Recurrent hepatitis C virus infection after liver transplantation: Immunohistochemical assessment of the viral antigen (original) (raw)
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Liver Transplantation, 2006
The aim of this study was to evaluate how the immunohistochemical detection of liver hepatitis C virus (HCV) antigens (HCV-Ag) could support the histologic diagnosis and influence the clinical management of post-liver transplantation (LT) liver disease. A total of 215 liver specimens from 152 HCV-positive patients with post-LT liver disease were studied. Histologic coding was: hepatitis (126), rejection (34), undefined (24; coexisting rejection grade I and hepatitis), or other (31). The percentage of HCV-Ag infected hepatocytes were evaluated, on frozen sections, by an immunoperoxidase technique. HCV-Ag were detectable early in 57% of cases within 30 days post-LT, 92% of cases between 31 and 180 days, and 74% of cases after more than 180 days. Overall, HCV-Ag were detected more frequently in histologic hepatitis as compared to rejection (P Ͻ 0.0001) with a higher percentage of positive hepatocytes (P Ͻ 0.00001). In 16 patients with a high number of HCV-Ag-positive hepatocytes (65%; range 40-90%) a clinical diagnosis of recurrent hepatitis (RHC) was made despite inconclusive histopathologic diagnosis. Multivariate analysis identified the percentage of HCV-Ag-positive hepatocytes and the time post-LT as independent predictors for RHC (P ϭ 0.008 and P ϭ 0.041, respectively) and the number of HCV-Ag-positive hepatocytes Ն50% as the only independent predictor for nonresponse (P Ͻ 0.001) in 26 patients treated with ␣-interferon plus ribavirin. In conclusion, HCV reinfection occurs early post-LT, reaching its peak within 6 months. Immunohistochemical detection of post-LT HCV reinfection support the diagnosis of hepatitis when the histologic features are not conclusive. A high number of infected cells, independently from the genotype, represents a negative predictive factor of response to antiviral treatment.
Liver International, 2008
Introduction: The mechanisms by which severe cholestatic hepatitis develops after liver transplantation are not fully understood. Reports on immunohistochemical distribution of hepatitis C virus (HCV) antigens are still scarce, but recently, HCV immunostaining was suggested for early diagnosis of cholestatic forms of recurrent hepatitis C in liver grafts. After purification, Rb246 pab anticore (aa1-68) yielded specific, granular cytoplasmic staining in hepatocytes. Signal amplification through the Envision-Alkaline Phosphatase System avoided endogenous biotin and peroxidase. Aims/Methods: Rb246 was applied to liver samples of explants of 12 transplant recipients, six with the most severe form of post-transplantation recurrence, severe cholestatic hepatitis (group 1) and six with mild recurrence (group 2). We also assessed immuno-reactivity at two time-points post-transplantation (median 4 and 22 months) in both groups. HCV-core Ag was semiquantified from 0 to 31 in each time point. Serum HCV-RNA was also measured on the different time points by branched DNA. Results: In the early post-transplant time point, one patient had a mild staining (11), two patients had a moderate staining (21) and the other three had no staining in group 1, compared with five patients with no staining (0) and one patient with mild staining (11) in group 2. Late post-transplant liver samples were available in nine patients, and two out of four samples in group 1 showed a mild staining, compared with no staining patients in five patients in group 2. Strikingly, on the explant samples, HCV immunostaining was strongly positive in group 1, and mildly positive in group 2. Two out of five samples showed 31 staining, and three samples showed 21 staining in group 1; two out of five samples showed no staining, two samples showed 11 staining and one sample showed 21 staining in group 2. Serum HCV-RNA was significantly higher in group 1, on both time-points posttransplantation. HCV-core Ag was not directly associated with serum HCV-RNA on the different time points. Conclusion: These preliminary results suggest that strong HCV immunostaining in the explant is predictive of more severe disease recurrence. Chronic hepatitis C virus (HCV) infection is the leading indication for liver transplantation, estimated at 35-45% (1). In that population, recurrent infection post-transplantation is almost universal, with a 10-to 20-fold increase in levels of viraemia (2). The majority of transplant recipients will develop histological evidence of disease as a result of recurrent HCV infection, if followed for up to 5 years, with a significant proportion progressing to chronic hepatitis and cirrhosis (3-7). The natural history of HCV infection post-transplantation is now beginning to be delineated, and we have limited understanding of the factors that influence disease progression. After transplantation, the majority of patients evolve to a slowly
Hepatology, 1997
Recurrence of hepatitis C after liver transplantation is com-the 5-year actuarial rate of HCV-related chronic active hepatitis is 75% 3 and the 7-year actuarial rate of HCV-related cir-mon and can lead to severe liver diseases. Although immunosuppression and high levels of viremia suggest a direct patho-rhosis is 15% (Feray C, unpublished data, March 1997). HCV infection in this setting is characterized by an intense viral genicity of hepatitis C virus (HCV), the relations between viral replication and long-term histological course are still replication. 4-6 Both intense replication and accelerated histological course suggest that HCV is able to directly induce unknown. Thirty-three patients with a mean histological follow-up of 3.5 years (3 months -8.6 years) were analyzed. liver damage after transplantation. However, the relations between the level of HCV replication and the severity of liver Nineteen patients were infected by genotype 1b. Liver HCV RNA was determined in parallel with the quantitation of an disease or the occurrence of histologically-defined hepatitis, 6,7 are far to be clear. 