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Generation and characterization of koi herpesvirus recombinants lacking viral enzymes of nucleotide metabolism

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Abstract

Koi herpesvirus (KHV) causes a fatal disease in koi and common carp, but no reliable and genetically characterized vaccines are available up to now. Therefore, we generated KHV recombinants possessing deletions within the viral ribonucleotide reductase (RNR), thymidine kinase (TK), dUTPase, or TK and dUTPase genes, and their corresponding rescuants. All KHV mutants were replication competent in cultured cells. Whereas plaque sizes and titers of RNR-negative KHV were reduced, replication of the other mutants was not affected. Experimental infection of carp indicated attenuation of TK- or dUTPase-deleted KHV, and PCR analysis of tissue samples permitted differentiation of mutant from wild-type virus.

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References

  1. Aoki T, Hirono I, Kurokawa K, Fukuda H, Nahary R, Eldar A, Davison AJ, Waltzek TB, Bercovier H, Hedrick RP (2007) Genome sequences of three koi herpesvirus isolates representing the expanding distribution of an emerging disease threatening koi and common carp worldwide. J Virol 81:5058–5065

    Article  PubMed  CAS  Google Scholar 

  2. Bercovier H, Fishman Y, Nahary R, Sinai S, Zlotkin A, Eyngor M, Gilad O, Eldar A, Hedrick RP (2005) Cloning of the koi herpesvirus (KHV) gene encoding thymidine kinase and its use for a highly sensitive PCR based diagnosis. BMC Microbiol 5:13

    Article  PubMed  Google Scholar 

  3. Bigarré L, Baud M, Cabon J, Antychowicz J, Bergmann SM, Engelsma M, Pozet F, Reichert M, Castric J (2009) Differentiation between Cyprinid herpesvirus type-3 lineages using duplex PCR. J Virol Methods 158:51–57

    Article  PubMed  Google Scholar 

  4. Bretzinger A, Fischer-Scherl T, Oumouna M, Hoffmann R, Truyen U (1999) Mass mortalities in koi carp, Cyprinus carpio, associated with gill and skin disease. Bull Eur Assoc Fish Pathol 19:182–185

    Google Scholar 

  5. Costes B, Fournier G, Michel B, Delforge C, Raj VS, Dewals B, Gillet L, Drion P, Body A, Schynts F, Lieffrig F, Vanderplasschen A (2008) Cloning of the koi herpesvirus genome as an infectious bacterial artificial chromosome demonstrates that disruption of the thymidine kinase locus induces partial attenuation in Cyprinus carpio koi. J Virol 82:4955–4964

    Article  PubMed  CAS  Google Scholar 

  6. Davison AJ, Eberle R, Ehlers B, Hayward GS, McGeoch DJ, Minson AC, Pellett PE, Roizman B, Studdert MJ, Thiry E (2009) The order Herpesvirales. Arch Virol 154:171–177

    Article  PubMed  CAS  Google Scholar 

  7. de Wind N, Berns A, Gielkens A, Kimman T (1993) Ribonucleotide reductase-deficient mutants of pseudorabies virus are avirulent for pigs and induce partial protective immunity. J Gen Virol 74:351–359

    Article  PubMed  Google Scholar 

  8. Efstathiou S, Kemp S, Darby G, Minson AC (1989) The role of herpes simplex virus type 1 thymidine kinase in pathogenesis. J Gen Virol 70:869–879

    Article  PubMed  CAS  Google Scholar 

  9. Fuchs W, Mettenleiter TC (1996) DNA sequence and transcriptional analysis of the UL1 to UL5 gene cluster of infectious laryngotracheitis virus. J Gen Virol 77:2221–2229

    Article  PubMed  CAS  Google Scholar 

  10. Fuchs W, Mettenleiter TC (1999) DNA sequence of the UL6 to UL20 genes of infectious laryngotracheitis virus and characterization of the UL10 gene product as a nonglycosylated and nonessential virion protein. J Gen Virol 80:2173–2182

    PubMed  CAS  Google Scholar 

  11. Fuchs W, Ziemann K, Teifke JP, Werner O, Mettenleiter TC (2000) The non-essential UL50 gene of avian infectious laryngotracheitis virus encodes a functional dUTPase which is not a virulence factor. J Gen Virol 81:627–638

    PubMed  CAS  Google Scholar 

  12. Gilad O, Yun S, Andree KB, Adkison MA, Zlotkin A, Bercovier H, Eldar A, Hedrick RP (2002) Initial characteristics of koi herpesvirus and development of a polymerase chain reaction assay to detect the virus in koi, Cyprinus carpio koi. Dis Aquat Organ 48:101–108

    Article  PubMed  CAS  Google Scholar 

  13. Gilad O, Yun S, Adkison MA, Way K, Willits NH, Bercovier H, Hedrick RP (2003) Molecular comparison of isolates of an emerging fish pathogen, koi herpesvirus, and the effect of water temperature on mortality of experimentally infected koi. J Gen Virol 84:2661–2668

