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

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

Author information

1
Joint R&D Division of Virology, Department of Virology, Immunobiology and Parasitology, The National Veterinary Institute (SVA), Uppsala, Sweden.
2
Division of Veterinary Diagnostics and Research, National Veterinary Institute, Technical University of Denmark, Copenhagen, Denmark.
3
Department of Biomedical Sciences and Veterinary Public Health, Division of Virology, Swedish University of Agricultural Sciences, Uppsala, Sweden.
4
The National Veterinary Institute, Department of Animal Health and Antimicrobial Resistance, Uppsala, Sweden.

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.

PMID:
24376619
PMCID:
PMC3871642
DOI:
10.1371/journal.pone.0082978
[Indexed for MEDLINE]
Free PMC Article

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