Hepatitis C subtype distribution in chronically infected patients with mild liver fibrosis in France: the GEMHEP study.

Treatment options for Hepatitis C infection have greatly improved with direct-acting antiviral (DAA) combinations achieving high cure rates. Nevertheless, the cost of this treatment is still high and access to treatment in many countries has been preferentially reserved for patients with more severe fibrosis (F3 and F4). In this French nationwide study, we investigated the epidemiological characteristics and genotype distribution of hepatitis C virus (HCV) in treatment-naive patients with METAVIR fibrosis stages between F0 and F2 in order to identify patient profiles that became eligible for unrestricted treatment in a second period. Between 2015 and 2016 we collected data from nine French university hospitals on a total of 584 HCV positive patients with absent, mild or moderate liver fibrosis. The most represented genotypes were genotype 1b (159/584; 27.2%), followed by genotype 1a (150/584; 25.7%); genotype 3 (87/584: 14.9%); genotype 4 (80/584; 13.7%). Among genotype 4: 4a was predominantly encountered with 22 patients (27.5% of genotype 4). Genotypes 1b and 1a are currently the most frequent virus types present in treatment-naive patients with mild fibrosis in France. They can be readily cured using the available DAA. Nevertheless, non-a/non-d genotype 4 is also frequent in this population and clinical data on the efficacy of DAA on these subtypes is missing. The GEMHEP is the French group for study and evaluation of viral hepatitis on a national scale. Data collection on epidemiological and molecular aspects of viral hepatitis is performed on a regular basis in all main French teaching hospitals and serves as a basis for surveillance of these infections. Analysis and trends are regularly published on behalf of the GEMHEP group. Data collection was performed retrospectively over the 2015-2016 period, covering nine main university hospitals in France. A total of 584 hepatitis C positive patients were included in this study. Genotyping of the circulating viruses showed a high prevalence of genotypes 1b and 1a in our population. The epidemiology of hepatitis C is slowly changing in France, particularly as a consequence of the rise of 'non-a non-d' genotype 4 viruses mainly originating from African populations. More data concerning treatment efficacy of these genotypes is needed in order to guide clinical care.

Human papillomavirus does not play a role in the Barrett esophagus: a French cohort.

The role of human papillomavirus (HPV) in Barrett's esophagus (BE) has been examined but remains unclear. The purpose of the study is to dispute the connection between HPV and BE in a prospective case-control study. Biopsies were performed above and inside the Barrett's segment for BE patients and in the distal third of the esophagus for control patients for histological interpretation and for virological analysis. Biopsies for virological analysis were placed in a virus transport medium and immediately frozen in liquid nitrogen. Virological analysis involved real-time PCR using the SyBr® green protocol with modified SPF10 general primers. A total of 180 patients (119 control and 61 BE, respectively) were included. In BE patients, 31, 18, and 12 patients had, respectively, no dysplasia, low-grade dysplasia, and high grade dysplasia. Overall, nine were found to be HPV positive: five were control patients and four BE patients. HPV positive status was not associated with BE. No factors were associated with HPV, in particular the degree of BE dysplasia. HPV infection appears unlikely to be significant in the etiology of BE compared with control patients. (ClinicalTrials.gov, Number NCT02549053).

[An experience of hepatitis B control in a rural area in Far North Cameroon]. / Une expérience de lutte contre l'hépatite B en zone rurale à l'extrême nord du Cameroun.

Experience of four years of control of the transmission of hepatitis B in a rural area in Far North Cameroon is presented: (i) prevention of mother to child transmission, (ii) HBsAg screening before blood transfusion, (iii) detection of HIV/HBV co-infections, (iv) protection of healthcare workers. The prevalence of HBsAg is very high in the four populations studied: 18.2% of pregnant women, 16.9% of candidate for blood donation, 14.4% of people living with HIV and 18 % of healthcare workers. Despite limited resources, effective actions are possible. Prevention of mother to child transmission of HBV with vaccination at birth has been set up, with bottlenecks - similar to those observed in HIV - but decreasing over the study. The screening of all potential blood donors has been reached over the years for HIV, HBsAg and HCV, which has led to the eviction of one out of five potential blood donors. Screening of healthcare workers reminded us that adult protection is based on a very early vaccination and not when hiring, even if it is possible to diagnose rare adults eligible for vaccination by research of anti-HBc antibody. A program of hepatitis B control, essential in Africa, appears feasible in rural areas in a framework of an overall improvement in care delivery.

