title: Establishment of stably transfected cells constitutively expressing the full-length and truncated antigenic proteins of two genetically distinct mink astroviruses.
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.
authors.affiliation: Joint R&D Division of Virology, Department of Virology, Immunobiology and Parasitology, The National Veterinary Institute (SVA), Uppsala, Sweden.
authors.forename: Mehdi R M
authors.initials: MR
authors.lastname: Bidokhti
authors.affiliation: Joint R&D Division of Virology, Department of Virology, Immunobiology and Parasitology, The National Veterinary Institute (SVA), Uppsala, Sweden.
authors.forename: Karin
authors.initials: K
authors.lastname: Ullman
authors.affiliation: Division of Veterinary Diagnostics and Research, National Veterinary Institute, Technical University of Denmark, Copenhagen, Denmark.
authors.forename: Trine H
authors.initials: TH
authors.lastname: Jensen
authors.affiliation: Division of Veterinary Diagnostics and Research, National Veterinary Institute, Technical University of Denmark, Copenhagen, Denmark.
authors.forename: Mariann
authors.initials: M
authors.lastname: Chriél
authors.affiliation: Joint R&D Division of Virology, Department of Virology, Immunobiology and Parasitology, The National Veterinary Institute (SVA), Uppsala, Sweden.
authors.forename: Amin
authors.initials: A
authors.lastname: Mottahedin
authors.affiliation: Department of Biomedical Sciences and Veterinary Public Health, Division of Virology, Swedish University of Agricultural Sciences, Uppsala, Sweden.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Munir
authors.affiliation: The National Veterinary Institute, Department of Animal Health and Antimicrobial Resistance, Uppsala, Sweden.
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Andersson
authors.affiliation: Joint R&D Division of Virology, Department of Virology, Immunobiology and Parasitology, The National Veterinary Institute (SVA), Uppsala, Sweden.
authors.forename: Olivier
authors.initials: O
authors.lastname: Detournay
authors.affiliation: Division of Veterinary Diagnostics and Research, National Veterinary Institute, Technical University of Denmark, Copenhagen, Denmark.
authors.forename: Anne Sofie
authors.initials: AS
authors.lastname: Hammer
authors.affiliation: Joint R&D Division of Virology, Department of Virology, Immunobiology and Parasitology, The National Veterinary Institute (SVA), Uppsala, Sweden.
authors.forename: Claudia
authors.initials: C
authors.lastname: Baule
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000097:blood; D019200:Astroviridae / Q000235:genetics / Q000276:immunology*; D019350:Astroviridae Infections / Q000276:immunology / Q000517:prevention & control* / Q000662:veterinary* / Q000821:virology; D036022:Capsid Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D006224:Cricetinae; D005260:Female; D005333:Fetus; D018703:Founder Effect; D015870:Gene Expression; D007110:Immunity, Active; D008907:Mink / Q000276:immunology* / Q000821:virology; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014162:Transfection
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins