1887

Abstract

Elimination of the RNase activity of classical swine fever virus (CSFV) glycoprotein E was previously shown to result in virus attenuation. Specific reduction of B cell numbers in the peripheral blood, a typical symptom of CSFV infection in pigs, was not detected on infection with the RNase-negative mutant C-H346Δ [ Meyers (1999). , 10224–10235 ]. The present report shows that this feature is restricted to this specific virus mutant, and does not represent a general property of RNase-negative CSFV. The effects induced by infection with two other RNase-negative and wild-type (wt) CSFV strains on the composition of peripheral blood cells have been further analysed. For all viruses, not only general leukopenia but also a reduction of different subsets of leukocytes (T-lymphocytes, monocytes and granulocytes) was detected. Similar to the results with B-cells, no significant differences with regard to changes in cell number were determined for RNase-negative mutants and wt virus during the initial phase of infection. Later, the values returned to pre-infection levels for the mutants, but stayed at low levels in the wt virus-infected animals. A major difference was reflected in the virus load of the infected animals, which was dramatically higher for pigs infected with wt CSFV, so that reduction of the virus load represents a further marker for attenuation resulting from RNase destruction. Attenuation was also detectable for the RNase-negative mutant C-W300G, which showed rapid reversion to the wt sequence within the infected pig. The prevention of fatal disease after infection with C-W300G is apparently determined during the short time between infection and reversion, as the virus revertant reisolated from infected pigs was shown to be virulent when used for infection in a follow-up study. Reversion of C-W300G was also detected in tissue culture during passage on swine testis epithelioid cells and porcine transformed kidney (MAX) cells, whereas the mutation was stable when SK6 or 38A1D cells were tested.

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2004-07-01
2019-11-20
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References

  1. Bruschke, C. J., Hulst, M. M., Moormann, R. J., van Rijn, P. A. & van Oirschot, J. T. ( 1997; ). Glycoprotein Erns of pestiviruses induces apoptosis in lymphocytes of several species. J Virol 71, 6692–6696.
    [Google Scholar]
  2. Davis, W. C., Brown, W. C., Hamilton, M. J., Wyatt, C. R., Orden, J. A., Khalid, A. M. & Naessens, J. ( 1996; ). Analysis of monoclonal antibodies specific for the γδ TCR. Vet Immunol Immunopathol 52, 275–283.[CrossRef]
    [Google Scholar]
  3. Donis, R. O., Corapi, W. & Dubovi, E. J. ( 1988; ). Neutralizing monoclonal antibodies to bovine viral diarrhoea virus bind to the 56K to 58K glycoprotein. J Gen Virol 69, 77–86.[CrossRef]
    [Google Scholar]
  4. Floegel-Niesmann, G., Bunzenthal, C., Fischer, S. & Moennig, V. ( 2003; ). Virulence of recent and former classical swine fever virus isolates evaluated by their clinical and pathological signs. J Vet Med B Infect Dis Vet Public Health 50, 214–220.[CrossRef]
    [Google Scholar]
  5. Hammerberg, C. & Schurig, G. G. ( 1986; ). Characterization of monoclonal antibodies directed against swine leukocytes. Vet Immunol Immunopathol 11, 107–121.[CrossRef]
    [Google Scholar]
  6. Heinz, F. X., Collett, M. S., Purcell, R. H., Gould, E. A., Howard, C. R., Houghton, M., Moormann, J. M., Rice, C. M. & Thiel, H.-J. ( 2000; ). Family Flaviviridae. In Virus Taxonomy. Seventh Report of the International Committee on Taxonomy of Viruses, pp. 859–878. Edited by M. H. V. van Regenmortel, C. M. Fauquet, D. H. L. Bishop, E. B. Carstens, M. K. Estes, S. M. Lemon, J. Maniloff, M. A. Mayo, D. J. McGeoch, C. R. Pringle & R. B. Wickner. San Diego: Academic Press.
  7. Hulst, M. M. & Moormann, R. J. ( 1997; ). Inhibition of pestivirus infection in cell culture by envelope proteins Erns and E2 of classical swine fever virus: Erns and E2 interact with different receptors. J Gen Virol 78, 2779–2787.
    [Google Scholar]
  8. Hulst, M. M., Himes, G., Newbigin, E. & Moormann, R. J. M. ( 1994; ). Glycoprotein E2 of classical swine fever virus: expression in insect cells and identification as a ribonuclease. Virology 200, 558–565.[CrossRef]
    [Google Scholar]
  9. Hulst, M. M., Panoto, F. E., Hoekman, A., van Gennip, H. G. & Moormann, R. J. ( 1998; ). Inactivation of the RNase activity of glycoprotein Erns of classical swine fever virus results in a cytopathogenic virus. J Virol 72, 151–157.
