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The CC5 and CXC3 chemokines (CK) pathways are involved in the pathogenesis and outcome of several disease states, including chronic hepatitis C (CHC). The kinetics of Regulated upon Activation Normal T cell Expressed and Secreted (RANTES) (CCL5) and IP-10 (CXCL10) during direct-acting antivirals (DAA) treatment was retrospectively analyzed in 18 liver transplant recipients (LT-R) compared with 20 patients with CHC and 49 healthy controls (HC). CK levels were determined at baseline, week 4, end of treatment, 24 weeks post-treatment (sustained virological response [SVR]), and later-on during follow-up (FU), 12 and 24 months post-DAA. At baseline, median RANTES levels were higher in HC than in both LT-R ( p  > 0.01) and CHC ( p  > 0.01), whereas IP-10 levels were higher in LT-R and CHC than in HC ( p  > 0.05 and p  = 0.01), respectively. Mean RANTES values increased during DAA therapy to peak at SVR and FU with significantly higher levels than at baseline in LT-R ( p  < 0.01) and in CHC, but only at FU ( p  < 0.003). A subsequent return to baseline or lower levels was observed at extended FU. On the contrary, IP-10 values showed a significant decrease from baseline to SVR and FU in both LT-R ( p  < 0.03) and CHC ( p  < 0.01). RANTES profiles during the first 4 weeks of DAA treatment showed an increase or decrease from baseline according to baseline RANTES levels. CCR5 genotyping in LT-R showed the presence of 1 homozygous Δ32/Δ32 and 2 heterozygous WT/Δ32 haplotypes with a prevalence of 5.5% and 11.1%, respectively. In conclusion, although IP-10 showed the expected kinetics, the CC5 pathway appears extensively altered during CHC infection: monitoring these patients may be indicated as they may be at risk of other infections or immune-mediated disorders.

Background & Aims: The duration of current standard dual and protease inhibitor-based triple therapies for chronic hepatitis C is determined by assessment of early viral kinetics. Little is known about differences between HCV RNA assays for the use in response guided therapy., Methods: HCV RNA was assessed by two widely used real-time PCR-based assays, Cobas Ampliprep/Cobas TaqMan (CAP), and Real-Time HCV (ART) in 903 samples of hepatitis C genotype 1 patients treated with dual (n=169) or telaprevir-based triple therapy (n=164) in three European countries., Results: Overall, CAP and ART were in excellent agreement for the determination of HCV-RNA concentrations (mean difference 0.21 log10 IU/ml). For treatment-naïve patients treated with peginterferon-alfa and ribavirin a lower rate of undetectable HCV-RNA at week 4 (RVR) was observed for ART (9%) vs. CAP (16%). Although 11/27 (41%) of patients with shortened treatment (24weeks) had detectable HCV-RNA <12IU/ml by ART at week 4 none of these patients experienced virologic relapse after treatment cessation. In patients who received triple therapy, 67% and 37% had undetectable HCV-RNA at week 4 by CAP and ART, respectively. However, 18/31 (58%) eligible patients for shortened treatment based on CAP had detectable HCV-RNA by ART at week 4. Again, relapse was not observed in these patients., Conclusions: Lower rates of undetectable HCV-RNA at week 4 were observed with ART compared to CAP in patients treated with dual and triple therapies. For ART, detectable <12IU/ml HCV-RNA levels at week 4 may be sufficient as part of the criteria used for selecting patients who receive a shortened treatment regimen., (Copyright © 2014 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

Dialysis patients remain a high-risk group for hepatitis C virus (HCV) infection. The current diagnosis of HCV infection among dialysis patients includes serological assays and nucleic acid amplification technology (NAT) for assessing serum anti-HCV antibody and HCV viremia, respectively. However, current NAT techniques are expensive and labor-intensive and often lack standardization. An assay prototype designed to detect and quantify total HCV core antigen (total HCV core Ag) protein in serum and plasma in the presence or absence of anti-HCV antibodies has been recently developed. A comparison between a total anti-HCV core Ag enzyme-linked immunosorbent assay (ELISA) and a quantitative HCV RNA assay based on reverse transcription-PCR (RT-PCR) (Amplicor HCV Monitor test) was performed using a large (n = 305) cohort of ELISA HCV 3.0 HCV-negative and -positive patients on maintenance dialysis. The concentrations of HCV core Ag and HCV RNA levels (measured by RT-PCR) were significantly correlated (r = 0.471, P = 0.0001) over a wide range of HCV RNA levels and were maintained among different HCV genotypes (HCV genotype 1, r = 0.862, P = 0.0001; HCV genotype 2, r = 0.691, P = 0.0001). We estimated that 1 pg of total HCV core Ag per ml is equivalent to approximately 19.952 IU of HCV RNA per ml, even if the wide range in the ratio of core Ag to HCV RNA (95% confidence intervals, 2.8 x 10(3) to 1.6 x 10(5) IU/ml) precluded definitive conclusions. In summary, total HCV core Ag proved to be useful for performing HCV RNA measurement among dialysis patients in routine laboratories without the need for special equipment or training. The present study supports the use of the total anti-HCV core Ag ELISA for assessing viral load among dialysis patients with HCV infection.

