S. Sauleda

Clinical outcome of acute and chronic hepatitis delta over time: a long-term follow-up study.

Summary.u2002 Long‐term changes in the frequency and outcome of hepatitis delta virus (HDV) infection have seldom been analysed. This retrospective, longitudinal study includes 398 consecutive hepatitis B surface antigen (HBsAg)‐positive patients with anti‐HDV antibodies who attended our institution between 1983 and 2008. At enrolment, 182 patients had acute and 216 chronic hepatitis. Patients were grouped into two periods. Those who attended between 1983 and 1995 and those between 1996 and 2008. The former group was significantly younger, mainly intravenous drugs users, and had a greater incidence of acute HDV and HIV and HCV coinfection. Patients with acute HBV/HDV coinfection cleared both infections in 90% of cases, while all patients with HDV superinfection evolved to chronic disease. One hundred and fifty‐eight patients with chronic HDV were followed for a median period of 158u2003months. Seventy‐two per cent of the patients remained stable, 18% had hepatic decompensation, 3% developed hepatocellular carcinoma, and 8% cleared HBsAg. Liver‐related death was observed in 13% of patients and mainly occurred in patients from the first period (Pu2003=u20030.012). These results indicate an outbreak of HDV at the end of the 1980s and the beginning of the 1990s, with a large number of acute HDV cases affecting predominately young, male intravenous drug users. Currently, patients with chronic HDV disease are older, and factors associated with worse prognosis include the presence of cirrhosis and age at the time of diagnosis.

High-Resolution Hepatitis C Virus Subtyping Using NS5B Deep Sequencing and Phylogeny, an Alternative to Current Methods

ABSTRACT Hepatitis C virus (HCV) is classified into seven major genotypes and 67 subtypes. Recent studies have shown that in HCV genotype 1-infected patients, response rates to regimens containing direct-acting antivirals (DAAs) are subtype dependent. Currently available genotyping methods have limited subtyping accuracy. We have evaluated the performance of a deep-sequencing-based HCV subtyping assay, developed for the 454/GS-Junior platform, in comparison with those of two commercial assays (Versant HCV genotype 2.0 and Abbott Real-time HCV Genotype II) and using direct NS5B sequencing as a gold standard (direct sequencing), in 114 clinical specimens previously tested by first-generation hybridization assay (82 genotype 1 and 32 with uninterpretable results). Phylogenetic analysis of deep-sequencing reads matched subtype 1 calling by population Sanger sequencing (69% 1b, 31% 1a) in 81 specimens and identified a mixed-subtype infection (1b/3a/1a) in one sample. Similarly, among the 32 previously indeterminate specimens, identical genotype and subtype results were obtained by direct and deep sequencing in all but four samples with dual infection. In contrast, both Versant HCV Genotype 2.0 and Abbott Real-time HCV Genotype II failed subtype 1 calling in 13 (16%) samples each and were unable to identify the HCV genotype and/or subtype in more than half of the non-genotype 1 samples. We concluded that deep sequencing is more efficient for HCV subtyping than currently available methods and allows qualitative identification of mixed infections and may be more helpful with respect to informing treatment strategies with new DAA-containing regimens across all HCV subtypes.

T cell responses and viral variability in blood donation candidates with occult hepatitis B infection.

BACKGROUND & AIMSnOccult HBV infection (OBI) is defined by the presence of HBV DNA in the liver and/or serum and negative HBsAg testing. Since the implementation of highly sensitive HBV DNA screening, OBI is also detected in healthy blood donors. The aims of this study were to investigate HBV-specific immune responses and genetic variability in donors with OBI, established by HBV DNA in serum.nnnMETHODSnHBV-specific T-cell responses to HBV antigens were tested in 34 OBI donors by IFN-γ ELISpot, cytometric bead array, and intracellular cytokine staining. As comparison populations, 36 inactive HBV carriers, 22 donors with spontaneously resolved HBV infection, 24 vaccinated donors, and 25 seronegative donors were also included. Surface, pre-S, and pre-c/core genes from 44 genotype D isolates (24 OBI and 20 HBsAg-positive) were sequenced.nnnRESULTSnThe immune response of OBI donors to the 3 HBV antigens was 29-41%, similar to the response in subjects with resolved HBV infection and higher than that in HBsAg-positive subjects. On sequence analysis, OBI donors presented a higher HBsAg mutation rate than HBsAg-positive subjects. Mutations were clustered in the major hydrophilic region of HBsAg, and no stop codons or relevant mutations that could affect antigen formation or detection were observed.nnnCONCLUSIONSnOur results suggest that immune response can suppress viral replication to low levels and HBsAg expression to undetectable levels in OBI blood donors. Relevant mutations were not found in the genomic HBsAg coding region. Hence, the fact that HBsAg was not detected in OBI is likely due to low HBsAg production, rather than to a failure of laboratory reagents.

