Archives of Virology

, Volume 160, Issue 10, pp 2503–2516 | Cite as

Establishment of persistent foot-and-mouth disease virus (FMDV) infection in MDBK cells

  • Lela Kopliku
  • Anthony Relmy
  • Aurore Romey
  • Kamila Gorna
  • Stephan Zientara
  • Labib Bakkali-Kassimi
  • Sandra Blaise-Boisseau
Original Article

Abstract

In addition to acute infection and disease, foot-and-mouth disease virus (FMDV) can cause persistent infection in ruminants. Such “carrier” animals represent a potential risk for FMDV transmission to susceptible animals. However, the mechanisms and the factors that determine FMDV persistence remain unknown. We describe here the establishment of FMDV type O persistent infection in a bovine epithelial cell line (Madin-Darby bovine kidney; MDBK). Preliminary experiments to assess the permissivity of MDBK cells to FMDV O infection revealed an unusual pattern of infection: after the initial phase of acute cell lysis, new monolayers formed within 48-72 h post-infection. We found that some cells survived cytolytic infection and subsequently regrew, thereby demonstrating that this bovine cell line can be persistently infected with FMDV type O. Further evidence that MDBK cells were persistently infected with FMDV includes: (i) detection of viral RNA in cells as well as in cell culture supernatants, (ii) detection of viral antigens in the cells by immunofluorescence analysis, and (iii) production of infectious viral particles for up to 36 cell passages. Furthermore, preliminary sequence analysis of persistent virus revealed a single nucleotide substitution within the VP1 coding region, resulting in the V50A amino acid substitution. This bovine model of FMDV persistence holds promise for the investigation of the viral and cellular molecular determinants that promote FMDV persistence.

Supplementary material

705_2015_2526_MOESM1_ESM.tif (4.9 mb)
S1 Immunofluorescence analysis of MDBK cells infected with FMDV O/FRA/1/2001. MDBK cells grown in 96-well tissue cultures plates were mock-infected or infected with FMDV O/FRA/1/2001 at an MOI of 0.005, 0.05 or 0.5. At 24 h p.i., cells were washed and fixed with 4 % PFA and incubated with a monoclonal antibody raised against the VP2 protein of FMDV type O (anti-VP2, mAb 13G11). Anti-mouse Alexa Fluor 546 (red fluorescence) was used as secondary antibody. Nuclei were counterstained with DAPI (blue fluorescence). All photographs are at the same magnification (x 10). Scale bar = 200 µm (TIFF 5012 kb)

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Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  • Lela Kopliku
    • 1
  • Anthony Relmy
    • 1
  • Aurore Romey
    • 1
  • Kamila Gorna
    • 1
  • Stephan Zientara
    • 1
  • Labib Bakkali-Kassimi
    • 1
  • Sandra Blaise-Boisseau
    • 1
  1. 1.Université Paris-EstANSES, Laboratoire de Santé Animale, UMR Virologie 1161Maisons-AlfortFrance

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