4 Some patients have high levels of internal control (28S ribosomal RNA) by competitive polymerase chain reaction (PCR). Lobular hepatitis (LH) and serum HCV RNA without evidence of liver damage whereas others have severe chronic hepatitis in spite of low levels of chronic active hepatitis (CAH) occurred in 27 and 19 patients, respectively. Levels of liver HCV RNA determined in 84 biop-serum HCV RNA. 4 One reason for these discrepancies may be that serum HCV RNA does not perfectly reflect the intra-sies were higher in cases of LH than in the other patterns (82 { 123 vs. 19 { 38; P õ .01) and were unrelated to the hepatic HCV replication, 8,9 either because of extrahepatic sources of HCV replication or because of an alteration of genotype. Progression from LH to CAH was associated with a highly significant decrease of liver HCV RNA (P Å .006), serum preservation with time. 10 Another important point is that most studies on HCV replication after transplantation which was not observed in patients with stable histology. Among patients with CAH, those infected by genotype 1b had are cross-sectional. For these reasons, it would be useful to longitudinally assess the level of intrahepatic HCV RNA and more severe liver damage and lower levels of liver HCV RNA than others (P Å .04). Multivariate analysis showed that high its relation with the long-term histological outcome of HCVrelated recurrent liver disease. levels of liver HCV RNA at the time of the first posttransplantation biopsy was an independent predictor of CAH (P Our hypothesis is that a longitudinal and quantitative analysis of intrahepatic HCV replication after liver transplanta-Å .01). After liver transplantation, the progression to CAH together with a decrease of liver HCV RNA suggests that a tion could show variations related to the histological course and could enlighten the pathogenesis of recurrent hepatitis host's response is involved in the long-term viral pathogenicity. This response may be stronger and liver disease more C. A positive relation between intrahepatic HCV replication and liver damage should suggest a direct pathogenicity of severe in patients with high levels of replication at the time of LH and in those infected by genotype 1b. (HEPATOLOGY HCV. Conversely, a decrease of intrahepatic HCV RNA together with a worsening of liver damage should suggest the 1997;26:1343-1350.)
Liver, 1994
We measured hepatitis C virus (HCV) RNA and antibodies against HCV recombinant proteins (C22/S1, El/S2, E2INS1, C33/NS3, C100iNS4, NS5) in serial serum samples from 22 interferon-treated patients with a long-term follow up (range: 36-44 months). Eleven of them showed persistently normal liver function tests and a significant histological amelioration or a complete resolution of chronic hepatitis (long-term responders, LTRs). In the remaining 1 1 patients (non-responders (NRs)) liver function tests normalized temporarily during therapy or remained unchanged. At the end of the follow up (3 years), viraemia was undetectable in six of 11 LTRs (54.6%). HCV-RNA was always detectable in the serum of NRs (p=O.O17). At admission, anti-C22/Sl, anti-E1/S2, anti-E2INS1, anti-C33/NS3, anti-CIOOiNS4 and anti-NS5 were detected ih 95.4%, 40.9%, 77.3%, 95.4%, 72.7% and 77.3% of the patients, respectively. Three years after suspension of therapy, anti-C100/NS4 was undetectable in five of six (83.3%) LTRs who cleared HCV-RNA and in only one with ongoing viraemia (20%). Anti-E2/NS1 was undetectable in 54.5% of LTRs and in no NRs (p=O.O67). Anti-EliS2 was detected more frequently in LTRs than in NRs (81.8% vs 45.5%). Serum levels of anti-C22lS1, C33lNS3 and NS5 did not change during therapy and the follow up in either group of patients. The clearance of viraemia in LTRs was associated with that of anti-C100/NS4 (p=O.O17). Serum HCV-RNA and anti-C100/NS4 appear suitable tools for monitoring patients who respond to therapy. More than 40% of LTRs remained HCV-RNApositive in spite of the biochemical remission of their liver diseases,
Hepatology, 2000
Approximately half of patients undergoing liver transplantation (LT) for hepatitis C virus (HCV) develop histologic evidence of recurrence within the first postoperative year. Early identification of recipients at risk for more severe recurrence of HCV may be useful in selecting patients for antiviral therapy. We determined whether recipients at greatest risk for more severe recurrence of HCV can be identified by pre-and/or early post-LT HCV-RNA levels in serum or tissue. Serum and tissue samples were prospectively collected pre-LT and at 7 days, 4 months, 1 year, and at 3 years posttransplantation from patients undergoing LT for HCV. Hepatitis activity index (HAI) and fibrosis stage (FS) were assessed in all liver biopsies. Forty-seven patients (32 men) were studied. Higher HCV-RNA levels at 4 months post-LT (>10 9 copies/mL, n ؍ 29) were associated with higher HAI at 1 year and at 3 years post-LT. The HAI seen on protocol biopsies at 4 months correlated significantly with fibrosis stage (FS) at 1 year (r ؍ .56, P < .001) and 3 years (r ؍ .53, P ؍ .002). Higher HCV-RNA levels at 7 days and 4 months post-LT were sensitive (66% and 84%, respectively) and specific (92% and 63%, respectively) in identifying recipients with an HAI greater than 3 at 3 years. Higher pre-and early post-LT HCV-RNA levels are associated with more severe recurrence of HCV. The correlation of early HAI with subsequent FS suggests that higher mean HAI will eventually translate into more advanced stages of fibrosis. Patients at risk for more severe post-LT recurrence of HCV can be identified by early posttransplant HCV-RNA levels. (HEPATOLOGY 2000;32: 1125-1130 Abbreviations: HCV, hepatitis C virus; LT, liver transplantation; EIA2, second generation enzyme-linked immunosorbent assay; RIBA, recombinant immunoblot assay; RT-PCR, reverse-transcriptase polymerase chain reaction; bDNA, branched DNA; HAI, hepatitis activity index; FS, fibrosis stage; ROC, receiver operating characteristic; AUC, area under the curve.