Identification of a duplicated V3 domain in NS5A associated with cirrhosis and hepatocellular carcinoma in HCV-1b patients.

BACKGROUND: The NS5A protein of the hepatitis C virus has been shown to be involved in the development of hepatocellular carcinoma. OBJECTIVES: In a French multicenter study, we investigated the clinical and epidemiological features of a new HCV genotype 1b strain bearing a wide insertion into the V3 domain. STUDY DESIGN: We studied NS5A gene sequences in 821 French patients infected with genotype 1b HCV. RESULTS: We identified an uncharacterized V3 insertion without ORF disruption in 3.05% of the HCV sequences. The insertion comprised 31 amino-acids for the majority of patients; 3 patients had 27 amino-acids insertions and 1 had a 12 amino-acids insertion. Sequence identity between the 31 amino-acids insertions and the V3 domain ranged from 48 to 96% with E-values above 4e(-5), thus illustrating sequence homology and a partial gene duplication event that to our knowledge has never been reported in HCV. Moreover we showed the presence of the duplication at the time of infection and its persistence at least during 12 years in the entire quasispecies. No association was found with extrahepatic diseases. Conversely, patients with cirrhosis were two times more likely to have HCV with this genetic characteristic (p=0.04). Moreover, its prevalence increased with liver disease severity (from 3.0% in patients without cirrhosis to 9.4% in patients with both cirrhosis and HCC, p for trend=0.045). CONCLUSIONS: We identified a duplicated V3 domain in the HCV-1b NS5A protein for the first time. The duplication may be associated with unfavorable evolution of liver disease including a possible involvement in liver carcinogenesis.

Correlation between the promoter basal core and precore mutations and HBsAg quantification in French blood donors infected with hepatitis B virus.

Hepatitis B virus (HBV) basal core promoter (BCP) and precore (PC) mutations, HBV viral load and HBV surface antigen (HBsAg) quantitation were screened to assess correlations between these HBV markers in asymptomatic chronic hepatitis B carriers in France. From January 2006 to July 2007, 200 sera were collected from patients who were discovered to be HBsAg-positive when they volunteered to give blood. Direct sequencing of precore/core gene was used to detect A1762T/G1764A mutations in the BCP and G1896A in the PC region. HBV viral load and HBsAg were quantified with two commercials assays. The prevalence of the BCP and PC mixed/mutants were 37% and 60% respectively (P = 0.0001). HBV DNA level and HBsAg titer were significantly lower in subjects harboring the mixed/mutant PC virus compared to those infected by the wild phenotype. No significant difference was observed in HBV viral loads of blood donors infected by wild or mixed/mutant BCP viruses. Mutant or mixed PC virus was associated with male gender, HBeAb-positive status and HBV/D and HBV/E genotypes. BCP mutations were associated with age, and both HBV/A-HBV/E genotypes.The genetic properties of HBV in this cohort showed that most of the blood donors had a negative HBeAg serological status and harbored the PC mutant phenotype in combination with low levels of both HBV DNA and HBsAg. As the study was conducted in healthy subjects who could be considered as asmptomatic carriers, these results suggest a possible protective effect of the G1896A mutation against severe liver lesions.

Impact of serodiagnosis on the management of Lyme borreliosis at Angers University Hospital.

INTRODUCTION: Lyme borreliosis (LB) is an emerging arthropod-borne disease the diagnosis of which is made on clinical and biological data. We assessed the Angers University Hospital physicians' management of LB, in case of positive serology, and estimated their compliance to European recommendations (EUCALB). METHODS: We retrospectively included 75 cases with positive ELISA serologies confirmed by Western-Blot, performed at the Angers University Hospital between 2008 and 2012. RESULTS AND DISCUSSION: There were 4 cases of early localized phase, 26 of early-disseminated phase (including 17 cases of neuroborreliosis), and one case of late phase. The curative management complied with EUCALB guidelines in 28 cases out of 31. CONCLUSION: Serology remains a reference diagnostic tool for LB, as long as the practitioner is aware of the main clinical and biological criteria.

