverse-transcription (RT) polymerase chain reaction (PCR) assay with primers from the 5'-untranslated region of the viral genome. The prevalence of HGV infection was similar in HD and CAPD patients (9.8% vs. 12.7%), while that of HCV infection was significantly higher in HD patients compared to CAPD patients (16.4% vs. 1.3%, p < 0.05). The mean age (49.2 +/- 13.4 vs. 46.7 +/- 13.0 years), male to female ratio (2.4:1 vs. 1.3:1), history of transfusion (62.3% vs. 49.4%), history of hepatitis (27.9% vs. 26.6%), mean ALT level during the previous 6 months (22.4 +/- 37.9 vs. 14.0 +/- 7.4 IU/L), and the prevalence of HBsAg (8.2% vs. 6.3%) showed no difference between HD and CAPD patients. In both HD and CAPD patients, the presence of HGV RNA was not related to age, sex, duration of dialysis, history of transfusion, history of hepatitis, or to the presence of HBV or HCV markers. There was no significant difference in the clinical and biochemical data between patients with isolated HGV infection (n = 12) and patients without viremia (n = 106). The clinical feature of patients coinfected with HGV and HBV (n = 2), or HGV and HCV (n = 2) seemed to be similar to those of patients with isolated HBV (n = 8) or HCV (n = 9) infection. In conclusion, the prevalence of HGV infection was not different between HD and CAPD patients, and HGV infections did not seem to be associated with clinically significant hepatitis. The routes of HGV transmission, other than transfusion or contamination during HD procedure, were suspected.
Female ; Hepatitis Agents, GB*/genetics ; Hepatitis C/genetics ; Hepatitis C-Like Viruses/genetics ; Hepatitis, Viral, Human/virology ; Hepatitis, Viral, Human/genetics ; Hepatitis, Viral, Human/etiology* ; Human ; Male ; Middle Age ; Peritoneal Dialysis, Continuous Ambulatory/adverse effects* ; Prevalence ; RNA, Viral/analysis ; Renal Dialysis/adverse effects* ; Viremia/genetics

Interferon-alpha (IFN-alpha) has been used to treat hepatitis C virus (HCV)-induced hepatitis, but it has been effective in only about half of the treated patients, with recurrence appearing in the other half. As a consequence of the possible complications associated with IFN-alpha and the high cost of treatment, it has become extremely important to select the proper patients for IFN-alpha treatment. In our previous study, we found that the quasispecies in the hypervariable region (HVR) 1 of HCV were various and that a new quasispecies can appear in non-responders and/or lead to deterioration in the patients' condition. The preliminary data we obtained in the process of our previous research led us to believe that the quasispecies of HVR 1 has something to do with the effect of IFN-alpha. Thus, in this investigation, we tried to determine the predictive factors of IFN-alpha therapy. Thirty patients with HCV infection were treated with IFN-alpha. Among them, 15 patients recovered after six months IFN-alpha treatment, but the remaining 15 patients showed no response after six months IFN-alpha treatment. We cloned HVR 1 DNA by reverse transcription-polymerase chain reaction (RT-PCR) and examined the quasispecies of HVR 1. As the quasispecies of HVR 1 in non-responders varied more than in the complete remission group, we concluded that the sequence variation in HVR 1 of HCV can be used to predict the effect of IFN-alpha.
Adult ; Aged ; Amino Acid Sequence ; Female ; Genotype ; Hepatitis C/virology ; Hepatitis C/drug therapy* ; Hepatitis C-Like Viruses/classification* ; Human ; Interferon-alpha/therapeutic use* ; Male ; Middle Age ; Molecular Sequence Data ; Viral Nonstructural Proteins/genetics ; Viral Nonstructural Proteins/chemistry* ; Viral Nonstructural Proteins/blood