Liver Hepatitis C Virus RNA Research Articles

Role of Therapeutic Drug Monitoring in a Patient With Human Immunodeficiency Virus Infection and End-Stage Liver Disease Undergoing Orthotopic Liver Transplantation

Pharmacological interactions between protease inhibitors and tacrolimus require careful monitoring to prevent toxicity in the posttransplantation period. A 42-year-old man with human immunodeficiency virus (HIV) infection and end-stage liver disease due to hepatitis C virus (HCV) received an orthotopic liver transplant. At the time of surgery the patient was on triple antiretroviral therapy (tenofovir, lamivudine, and lopinavir/ritonavir) with a stable CD4 + count (>500 cells/mm 3) and HIV-1 RNA (<50 copies/mL). Immunosuppression was maintained with tacrolimus (0.5 mg at a single dose once per week). One month after surgery HCV recurrence was documented. Pharmacokinetic evaluation of lopinavir/ritonavir showed a rapid increase in the area under the curve. Drug concentrations returned to normal levels, with reduction in liver enzymes. At the same time, tacrolimus dosages were reduced to a maintenance dose of 0.5 mg every 2 weeks. The patient, at 17 months postoperatively, is alive in good health with normal liver function and HCV RNA load levels. This is the first case in which a profound change in the pharmacokinetics of a protease inhibitor caused by a drug-drug interaction was observed during transient liver damage. Because this clinical event is particularly common in HIV-infected patients, our findings suggest that therapeutic drug monitoring should be performed to determine the impact of potential drug interactions in the early posttransplantation period, at the time of resumption of therapy or introduction of new anti-retroviral therapy and during HCV recurrence in order to optimize both tacrolimus and protease inhibitor treatment.

Lack of Evidence for Ribavirin Monotherapy Efficacy on Liver Fibrosis in Hepatitis C Virus Positive Renal Transplant Patients

To date there is still no efficient and safe treatment for chronic hepatitis C virus (HCV) infection after renal transplantation. Recently, Fontaine et al. (1) reported on the treatment of 13 renal transplant (RT) patients with ribavirin monotherapy. They found that ribavirin monotherapy was associated with a significant decrease in serum alanine aminotransferase levels without any effect upon either HCV RNA viremia or quantitative intrahepatic HCV RNA viral load. They also suggested that ribavirin monotherapy improved liver histopathology. We have previously observed similar findings with respect to liver enzyme levels and HCV RNA viremia in HCV-positive RT patients treated by ribavirin monotherapy (2). However, after 12 months of ribavirin therapy, our patients experienced a significant worsening in liver fibrosis; the mean time between the two liver biopsies (LB) was 17 (11–27) months. Conversely, in the study by Fontaine et al. (1), although they found a significant decrease in the mean activity score, there was no significant change in the grade of liver fibrosis (P= 0.1). The absence of a significant change in liver fibrosis grade might have been related to the fact that 5 of their 13 patients had already had cirrhosis before treatment and, consequently, had no potential to deteriorate their liver fibrosis grade further. The time between the two LBs was as long as 5.7 ± 9.3 years. Hence, the actual liver activity and fibrosis scores just before treatment are unknown and, consequently, we might wonder whether the modifications in liver histology observed in their patients were related to ribavirin therapy and/or to the natural history of HCV infection after renal transplantation. We, and others, have recently reported that liver fibrosis might regress in untreated HCV-positive RT patients despite the persistence of a high HCV RNA viral load (3, 4). Finally, in the study of Fontaine et al. (1), there was no control group; thus, the authors compared the yearly liver fibrosis progression rate of patients treated by ribavirin with the data obtained previously from a cohort of 28 RT patients who had undergone two consecutive LBs (5). In the study by Zylbergberg et al. (5), the yearly liver fibrosis progression rate was abnormally high (i.e., 0.26 ± 0.35) compared with the observations of Alric et al. (6) in HCV RNA-positive RT patients (i.e., 0.067 [−0.05–0.18]). In this prospective controlled study, we observed a slow progression of liver fibrosis in patients with early moderate liver disease. More recently, Fehr et al. (7) found no cases of cirrhosis amongst 23 RT patients infected with HCV for more than 15 years. The discrepancy between the results observed by Zylbergberg et al. (5) and the two other studies, which were not quoted in the paper of Fontaine et al. (1), might be related to the use of different immunosuppressive treatments because almost none of their patients were on cyclosporine-A-based immunosuppression. Conversely, in the studies of Alric et al. (6) and Fehr et al. (7), nearly all the patients were receiving cyclosporine A, which is known to inhibit HCV replication in vitro. Hence, to date, there is still no evidence for a beneficial effect of ribavirin monotherapy upon liver histology in RT patients. Further randomized studies in larger cohorts of renal transplant patients, including the analysis of intrahepatic cytokine profiles before and during ribavirin monotherapy, are required to evaluate the impact of ribavirin upon liver histology. Nassim Kamar Department of Nephrology, Dialysis and Multiorgan Transplantation CHU Rangueil Department of Virology CHU Purpan Toulouse, France Jacques Izopet Department of Virology CHU Purpan Toulouse, France Laurent Alric Department of Internal Medicine CHU Purpan Toulouse, France Lionel Rostaing Department of Nephrology, Dialysis and Multiorgan Transplantation CHU Rangueil Toulouse, France

