Establishment of stably transfected cells constitutively expressing the full-length and truncated antigenic proteins of two genetically distinct mink astroviruses

PLoS One. 2013 Dec 23;8(12):e82978. doi: 10.1371/journal.pone.0082978. eCollection 2013.

Abstract

Astroviruses are becoming a growing concern in veterinary and public health. To date there are no registered vaccines against astrovirus-induced disease, mostly due to the difficulty to cultivate astroviruses to high titer for vaccine development using conventional techniques. As means to circumvent this drawback, we have developed stably transfected mink fetal cells and BHK21 cells constitutively expressing the full-length and truncated capsid proteins of two distinct genotypes of mink astrovirus. Protein expression in these stably transfected cells was demonstrated by strong signals as evaluated by in-situ PLA and IFA, and confirmed by Western blotting. The recombinant full-length and truncated proteins induced a high level of antibodies in mink, evaluated by ELISA, demonstrating their immunogenicity. In a challenge experiment in mink, a reduction in presentation clinical signs and virus shedding was observed in mink kits born from immunized females. The gene integration and protein expression were sustained through cell passage, showing that the used approach is robust and reliable for expression of functional capsid proteins for vaccine and diagnostic applications.

MeSH terms

  • Animals
  • Antibodies, Viral / biosynthesis*
  • Antibodies, Viral / blood
  • Astroviridae / genetics
  • Astroviridae / immunology*
  • Astroviridae Infections / immunology
  • Astroviridae Infections / prevention & control*
  • Astroviridae Infections / veterinary*
  • Astroviridae Infections / virology
  • Capsid Proteins / administration & dosage
  • Capsid Proteins / genetics
  • Capsid Proteins / immunology*
  • Cell Line
  • Cricetinae
  • Female
  • Fetus
  • Founder Effect
  • Gene Expression
  • Immunity, Active
  • Mink / immunology*
  • Mink / virology
  • Recombinant Proteins / administration & dosage
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Transfection

Substances

  • Antibodies, Viral
  • Capsid Proteins
  • Recombinant Proteins

Grants and funding

This study was funded by a Grant of the Danish Fur Breeder’s Association and by internal funds from the Department of Virology, Immunobiology and Parasitology of the National Veterinary Institute (SVA) and from the Division of Veterinary Diagnostics and Research of the Danish Technical University (DTU-VET). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.