    Article  PubMed  CAS  Google Scholar 

  14. Gilad O, Yun S, Zagmutt-Vergara FJ, Leutenegger CM, Bercovier H, Hedrick RP (2004) Concentrations of a Koi herpesvirus (KHV) in tissues of experimentally infected Cyprinus carpio koi as assessed by real-time TaqMan PCR. Dis Aquat Organ 60:179–187

    Article  PubMed  CAS  Google Scholar 

  15. Goldstein DJ, Weller SK (1988) Herpes simplex virus type 1-induced ribonucleotide reductase activity is dispensable for virus growth and DNA synthesis: isolation and characterization of an ICP6 lacZ insertion mutant. J Virol 62:196–205

    PubMed  CAS  Google Scholar 

  16. Hedrick RP, Gilad O, Yun S, Spangenberg JV, Marty GD, Nordhausen RW, Kebus MJ, Bercovier H, Eldar A (2000) A herpesvirus associated with mass mortality of juvenile and adult koi, a strain of common carp. J Aquat Anim Health 12:44–57

    Article  Google Scholar 

  17. Heineman TC, Cohen JI (1994) Deletion of the varicella-zoster virus large subunit of ribonucleotide reductase impairs growth of virus in vitro. J Virol 68:3317–3323

    PubMed  CAS  Google Scholar 

  18. Jacobson JG, Leib DA, Goldstein DJ, Bogard CL, Schaffer PA, Weller SK, Coen DM (1989) A herpes simplex virus ribonucleotide reductase deletion mutant is defective for productive acute and reactivatable latent infections of mice and for replication in mouse cells. Virology 173:276–283

    Article  PubMed  CAS  Google Scholar 

  19. Jöns A, Mettenleiter TC (1996) Identification and characterization of pseudorabies virus dUTPase. J Virol 70:1242–1245

    PubMed  Google Scholar 

  20. Jöns A, Gerdts V, Lange E, Kaden V, Mettenleiter TC (1997) Attenuation of dUTPase-deficient pseudorabies virus for the natural host. Vet Microbiol 56:47–54

    Article  PubMed  Google Scholar 

  21. Kit S (1989) Recombinant-derived modified-live herpesvirus vaccines. Adv Exp Med Biol 251:219–236

    PubMed  CAS  Google Scholar 

  22. Neukirch M, Böttcher K, Bunnajirakul S (1999) Isolation of a virus from koi with altered gills. Bull Eur Assoc Fish Pathol 19:221–224

    Google Scholar 

  23. Perelberg A, Ronen A, Hutoran M, Smith Y, Kotler M (2005) Protection of cultured Cyprinus carpio against a lethal viral disease by an attenuated virus vaccine. Vaccine 23:3396–3403

    Article  PubMed  CAS  Google Scholar 

  24. Perelberg A, Ilouze M, Kotler M, Steinitz M (2008) Antibody response and resistance of Cyprinus carpio immunized with cyprinid herpes virus 3 (CyHV-3). Vaccine 26:3750–3756

    Article  PubMed  CAS  Google Scholar 

  25. Ronen A, Perelberg A, Abramowitz J, Hutoran M, Tinman S, Bejerano I, Steinitz M, Kotler M (2003) Efficient vaccine against the virus causing a lethal disease in cultured Cyprinus carpio. Vaccine 21:4677–4684

    Article  PubMed  CAS  Google Scholar 

  26. Rosenkranz D, Klupp BG, Teifke JP, Granzow H, Fichtner D, Mettenleiter TC, Fuchs W (2008) Identification of envelope protein pORF81 of koi herpesvirus. J Gen Virol 89:896–900

    Article  PubMed  CAS  Google Scholar 

  27. Schnitzlein WM, Winans R, Ellsworth S, Tripathy DN (1995) Generation of thymidine kinase-deficient mutants of infectious laryngotracheitis virus. Virology 209:304–314

    Article  PubMed  CAS  Google Scholar 

  28. Zhang HG, Hanson LA (1995) Deletion of thymidine kinase gene attenuates channel catfish herpesvirus while maintaining infectivity. Virology 209:658–663

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The authors thank R. P. Hedrick and M. Neukirch for providing viruses and cells, and J. P. Teifke for help with pathological examinations. Carp were provided by S. H. Leenstra, Wageningen, The Netherlands. The technical assistance of C. Ehrlich and M. Legien is greatly appreciated.

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Authors and Affiliations

  1. Institute of Molecular Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493, Greifswald-Insel Riems, Germany

    Walter Fuchs & Thomas C. Mettenleiter

  2. Institute of Infectology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493, Greifswald-Insel Riems, Germany

    Dieter Fichtner & Sven M. Bergmann

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  1. Walter Fuchs
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  2. Dieter Fichtner
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  3. Sven M. Bergmann
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  4. Thomas C. Mettenleiter
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Correspondence to Walter Fuchs.

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Fuchs, W., Fichtner, D., Bergmann, S.M. et al. Generation and characterization of koi herpesvirus recombinants lacking viral enzymes of nucleotide metabolism. Arch Virol 156, 1059–1063 (2011). https://doi.org/10.1007/s00705-011-0953-8

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  • DOI: https://doi.org/10.1007/s00705-011-0953-8

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