Long-term propagation of serum hepatitis C virus (HCV) with production of enveloped HCV particles in human HepaRG hepatocytes.

International audienceUNLABELLED: HepaRG human liver progenitor cells exhibit morphology and functionality of adult hepatocytes. We investigated the susceptibility of HepaRG hepatocytes to in vitro infection with serum-derived hepatitis C virus (HCV) particles (HCVsp) and the potential neutralizing activity of the E1E2-specific monoclonal antibody (mAb) D32.10. The infection was performed using HCVsp when the cells actively divided at day 3 postplating. HCV RNA, E1E2, and core antigens were quantified in HCV particles recovered from culture supernatants of differentiated cells for up to 66 days. The density distributions of particles were analyzed on iodixanol or sucrose gradients. Electron microscopy (EM) and immune-EM studies were performed for ultrastructural analysis of cells and localization of HCV E1E2 proteins in thin sections. HCV infection of HepaRG cells was documented by increasing production of E1E2-core-RNA(+) HCV particles from day 21 to day 63. Infectious particles sedimented between 1.06 and 1.12 g/mL in iodixanol gradients. E1E2 and core antigens were expressed in 50% of HCV-infected cells at day 31. The D32.10 mAb strongly inhibited HCV RNA production in HepaRG culture supernatants. Infected HepaRG cells frozen at day 56 were reseeded at low density. After only 1-3 subcultures and induction of a cell differentiation process the HepaRG cells produced high titer HCV RNA and thus showed to be sustainably infected. Apolipoprotein B-associated empty E1E2 and complete HCV particles were secreted. Characteristic virus-induced intracellular membrane changes and E1E2 protein-association to vesicles were observed. CONCLUSION: HepaRG progenitor cells permit HCVsp infection. Differentiated HepaRG cells support long-term production of infectious lipoprotein-associated enveloped HCV particles. The E1E2-specific D32.10 mAb neutralizes the infection and this cellular model could be used as a surrogate infection system for the screening of entry inhibitors

Acceptability and perception of the herpes zoster vaccine in the 65 and over population: A French observational study

International audienceL’objectif de l’étude était d’évaluer l’acceptabilité et de décrire la perception de la vaccination contre l’herpès zoster (HZ) chez les patients ambulatoires et hospitalisés à Lyon, en France, âgés de 65 ans et plus. Une étude observationnelle était basée sur un questionnaire rempli lors d’une entrevue en personne de janvier 2018 à mars 2019. Les patients externes bénévoles qui ont fréquenté des laboratoires médicaux privés ou qui ont été hospitalisés dans le service de gériatrie, ou qui étaient à la clinique médicale ambulatoire pour une consultation ont été invités à participer. Au total, 907 personnes ont été interrogées, avec un âge moyen de 75,8 ans. Une grande majorité de 87,6 % (795) connaissaient le ZONA et 68,9 % (625) accepteraient d’être vaccinés contre l’AS S’ils présentaient des facteurs de risque. Les participants connaissaient le HZ en tant que maladie, mais la sensibilisation au vaccin fait encore défaut dans le grand public

GEMHEP multicenter quality control study of PCR detection of GB virus C/hepatitis G virus RNA in serum

International audiencePCR is, to date, the only available tool for the detection of GB virus C (GBV-C) and hepatitis G virus (HGV) RNAs. Twenty-two French laboratories participated in a quality control study to assess the sensitivity and specificity of their procedures. The panel included 13 positive controls and 7 negative controls. The laboratories used either in-house PCR techniques adapted from the literature or partly standardized commercial tests. Three laboratories performed faultlessly with the entire panel. Most laboratories had excellent specificity (100% in 20 of 22 laboratories). Sensitivity was acceptable (85 to 100%) in 15 centers and insufficient (38 to 77%) in 7. As with nonstandardized in-house PCR, the commercial assays gave discrepant performances in different laboratories. These results suggest that laboratories willing to use PCR for detection of GBV-C/HGV RNA for research or diagnostic purposes should participate in multicenter quality control trials