A Novel Immunocompetent Rat Model of HCV Infection and Hepatitis

Hepatitis C virus (HCV) infects millions of people worldwide. Therapy is limited, and treatment does not produce a sustained response in the majority of patients. Development of new agents has been hampered by the lack of a convenient animal model. The aim of this study was to determine whether an immunocompetent rat, tolerized and transplanted with a human hepatoma cell line (Huh 7 cells), could be used to sustain an HCV infection. Fetal rats were tolerized in utero with 10(5) Huh 7 cells. One day after birth, rats were transplanted with 5 x 10(6) Huh 7 cells and, a week later, inoculated with HCV, genotype 1. In tolerized, transplanted, and HCV-infected rats, Huh 7 cells were found in the liver, and HCV viral replication was detected by the presence of negative strand HCV RNA. HCV levels in serum were measured at 11,000 copies/mL at week 4, peaked at 22,500 copies/mL by week 12. In tolerized, transplanted, inoculated rats, but not controls, serum alanine aminotransferase (ALT) values increased to 60 IU/L by week 4 and reached a peak of approximately 120 IU/L by week 13. Histology showed foci of mononuclear infiltrates in portal and central regions. HCV-inoculated immunocompetent rats tolerized and transplanted with Huh 7 cells support HCV gene expression, viral replication, and develop biochemical and histologic evidence of hepatitis.

Role of Liver Biopsy in the Evaluation of Hepatitis C Virus Infection in HIV Coinfection

Hepatitis C virus (HCV) coinfection is common in patients infected with human immunodeficiency virus (HIV), because the viruses share common routes of transmission. With the marked increase in life expectancy of HIV-infected patients associated with the use of highly active antiretroviral therapy, HCV infection has become a significant cause of morbidity and mortality in coinfected patients. As a result, there has been increasing attention to adequate assessment of HCV infection during the last several years. Unlike liver enzymes and HCV RNA levels, which can fluctuate widely and do not correlate with the severity of disease, liver biopsy has become the cornerstone in the evaluation of chronic HCV infection. However, there remain important questions and controversies related to adequately determining the histological severity of liver disease and the role of liver biopsy in HIV-HCV-coinfected patients.

Early Hepatitis C Viral Response (EVR) to Peginterferon Alfa 2a and Ribavirin in Patients with β Thalassemia.

