title: Assembly of hepatitis delta virus: particle characterization, including the ability to infect primary human hepatocytes.
abstract: Efficient assembly of hepatitis delta virus (HDV) was achieved by cotransfection of Huh7 cells with two plasmids: one to provide expression of the large, middle, and small envelope proteins of hepatitis B virus (HBV), the natural helper of HDV, and another to initiate replication of the HDV RNA genome. HDV released into the media was assayed for HDV RNA and HBV envelope proteins and characterized by rate-zonal sedimentation, immunoaffinity purification, electron microscopy, and the ability to infect primary human hepatocytes. Among the novel findings were that (i) immunostaining for delta antigen 6 days after infection with 300 genome equivalents (GE) per cell showed only 1% of cells as infected, but this was increased to 16% when 5% polyethylene glycol was present during infection; (ii) uninfected cells did not differ from infected cells in terms of albumin accumulation or the presence of E-cadherin at cell junctions; and (iii) sensitive quantitative real-time PCR assays detected HDV replication even when the multiplicity of infection was 0.2 GE/cell. In the future, this HDV assembly and infection system can be further developed to better understand the mechanisms shared by HBV and HDV for attachment and entry into host cells.
authors.affiliation: Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, PA 19111-2497, USA.
authors.forename: Severin
authors.initials: S
authors.lastname: Gudima
authors.forename: Yiping
authors.initials: Y
authors.lastname: He
authors.forename: Anja
authors.initials: A
authors.lastname: Meier
authors.forename: Jinhong
authors.initials: J
authors.lastname: Chang
authors.forename: Rongji
authors.initials: R
authors.lastname: Chen
authors.forename: Michal
authors.initials: M
authors.lastname: Jarnik
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Nicolas
authors.forename: Volker
authors.initials: V
authors.lastname: Bruss
authors.forename: John
authors.initials: J
authors.lastname: Taylor
mesh_terms: D000956:Antigens, Viral / Q000276:immunology / Q000378:metabolism; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003698:Hepatitis Delta Virus / Q000502:physiology* / Q000648:ultrastructure; D022781:Hepatocytes / Q000378:metabolism* / Q000821:virology*; D006801:Humans; D046529:Microscopy, Electron, Transmission; D016253:Microscopy, Immunoelectron; D012367:RNA, Viral / Q000378:metabolism; D013997:Time Factors; D014759:Viral Envelope Proteins / Q000276:immunology / Q000378:metabolism; D014771:Virion / Q000276:immunology / Q000378:metabolism*; D019065:Virus Assembly / Q000276:immunology; D014779:Virus Replication
chemical_list: D000956:Antigens, Viral; D012367:RNA, Viral; D014759:Viral Envelope Proteins