Hepatitis C virus (HCV) infection remains frequent among dialysis patients. The relationship between viral load and liver disease progression is currently a matter of debate; however, low HCV viral load (HCV RNA) is a well established predictor of successful antiviral treatment. Dialysis patients have immune compromise due to uremia; in spite of this, HCV viral load is not high and does not increase over time. A number of studies support the notion that this dynamics of HCV viral load may be related to lowering of HCV RNA titers during the HD procedure. It has been suggested that the intradialytic reduction of HCV is membrane-dependent; polysulphone (PS) and hemophan membranes appear more effective. Various mechanisms have been mentioned to explain the reduction of HCV RNA during hemodialysis (HD): adsorption of HCV onto dialysis membrane, HCV escape into spent dialysate, destruction of HCV particles or increased interferon (IFN) activity. Several investigators have noted that, at the end of the HD procedure, a virological rebound of HCV viremia occurs. Some suggestions have been made to minimize the risk for HCV escape into spent dialysate. However, controversial evidence on these issues exists: the HCV RNA reduction during HD procedures has not been seen in some studies and the relationship between the intra-dialytic reduction of HCV RNA titers and dialysis membrane is not completely understood. It is evident that additional long-term longitudinal studies by serial viral load estimations are needed to better define the dynamics of HCV viral load in the dialysis population.

Mixed cryoglobulinemia is a systemic disease, almost always associated with hepatitis C virus infection and characterized by purpura and cutaneous vasculitis, asthenia, arthralgias, and often renal and neurological involvement. No significant differences have been described to date in mixed cryoglobulinemia patients with type 1, 2, or 3 hepatitis C virus infection with respect to symptoms, while a higher prevalence of genotype 2a has been reported in patients without clinical and biochemical signs of liver disease or with serum autoantibodies. We examined 33 hepatitis C virus-positive patients with mixed cryoglobulinemia to assess if any clinical or serological feature is related to infection with different genotypes. All subjects underwent viral genotype determination by means of a single-step polymerase chain reaction. Thirteen patients (39%) were infected with hepatitis C virus type 1b, 17 (52%) with type 2a or 2a/c, and 3 (9%) with type 3. There was a significant difference in the frequency of peripheral nervous system involvement: paresthesias or other symptoms of peripheral neuropathy were less frequent in patients with 2a or 2a/c infection (29%) than in patients with type 1b or type 3 infection (88%, P = 0.003). Only patients with hepatitis C virus type 2 had urticaria or cutaneous ulcers. These patients also had a lower frequency of arthralgias, lower cryocrit values (P = 0.02), and lower serum levels of alanine-aminotransferase and gamma-glutamyl-transpeptidase (P < 0.04) than patients with type 1 and type 3 infection. The prevalence of antinuclear antibody positivity was similar in the three groups.

Purpose of the Study: We evaluated the presence of HCV in the peritoneal effluents of viraemic patients treated with continuous ambulatory peritoneal dialysis (CAPD) to evaluate the risk of transmitting the infection with this procedure., Procedure: Fifteen of 81 CAPD patients (18.5%) had anti-HCV antibodies and eight were viraemic. At the beginning of CAPD two of the viraemic patients had ascites with a clinical picture of chronic active hepatitis and cirrhosis. Peritoneal dialysates were collected after an overnight exchange with 1.36% glucose and after a 4-h exchange with 3.86% glucose. Fluids from the overnight exchange were spun to obtain a cellular pellet and the supernatant 100-fold concentrated., Results: No viral genome could be detected in unconcentrated samples and in cellular pellets, while HCV-RNA at low titre was detected in concentrated dialysates from the two patients with active liver disease., Conclusions: Our findings confirm that HCV may be present in the CAPD effluent of some patients; however, the titre of virus in the effluent was extremely low, at the limit of detection of the PCR assay. Peritoneal fluids originating from patients with HCV associated severe liver disease may be a potential source of infection.