Hepatitis C virus (HCV) infection is an important cause of liver failure and death in patients with thalassemia who received frequent blood transfusions before HCV screening became routine. Severe transfusion-related iron overload may accelerate the progression of liver disease, influence response to therapy, and increase the risk of antiviral treatment in these patients. An open-label, single arm trial of peginterferon alfa 2a (180 ug weekly) and ribavirin (1200 mg/day for > 75 kg, 1000 mg/day 50 to 75 kg, 800 mg for < 50kg) was initiated in August, 2003 in the NIH-sponsored Thalassemia Clinical Trials Network to examine safety and efficacy in adult patients. Although the study is still enrolling patients, we are presenting viral response data for the first 16 patients (mean age: 33 yrs, range 22 to 44, 50% female, 75% genotype 1). Iron burden was assessed by chemical analysis of liver biopsy and HCV RNA levels were measured using the Roche COBAS HCV Tests (qualitative and quantitative). These analyses were performed by a central laboratory. HCV levels were measured at baseline, weeks 1, 2, 3, 4, 12 and 24.Early Virologic ResponsesWeek 1Week 2Week 3Week 4Week 12Week 24N*16161616116Non-d**122375>2-log567731% >2 log38%50%56%63%91%100%*number with result available; **non-detectable; ***greater than 2 log decreaseFour patients discontinued between weeks 4 through 12 (1 death, 1 persistent cough, 1 extreme fatigue, and 1 pregnancy (spouse). These patients had all achieved > 2 log drop by week 4. The mean liver iron at baseline was 10.1 mg/gm dry wt (range: 1.1 to 21.2) and the mean log HCV RNA level was 5.8 (range 4.0 to 6.8). The median time to > 2 log decrease was 1 week for patients with genotype 2 and 4 weeks for patients with genotype 1 (p=0.004). There was no correlation either between baseline iron or baseline HCV level and time to > 2 log decrease. Transfusion requirement increased 52% as assessed at 12 weeks and did not predict earlier viral response. In summary, there is EVR (>2 log drop) in 63% at 4 weeks and 91 % at 12 weeks, comparable to those seen in patients without thalassemia. Quantitative hepatic iron concentration at baseline was not a predictor of EVR. Completion of the study is planned to determine whether the EVR predicts sustained viral response and whether the iron overload or increased transfusion requirements during therapy pose significant safety issues.

EVOLUTION OF THE HYPERVARIABLE REGION-1 OF HEPATITIS C VIRUS AND LIVER FIBROSIS PROGRESSION IN RENAL-TRANSPLANT RECIPIENTS

P793 Aim: The aim of our study was to assess hepatitis C virus (HCV) evolution and liver histology outcome in HCV(+)/RNA(+) renal-transplant (RT) patients. Methods: A total of 55 patients (34 men, 21 women, median age 38 years) underwent serial liver biopsies (LB) after transplantation (2 LB, n=11; 3 LB, n=35; 4 LB, n=9). The median times between transplantation and the first LB was 30 (9−145) months. The median time between two consecutive biopsies was 37 months. Results: Overall, the rate of liver fibrosis progression per year, defined as the ratio of the difference in fibrosis stage between two consecutive LBs and the time between these two biopsies, was 0.07 ± 0.03 Metavir units/year. We identified three groups of patients: those in whom liver fibrosis remained stable (group I, n=21), those with progressing liver fibrosis (group II, n=21, 0.34 ± 0.05 units/year), and those with a regression in liver fibrosis (group III, n=13, −0.25 ± 0.24 units/year). In the last two groups, the progression and the regression of liver fibrosis were progressive during follow-up. Demographic data, immunosuppressive treatment, the duration of HCV infection, HCV genotype, as well as liver enzyme (AST and ALT) levels, renal parameters, and HCV RNA concentrations before transplantation and at each LB, were similar in all groups. After transplantation, ALT only increased significantly in group II. In contrast, AST and HCV viremia increased significantly in all groups. At the third LB, ferritin levels and hepatosiderosis were significantly higher in group II compared with the other groups. Overall, liver activity scores were similar in the three groups. Sequence analysis of the hypervariable region-1 of the HCV genome revealed a significantly lower diversification between transplantation and the first biopsy in patients with stable or progressing liver fibrosis compared with those with regressing liver fibrosis. Conclusion: In conclusion, after renal transplantation, HCV infection is not harmful upon liver fibrosis in all patients. Sequencing of the hypervariable region-1 of HCV might predict the long-term liver histology outcome.

Lack of in vivo blockade of Fas- and TNFR1-mediated hepatocyte apoptosis by the hepatitis C virus.