    [Google Scholar]
  10. Iqbal, M., Poole, E., Goodbourn, S. & McCauley, J. W. ( 2004; ). Role for bovine viral diarrhea virus Erns glycoprotein in the control of activation of beta interferon by double-stranded RNA. J Virol 78, 136–145.[CrossRef]
    [Google Scholar]
  11. Jonjic, S. & Koszinowski, U. H. ( 1984; ). Monoclonal antibodies reactive with swine lymphocytes. I. Antibodies to membrane structures that define the cytolytic T lymphocyte subset in the swine. J Immunol 133, 647–652.
    [Google Scholar]
  12. Kunkel, T. A., Roberts, J. D. & Zakour, R. A. ( 1987; ). Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol 154, 367–382.
    [Google Scholar]
  13. Langedijk, J. P. ( 2002; ). Translocation activity of C-terminal domain of pestivirus Erns and ribotoxin L3 loop. J Biol Chem 277, 5308–5314.[CrossRef]
    [Google Scholar]
  14. Lindenbach, B. D. & Rice, C. M. ( 2001; ). Flaviviridae: the viruses and their replication. In Fields Virology, 4th edn, vol. 1, pp. 991–1041. Edited by D. M. Knipe & P. M. Howley. Philadelphia: Lippincott Williams & Wilkins.
  15. Meyer, C., Von Freyburg, M., Elbers, K. & Meyers, G. ( 2002; ). Recovery of virulent and RNase-negative attenuated type 2 bovine viral diarrhea viruses from infectious cDNA clones. J Virol 76, 8494–8503.[CrossRef]
    [Google Scholar]
  16. Meyers, G., Thiel, H.-J. & Rümenapf, T. ( 1996; ). Classical swine fever virus: recovery of infectious viruses from cDNA constructs and generation of recombinant cytopathogenic defective interfering particles. J Virol 70, 1588–1595.
    [Google Scholar]
  17. Meyers, G., Saalmüller, A. & Büttner, M. ( 1999; ). Mutations abrogating the RNase activity in glycoprotein Erns of the pestivirus classical swine fever virus lead to virus attenuation. J Virol 73, 10224–10235.
    [Google Scholar]
  18. Moennig, V. & Plagemann, P. G. W. ( 1992; ). The pestiviruses. Adv Virus Res 41, 53–98.
    [Google Scholar]
  19. Paton, D. J., Lowings, J. P. & Barrett, A. D. T. ( 1992; ). Epitope mapping of the gp53 envelope protein of bovine viral diarrhea virus. Virology 190, 763–772.[CrossRef]
    [Google Scholar]
  20. Pauly, T., Elbers, K., König, M., Lengsfeld, T., Saalmüller, A. & Thiel, H.-J. ( 1995; ). Classical swine fever virus-specific cytotoxic T lymphocytes and identification of a T cell epitope. J Gen Virol 76, 3039–3049.[CrossRef]
    [Google Scholar]
  21. Pescovitz, M. D., Lunney, J. K. & Sachs, D. H. ( 1984; ). Preparation and characterization of monoclonal antibodies reactive with porcine PBL. J Immunol 133, 368–375.
    [Google Scholar]
  22. Pescovitz, M. D., Lunney, J. K. & Sachs, D. H. ( 1985; ). Murine anti-swine T4 and T8 monoclonal antibodies: distribution and effects on proliferative and cytotoxic T cells. J Immunol 134, 37–44.
    [Google Scholar]
  23. Saalmüller, A., Reddehase, M. J., Buhring, H. J., Jonjic, S. & Koszinowski, U. H. ( 1987; ). Simultaneous expression of CD4 and CD8 antigens by a substantial proportion of resting porcine T lymphocytes. Eur J Immunol 17, 1297–1301.[CrossRef]
    [Google Scholar]
  24. Saalmüller, A., Hirt, W. & Reddehase, M. J. ( 1990; ). Porcine gamma/delta T lymphocyte subsets differing in their propensity to home to lymphoid tissue. Eur J Immunol 20, 2343–2346.[CrossRef]
    [Google Scholar]
  25. Saalmüller, A., Hirt, W., Maurer, S. & Weiland, E. ( 1994; ). Discrimination between two subsets of porcine CD8+ cytolytic T lymphocytes by the expression of CD5 antigen. Immunology 81, 578–583.