[Results of a novel real-time PCR, sequence analysis, Inno-LiPA line probe assays in the detection of hepatitis B virus G1896A precore mutation in French blood donors]. / Résultats de trois méthodes pour la détection de la mutation précore G1896A du virus de l'hépatite B chez les donneurs de sang français : PCR temps réel, séquençage et test Inno-LIPA.

AIM: To screen hepatitis B virus (HBV) genotypes and associated basal core promoter (BCP; T1762A/A1764) and precore (PC; A1896) mutations among the 100 HBV surface antigen (HBsAg) positive voluntary blood donors in France. METHODS: HBV genotypes were determined by using direct sequence analysis. Three methods were used to detect G1896A mutation: non-commercial real-time PCR (PCRTR°, line probe assay (InnoLiPA HBV PreCore, INNOGENETICS(®)) and direct sequencing of precore gene. HBV viral load was quantified with two commercial real-time PCR (COBAS(®) AmpliPrep/COBAS(®) TaqMan(®) HBV Test/Roche and Real Time HBV/M2000/Abbott). RESULTS: The mean age of donors was 30 (18-64). Patients were from Africa (42%), Europa (50%), and Asia (8%). HBV/D was the most predominant (37%) genotype followed by HBV/A (31%) and HBV/E (22%). PC and BCP mutants were found in 57% with Inno-LIPA HBV test and 59% with both PCRTR and sequencing methods. A significant difference in the viral load of blood donors with wild and PC mutants was observed with the Taqman Cobas real time PCR (3,19 Log(10) UI/ml versus 4,93 Log(10) UI/ml, p < 0.05). Precore phenotype determination was in agreement with the three PC mutation detection methods in 56% of cases. CONCLUSIONS: Non-Caucasian genotype E was present in the French blood donors. PC mutation was more common than BCP mutations in this study. As HBV infected blood donors were more often asymptomatic carriers, we could speculate that the G1896A mutation may favour the asymptomatic state, supporting previous observations.

Genetic diversity of the hepatitis C virus: impact and issues in the antiviral therapy.

The hepatitis C Virus (HCV) presents a high degree of genetic variability which is explained by the combination of a lack of proof reading by the RNA dependant RNA polymerase and a high level of viral replication. The resulting genetic polymorphism defines a classification in clades, genotypes, subtypes, isolates and quasispecies. This diversity is known to reflect the range of responses to Interferon therapy. The genotype is one of the predictive parameters currently used to define the antiviral treatment strategy and the chance of therapeutic success. Studies have also reported the potential impact of the viral genetic polymorphism in the outcome of antiviral therapy in patients infected by the same HCV genotype. Both structural and non structural genomic regions of HCV have been suggested to be involved in the Interferon pathway and the resistance to antiviral therapy. In this review, we first detail the viral basis of HCV diversity. Then, the HCV genetic regions that may be implicated in resistance to therapy are described, with a focus on the structural region encoded by the E2 gene and the non-structural genes NS3, NS5A and NS5B. Both mechanisms of the Interferon resistance and of the new antiviral drugs are described in this review.

[Use of erythropoietin in the treatment of anemia induced by ribavirin/interferon in patients with hepatitis C]. / Traitement de l'anémie des patients atteints d'une hépatite C chronique sous interféron/ribavirine par l'érythropoïétine.

We are presenting 20 patients with hepatitis C, who developed anemia on interferon alpha-2b/ribavirin treatment and were treated with recombinant human c alpha. Median age was 43 years (range 25-72). Four patients received previous treatment. Interferon-alpha-2b was given at six million units three times a week to 10 patients and at three million units three times a week to five patients. PEG-interferon-alpha-2b (80-120 mug/week) was given to five patients. The dose of ribavirin was 800-1200 mg/day (19 patients) and 200 mg/day (one patient with renal failure). Duration of an interferon/ribavirin treatment was 6-12 months. Baseline median hemoglobin was 13.3 g/dl (range 12.2-15.8); median hemoglobin nadir: 9.8 g/dl (range 8.4-11.2). On erythropoietin, the hemoglobin increased to median 11.7 g/dl (range 9.6-12.8). The ribavirin dose had been decreased to 800 mg in four patients, to 600 mg in four patients, to 400 mg in one patient. Thirteen patients responded to interferon/ribavirin treatment, six patients (all genotype 1) did not. Of the 13 initial responders 11 had sustained response, one still under treatment and two patients relapsed. In conclusion, in our patients with chronic hepatitis C treated with interferon/ribavirin combination therapy, erythropoietin was beneficial in the treatment of ribavirin-induced anemia.