In vitro data have shown that the hepatitis C virus (HCV) core protein binds to protein members of the tumour necrosis factor receptor (TNFR) superfamily. Since this interaction could be relevant to HCV persistence and oncogenesis, this study assessed whether HCV may interfere with the apoptotic cascade in vivo. Apoptosis (by TUNEL) and Fas and TNFR1 expression (by immunohistochemistry) were scored in the liver of 60 chronic hepatitis C patients. Results were compared with the liver disease grading and staging scores and the HCV replication level in serum and liver. Apoptotic hepatocytes were stained in 29 cases. Fas was expressed in 35 cases and TNFR1 in 21, 15 patients (25%) being negative for both receptors. Overall, the numbers of TUNEL-, Fas- and TNFR-positive hepatocytes did not correlate with the extent of intrahepatic CD8+ T-lymphocyte infiltration, the grading and staging of liver disease, or the serum or liver HCV RNA levels. Furthermore, when patients expressing either Fas or TNFR1 were stratified according to serum HCV RNA levels, cases with detectable hepatocyte apoptosis had higher HCV viraemias. In conclusion, an HCV-mediated, in vivo blockade of hepatocyte apoptosis via the Fas- or TNFR1-dependent pathways seems unlikely.

Intrahepatic HCV RNA loads in 37 HIV-HCV co-infected patients with controlled HIV infection.

Serum and intrahepatic hepatitis C virus (HCV) RNA were measured in 37 HIV-HCV co-infected patients with controlled human immunodeficiency virus (HIV) infection and correlated with clinical, biological, and histological parameters. Thirty-seven interferon-naive patients underwent liver biopsy. HCV-induced activity (A) and fibrosis (F) were evaluated with METAVIR score. The 37 patients included had HIV plasma loads < 10,000 copies/ml, CD4(+) count > 250/microl. All the patients but two were receiving antiretroviral treatment. Liver tissue and sera were used for measurement of HCV RNA by the Cobas Amplicor HCV Monitor. All patients had serum and liver HCV RNA, and both levels were correlated (r = 0.47; P = 0.003). Intrahepatic HCV load did not depend on age, sex, duration of HCV infection, CD4(+), HCV genotype, or fibrosis. AST levels correlated with intrahepatic HCV load (r = 0.52; P = 0.001). Patients with METAVIR A1/A2 had significantly lower levels of liver HCV-RNA than were found in patients with METAVIR A3 (P = 0.026). Highly active antiretroviral therapy (HAART) including protease inhibitors(PI)-treated patients had significantly lower intrahepatic HCV load (P = 0.04). A weak but significant correlation between serum and liver HCV RNA was found. The amount of hepatic HCV RNA was correlated with AST levels, histological activity, but not with HCV genotype or fibrosis. The immune improvement associated with PI regimens could help reduce HCV load, supporting a protective effect of PI-induced immune restoration.

Lack of Association Between Hepatitis C Viral RNA in Serum and Liver and Histologic Gradings: A Study on Irish Anti-D-Treated Patients

In this study the authors applied a reverse transcription-polymerase chain reaction (RT-PCR) assay to detect hepatitis C virus (HCV) RNA in 15 frozen liver biopsy samples from anti-D-treated patients. They also correlated the presence or absence of HCV RNA in the serum and liver of each patient with their histologic gradings. RNA was extracted from 36 frozen liver biopsy samples. These included 15 liver biopsy samples from patients infected with HCV through contamination of anti-D blood products. Three of these 15 anti-D-treated patients were receiving alpha-interferon treatment at the time of liver biopsy. Nine frozen liver biopsy samples from patients with a history of intravenous drug abuse were included as positive controls. HCV-negative frozen liver biopsy samples from 12 noninfected patients were used as negative controls. RNA was also extracted from six frozen skin biopsy specimens to check for cross-contamination of samples. Eleven of 15 anti-D-treated patients were HCV RNA positive by RT-PCR, with 100% correlation between HCV RNA in the serum and liver. The nine frozen liver biopsy samples from the intravenous drug abuse patients (positive controls) were also RT-PCR positive for HCV RNA. The 12 noninfected samples and the negative control biopsy samples were negative for HCV. Twenty-seven percent of the recombinant immunoblot assay-positive patients were serum and liver HCV RNA negative. HCV-positive patients receiving alpha-interferon therapy at the time of biopsy had cleared the virus from the serum and the liver. There was no correlation between the presence or absence of serum and liver HCV RNA with the histologic grading. This lack of correlation shows clearly the importance of histopathologic evaluation of liver biopsy samples in monitoring HCV-associated liver disease progression. In addition, this finding indicates that one cannot rely only on the presence or absence of HCV RNA in either serum or liver tissue as a parameter in monitoring HCV-associated liver disease progression in this unique cohort of anti-D-treated patients.

