1887

Abstract

A panel of six neutralizing murine monoclonal antibodies (MAbs) to equine arteritis virus (EAV) was produced. The MAbs were characterized by Western immunoblotting assay and competitive ELISA. The six MAbs identify a single neutralization site on a 29K envelope glycoprotein. Deglycosylation of viral proteins prior to immunoblotting showed that the 29K protein is the glycosylated form of a 20K protein. Equine anti-EAV serum also strongly bound the 29K glycoprotein, as well as an unglycosylated protein of 17K. The equine antisera to EAV blocked the binding of a selected MAb to EAV, whereas normal equine serum did not. Two neutralization-resistant escape mutant (EM) variants of the EAV prototype were produced using MAb 6D10. The phenotypic properties of the EM viruses were characterized by neutralization and immunoblotting assays with two MAbs (6D10 and 5G11). The two MAbs failed to neutralize either EM virus, and they did not react in an immunoblot assay with any proteins of the EM viruses. In contrast, binding of the equine antiserum to viral proteins was equivalent with prototype and EM virus strains. These data clearly indicate that a 29K envelope glycoprotein expresses at least one neutralization determinant of EAV.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-74-11-2525
1993-11-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/74/11/JV0740112525.html?itemId=/content/journal/jgv/10.1099/0022-1317-74-11-2525&mimeType=html&fmt=ahah

References

  1. Chirnside E. D. 1992; Equine arteritis virus: an overview. British Veterinary Journal 148:181–197
    [Google Scholar]
  2. Den Boon J. A., Snijder E. J., Chirnside E. D., De Vries A. A. F., Horzinek M. C., Spaan W. J. M. 1991; Equine arteritis virus is not a togavirus but belongs to the coronavirus like superfamily. Journal of Virology 65:2910–2920
    [Google Scholar]
  3. De Vries A. A. F., Chirnside E. D., Horzinek M. C., Rottier P. J. M. 1992; Structural proteins of equine arteritis virus. Journal of Virology 66:6294–6303
    [Google Scholar]
  4. Doll E. R., Bryans E. T., McCollum W. H., Crow M. E. W. 1957; Isolation of a filterable agent causing arteritis of horses and abortion by mares. Its differentiation from the equine abortion (influenza) virus. Cornell Veterinarian 47:3–41
    [Google Scholar]
  5. Goding J. W. 1986 Monoclonal Antibodies: Principles and Practices 2nd edn Orlando: Academic Press;
    [Google Scholar]
  6. Harlow E., Lane D. 1988; Immunoblotting. In Antibodies A Laboratory Manual pp 471–540 New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  7. 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]
  8. Hyllseth B. 1973; Structural proteins of equine arteritis virus. Archiv für die gesamte Virusforschung 40:177–188
    [Google Scholar]
  9. Iwashita O., Harasawa R. 1987a; Translation of equine arteritis virus RNA in rabbit reticulocyte lysates. Japanese Journal of Veterinary Science 49:387–389
    [Google Scholar]
  10. Iwashita O., Harasawa R. 1987b; Structural polypeptides of equine arteritis virus. Japanese Journal of Veterinary Science 49:923–925
    [Google Scholar]
  11. Kearney J. F., Radbruch A., Liesegang B., Rajewsky K. 1979; A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. Journal of Immunology 123:1548–1550
    [Google Scholar]
  12. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  13. Liddell J. E., Cryer A. 1991; Labelling of monoclonal antibodies. In A Practical Guide to Monoclonal Antibodies pp 122–138 Chichester: John Wiley & Sons;
    [Google Scholar]
  14. Lietzke R., Unsicker K. 1985; A statistical approach to determine monoclonality after limiting cell plating of a hybridoma clone. Journal of Immunological Methods 76:223–228
    [Google Scholar]
  15. McCollum W. H. 1976; Studies of passive immunity in foals to equine viral arteritis. Veterinary Microbiology 1:45–54
    [Google Scholar]
  16. Ouchterlony Ö, Nilsson L.-Å. 1986; Immunodiffusion and immunoelectrophoresis. In Handbook of Experimental Immunology vol 1 Immunohistochemistry, pp. 32 1–32 50 Edited by Weir D. M. Oxford: Blackwell Scientific Publications;
    [Google Scholar]
  17. Reddington J., Reddington G., MacLachlan N. J. 1991; A competitive ELISA for detection of antibodies to the five serotypes of bluetongue virus present in the United States. Journal of Veterinary Diagnostic Investigation 3:144–147
    [Google Scholar]
  18. Reed L. J., Muench H. 1938; A simple method of estimating fifty percent endpoints. American Journal of Hygiene 27:493–497
    [Google Scholar]
  19. 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]
  20. Timoney P. J., McCollum W. H. 1988; Equine viral arteritis–epidemiology and control. Equine Veterinary Science 8:54–59
    [Google Scholar]
  21. Timoney P. J., McCollum W. H., Murphy T. W., Roberts A. W., Willard J. G., Carswell G. D. 1987; The carrier state in equine viral arteritis virus infection in the stallion with specific emphasis on the venereal mode of virus transmission. Journal of Reproduction and Fertility Supplement 3595–102
    [Google Scholar]
  22. Van Berlo M. F., Horzinek M. C., Zeijst B. A. M. 1982; Equine arthritis virus-infected cells contain six virus-specific RNAs. Virology 118:345–352
    [Google Scholar]
  23. Van Berlo M. F., Rottier P. J. M., Spaan W. J. M., Horzinek M. C. 1986; Equine arteritis virus-induced polypeptide synthesis. Journal of General Virology 67:1543–1549
    [Google Scholar]
  24. Westaway E. G., Brinton M. A., Gaidamovich S. Ya., Horzinek M. C., Igarashi A., Kääriäinen L., Lvov D. K., Porterfield J. S., Russell P. K., Trent D. W. 1985; Togaviridae. Intervirology 24:125–139
    [Google Scholar]
  25. 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 70:1663–1676
    [Google Scholar]
  26. Zeegers J. J. W., Van Der Zeijst B. A., Horzinek M. C. 1976; The structural proteins of equine arteritis virus. Virology 73:200–205
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-74-11-2525
Loading
/content/journal/jgv/10.1099/0022-1317-74-11-2525
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