1887

Abstract

Glycoprotein 5 (GP) is the major glycoprotein of porcine reproductive and respiratory syndrome virus (PRRSV). Expression of GP has been improved by removing the transmembrane regions. Vectors were constructed encoding complete GP plus three mutants: GP ΔNs (residues 28–201), GP[30–67] (residues 30–67) and GP[30–201] (residues 30–67/130–201). The three deletion mutants were expressed at levels 20–30 times higher than complete GP. GP[30–201] was well recognized in ELISA or immunoblotting by a collection of pig sera. All the fragments were tested for the generation of MAbs, but only the polyhistidine-tagged fragment GP[30–201]H elicited an antibody response sufficient to produce MAbs. The two MAbs were positive for PRRSV in ELISA and immunoblotting, but negative for virus neutralization. MAb 4BE12 reacted with residues 130–170 and MAb 3AH9 recognized residues 170–201. This region was recognized strongly in immunoblotting by a collection of infected-pig sera. These results indicate diagnostic potential for this epitope.

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2001-05-01
2024-04-26
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References

  1. Balasuriya U. B., Patton J. F., Rossitto P. V., Timoney P. J., McCollum W. H., MacLachlan N. J. 1997; Neutralization determinants of laboratory strains and field isolates of equine arteritis virus: identification of four neutralization sites in the amino-terminal ectodomain of the GL envelope glycoprotein. Virology 232:114–128
    [Google Scholar]
  2. Benfield D. A., Nelson E., Collins J. E., Harris L., Goyal S. M., Robison D., Christianson W. T., Morrison R. B., Gorcyca D., Chladek D. 1992; Characterization of swine infertility and respiratory syndrome (SIRS) virus (isolate ATCC VR-2332). Journal of Veterinary Diagnostic Investigation 4:127–133
    [Google Scholar]
  3. Chirnside E. D., de Vries A. A. F., Mumford J. A., Rottier P. J. M. 1995; Equine arteritis virus-neutralizing antibody in the horse is induced by a determinant on the large envelope glycoprotein GL. Journal of General Virology 76:1989–1998
    [Google Scholar]
  4. Conzelmann K. K., Visser N., Van Woensel P., Thiel H. J. 1993; Molecular characterization of porcine reproductive and respiratory syndrome virus, a member of the arterivirus group. Virology 193:329–339
    [Google Scholar]
  5. Dea S., Gagnon C. A., Mardassi H., Pirzadeh B., Rogan D. 2000; Current knowledge on the structural proteins of porcine reproductive and respiratory syndrome (PRRS) virus: comparison of the North American and European isolates. Archives of Virology 145:659–688
    [Google Scholar]
  6. Goldberg T. L., Hahn E. C., Weigel R. M., Scherba G. 2000; Genetic, geographical and temporal variation of porcine reproductive and respiratory syndrome virus in Illinois. Journal of General Virology 81:171–179
    [Google Scholar]
  7. Hulst M. M., Westra D. F., Wensvoort G., Moormann R. J. 1993; Glycoprotein E1 of hog cholera virus expressed in insect cells protects swine from hog cholera. Journal of Virology 67:5435–5442
    [Google Scholar]
  8. Kreutz L. C., Mengeling W. L. 1997; Baculovirus expression and immunological detection of the major structural proteins of porcine reproductive and respiratory syndrome virus. Veterinary Microbiology 59:1–13
    [Google Scholar]
  9. Loemba H. D., Mounir S., Mardassi H., Archambault D., Dea S. 1996; Kinetics of humoral immune response to the major structural proteins of the porcine reproductive and respiratory syndrome virus. Archives of Virology 141:751–761
    [Google Scholar]
  10. Martinez-Torrecuadrada J. L., Casal J. I. 1995; Identification of a linear neutralization domain in the protein VP2 of African horse sickness virus. Virology 210:391–399
