1887

Abstract

Cultures of adherent and non-adherent bovine peripheral blood mononuclear (PBM) cells were inoculated with bluetongue virus (BTV) serotype 10. Some cultures of non-adherent cells were stimulated with interleukin 2 (IL-2) and concanavalin A for 24 h prior to virus inoculation. Cells were harvested at various intervals up to 72 h after inoculation. A panel of leukocyte differentiation antigen-specific monoclonal antibodies (MAbs), specific for bovine CD2, CD4 or CD8, monocytes and granulocytes, B cells, T cells or the IL-2 receptor (IL-2r), was directly conjugated to fluorescein isothiocyanate, and a MAb specific for the BTV major core protein VP7 was directly conjugated to phycoerythrin. Cells were labelled with conjugated MAbs in single- and double-label immunofluorescence studies to identify specifically the BTV-infected cells in inoculated cultures. The viability of cells was determined by propidium iodide exclusion, and all analyses were done using flow cytometry. Productive infection of cultures of PBM cells was confirmed by virus titration. The data revealed a clear difference between subsets of bovine PBM cells in susceptibility to infection with BTV . Monocytes were readily infected with BTV, as were stimulated CD4 cells, and infection was cytopathic to monocytes and stimulated lymphocytes. The proportion of infected cells decreased after 24 h and virus titres dropped markedly by 72 h in all cultures. CD4 cells in cultures of unstimulated non-adherent cells inoculated with BTV showed increased expression of IL-2r. The possible relevance of these findings to the pathogenesis of BTV infection of cattle is discussed.

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1992-08-01
2021-10-24
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References