Hepatitis C virus infection and disease
Journal of Hepatology, 1993
Hepatitis C virus (HCV) is a lipid-enveloped single-stranded RNA virus with an unknown physical structure as only putative HCV particles have been identified by electron microscopy. Although HCV lacks the retroviral properties of being able to integrate into host DNA, it causes chronic infection in a considerable number of infected individuals (40-60%). Chronic infection is associated with a wide spectrum of liver diseases ranging from normal presentation to the different forms of chronic hepatitis, cirrhosis (about 20% of cases) and hepatocellular carcinoma. HCV therefore is not invariably and equally pathogenic, and genetic heterogeneity could be a major cause of such variability. Diagnosis of HCV infection relies on anti-HCV and HCV-RNA detection. Using second-generation assays, diagnostic sensitivity has increased to about 95%, but detection of anti-HCV does distinguish past from present infections. Only rising anti-HCV titres or anti-HCV seroconversion confirm a recent HCV infection. In anti-HCV-negative infections and cases of early acute hepatitis, HCV-RNA detection by RT-PCR represents a valid diagnostic alternative. In patients undergoing interferon therapy, testing for anti-HCV by immunoblotting represents a valid routine tool to monitor response. Anti-C-22 has the highest titre and persists longer while anti-C-100 is the earliest antibody to disappear in responders. The significant association between serum anti-C-100, HCV-RNA and liver disease suggests that anti-C-100 is an indirect marker of hepatitis C, but true markers of HCV-induced liver disease are still lacking.
Liver Transplantation and Surgery, 1997
The reasons for the wide variation of incidence and severity of recurrent hepatitis C after liver transplantation are not clear. We have studied liver transplant recipients to assess the impact of hepatitis C virus (HCV) genotype and HCV RNA quantification on HCV recurrence after transplantation. Twenty-two patients received transplants for HCV cirrhosis and were followed up with virological and histological assessments. Mean follow-up was 39 months. HCV genotype was determined with line probe assay (Inno-Lipa). HCV RNA quantity was determined in serum samples by use of polymerase chain reaction nested assay. HCV genotype 1 was detected in 13 patients and other genotypes in 9. Histological recurrence rates were 69% in patients with geno-type 1 and 66% in patients with other genotypes. All cases of severe histological injury (chronic active hepatitis or cirrhosis) were observed in patients with genotype 1. HCV RNA quantity was significantly higher in patients with genotype 1 (mean, 2.023 ؋ 10 3 copies/mL) than in patients with other genotypes (mean, 27,403 copies/mL). In conclusion, the severity of histological recurrence after liver transplantation for HCV disease was higher in patients infected by HCV genotype 1 than in those infected with other genotypes. The levels of viral replication were higher in patients with HCV genotype 1 than in those with other genotypes.
Prospective study of hepatitis C virus infection after orthotopic liver transplantation
Transplantation Proceedings, 1997
The aims of this study were to (i) evaluate the prevalence and the incidence of hepatitis C virus (HCV) infection in hemodialysis patients in two different centers in São Paulo (Brazil), (ii) determine the time required to detect HCV infection among these patients by serology or PCR, (iii) establish the importance of alanine aminotransferase determination as a marker of HCV infection, and (iv) identify the HCV genotypes in this population. Serum samples were collected monthly for 1 year from 281 patients admitted to hospital for hemodialysis. Out of 281 patients, 41 patients (14.6%) were HCV positive; six patients seroconverted during this study (incidence = 3.1/1000 person-month). In 1.8% (5/281) of cases, RNA was detected before the appearance of antibodies (up to 5 months), and in 1.1% (3/281) of cases, RNA was the unique marker of HCV infection. The genotypes found were 1a, 1b, 3a, and 4a. The presence of genotype 4a is noteworthy, since it is a rare genotype in Brazil. These data pointed out the high prevalence and incidence of HCV infection at hemodialysis centers in Brazil and showed that routine PCR is fundamental for improving the detection of HCV carriers among patients undergoing hemodialysis.