1. Cycle infectieux du virus de l'hépatite E et identification de 3 formes de la protéine de capside ORF2
- Author
-
Montpellier, Claire, Wychowski, Czeslaw, Sayed, Ibrahim, Meunier, Jean-Christophe, Saliou, Michel, Ankavay, Maliki, Bull, Anne, Pillez, André, Legrand-Abravanel, Florence, Helle, Francois, Brochot, Etienne, Drobecq, Hervé, Farhat, Rayan, Aliouat-Denis, Cécile-Marie, Haddad, Juliano, Izopet, Jacques, Meuleman, Philip, Goffard, Anne, Dubuisson, Jean, Cocquerel, Laurence, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Universiteit Gent = Ghent University [Belgium] (UGENT), Assiut University, Morphogénèse et antigénicité du VIH et du virus des Hépatites (MAVIVH - U1259 Inserm - CHRU Tours ), Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire de Tours (CHRU TOURS), Laboratoire de Virologie [CHU Purpan, Toulouse], Institut Fédératif de Biologie (IFB) - Hôpital Purpan, Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse]-Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse], Unité de Virologie clinique et fondamentale (UVCF), Université de Picardie Jules Verne (UPJV)-CHU Amiens-Picardie, Mécanismes de la Tumorigénèse et Thérapies Ciblées - UMR 8161 (M3T), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Virologie [Toulouse], CHU Toulouse [Toulouse], This work was supported by the French 'Agence Nationale de la Recherche sur le Sida et les hépatites virales' (ANRS) and the University of Lille. M.A. was supported by a fellowship from the ANRS. P.M. was supported by grants from the Research Foundation Flanders (FWO Vlaanderen), Ghent University (BOF GOA and IRO), and the Belgian State (BELSPO IAP HEPRO-2). I.M.S. was supported with PhD fellowships from the Egyptian Government and Ghent University. J.G.H. was supported by a fellowship from the Lebanese Association for Development., We thank Sandrine Belouzard, Karin Seron, and Sophana Ung for their technical contribution. We thank Suzanne U. Emerson (National Institutes of Health) and Shoshana Levy (Stanford University) for providing us with reagents. We thank Lydia Linna for proofreading the manuscript., Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), Universiteit Gent = Ghent University (UGENT), Centre Hospitalier Régional Universitaire de Tours (CHRU Tours)-Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire Virologie [CHU Toulouse], Institut Fédératif de Biologie (IFB), Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Pôle Biologie [CHU Toulouse], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), and CHU Amiens-Picardie-Université de Picardie Jules Verne (UPJV)
- Subjects
MESH: Cell Culture Techniques ,MESH: Humans ,MESH: Viral Proteins/isolation & purification ,MESH: Hepatitis E/pathology ,viruses ,ORF2 Products ,MESH: Capsid Proteins/isolation & purification ,virus diseases ,Hepatitis E ,MESH: Hepatocytes ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,Infectious Particles ,PLC/PRF/5 Cells ,MESH: Hepatitis E/metabolism ,MESH: Animals ,MESH: Disease Models, Animal ,MESH: Mice ,MESH: Hepatitis E/etiology ,MESH: Hepatitis E virus/physiology - Abstract
Comment in : New Models to Study Hepatitis E Virus Replication and Particular Characteristics of Infection: The Needle Hides in the Hay Stack. [Gastroenterology. 2018]; International audience; BACKGROUND & AIMS: Hepatitis E virus (HEV) infection is a major cause of acute hepatitis worldwide. Approximately 2 billion people live in areas endemic for HEV and are at risk of infection. The HEV genome encodes 3 proteins, including the ORF2 capsid protein. Detailed analyses of the HEV life cycle has been hampered by the lack of an efficient viral culture system.METHODS: We performed studies with gt3 HEV cell culture-produced particles and patient blood and stool samples. Samples were fractionated on iodixanol gradients and cushions. Infectivity assays were performed in vitro and in human liver chimeric mice. Proteins were analyzed by biochemical and proteomic approaches. Infectious particles were analyzed by transmission electron microscopy. HEV antigen levels were measured with the Wantaï enzyme-linked immunosorbent assay.RESULTS: We developed an efficient cell culture system and isolated HEV particles that were infectious in vitro and in vivo. Using transmission electron microscopy, we defined the ultrastructure of HEV cell culture-produced particles and particles from patient sera and stool samples. We also identified the precise sequence of the infectious particle-associated ORF2 capsid protein. In cultured cells and in samples from patients, HEV produced 3 forms of the ORF2 capsid protein: infectious/intracellular ORF2 (ORF2i), glycosylated ORF2 (ORF2g), and cleaved ORF2 (ORF2c). The ORF2i protein associated with infectious particles, whereas the ORF2g and ORF2c proteins were massively secreted glycoproteins not associated with infectious particles. ORF2g and ORF2c were the most abundant antigens detected in sera from patients.CONCLUSIONS: We developed a cell culture system and characterized HEV particles; we identified 3 ORF2 capsid proteins (ORF2i, ORF2g, and ORFc). These findings will advance our understanding of the HEV life cycle and improve diagnosis.
- Published
- 2017
- Full Text
- View/download PDF