    [Google Scholar]
  11. Meulenberg J. J., Hulst M. M., de Meijer E. J., Moonen P. L., den Besten A., de Kluyver E. P., Wensvoort G., Moormann R. J. 1993; Lelystad virus, the causative agent of porcine epidemic abortion and respiratory syndrome (PEARS), is related to LDV and EAV. Virology 192:62–72
    [Google Scholar]
  12. Meulenberg J. J., Petersen-den Besten A., De Kluyver E. P., Moormann R. J., Schaaper W. M., Wensvoort G. 1995; Characterization of proteins encoded by ORFs 2 to 7 of Lelystad virus. Virology 206:155–163
    [Google Scholar]
  13. Morrison R. B., Collins J. E., Harris L., Christianson W. T., Benfield D. A., Chladek D. W., Gorcyca D. E., Joo H. S. 1992; Serologic evidence incriminating a recently isolated virus (ATCC VR-2332) as the cause of swine infertility and respiratory syndrome (SIRS). Journal of Veterinary Diagnostic Investigation 4:186–188
    [Google Scholar]
  14. Pirzadeh B., Dea S. 1997; Monoclonal antibodies to the ORF5 product of porcine reproductive and respiratory syndrome virus define linear neutralizing determinants. Journal of General Virology 78:1867–1873
    [Google Scholar]
  15. Pirzadeh B., Dea S. 1998; Immune response in pigs vaccinated with plasmid DNA encoding ORF5 of porcine reproductive and respiratory syndrome virus. Journal of General Virology 79:989–999
    [Google Scholar]
  16. Pirzadeh B., Gagnon C. A., Dea S. 1998; Genomic and antigenic variations of porcine reproductive and respiratory syndrome virus major envelope GP5 glycoprotein. Canadian Journal of Veterinary Research 62:170–177
    [Google Scholar]
  17. Plana Duran J., Climent I., Sarraseca J., Urniza A., Cortes E., Vela C., Casal J. I. 1997; Baculovirus expression of proteins of porcine reproductive and respiratory syndrome virus strain Olot/91. Involvement of ORF3 and ORF5 proteins in protection. Virus Genes 14:19–29
    [Google Scholar]
  18. Pont-Kingdon G. 1994; Creation of chimeric junctions, deletions and insertions by PCR. Methods in Molecular Biology 67:167–172
    [Google Scholar]
  19. Rodriguez M. J., Sarraseca J., Garcia J., Sanz A., Plana-Durán J., Casal J. I. 1997; Epitope mapping of the nucleocapsid protein of European and North American isolates of porcine reproductive and respiratory syndrome virus. Journal of General Virology 78:2269–2278
    [Google Scholar]
  20. Rost B. 1996; PHD: predicting one-dimensional protein structure by profile-based neural networks. Methods in Enzymology 266:525–539
    [Google Scholar]
  21. Rost B., Casadio R., Fariselli P., Sander C. 1995; Transmembrane helices predicted at 95% accuracy. Protein Science 4:521–533
    [Google Scholar]
  22. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. 1990; Use of T7 RNA polymerase to direct expression of cloned genes. Methods in Enzymology 185:60–89
    [Google Scholar]
  23. Wensvoort G., Terpstra C., Pol J. M., ter Laak E. A., Bloemraad M., de Kluyver E. P., Kragten C., van Buiten L., den Besten A., Wagenaar F. and others 1991; Mystery swine disease in The Netherlands: the isolation of Lelystad virus. Veterinary Quarterly 13:121–130
    [Google Scholar]
  24. Wieczorek-Krohmer M., Weiland F., Conzelmann K., Kohl D., Visser N., van Woensel P., Thiel H. J., Weiland E. 1996; Porcine reproductive and respiratory syndrome virus (PRRSV): monoclonal antibodies detect common epitopes on two viral proteins of European and US isolates. Veterinary Microbiology 51:257–266
    [Google Scholar]
  25. Zhang Y., Sharma R. D., Paul P. S. 1998; Monoclonal antibodies against conformationally dependent epitopes on porcine reproductive and respiratory syndrome virus. Veterinary Microbiology 63:125–136
    [Google Scholar]
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