1887

Abstract

SUMMARY

Cultures of adherent and non-adherent bovine blood mononuclear cells were infected with bluetongue virus (BTV) serotype 10. Production of BTV proteins in mononuclear cell cultures was detected by immune precipitation of viral proteins from [S]methionine-labelled extracts of these cells, by immunofluorescence staining of cells using monoclonal antibodies (MAbs) to BTV proteins VP7 and NS2, and by flow cytometry with MAbs to VP2, VP7, NS1 and NS2. BTV-infected cells were most numerous in cultures of adherent mononuclear cells; infected cells were initially identified as monocytes on the basis of their morphology, and size and scatter characteristics as determined by analysis with a fluorescence-activated cell sorter (FACS). The majority of adherent mononuclear cells with these scatter characteristics were confirmed to be monocytes by FACS analysis with a MAb specific for bovine monocytes. Identification of BTV-infected adherent mononuclear cells as monocytes was further established by double immunofluorescent labelling, as infected adherent cells reacted with the MAb specific for bovine monocytes, and with another MAb specific for class II antigen. Infection of adherent mononuclear cells was also confirmed by transmission electron microscopy, as BTV virions and tubules were present in lysates of cultures of BTV-infected adherent mononuclear cells and within the cytoplasm of adherent cells. In contrast, BTV proteins were detected in few cells identified as lymphocytes on the basis of their scatter characteristics, and mean fluorescence of such cells was considerably less than that of BTV-infected monocytes. Viraemia persisted until 35 days after inoculation of a colostrum-deprived calf inoculated with BTV. Virus was isolated from blood mononuclear cells at 1 week after infection of the calf, but not thereafter. BTV infection of blood mononuclear cells was demonstrated until 9 days after inoculation by indirect immunofluorescence staining of mononuclear cells. In contrast, virus was consistently isolated from erythrocyte-enriched preparations throughout viraemia in titres comparable to those in whole blood. These results indicate that although bovine monocytes are readily infected with this strain of BTV serotype 10, infection of blood monocytes is unlikely to be responsible for the prolonged viraemia that consistently occurs in BTV-infected cattle.

Keyword(s): BTV , cattle and monocytes
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-70-7-1663
1989-07-01
2024-11-08
Loading full text...

Full text loading...

/deliver/fulltext/jgv/70/7/JV0700071663.html?itemId=/content/journal/jgv/10.1099/0022-1317-70-7-1663&mimeType=html&fmt=ahah

References

  1. Appleton J. A., Letchworth G. I. 1983; Monoclonal antibody analysis of serotype-restricted and unrestricted bluetongue viral antigenic determinants. Virology 124:286–299
    [Google Scholar]
  2. Baldwin C. L., Morrison W. I., Naessens J. 1988; Differentiation antigens and functional characteristics of bovine leukocytes. In Differentiation Antigens in Lymphohemopoietic Tissues455–498 Miyasaka M., Trnka Z. New York: Marcel Dekker;
    [Google Scholar]
  3. Collisson E. W., Barber T. L. 1983; Blood cells associated with bluetongue virus infection in cattle. American Association of Veterinary Laboratory Diagnosticians, 26th Annual Proceedings287–300
    [Google Scholar]
  4. Davis W. C, Marusic S., Lewin H. A., Splitter G. A., Perryman L. E., Mcguire T. C., Gorham J. R. 1987; The development and analysis of species specific and cross reactive monoclonal antibodies to leukocyte differentiation antigens and antigens of the major histocompatibility complex for use in the study of the immune system in cattle and other species. Veterinary Immunology and Immunopathology 15:337–376
    [Google Scholar]
  5. Doyle M. V., Oldstone M. B. A. 1979; Persistence of virus in lymphocytes during infection. In Virus-Lymphocyte Interactions: Implications for Disease37–52 Profitt M. R. Amsterdam: Elsevier/North-Holland;
    [Google Scholar]
  6. 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]
  7. Eaton B. T., Hyatt A. D., White J. R. 1987; Association of bluetongue virus with the cytoskeleton. Virology 157:107–116
    [Google Scholar]
  8. Emmons R. W., Oshiro L. S., Johnson H. N., lennette E. H. 1972; Intra-erythrocytic location of Colorado tick fever virus. Journal of General Virology 17:185–195
    [Google Scholar]
  9. Heidner H. W., MacLachlan N. J., Fuller F. J., Richards R. G., Whetter L. E. 1988; Bluetongue virus genome remains stable throughout prolonged infection of cattle. Journal of General Virology 69:2629–2636
    [Google Scholar]
  10. Huismans H., Els H. J. 1979; Characterization of the tubules associated with the replication of three different Orbiviruses. Virology 92:397–406
    [Google Scholar]
  11. Huismans H., Van Der Walt N. T., Cloete M., Erasmus B. J. 1987; Isolation of a capsid protein of bluetongue virus that induces a protective immune response in sheep. Virology 157:172–179
    [Google Scholar]
  12. Luedke A. J. 1970; Distribution of virus in blood components during viremia of bluetongue. United States Animal Health Association, 74th Annual Proceedings9–21
    [Google Scholar]
  13. Luedke A. I., Jochim M. M., Tones R. H. 1969; Bluetongue in cattle: viremia. American Journal of Veterinary Research 30:511–516
    [Google Scholar]
  14. 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]
  15. 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]
  16. 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]
  17. Mitchell K. F., Fuhrer J. P., Steplewski Z., Koprowski H. 1980; Biochemical characterization of human melanoma cell surfaces: dissection with monoclonal antibodies. Proceedings of the National Academv of SciencesU.S.A. 77:7287–7291
    [Google Scholar]
  18. Moore P. F., Ellingsworth L. R., Toedter G. P. 1986; Development and characterization of a monoclonal antibody to class II MHC antigens in rhesus macaques. Tissue Antigens 28:88–99
    [Google Scholar]
  19. Morahan P. S., Morse S. S. 1979; Macrophage-virus interactions. In Virus-Lymphocyte Interactions:Implications for Disease17–35 Profitt M. R. AmsterdamElsevier/North Holland
    [Google Scholar]
  20. Morrill J. C., McConnell S. 1985; An electron microscopic study of blood cells from calves experimentally infected with bluetongue virus. In Bluetongue and Related Orbiviruses279–287 Barber T. L., Jochim. M. M. New YorkAlan R. Liss
    [Google Scholar]
  21. Natuk R. J., Holowczak J. A. 1985; Vaccinia virus proteins on the plasma membrane of infected cells. III.Infection of peritoneal macrophages. Virology 147:354–372
    [Google Scholar]
  22. Nowakowski M. 1985; Macrophage surface and intracellular changes caused by influenza virus and interferon. In Macrophage Biology463–171 Reichard S., Kojima M. New YorkAlan R. Liss
    [Google Scholar]
  23. 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]
  24. Pennline K. J. 1981; Adherence to plastic or glass surfaces. In Manual of Macrophage Methodology. 63–71 Herscowitz H. B., Holden H. T., Ballanti J. A., Ghaffer A. New YorkMarcel Decker
    [Google Scholar]
  25. 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]
  26. Unanue E. R., Allen P. M. 1987; The basis for the immunoregulatory role of macrophages and other accessory cells. Science 236:551–557
    [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-70-7-1663
Loading
/content/journal/jgv/10.1099/0022-1317-70-7-1663
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error