Failure to infect rhesus monkeys with hepatitis C virus strains of genotypes 1a, 2a or 3a.

The chimpanzee is the only recognized animal model for the study of hepatitis C virus (HCV). However, recently it was reported that rhesus monkeys were susceptible to HCV and developed hepatitis during infection. In the present study, we inoculated two rhesus monkeys each with HCV strain H77 (genotype 1a), strain HC-J6 (genotype 2a) or strain S52 (genotype 3a). Weekly serum samples were tested for liver enzyme values, HCV antibodies and HCV RNA. We did not find evidence of HCV infection in any of the monkeys during 24 weeks of follow-up. Our study demonstrates that rhesus monkeys are not readily infected with HCV and apparently do not represent a useful animal model for the study of HCV.

Serum hepatitis C virus RNA levels and histologic findings in liver allografts with early recurrent hepatitis C.

Histopathologic features of early recurrent hepatitis C after orthotopic liver transplantation (OLTx) may be modified by immunosuppressive therapy or complicated by other conditions. Hepatitis C virus (HCV) RNA level usually increases after OLTx, but its correlation to histologic findings is not clear. To evaluate the histologic findings of early recurrent hepatitis C in liver allografts and its correlation to serum HCV RNA level. We studied 14 patients who underwent OLTx for chronic HCV infection. Thirty liver biopsy specimens and HCV RNA levels of 22 corresponding plasma samples obtained during the first 6 months following OLTx were analyzed. The control group (9 patients, 25 biopsy specimens) was chosen at random from patients with chronic liver disease other than HCV who were undergoing OLTx, and all tested negative for HCV RNA by polymerase chain reaction after OLTx. Statistically significant pathological features of early recurrent HCV infection were the number of acidophilic bodies, piecemeal necrosis, lymphocyte predominance in the portal tracts, and fibrous septum. These findings and histologic activity index scores increased with time after OLTx. The HCV RNA levels determined by branched DNA assay showed no significant correlation with histologic features. However, patients with higher histologic activity index scores tended to have higher RNA levels. Liver biopsy specimens are helpful for the diagnosis or confirmation of early recurrent hepatitis C in liver allografts, but serial biopsy specimens are sometimes required for definite diagnosis. The HCV RNA levels are usually higher in patients who display signs of more severe liver damage.

Influence of liver hepatitis C virus RNA and hepatitis C virus genotype on FAS-mediated apoptosis after liver transplantation for hepatitis C.

Recurrent hepatitis C virus (HCV) infection after liver transplantation is characterized by a high level of intrahepatic HCV replication and more severe liver damage in case of genotype 1b infection. We investigated the involvement of apoptosis in recurrent HCV liver disease, and its possible links with histological findings, HCV genotype, liver HCV RNA level, and liver Fas mRNA level. We studied 61 liver graft biopsy specimens from 25 patients transplanted for HCV-related cirrhosis. DNA fragmentation was determined semi-quantitatively by in situ end labeling. HCV RNA and liver Fas mRNA were determined in parallel by quantitative polymerase chain reaction, with ribosomal 28S RNA as internal standard. Apoptotic lesions were predominantly portal (nonhepatocytic) or lobular (hepatocytic). Both were correlated with serum aminotransferase levels. The degree of portal apoptosis correlated with acute rejection (P<0.001), although lobular apoptosis was associated with lobular hepatitis (P<0.02), and HCV genotype 1b (P=0.04). In multivariate analysis, liver Fas mRNA level independently correlated with HCV-related chronic active hepatitis (P=0.04), age (P=0.01), and liver HCV RNA level (P=0.0007). After liver transplantation, 1) apoptosis is involved in HCV-related liver damage; 2) HCV type 1b may induce more severe apoptotic lesions than other genotypes; and 3) Fas transcription may be up-regulated by intrahepatic HCV replication.