    [Google Scholar]
  26. Saalmüller, A., Pauly, T., Höhlich, B. J. & Pfaff, E. ( 1999; ). Characterization of porcine T lymphocytes and their immune response against viral antigens. J Biotechnol 73, 223–233.[CrossRef]
    [Google Scholar]
  27. Saalmüller, A., Werner, T. & Fachinger, V. ( 2002; ). T-helper cells from naive to committed. Vet Immunol Immunopathol 87, 137–145.[CrossRef]
    [Google Scholar]
  28. Sambrook, J. & Russell, D. W. ( 2001; ). Molecular Cloning: a Laboratory Manual, 3rd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  29. Schneider, R., Unger, G., Stark, R., Schneider-Scherzer, E. & Thiel, H.-J. ( 1993; ). Identification of a structural glycoprotein of an RNA virus as a ribonuclease. Science 261, 1169–1171.[CrossRef]
    [Google Scholar]
  30. Summerfield, A., Knötig, S. M. & McCullough, K. C. ( 1998; ). Lymphocyte apoptosis during classical swine fever: implication of activation-induced cell death. J Virol 72, 1853–1861.
    [Google Scholar]
  31. Summerfield, A., Knoetig, S. M., Tschudin, R. & McCullough, K. C. ( 2000; ). Pathogenesis of granulocytopenia and bone marrow atrophy during classical swine fever involves apoptosis and necrosis of uninfected cells. Virology 272, 50–60.[CrossRef]
    [Google Scholar]
  32. Summerfield, A., McNeilly, F., Walker, I., Allan, G., Knoetig, S. M. & McCullough, K. C. ( 2001a; ). Depletion of CD4+ and CD8high+ T-cells before the onset of viraemia during classical swine fever. Vet Immunol Immunopathol 78, 3–19.[CrossRef]
    [Google Scholar]
  33. Summerfield, A., Zingle, K., Inumaru, S. & McCullough, K. C. ( 2001b; ). Induction of apoptosis in bone marrow neutrophil-lineage cells by classical swine fever virus. J Gen Virol 82, 1309–1318.
    [Google Scholar]
  34. Susa, M., König, M., Saalmüller, A., Reddehase, M. J. & Thiel, H. J. ( 1992; ). Pathogenesis of classical swine fever: B-lymphocyte deficiency caused by hog cholera virus. J Virol 66, 1171–1175.
    [Google Scholar]
  35. Thiel, H.-J., Stark, R., Weiland, E., Rümenapf, T. & Meyers, G. ( 1991; ). Hog cholera virus: molecular composition of virions from a pestivirus. J Virol 65, 4705–4712.
    [Google Scholar]
  36. Thiel, H.-J., Plagemann, P. G. W. & Moennig, V. ( 1996; ). Pestiviruses. In Fields Virology, 3rd edn, vol. 1, pp. 1059–1073. Edited by B. N. Fields, D. M. Knipe & P. M. Howley. Philadelphia & New York: Lippincott–Raven.
  37. van Rijn, P. A., van Gennip, H. G., de Meijer, E. J. & Moormann, R. J. ( 1993; ). Epitope mapping of envelope glycoprotein E1 of hog cholera virus strain Brescia. J Gen Virol 74, 2053–2060.[CrossRef]
    [Google Scholar]
  38. Weiland, E., Stark, R., Haas, B., Rümenapf, T., Meyers, G. & Thiel, H.-J. ( 1990; ). Pestivirus glycoprotein which induces neutralizing antibodies forms part of a disulfide linked heterodimer. J Virol 64, 3563–3569.
    [Google Scholar]
  39. Weiland, E., Ahl, R., Stark, R., Weiland, F. & Thiel, H.-J. ( 1992; ). A second envelope glycoprotein mediates neutralization of a pestivirus, hog cholera virus. J Virol 66, 3677–3682.
    [Google Scholar]
  40. Widjojoatmodjo, M. N., van Gennip, H. G., Bouma, A., van Rijn, P. A. & Moormann, R. J. ( 2000; ). Classical swine fever virus Erns deletion mutants: trans-complementation and potential use as nontransmissible, modified, live-attenuated marker vaccines. J Virol 74, 2973–2980.[CrossRef]
    [Google Scholar]
  41. Windisch, J. M., Schneider, R., Stark, R., Weiland, E., Meyers, G. & Thiel, H.-J. ( 1996; ). RNase of classical swine fever virus: biochemical characterization and inhibition by virus-neutralizing monoclonal antibodies. J Virol 70, 352–358.
    [Google Scholar]
  42. Youle, R. J. & D'Allessio, G. ( 1997; ). Antitumor RNases. New York: Academic Press.
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