Background: Hepatitis C virus (HCV) infection is highly prevalent in dialysis patients. To further characterize HCV infection in this patient population, we studied the distribution of viral genotypes in 55 patients undergoing chronic dialysis treatment with seropositivity for HCV markers., Methods: Thirty-two of 55 (58%) patients showed HCV RNA in the serum using reverse transcription polymerase chain reaction (RT-PCR) in the 5'-untranslated region (UTR) of the viral genome. HCV genotyping was performed using biotinylated type-specific oligonucleotide probes after hybridization with amplified sample material., Results: HCV subtype 2a was dominant (56%), followed by HCV subtype 1b (31%), type 3 (3%) and 4 (3%). There was no association between demographic or clinical features of this cohort and HCV genotype. Genotype dependence was observed for antibody response toward the NS4 (c 100-3 and 5-1-1) proteins, which was infrequent in genotype 2a carriers compared with genotype 1b (p = 0.004)., Conclusions: HCV subtype 2a was dominant in our cohort of anti-HCV-positive dialysis patients; there was no association between HCV genotypes and demographic or clinical features of patients; the absence of antibody response toward the NS4-related antigens was frequent in genotype 2a carriers and may serve to predict responses to interferon therapy. The relative homogeneity of the viral population in dialysis patients attending our unit suggests a nosocomial transmission of HCV in this clinical setting.

Background: In HBV infection, as in other viral diseases, antibodies of the IgM class are associated with acute or ongoing infection. In contrast, the significance of this antibody in HCV infection is unclear and data regarding end-stage renal disease (ESRD) patients are lacking., Methods: We tested sera from 78 ESRD patients (66 chronic dialysis patients, 12 renal allograft recipients) showing anti-HCV IgG antibody, for serum anti-HCV IgM core antibody. A specific solid-phase enzyme-linked immunoassay (HCV IgM EIA, Abbot) was used. In all patients serum HCV RNA was detected by RT-PCR technique, with primers from the 5' untranslated region of the viral genome., Results: Prevalence of anti-HCV IgM core antibody was 22% (17/78). We observed association between prevalence of anti-HCV IgM core antibody and frequency of HCV RNA in the serum. Thus 15 of 45 (33%) sera containing HCV RNA also contained anti-HCV IgM, while two of 33 (6%) HCV RNA negative sera showed anti-HCV IgM core antibody (P = 0.01). There was a significant relationship between absorbance values of anti-HCV IgM core antibody and NS3 (P = 0.01), NS5 (P = 0.04), and core (P = 0.0001) reactivity in RIBA-2 and RIBA-3 assays. Optical density values of anti-HCV IgM were significantly associated with the number of reactive bands in RIBA-2 (P = 0.04) and RIBA-3 (P = 0.03) assays., Conclusions: 22% of ESRD patients with anti-HCV IgG activity showed anti-HCV IgM core antibody in the serum. Anti-HCV IgM activity seems to correlate positively with HCV viraemia in ESRD population. Testing for this antibody may be useful as a serological marker to indicate the presence of ongoing HCV infection.

Background: Although there are some reports regarding prevalence of anti-HCV antibodies in kidney transplant patients, there are scarce data about viraemia, genotyping and liver histology of HCV infection in kidney transplant recipients., Methods: We studied the prevalence of anti-HCV antibodies by second-generation screening and confirmatory assays in a cohort of 73 renal allograft recipients. All patients were tested for serum HCV RNA using reverse transcription polymerase chain reaction in the 5'-untranslated region (UTR) of the viral genome. HCV RNA positive patients were subjected to genotype analysis using biotinylated type-specific oligonucleotide probes after hybridization with amplified sample material. Eleven of 73 patients showing raised aminotransferase levels underwent hepatic biopsy., Results: Fifteen of 73 (20%) patients were determined anti-HCV positive. Eleven of 73 (15%) showed detectable serum HCV RNA; no viraemic, seronegative patients were identified. Genotyping showed that HCV subtype 2a was dominant (64%), followed by HCV subtypes 1b (27%) and 1a (9%). Six of 11 (54%) HCV RNA patients and 12 of 62 (19%) HCV RNA negative patients showed raised aminotransferase levels (P = 0.03). Liver biopsies showed histological features of chronic hepatitis with mild or moderate degrees of activity., Conclusions: The prevalence of anti-HCV antibodies and HCV viraemia was 20% and 15% respectively; there was a good association between anti-HCV anti-bodies and HCV viraemia; hepatic enzyme levels were good indicators of ongoing HCV infection; HCV subtype 2a was prevalent; liver histology showed histological characteristics of chronic hepatitis with mild or moderate degrees of activity.

