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Binding of infectious pancreatic necrosis virus (IPNV) to membrane proteins from different fish cell lines

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Abstract

The early interactions between infectious pancreatic necrosis virus (IPNV) from Atlantic salmon and susceptible cell lines were studied using a virus overlay protein binding assay (VOPBA). Membrane preparations from four different cell lines, bluegill fry (BF)-2 cells, Chinook salmon embryo (CHSE)-214 cells, salmon head kidney (SHK)-1 cells and Atlantic salmon kidney (ASK) cells were separated by SDS-PAGE, blotted to nitrocellulose membranes and incubated with either a highly virulent IPNV field isolate or different recombinant IPNV strains exhibiting variations in virulence. Binding of virus to the respective membrane fractions was detected using either polyclonal or monoclonal antibodies. Independent of variations in virulence, IPNV bound in a specific manner to an approximately 220-kDa membrane protein from the salmonid cell lines CHSE-214, SHK-1 and ASK, whereas the size of the binding protein from the non-salmonid cell line BF-2 was approximately 190 kDa. Neutralization of IPNV prior to incubation with the blotted membrane fractions inhibited binding to the indicated proteins. Complete deglycosylation of the membrane proteins did not interfere with the binding of IPNV, suggesting that the interaction between virus and cells is not carbohydrate specific. The described binding proteins may represent putative cellular binding sites or receptors for IPNV.

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Acknowledgments

This study was supported by grant 143286/140 from the Research Council of Norway. We thank Nina Santi at the Norwegian School of Veterinary Science (NSVS), for providing the recombinant IPNV strains, and Espen Rimstad (NSVS) and Thomas Küntziger (University of Oslo, Institute of Medical Biochemistry) for critical reading of the manuscript. The skilled technical assistance of Ingebjørg Modahl and Lone Thiel Engerdahl at the National Veterinary Institute is gratefully acknowledged.

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Authors and Affiliations

  1. National Veterinary Institute, P.O. Box 8156, Dep, 0033, Oslo, Norway

    Irene Ørpetveit, Hilde Sindre & Birgit H. Dannevig

  2. Department of Pharmaceutical Biosciences, School of Pharmacy, University of Oslo, P.O. Box 1068, Blindern, 0316, Oslo, Norway

    Tor Gjøen

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  1. Irene Ørpetveit
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  2. Tor Gjøen
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Correspondence to Irene Ørpetveit.

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Ørpetveit, I., Gjøen, T., Sindre, H. et al. Binding of infectious pancreatic necrosis virus (IPNV) to membrane proteins from different fish cell lines. Arch Virol 153, 485–493 (2008). https://doi.org/10.1007/s00705-007-0018-1

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  • DOI: https://doi.org/10.1007/s00705-007-0018-1

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