Hepatocellular injury determinants

The pathogenesis of liver injury resulting from chronic hepatitis C virus (HCV) infection is not well understood. HCV replication is thought to occur via the production of an antisense or replicative intermediate (RI) RNA, from which progeny genomic strands are transcribed. One approach to studying HCV pathogenesis is to detect and quantify genomic and RI HCV RNAs in human liver biopsy specimens using in situ hybridization or in situ PCR. However, results from previous studies vary greatly. For example, examination of the relationship between levels of HCV RNA in liver and the degree of hepatocellular injury have provided data both for and against HCV-induced cytopathic effects. However, a recent report has described a novel, quantitative, highly sensitive, strand-specific in situ hybridization assay for detecting HCV RNAs in liver tissue 1 Chang M. et al. In situ distribution of the replicative intermediate RNA of hepatitis C virus in hepatic tissue and its correlation with liver disease. J. Virol. 2000; 74: 944-955 Crossref PubMed Scopus (55) Google Scholar . Using this assay, HCV genomic RNA was found to be variably distributed throughout infected livers and located primarily in the cytoplasm of hepatocytes. By contrast, HCV RI RNA showed a more focal pattern of distribution, and was exclusively localized in the cytoplasm of hepatocytes. There was no significant relationship between the distribution pattern of HCV genomic RNA and any indices of liver injury. However, a significant correlation was observed between the percentage of cells staining positive for RI RNA and the degree of hepatic inflammatory activity. Furthermore, the ratio of cells staining positive for HCV replicative vs genomic RNA correlated with the histological severity of liver injury, supporting the hypothesis that active replication of HCV in liver tissue could be a significant determinant of hepatocellular injury.

Hemochromatosis gene mutations in chronic hepatitis C patients with and without liver siderosis

Chronic hepatitis C is often associated with liver iron overload, which may affect the long-term prognosis and the response to antiviral treatment. The occurrence of hemochromatosis (HFE) mutations were studied to determine whether may contribute to the liver iron overload of chronic hepatitis C patients. The prevalence of two HFE mutations (C282Y and H63D) in 120 chronic hepatitis C patients was determined and the findings were correlated with clinical, histological and virological features. Hepatic iron was determined semiquantitatively by a histochemical hepatic iron index, defined as the ratio of a histochemical staining score to the patient's age, after correction for heterogeneous lobular iron distribution. Serum hepatitis C virus (HCV) RNA was measured by bDNA assay and typed by restriction fragment length polymorphism. Liver HCV RNA was measured by a semi-quantitative strand-specific reverse transcription-polymerase chain reaction (RT-PCR). Excess liver iron was stained in the liver of 36 patients (30%). Siderotic patients had the same geographic origin, serum and liver HCV RNA levels and H63D and C282Y mutations frequency as non-siderotic patients. However, siderotic patients were older (P = 0.015), more frequently males (P = 0.02), less frequently infected with HCV genotype 3 (P = 0.037) and had a higher liver fibrosis score (P = 0.008). The liver iron content did not correlate with the serum or liver HCV RNA titers. Ten of the 36 patients with liver siderosis had neither a history of excess alcohol intake, multiple transfusions, or HFE mutations. In conclusion, the pathogenesis of the liver iron overload in chronic hepatitis C patients cannot be fully explained by the occurrence of HFE mutations. The exact mechanism of iron accumulation in these patients therefore remains unexplained.

Patients co-infected with human immunodeficiency virus and hepatitis C virus demonstrate higher levels of hepatic HCV RNA.

Serum and liver hepatitis C virus (HCV) RNA levels in patients with hepatitis C have previously been quantified using different techniques. In this work, we used an automated, multicycle, polymerase chain reaction (PCR)-based technique to quantify HCV RNA in 1-2 mm of frozen liver tissue, and in serum, from 70 patients with antibodies to HCV (anti-HCV), with and without human immunodeficiency virus (HIV) co-infection. Stored liver tissue and sera collected at the time of liver biopsy were used for measurement of HCV RNA. Forty-eight HCV patients and 22 HIV/HCV co-infected patients were studied. Co-infected patients had significantly higher median serum and liver HCV RNA (6.7 log copies ml-1 serum and 2.90 log copies microg-1 liver nucleic acids) than patients with HCV alone (6.2 log copies ml-1 serum and 2.19 log copies microg-1 liver nucleic acids). There was only a weak correlation between serum and liver HCV RNA (r = 0.43). There was no correlation between liver and serum HCV RNA and host factors such as duration of disease, CD4 counts, alanine aminotransferase levels or histological score. There was no correlation with HCV genotype. Co-infected patients were more likely to harbour HCV genotype 1 (85%) when compared to patients with HCV alone (58%). An identical genotype was found in liver and serum in 89% of those tested; in 11%, a mixed genotype was present in serum. Patients with HCV genotypes 1 and non-1 had similar histological scores. Hence, an automated PCR-based technique is useful for measuring both liver and serum HCV RNA. Serum HCV genotypes closely paralleled those found in liver tissue. HIV co-infection was associated with higher serum, as well as intrahepatic, HCV RNA levels, by mechanisms not directly related to CD4 counts. The lack of correlation between liver HCV RNA and histology suggests that HCV is not directly cytopathic.

