Archives of Virology

, 153:1703

Monoclonal antibodies raised against the ORF3 protein of hepatitis E virus (HEV) can capture HEV particles in culture supernatant and serum but not those in feces

  • Masaharu Takahashi
  • Kentaro Yamada
  • Yu Hoshino
  • Hideyuki Takahashi
  • Koji Ichiyama
  • Toshinori Tanaka
  • Hiroaki Okamoto
Original Article

DOI: 10.1007/s00705-008-0179-6

Cite this article as:
Takahashi, M., Yamada, K., Hoshino, Y. et al. Arch Virol (2008) 153: 1703. doi:10.1007/s00705-008-0179-6

Abstract

Ten murine monoclonal antibodies (MAbs) against a synthetic peptide corresponding to the well-conserved, C-terminal 24-amino acid portion of ORF3 protein of hepatitis E virus (HEV) were produced and characterized. Immunofluorescent assays using the anti-ORF3 MAbs revealed accumulation of ORF3 protein in the cytoplasm of PLC/PRF/5 cells transfected with ORF3-expressing plasmid or inoculated with cell-culture-generated HEV. The anti-ORF3 MAbs could capture HEV particles in culture medium and serum at variable efficiency of up to 61 and 49%, respectively, but not those in feces. By sandwiching between immobilized and enzyme-labeled anti-ORF3 MAbs in ELISA, ORF3 antigen was detected in the culture media with an HEV RNA titer of >106 copies/ml and increased in parallel with the increase in HEV load. HEV progenies in the culture supernatant, with ORF3 protein on the surface, banded at a low buoyant density of 1.15 g/cm3 in sucrose. A representative anti-ORF3 MAb (TA0536) could partially neutralize the infection of cell-culture-generated HEV in a cell culture system. These results indicate that ORF3 protein, at least its C-terminal portion, is present on the surface of HEV virions released from infected cells and support a previously proposed assumption that ORF3 protein is associated with virus release from infected cells.

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Masaharu Takahashi
    • 1
  • Kentaro Yamada
    • 1
  • Yu Hoshino
    • 1
  • Hideyuki Takahashi
    • 1
  • Koji Ichiyama
    • 1
  • Toshinori Tanaka
    • 1
  • Hiroaki Okamoto
    • 1
  1. 1.Division of Virology, Department of Infection and ImmunityJichi Medical University School of MedicineShimotsuke-ShiJapan