Advertisement

Virus Genes

, Volume 32, Issue 1, pp 27–35 | Cite as

Differential Transcription of Ovine Herpesvirus 2 Genes in Lymphocytes from Reservoir and Susceptible Species

  • Leenadevi Thonur
  • George C. Russell
  • James P. Stewart
  • David M. Haig
Original Article

Abstract

Ovine herpesvirus 2 (OvHV-2) is a lymphotropic gammaherpesvirus that asymptomatically infects most sheep, but causes malignant catarrhal fever in cattle, bison, pigs and deer. There is no permissive cell culture system but OvHV-2-infected T lymphocytes can be cultured from diseased animals. We showed that the OvHV-2 genome was in a circular conformation in sheep peripheral blood mononuclear cells and that the latency-associated ORF73 was transcribed, while expression of the productive cycle genes ORF9 (DNA polymerase) and ORF50 (R-transactivator) was barely detectable, suggestive of latency. Doxorubicin treatment of these cells induced the appearance of linear viral DNA and transcription of productive cycle genes along with several viral unique genes. In contrast, cultured T cells from diseased cattle and rabbits contained a mixture of circular and linear genome configurations indicative of a mixture of latently- and productively-infected cells. Most of the OvHV-2 unique genes were transcribed in these cells but ORF50 expression was only seen after doxorubicin treatment indicating a ‘leaky’ latent pattern of gene expression. 5-azacytidine treatment increased the proportion of circular DNA and inhibited the expression of most of the OvHV-2 unique genes except Ov2.5 (vIL-10) and Ov4.5 (Bcl-2 homologue) in the cattle cell line. These studies provide key insights into the differences in OvHV-2 gene expression in cells from reservoir and susceptible species and, for the first time, an in vitro system for studying the latent and productive phases of the OvHV-2 virus life cycle.

Keywords

Malignant Catarrhal Fever (MCF) Latency Genome conformation Gene expression Doxorubicin 5-azacytidine 

Notes

Acknowledgements

This work was supported by a joint BBSRC/SEERAD grant (26/S16844). We thank Iris Campbell for help with the cell culture, Jim Williams for help with blood collection and Colin J McInnes for useful discussions. JPS is the recipient of a Royal Society University Research Fellowship.