[Quantitative antibody analysis: use for the diagnosis of hepatitis C virus chronic infection]. / Analyse quantitative des anticorps dirigés contre le virus de l'hépatite C (VHC): application au diagnostic d'infection chronique par le VHC.

Hepatitis C virus (HCV) infection has been estimated in 600,000 subjects in France, with about 80 % of chronic infection. In the latter, anti-HCV antibodies and viral RNA are found together in patients blood. Today, only the use of polymerase chain reaction (PCR) technology allows the diagnosis of HCV chronic infection, confirmed by a positive PCR. However, PCR is a laborious and cost effective method. The aim of this study was to distinguish HCV chronic infection to past-infection or false reactivity only using the serology testing. Therefore, we looked for a correlation between the results of PCR, using the HCV Cobas Amplicor 2.0 assay, and the level of anti-HCV antibodies, assessed by the AxSYM HCV v.3.0 and expressed in signal/cutoff (s/co) ratio. We found using a panel of 200 sera issued from 181 patients, a significant variation of s/co ratios between PCR positive and negative patients (respectively, 87.76 +/- 27.18 vs 10.13 +/- 13.68 s/co, p < 0.0001), only in non treated or previously treated patients, non HIV coinfected, non renal transplanted or haemodialysis patients. An anti-HCV cutoff value at 34 s/co allows a predictive PCR results with 100 % sensitivity and 93.3 % specificity. Thus, for patients having a s/co equal or over 34, a positive PCR was found in 98.1 % of cases, allowing the diagnosis of HCV chronic infection (positive predictive value). Conversely, in patients with less than 34, HCV chronic infection can be excluded in 100 % of cases (negative predictive value). In conclusion, in most cases, the use of anti-HCV quantitative analysis in the AxSYM HCV v.3.0 assay could avoid PCR testing and facilitate the diagnosis of HCV chronic infection.

Ribavirin in hepatitis C related cryoglobulinemia.

OBJECTIVE: An open, uncontrolled trial of ribavirin, an oral guanosine nucleoside analog for treatment of hepatitis C, in patients with hepatitis C virus (HCV) associated cryoglobulinemia intolerant to interferon. METHODS: Five patients with cryoglobulinemia related to HCV infection unresponsive to interferon therapy received oral ribavirin (100 to 1200 mg daily) for 10 to 36 months. RESULTS: Patients treated with ribavirin had prompt decrease in serum aminotransferase levels and marked improvement of manifestations of cryoglobulinemia within a few weeks. Ribavirin did not eradicate HCV RNA from the sera, but a decrease in viral load was observed in 3 patients, from 232 to 86 x 10(5) copies HCV/ml. Relapse occurred within 3 months once therapy was discontinued. The drug was well tolerated, but mild dose related hemolysis was common. CONCLUSION: Ribavirin monotherapy may be effective in patients with symptomatic cryoglobulinemia related to HCV infection, but this effect is not sustained when ribavirin therapy is discontinued.

[Human herpesvirus-6 infection and hepatitis following heart transplantation]. / Infection à herpèsvirus humain-6 et hépatite chez les transplantés cardiaques.