Comparative evaluation of hepatitis C virus infection in serum and liver tissue of patients with chronic liver disease by reverse transcription-polymerase chain reaction.

OBJECTIVE: To undertake a comparative evaluation of diagnostic modalities used in the detection of hepatitis C virus (HCV) infection by serum antibody in ELISA and HCV-RNA in the serum as well as in the liver tissue of the same patients with chronic liver disease by reverse transcription-polymerase chain reaction (RT-PCR). METHODS: Thirty-five subjects comprising 30 cases of cirrhosis of the liver and five cases of chronic hepatitis were included in the study group. The criteria for including the subjects in the study were the availability and quality of serum and liver samples. Serologic ELISA tests for HCV antibody as well as HBsAg and anti-HBc IgG for hepatitis B virus (HBV) were performed on the patient's sera, while RT-PCR was carried out with RNA extracted from both serum and liver biopsies of the patients. Primer sequences selected from the conserved NS5 region of the HCV genome were used in RT-PCR. RESULTS: Of 30 cases of cirrhosis studied, five (16.6%) were anti-HCV antibody positive, while seven (23.3%) were positive for HCV RNA in serum and 10 (33.3%) were positive in liver tissue. One of the anti-HCV antibody-positive patients was negative for HCV RNA in both liver and serum by PCR. All seven patients who showed positivity for HCV RNA in serum were also positive for HCV RNA in the liver. Another three cirrhosis cases (10%) found to be HCV positive in liver tissue were negative for both antibody and HCV RNA in serum. Of the five cases of chronic hepatitis, two (40%) were found to be positive by all three tests. HCV was found to be more prevalent in HBV-infected cases (8/18; 44.5%). However, two of the three cirrhotic patients who showed HCV positivity only in liver PCR tested negative for HBV surface antigen. CONCLUSIONS: RT-PCR detection of HCV RNA in liver tissue is of significant clinical importance and is more reliable than RT-PCR in serum or antibody tests by ELISA. RT-PCR can effectively complement antibody tests for reliable diagnosis, since assessment of HCV positivity by a single test seems to be inadequate. The frequent association of HCV infection with HBV suggests that hepatitis B and C viruses may share similar modes of transmission in India.

Varying incidence of cirrhosis and hepatocellular carcinoma in patients with chronic hepatitis C responding differently to interferon therapy

To determine whether interferon (IFN) therapy can reduce incidence of the development of cirrhosis and hepatocellular carcinoma equally in patients with chronic hepatitis C virus (HCV) who responded differently to therapy, a retrospective analysis of 250 patients treated with IFN was conducted. Two hundred and fifty patients with chronic HCV who were treated with IFN were classified into 3 groups based on serum aminotransferase levels during and after therapy with IFN-alpha: long term responders (n=93), short term responders (n=70), and nonresponders (n=87). Eighty-nine untreated patients served as a control group. The follow-up period was 4 years. Liver function tests, HCV RNA levels, and alpha-fetoprotein were measured each month. Imaging diagnosis was made every 6 months with ultrasonography and yearly with dynamic computed tomography. Serum and liver HCV RNA was assayed by reverse transcriptase-polymerase chain reaction and branched DNA probe. Of the 93 long term responders, 67 (72%) remained HCV RNA negative whereas 26 (28%) were HCV RNA positive. Further histologic improvement occurred only in HCV RNA negative long term responders. During the follow-up period, the overall annual incidence of cirrhosis in the 250 patients was decreased significantly compared with the control group, whereas the overall annual incidence of hepatocellular carcinoma was not. None of the long term responders but 5 (7%) short term responders and 33 (38%) nonresponders had disease progression to cirrhosis. No HCV RNA negative long term responders but 1 (3.8%) HCV RNA positive long term responder, 1 (1.4%) short term responder, and 14 (16%) nonresponders developed hepatocellular carcinoma. Long term and short term responders had significantly lower incidence of cirrhosis whereas nonresponders had significantly higher incidence of cirrhosis and hepatocellular carcinoma than the control group. Multivariate analysis showed that staging score, the presence of high viral levels, and lack of response to therapy significantly influenced the incidence of these events. These findings suggest that patients with chronic HCV responding differently to IFN therapy had different incidence of disease progression to cirrhosis and of the development of hepatocellular carcinoma, and that nonresponders should be regarded as a group at high risk for these events.