  1. Baldwin C. L., Teale A. J., Naessens J. G., Goddeeris B. M., MacHugh N. D., Morrison W. I. 1986; Characterization of a subset of bovine T lymphocytes that express BoT4 by monoclonal antibodies and function: similarity to lymphocytes defined by human T4 and murine L3T4. Journal of Immunology 136:4385–4391
    [Google Scholar]
  2. Bowne J. G. 1971; Bluetongue disease. Advances in Veterinary Science and Comparative Medicine 15:1–46
    [Google Scholar]
  3. Clevers H., MacHugh N. D., BenSaid A., Dunlap S., Baldwin C. L., Kaushal A., Iams K., Howard C. J., Morrison W. I. 1990; Identification of a bovine surface antigen uniquely expressed on CD4CD8 T cell receptor γδ+ T lymphocytes. European Journal of Immunology 20:809–817
    [Google Scholar]
  4. Collisson E. W., Barber T. L. 1983; Blood cells associated with bluetongue virus infection in cattle. American Association of Veterinary Laboratory Diagnosticians. 26th Annual Proceedings pp. 287–300
    [Google Scholar]
  5. DuToit R. M. 1962; The role played by bovines in the transmission of bluetongue in sheep. Journal of the South African Veterinary Medical Association 33:483–490
    [Google Scholar]
  6. Ellis J. A., Baldwin C. L., MacHugh N. D., BenSaid A., Teale A. J., Goddeeris B. M., Morrison W. I. 1986; Characterization by a monoclonal antibody and functional analysis of a subset of bovine T lymphocytes that express BoT8, a molecule analogous to human CD8. Immunology 58:351–358
    [Google Scholar]
  7. Ellis J. A., Morrison W. I., Goddeeris B. M., Emery D. L. 1987; Bovine mononuclear phagocytic cells: identification by monoclonal antibodies and analysis of functional properties. Veterinary Immunology and Immunopathology 17:125–134
    [Google Scholar]
  8. Ellis J. A., Davis W. C., MacHugh N. D., Emery D. L., Kaushal A., Morrison W. I. 1988; Differentiation antigens on bovine mononuclear phagocytes identified by monoclonal antibodies. Veterinary Immunology and Immunopathology 19:325–340
    [Google Scholar]
  9. Emery D. L., MacHugh N. D., Ellis J. A. 1987; The properties and functional activity of non-lymphoid cells from bovine afferent (peripheral) lymph. Immunology 62:177–183
    [Google Scholar]
  10. Erasmus B. J. 1975; Bluetongue in sheep and goats. Australian Veterinary Journal 51:165–170
    [Google Scholar]
  11. Goding J. W. 1986; Monoclonal Antibodies: Principles and Practices. 2nd edn Orlando: Academic Press;
    [Google Scholar]
  12. Hall J. G., Morris B. 1963; The lymph-borne cells of the immune response. Quarterly Journal of Experimental Physiology 48:235–247
    [Google Scholar]
  13. Hein W. R., MacKay C. R. 1991; Prominence of γδ T cells in the ruminant immune system. Immunology Today 12:30–34
    [Google Scholar]
  14. Hourrigan J. L., Klingsporn A. L. 1975; Bluetongue: the disease in cattle. Australian Veterinary Journal 51:170–174
    [Google Scholar]
  15. Krishan A. 1975; Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining. Journal of Cell Biology 66:188–193
    [Google Scholar]
  16. Luedke A. J. 1970; Distribution of virus in blood components during viremia of bluetongue. United States Animal Health Association, 74th Annual Proceedings pp. 9–21
    [Google Scholar]
  17. Luedke A. J., Jochim M. M., Jones R. H. 1969; Bluetongue in cattle: viremia. American Journal of Veterinary Research 30:511–516
    [Google Scholar]
  18. MacKay C. R., Kimpton W. G., Brandon M. R., Cahill R. N. P. 1988; Lymphocyte subsets show marked differences in their distribution between blood and the afferent and efferent lymph and peripheral lymph nodes. Journal of Experimental Medicine 167:1755–1765
    [Google Scholar]
  19. McKeever D. J., MacHugh N. D., Goddeeris B. M., Awino E., Morrison W. I. 1991; Bovine afferent lymph veiled cells differ from blood monocytes in phenotype and accessory function. Journal of Immunology 147:3703–3709
    [Google Scholar]
  20. MacLachlan N. J., Fuller F. J. 1986; Genetic stability in calves of a single strain of bluetongue virus. American Journal of Veterinary Research 47:762–764
    [Google Scholar]
  21. MacLachlan N. J., Schore C. E., Osburn B. I. 1984; Antiviral responses of bluetongue virus-inoculated bovine fetuses and their dams. American Journal of Veterinary Research 45:1469–1473
    [Google Scholar]
  22. MacLachlan N. J., Heidner H. W., Fuller F. J. 1987; Humoral immune response of calves to bluetongue virus infection. American Journal of Veterinary Research 48:1031–1035
    [Google Scholar]
  23. MacLachlan N. J., Jagels G., Rossitto P. V., Moore P. F., Heidner H. W. 1990; The pathogenesis of experimental blue-tongue virus infection of calves. Veterinary Pathology 27:223–229
    [Google Scholar]
  24. Morrill J. C., McConnell S. 1985; An electron microscopic study of blood cells from calves experimentally infected with bluetongue virus. In Bluetongue and Related Orbiviruses pp. 279–287 Edited by Barber T. L., Jochim M. M. New York: Alan R. Liss;
    [Google Scholar]
  25. Moulton J. E. 1961; Pathology of bluetongue of sheep. Journal of the American Veterinary Medical Association 138:493–498
    [Google Scholar]
  26. Naessens J., Newson J., MacHugh N., Howard C. J., Parsons K., Jones B. 1990; Characterization of a bovine leucocyte differentiation antigen of 145, 000 MW restricted to B lymphocytes. Immunology 69:525–530
    [Google Scholar]
  27. Oi V. T., Glazer A. N., Stryer L. 1982; Fluorescent phycobiliprotein conjugates for analyses of cells and molecules. Journal of Cell Biology 93:981–986
    [Google Scholar]
  28. Parsonson I. M., Della-Porta A. J., McPhee D. A., Cybinski D. H., Squire K. R. E., Uren M. F. 1987; Experimental infection of bulls and cows with bluetongue virus serotype 20. Australian Veterinary Journal 64:10–13
    [Google Scholar]
  29. Reed L. J., Muench H. 1938; A simple method of estimating fifty percent endpoints. American Journal of Hygiene 27:493–497
    [Google Scholar]
  30. Richards R. G., MacLachlan N. J., Heidner H. W., Fuller F. J. 1988; Comparison of virologic and serologic responses of lambs and calves infected with bluetongue virus serotype 10. Veterinary Microbiology 18:233–242
    [Google Scholar]
  31. Spruell J. 1905; Malarial catarrhal fever (bluetongue) of sheep in South Africa. Journal of Comparative Pathology and Therapeutics 18:321–337
    [Google Scholar]
  32. Stott J. L., Blanchard-Channell M., Scibienski R. J., Stott M. L. 1990; Interaction of bluetongue virus with bovine lymphocytes. Journal of General Virology 71:363–368
    [Google Scholar]
  33. Taylor B. C., Stott J. L., Scibienski R. J., Redelman D. 1992; Development and characterization of a monoclonal antibody specific for the bovine low affinity IL-2 receptor, BoCD25. Immunology (in press)
    [Google Scholar]
  34. Teale A. J., Baldwin C. L., Morrison W. I., Ellis J., MacHugh N. D. 1987; Phenotypic and functional characteristics of bovine T lymphocytes. Veterinary Immunology and Immunopathology 17:113–123
    [Google Scholar]
  35. Whetter L. E., MacLachlan N. J., Gebhard D. H., Heidner H. W., Moore P. F. 1989; Bluetongue virus infection of bovine monocytes. Journal of General Virology 701663–1676
    [Google Scholar]
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