OBJECTIVES: Human Herpesvirus-6 (HHV-6) has been implicated in hepatitis and HHV-6 infections have been reported in patients receiving heart transplants. We investigated the occurrence of such infections and their possible relation with post-transplantation hepatitis in 58 heart transplanted patients. METHODS: The titre of HHV-6 antibodies was determined by anticomplement immunofluorescence assay and compared with the titre of CMV antibodies determined by using ELISA. RESULTS: HHV-6 seroprevalence after transplantation did not differ significantly between hepatitis, control patients (without hepatitis) and the healthy general population. In three controls and five hepatitis patients we observed, after transplantation, a seroconversion or a significant increase of antibody titre which suggested active HHV-6 infection. In six cases, HHV-6-specific IgM were found whereas CMV-specific IgM were not detected. CONCLUSION: These results indicate that HHV-6 infection is frequent after heart transplantation but not in relation with hepatitis. The mechanism of this infection needs to be clarified (either primo-infection or reactivation) and the question remains whether this infection is totally asymptomatic or could induce serious illness.

[Screening tests of anti-HVC antibodies used in France. Analysis of sensitivity]. / Les trousses de dépistage des anticorps anti-VHC utilisées en France. Analyse de la sensibilité. Groupe de Travail "Hépatites Virales" de la Société Française de Transfusion Sanguine.

The results concerning the hepatitis C antibodies screening tests sensitivity are presented by Viral Hepatitis Study Group of the French Society of Blood Transfusion. This evaluation was carried out in 1993, with five tests available for Hepatitis C Viral (HCV) antibodies screening. Several panels of human blood samples, collected by members of Viral Hepatitis Study Group, were used. For those specimens the data about whole serology (various reactivities against viral antigens) clinical history and other explorations (molecular biology, liver histology, other viral infections) are described. If possible all those results are used to describe some interpretation criteria for the HCV serology and sensitivity obtained for each screening test.

Hepatitis C virus and essential mixed cryoglobulinaemia.

Clinical and laboratory evidence of liver involvement are frequently found in essential mixed cryoglobulinaemia (EMC). We looked for evidence of hepatitis C virus (HCV) infection in 37 patients with EMC. Anti-HCV antibodies (Ab) were found in 16/37 (43%) patients with EMC using the ELISA 2 test and the RIBA 2 test. The 16 anti-HCV-Ab positive patients (group 1) were compared to the 21 anti-HCV-Ab negative patients (group 2). Group 1 patients had more frequent cutaneous involvement (P = 0.02), clinical, biological and histologic hepatic involvement (P < 0.01), higher serum cryoglobulin and lower CH50 levels (P < 0.001). Serum hepatitis B virus markers were infrequent in both groups and no patient from either group had detectable serum HBV DNA. These preliminary results suggest that HCV may be another cause of mixed cryoglobulinaemia.

[Diagnosis of viral hepatitis in 1991]. / Le diagnostic des hépatites virales en 1991.

Viral hepatitis is a disease which is frequent but often asymptomatic and therefore remains unrecognized until a late stage. Hence the importance of biochemical and, chiefly, virological detection in populations at risk such as transfusion recipients, drug addicts or exposed professionals. Detection is based on systematic transaminase assays during check-ups or in the presence of warning signs. Any rise in transaminase level must lead to serological examinations in search of the A, B, C and Delta markers. Routine vaccination of subjects at risk against hepatitis B should considerably reduce the prevalence of this disease and of Delta hepatitis in the populations exposed.

Protein F. A novel F(ab)-binding factor, present in normal liver, and largely released in the digestive tract during hepatitis.

Significant percentages of patients suffering from non-A non-B hepatitis (43%) and B hepatitis (35%) were found to release an Ig-binding factor in their stools. This factor, which we called "protein F" was less frequently observed (20%) in patients suffering from other liver disorders, and was found in only 6.7% of healthy subjects (p less than 10(-7), less than 10(-4), and less than 0.03, respectively). The specificity of the detection test (a nonimmune ELISA-like assay) was confirmed by inhibition experiments. Binding was located on the F(ab) fragment of Ig, irrespectively of their isotype. Protein F was inactivated by pepsin, neuraminidase, and high concentrations of subtilisin, whereas it was resistant to trypsin and chymotrypsin. Molecular sieving by HPLC indicated an apparent molecular mass of 175 kDa. In preparative SDS-PAGE, the molecular mass was 85 kDa in favor of a dimer disrupted under dissociating conditions. Preparative IEF showed the isoelectric charge to lie between 3.9 and 4.1. Analysis of liver extracts from two patients suffering fron non-A non-B hepatitis, and from a transplant donor, revealed the presence of the factor in the three cases.