To understand the clinical outcome of hepatitis C virus (HCV) recurrence, data from 35 liver transplant recipients who survived more than 6 months were reviewed. The presence of HCV-RNA was evaluated and genotyping was performed. On the basis of alanine aminotransferase (ALT) levels, patients were sorted into four groups. In 20 patients, a chronic elevation in ALT was found; HCV-RNA detection was positive in 17/17 and the following genotypes were found in 15 of them: 1b in ten patients, 2a in four patients, and 3a in one patient. In 11 patients, ALT levels remained normal throughout follow-up; in nine of them HCV-RNA was positive; HCV genotyping was available in eight patients and identified type 1b in two, type 2a in five, and type 3a in another patient. In two patients, ALT fluctuated above and below the upper limits of normality; type 1b HCV-RNA was found in one of them. In two patients, after an initial period of normality, ALT levels showed an abrupt rise; HCV-RNA was positive and type 1b was identified in both patients. Eight patients developed HCV-related deep jaundice and three of them spontaneously recovered. Progressive hepatic injury occurred in eight patients, six with chronic ALT elevation and two showing a late ALT elevation; genotype 1b was present in seven patients while in one, genotype 3a was found; sub-acute graft failure developed in five of them, leading to death in two and retransplantation in the others; the other three patients are alive with recurrent overt cirrhosis. The 1, 3, and 5 year actuarial survivals were 89%, 79%, and 63% respectively. The 1, 3, and 5 year actuarial risks of progressive graft damage were 6%, 7%, and 15%, respectively. In conclusion, HCV reinfection causes a slow decrease in the long-term patients' survival. Persistent elevation of ALT is more frequently observed in patients with genotype 1b infection.

Objective: To evaluate the relationship between hepatitis C virus antibodies (HCV-Ab) and viremia and to compare the prevalence of HCV-Ab and HCV viremia in hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) patients., Design: Cross-sectional study., Setting: Dialysis unit of a nephrology division in a public university hospital., Patients: All dialysis patients who came for routine clinic visits during the study period. None denied informed consent. Forty-eight patients on HD and 79 on CAPD were examined., Intervention: Blood samples were tested by second-generation enzyme-linked immunosorbent assay (ELISA II) and recombinant immunoblot assay (RIBA II) to look for HCV-Ab and by the polymerase chain reaction (PCR) to look for HCV viremia., Results: ELISA II was positive in 52% of HD patients and in 14% of CAPD patients. RIBA II was positive in 48% of HD patients and in 11% of CAPD patients. HCV viremia was positive by PCR in 41.6% of HD patients and in 12% of CAPD patients. Two of these PCR-positive patients did not show HCV-Ab by ELISA II and RIBA II. The sensitivity and specificity of ELISA II were 93% and 92%, the sensitivity and specificity of RIBA II were 86% and 94%., Conclusions: Our data confirm a higher prevalence of HCV viremia in HD than in CAPD patients. The absence of Ab against virus C in 2 patients positive with PCR might be due to recent HCV infection or to weak virus replication or to a poor immune response.

Detection of hepatitis C virus viremia (HCV RNA) in serum of hemodialysis (HD) patients is crucial for documenting ongoing infection because the clinical and epidemiological importance of anti-HCV positivity is not clear. HCV viremia was studied in 104 HD patients by reverse transcription polymerase chain reaction (RT PCR) using primers localized in the 5' non-coding region of the viral genome. We used two different methods to detect HCV RNA: a direct PCR amplification of HCV RNA from human serum, and a standard RT PCR procedure (requiring the RNA extraction step). There were 50 (48%) anti-HCV positive patients in this population. Twenty-two (21.1%) out of 104 patients showed HCV RNA in serum by standard RT PCR technique: they belonged to the anti-HCV positive patient group, whereas all anti-HCV negative patients were HCV RNA negative. Prevalence of HCV RNA was more than doubled when standard RT PCR was used compared to direct RT PCR protocol. There was a good association between serum HCV RNA and circulating anti-HCV antibodies, tested by second-generation ELISA and RIBA assays. HCV viremia was not associated with either the presence or the absence of a particular RIBA antibody specificity. AST and ALT levels had no predictive value for HCV viremia, because they were repeatedly normal in the majority of viremic patients (16/22: 73%).(ABSTRACT TRUNCATED AT 250 WORDS)

We conducted a prospective study in HD patients of our unit to evaluate the incidence of seroconversion for HCV in this high-risk group. Two hundred and thirty-five patients were observed during the average follow-up of 29.4 months: 183 were seronegative and 52 seropositive for anti-HCV antibodies at the start of the study. During the observation period two of 183 patients developed anti-HCV antibodies late in the study, while the other 181 patients remained seronegative throughout the observation period; anti-HCV antibodies persisted through the follow-up in the 52 HCV-positive patients at the beginning of the study. Our results showed a very low incidence of HCV seropositivity (0.44% per year) after implementation of our operative protocol including 'universal precautions' and other infection control procedures. Once infected, there is no disappearance rate of anti-HCV. The 4-RIBA results did not change during the follow-up period. Prevalence of HCV RNA by PCR technique was 41% (22 of 54) among anti-HCV-positive patients. Future investigations are warranted to clarify the exact route of transmission of HCV among HD patients and to reduce the rate of HCV transmission in this clinical setting.