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

Mehdi R. M. Bidokhti; Karin Ullman; Trine Hammer Jensen; Mariann Chriél; Amin Mottahedin; Muhammad Munir; Anna Maria Andersson; Olivier Detournay; Anne Sofie Vedsted Hammer; Claudia Baule

doi:10.1371/journal.pone.0082978

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

Mehdi R. M. Bidokhti, Karin Ullman, Trine Hammer Jensen, Mariann Chriél, Amin Mottahedin, Muhammad Munir, Anna Maria Andersson, Olivier Detournay, Anne Sofie Vedsted Hammer, Claudia Baule

5 Citations (Scopus)

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.

Original language	English
Article number	e82978
Journal	P L o S One
Volume	8
Issue number	12
Number of pages	13
ISSN	1932-6203
DOIs	https://doi.org/10.1371/journal.pone.0082978
Publication status	Published - 23 Dec 2013
Externally published	Yes

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Bidokhti, M. R. M., Ullman, K., Jensen, T. H., Chriél, M., Mottahedin, A., Munir, M., Andersson, A. M., Detournay, O., Hammer, A. S. V., & Baule, C. (2013). Establishment of stably transfected cells constitutively expressing the full-length and truncated antigenic proteins of two genetically distinct mink astroviruses. P L o S One, 8(12), [e82978]. https://doi.org/10.1371/journal.pone.0082978

Establishment of stably transfected cells constitutively expressing the full-length and truncated antigenic proteins of two genetically distinct mink astroviruses. / Bidokhti, Mehdi R. M.; Ullman, Karin; Jensen, Trine Hammer et al.

In: P L o S One, Vol. 8, No. 12, e82978, 23.12.2013.

Research output: Contribution to journal › Journal article › Research › peer-review

Bidokhti, MRM, Ullman, K, Jensen, TH, Chriél, M, Mottahedin, A, Munir, M, Andersson, AM, Detournay, O, Hammer, ASV & Baule, C 2013, 'Establishment of stably transfected cells constitutively expressing the full-length and truncated antigenic proteins of two genetically distinct mink astroviruses', P L o S One, vol. 8, no. 12, e82978. https://doi.org/10.1371/journal.pone.0082978

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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.",

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AU - Bidokhti, Mehdi R. M.

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AU - Chriél, Mariann

AU - Mottahedin, Amin

AU - Munir, Muhammad

AU - Andersson, Anna Maria

AU - Detournay, Olivier

AU - Hammer, Anne Sofie Vedsted

AU - Baule, Claudia

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AB - 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.

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

Abstract

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