Early detection of nonresponse to interferon plus ribavirin combination treatment of chronic hepatitis C

We have investigated the value of early hepatitis C virus (HCV) RNA decline (ΔHCV RNA) to predict response to combination therapy in 66 chronic hepatitis C patients treated with IFN‐α2b (3u2003MU thrice weekly) and ribavirin (800u2003mg daily) for 12 months [25 sustained responders (SR) and 41 nonresponders or relapsers (NR)]. Serum HCV RNA was retrospectively measured in samples obtained at baseline and 4, 8 and 12u2003weeks after treatment onset, using a commercially available quantitative RT‐PCR assay. At 4u2003weeks, serum HCV RNA had decreased a mean of 2.6u2003±u20030.8 logs among SR as compared with only 0.5u2003±u20030.8 logs in NR (Pu2003<u20030.001), and was already undetectable (<u2003600u2003IU/mL) in 12 (48%) of the SR but in none of the NR. At 8u2003weeks, HCV RNA was undetectable in 21 SR and in 2 NR and mean ΔHCV RNA were 4.2u2003±u20031.3 and 0.8u2003±u20031.0 logs, respectively (Pu2003<u20030.001). At week 12 all SR had undetectable HCV RNA as compared with only five NR (Pu2003<u20030.001). Stepwise logistic regression analysis identified ΔHCV RNA at 12u2003weeks as the strongest predictor of sustained response. Receiver operating characteristic (ROC) curves of ΔHCV RNA for sustained response prediction identified sensitivity peaks with 100% negative predictive value corresponding to ΔHCV RNAu2003>u20031 log at 4u2003weeks, >u20032 logs at 8u2003weeks and >u20033 logs at 12u2003weeks. Our results show that early changes in the HCV RNA level may reliably identify patients having no chance of a sustained virological response during the first 3u2003months of combination therapy, thus providing an excellent tool for optimizing antiviral treatment of chronic hepatitis C.

Evaluation of RNA and E2 antibodies in prospectively followed recipients of hepatitis G virus-infected blood.

BACKGROUND: Hepatitis G virus (HGV) has recently been cloned and tests for HGV RNA and envelope antibodies (anti‐E2) have been developed. HGV infection is widespread among blood donors worldwide, but the clinical and serologic outcome of transfusion‐associated HGV infection has not been fully characterized.

Serum hepatitis B core-related antigen is more accurate than hepatitis B surface antigen to identify inactive carriers, regardless of hepatitis B virus genotype

OBJECTIVESnTo investigate whether hepatitis B surface antigen (HBsAg) and hepatitis B core-related antigen (HBcrAg) levels are useful to identify inactive carriers among HBeAg-negative patients infected by different hepatitis B virus (HBV) genotypes.nnnMETHODSnIn all, 202 consecutive HBeAg-negative patients with chronic hepatitis B, 135 inactive carriers and 67 with HBV activity, were prospectively followed for 1 year.nnnRESULTSnIn HBeAg-negative patients, HBsAg levels differed across the different genotypes (p <0.001). The highest levels were observed in genotypes F or H (4.2xa0±xa00.6xa0logIU/mL), followed by genotype E (3.4xa0±xa01.1xa0logIU/mL), genotype A (3.4xa0±xa00.8xa0logIU/mL), and the lowest in genotype D (2.7xa0±xa01.1xa0logIU/mL). Variations in HBsAg levels were similar in inactive carriers and patients with HBV activity. HBsAg <3xa0logIU/mL showed good performance for identifying genotype D inactive carriers: 76% of genotype D inactive carriers met this cut-off versus ≤31% for genotypes A, E, F or H. However, in patients with genotype A, HBsAg levels ≤3.7xa0logIU/mL better classified inactive carriers. The combination of a single measurement of HBcrAg ≤3xa0logU/mL plus HBV DNA ≤2000xa0IU/mL yielded a positive predictive value and diagnostic accuracy >85% in all HBV genotypes, except genotype H or F, with values of 62.5% and 72.7%, respectively, for the two parameters.nnnCONCLUSIONSnHBsAg levels varied across genotypes in HBeAg-negative patients. HBsAg levels <3xa0logIU/mL were only useful for identifying genotype D inactive carriers. A single HBcrAg measurement ≤3xa0logU/mL plus HBV DNA ≤2000xa0IU/mL was highly accurate for identifying inactive carriers, regardless of their HBV genotype.

Red blood cell transfusion‐transmitted acute hepatitis E in an immunocompetent subject in Europe: a case report

Acute hepatitis E in industrialized countries is usually related to intake or manipulation of undercooked or raw meat. Cases of transfusion‐transmitted hepatitis E have rarely been documented in immunosuppressed patients, mainly after receiving frozen plasma.

Identification of host and viral factors involved in a dissimilar resolution of a hepatitis C virus infection.

Hepatitis C virus (HCV) transmission from a chronic patient to a susceptible individual is a good opportunity to study viral and host factors that may influence the natural course of hepatitis C infection towards either spontaneous recovery or chronicity. To compare a documented case of a bottleneck event in the sexual transmission of HCV from a chronically infected patient to a recipient host that cleared infection.

Reversal of nonstructural protein 3‐specific CD4+ T cell dysfunction in patients with persistent hepatitis C virus infection

Summary.u2002 Hepatitis C virus (HCV)‐specific T cell responses are essential for HCV control, and chronic infection is characterized by functionally altered antigen‐specific T cells. It has been proposed that the early inactivation of specific CD4+ T cell responses may be involved in establishment of HCV persistence. We have investigated whether HCV‐specific CD4+ T cells dysfunction can be reversed in vitro. Nonstructural protein 3 (NS3) and core‐specific CD4+ T cells from eight chronically infected and eight spontaneously resolved HCV individuals were selected through transient CD154 (CD40 ligand) expression, and their functional profile (IFN‐γ, IL‐2, TNF‐α, IL‐10 and IL‐4 production by enzyme‐linked immunospot assay, cytometric bead array and intracellular cytokine staining, and proliferation by carboxy‐fluorescein diacetate succinimidyl ester dilution assay) was determined both ex vivo and after in vitro expansion of sorted CD154‐expressing cells in the absence of specific antigen in IL‐7/IL‐15‐supplemented medium. Ex vivo bulk CD4+ T cells from chronic patients expressed CD154 in most cases, albeit at lower frequencies than those of resolved patients (0.11%vs 0.41%; Pu2003=u20030.01), when stimulated with NS3, but not core, although they had a markedly impaired capacity to produce IL‐2 and IFN‐γ. Antigen‐free in vitro expansion of NS3‐specific CD154+ cells from chronic patients restored IFN‐γ and IL‐2 production and proliferation to levels similar to those of patients with spontaneously resolved infection. Hence, NS3‐specific CD4+ T cell response can be rescued in most chronic HCV patients by in vitro expansion in the absence of HCV‐specific antigen. These results might provide a rationale for adoptive immunotherapy.

IL28B genetic variation and hepatitis C virus–specific CD4+ T-cell responses in anti-HCV-positive blood donors

Summary.u2002 Epidemiological, viral and host factors are associated with the outcome of hepatitis C virus (HCV) infection, and strong host immune responses against HCV favour viral clearance. Recently, genome‐wide association studies have shown a strong correlation between single‐nucleotide polymorphisms (SNPs) near the interleukin‐28B (IL28B) gene and spontaneous or treatment‐induced HCV clearance. We have investigated whether protective IL28B genetic variants are associated with HCV‐specific T‐cell responses among Spanish blood donors. The rs12979860 IL28B haplotype was determined in 69 anti‐HCV‐positive blood donors (21 HCV RNA negative and 48 HCV RNA positive) and 30 seronegative donors. In all cases, HCV‐specific CD4+ T‐cell responses to HCV recombinant proteins (core, NS3 and NS3 helicase) were assessed by ex vivo interferon‐γ ELISpot assay. The rs12979860‐CC genotype was highly overrepresented in donors with spontaneous HCV clearance when compared to those with chronic infection (76.2%vs 29.2%, Pu2003<u20030.001; odds ratio, 7.77; 95% confidence interval, 2.4–25.3, Pu2003<u20030.001). HCV‐specific CD4+ T‐cell responses were detected in 16 (76.2%) spontaneous resolvers especially towards nonstructural proteins, but with no correlation with IL28B genotype. Chronic individuals had a significantly lower overall T‐cell response again irrespective of IL28B genotype. When spontaneous resolvers and chronic individuals were stratified according to their IL28B genotype, significantly stronger T‐cell responses were only observed among those with non‐CC haplotypes. Although the protective rs12979860 IL28B CC genotype is associated with spontaneous HCV clearance, stronger CD4+ T‐cell responses towards NS3 were only evident among those with non‐CC haplotypes.