Varying incidence of cirrhosis and hepatocellular carcinoma in patients with chronic hepatitis C responding differently to interferon therapy

BACKGROUND. To determine whether interferon (IFN) therapy can reduce incidence of the development of cirrhosis and hepatocellular carcinoma equally in patients with chronic hepatitis C virus (HCV) who responded differently to therapy, a retrospective analysis of 250 patients treated with IFN was conducted. METHODS. Two hundred and fifty patients with chronic HCV who were treated with IFN were classified into 3 groups based on serum aminotransferase levels during and after therapy with IFN-α: long term responders (n = 93), short term responders (n = 70), and nonresponders (n = 87). Eighty-nine untreated patients served as a control group. The follow-up period was 4 years. Liver function tests, HCV RNA levels, and α-fetoprotein were measured each month. Imaging diagnosis was made every 6 months with ultrasonography and yearly with dynamic computed tomography. Serum and liver HCV RNA was assayed by reverse transcriptase-polymerase chain reaction and branched DNA probe. RESULTS. Of the 93 long term responders, 67 (72%) remained HCV RNA negative whereas 26 (28%) were HCV RNA positive. Further histologic improvement occurred only in HCV RNA negative long term responders. During the follow-up period, the overall annual incidence of cirrhosis in the 250 patients was decreased significantly compared with the control group, whereas the overall annual incidence of hepatocellular carcinoma was not. None of the long term responders but 5 (7%) short term responders and 33 (38%) nonresponders had disease progression to cirrhosis. No HCV RNA negative long term responders but 1 (3.8%) HCV RNA positive long term responder, 1 (1.4%) short term responder, and 14 (16%) nonresponders developed hepatocellular carcinoma. Long term and short term responders had significantly lower incidence of cirrhosis whereas nonresponders had significantly higher incidence of cirrhosis and hepatocellular carcinoma than the control group. Multivariate analysis showed that staging score, the presence of high viral levels, and lack of response to therapy significantly influenced the incidence of these events. CONCLUSIONS. These findings suggest that patients with chronic HCV responding differently to IFN therapy had different incidence of disease progression to cirrhosis and of the development of hepatocellular carcinoma, and that nonresponders should be regarded as a group at high risk for these events.

Serum and liver HCV RNA levels in patients with chronic hepatitis C: correlation with clinical and histological features

Background—Liver disease in chronic hepatitis C virus (HCV) infection ranges from minimal lesions to liver cirrhosis, eventually evolving to hepatocellular carcinoma. Whether and how HCV determines the different clinical and...

Clinical significance of hepatic HCV RNA in patients with chronic hepatitis C demonstrating long-term sustained response to interferon-alpha therapy

Whether sustained biochemical response and absence of serum HCV RNA in the 6-12 months following suspension of interferon-alpha (IFN-alpha) therapy reflect definitive viral clearance in patients with chronic hepatitis C virus (HCV) infection is controversial. To obtain more information on this topic, HCV RNA was sought in both liver and serum samples of 25 long-term responders who were followed for a median period of 39 months (range 21-79) after discontinuation of IFN-alpha. Liver biopsy was undertaken before and 6 to 12 months after IFN-alpha withdrawal. Liver and serum HCV RNA were tested by a nested polymerase chain reaction. Twenty-two patients (88%) tested negative for both liver and serum HCV RNA, two patients had detectable HCV RNA in both liver and serum, and one patient showed persistent HCV RNA only in the liver. Post-treatment liver histology improved markedly in all patients, including those with viral persistence. During further follow-up, biochemical remission was maintained in all patients except one in whom both serum and liver specimens remained HCV RNA positive. The data indicate that the large majority of long-term responders test negative for HCV RNA in the liver, which suggests definitive eradication of HCV RNA infection.