References

  1. 1.
    H. Li, N. Dyer, J. Keller, T.B. Crawford, J. Clin. Microbiol. 38, 1313–1318 (2000)PubMedGoogle Scholar
  2. 2.
    E.M. Milne, H.W. Reid, Vet. Rec. 126, 640–641 (1990)PubMedGoogle Scholar
  3. 3.
    D. O’Toole, H. Li, D. Miller, W.R. Williams, T.B. Crawford, Vet. Rec. 140, 519–524 (1997)PubMedGoogle Scholar
  4. 4.
    A. Bridgen, H.W. Reid, Res. Vet. Sci. 50, 38–44 (1991)PubMedGoogle Scholar
  5. 5.
    W. Plowright, R.D. Ferris, G.R. Scott, Nature 188, 1167–1169 (1960)PubMedGoogle Scholar
  6. 6.
    A. Bridgen, R. Munro, H.W. Reid, J. Comp. Pathol. 106, 351–359 (1992)CrossRefPubMedGoogle Scholar
  7. 7.
    S. Simon, H. Li, D. O’Toole, T.B. Crawford, J.L. Oaks, J. Gen. Virol. 84, 2009–2013 (2003)CrossRefPubMedGoogle Scholar
  8. 8.
    H.W. Reid, D. Buxton, I. Pow, J. Finlayson, E.L. Berrie, Res. Vet. Sci. 34, 109–113 (1983)PubMedGoogle Scholar
  9. 9.
    H.W. Reid, D. Buxton, I. Pow, J. Finlayson, Res. Vet. Sci. 47, 90–96 (1989)PubMedGoogle Scholar
  10. 10.
    A. Schock, H.W. Reid, Vet. Microbiol. 53, 111–119 (1996)CrossRefPubMedGoogle Scholar
  11. 11.
    S. Swa, H. Wright, J. Thomson, H. Reid, D. Haig, Immunology 102, 44–52 (2001)CrossRefPubMedGoogle Scholar
  12. 12.
    S.I. Baxter, I. Pow, A. Bridgen, H.W. Reid, Arch. Virol. 132, 145–159 (1993)CrossRefPubMedGoogle Scholar
  13. 13.
    D. Buxton, H.W. Reid, J. Finlayson, I. Pow, Res. Vet. Sci. 36, 205–211 (1984)PubMedGoogle Scholar
  14. 14.
    D. Buxton, R.O. Jacoby, H.W. Reid, P.A. Goodall, J. Comp. Pathol. 98, 155–166 (1988)PubMedGoogle Scholar
  15. 15.
    A. Schock, R.A. Collins, H.W. Reid, Vet. Immunol. Immunopathol. 66, 67–81 (1998)PubMedGoogle Scholar
  16. 16.
    E. Kieff, D. Knipe, P. Howley, D. Griffin, M. Martin, R. Lamb, B. Roizman, S. Straus (eds.), Fields Virology, 4th edn., vol. 2 (Lippincott Williams and Wilkins, Philadelphia, USA, 2001), pp. 2511–2573Google Scholar
  17. 17.
    A. Rickinson, E. Kie, D. Knipe, P. Howley, D. Griffin, M. Martin, R. Lamb, B. Roizman, S. Straus (eds.), Fields Virology, 4th edn., vol. 2 (Lippincott Williams and Wilkins, Philadelphia, USA, 2001), pp. 2575–2628Google Scholar
  18. 18.
    J. Rosbottom, R.G. Dalziel, H.W. Reid, J.P. Stewart, J. Gen. Virol. 83, 2999–3002 (2002)PubMedGoogle Scholar
  19. 19.
    G. Bauer, J. Gen. Virol. 64, 1337–1346 (1983)PubMedGoogle Scholar
  20. 20.
    W.E. Feng, G. Hong, H.J. Delecluse, S.C. Kenney, J. Virol. 78, 1893–1902 (2004)PubMedGoogle Scholar
  21. 21.
    S.A. Ben Sasson, G. Klein, Int. J. Cancer 28, 131–135 (1981)PubMedGoogle Scholar
  22. 22.
    M.G. Masucci, B. Contreras-Salazar, E. Ragnar, K. Falk, J. Minarovits, I. Ernberg, G. Klein, J. Virol. 63, 3135–3141 (1989)PubMedGoogle Scholar
  23. 23.
    P. Chomczynski, N. Sacchi, Anal. Biochem. 162, 156–159 (1987)CrossRefPubMedGoogle Scholar
  24. 24.
    T. Gardella, P. Medveczky, T. Sairenji, C. Mulder, J. Virol. 50, 248–254 (1984)PubMedGoogle Scholar
  25. 25.
    S.P. Mossman, D.A. Leib, K. McCarthy, C.A. Hart, J. Gen. Virol. 70, 2507–2512 (1989)PubMedGoogle Scholar
  26. 26.
    N. Raab-Traub, R. Hood, C.S. Yang, B. Henry, J.S. Pagano, J. Virol. 48, 580–590 (1983)PubMedGoogle Scholar
  27. 27.
    J.P. Simas, D. Swann, R. Bowden, S. Efstathiou, J. Gen. Virol. 80, 75–82 (1999)PubMedGoogle Scholar
  28. 28.
    C. Burrells, H.W. Reid, Vet. Immunol. Immunopathol. 29, 151–161 (1991)PubMedGoogle Scholar
  29. 29.
    W. Feng, B. Israel, N. Raab-Traub, P. Busson, S.C. Kenney, Cancer Res. 62, 1920–1926 (2002)PubMedGoogle Scholar
  30. 30.
    S.M. Moore, J.S. Cannon, Y.C. Tanhehco, F.M. Hamzeh, R.F. Ambinder, Antinmicrob. Agents Chemother. 45, 2082–2091 (2001)Google Scholar
  31. 31.
    B.C. Schaefer, J.L. Strominger, S.H. Speck, Mol. Cell Biol. 17, 364–377 (1997)PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Leenadevi Thonur
    • 1
  • George C. Russell
    • 1
  • James P. Stewart
    • 2
  • David M. Haig
    • 1
  1. 1.Moredun Research InstitutePenicuikUK
  2. 2.School of Infection and Host DefenceUniversity of LiverpoolLiverpoolUK

Personalised recommendations