1887

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Volume 78, Issue 8

Monoclonal antibodies to the ORF5 product of porcine reproductive and respiratory syndrome virus define linear neutralizing determinants Free

Abstract

Complementary DNA encoding the ORF5 gene of a Quebec reference isolate (IAF-Klop) of porcine reproductive and respiratory syndrome virus (PRRSV) was cloned into the prokaryotic expression vectors pGEX-4T and pET21a to produce ORF5- glutathione S-transferase and ORF5-polyhistidine fusion proteins. Five hybridoma cell lines producing monoclonal antibodies (MAbs) to the 25 kDa viral envelope glycoprotein (GP) were obtained from BALB/c mice immunized with the affinity chromatography-purified GST-ORF5 fusion protein. The polypeptide specificity of these anti-PRRSV MAbs, belonging to the IgG1 isotype, was confirmed by Western immunoblotting assays with recombinant and native viral proteins, and by radioimmuno- precipitation using [S]methionine-labelled concentrated extracellular virus. All these MAbs showed virus-neutralizing (VN) activity, with VN titres ranging from 1:32 to 1:128. Two MAbs (IAF- 1B8 and IAF-8A8) reacted with similar titres with the modified live attenuated vaccine strain ATCC VR-2332, but all five failed to react to the prototype European strain, the Lelystad virus, in VN and indirect immunofluorescence tests. The results obtained suggest that these five anti-PRRSV MAbs are directed to serotype-specific linear neutralizing epitopes which are not affected by the absence of carbohydrate residues.

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© Society for General Microbiology Ltd 1997
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1997-08-01
2023-09-28
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References

  1. Balasuriya U. B. R., MacLachlan N. J., de Vries A. A. F., Rossito P. V., Rottier P. J. M. 1995; Identification of a neutralization site in the major envelope glycoprotein (GL) of equine arteritis virus. Virology 207:518–527
     [Google Scholar]
  2. 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]
  3. Chomczynski P., Sacchi N. 1987; Single-step method of RNA isolation by acid guanidium thiocyanate phenol-chloroform extraction. Analytical Biochemistry 162:156–159
     [Google Scholar]
  4. Collins J. E., Benfield D. A., Christianson W. T., Harris L., Christopher-Hennings J., Shaw D. P., Goyal S. M., Cullough S., Morrison R. B., Joo H. S., Gorcyca D., Chladek D. 1992; Isolation of swine infertility and respiratory syndrome virus (isolate ATCC VR- 2332) in North America and experimental reproduction of the disease in gnotobiotic pigs. Journal Veterinary Diagnostic Investigation 4:17–126
     [Google Scholar]
  5. Dea S., Tijssen P. 1989; Antigenic and polypeptide structure of turkey enteric coronaviruses as defined by monoclonal antibodies. Journal General Virology 70:1725–1741
     [Google Scholar]
  6. Dea S., Gagnon C. A., Mardassi H., Milane G. 1996; Antigenic variability among North-American and European strains of PRRSV as defined by monoclonal antibodies to the matrix protein. Journal ofClinical Microbiology 34:1488–1493
     [Google Scholar]
  7. Deregt D., de Vries A. A. F., Raamsman M. J. B., Elmgren L. D., Rottier P. J. M. 1994; Monoclonal antibodies to equine arteritis virus proteins identify the GLprotein as a target for virus neutralization. Journal of General Virology 75:2439–2444
     [Google Scholar]
  8. de Vries A. A. F., Post S. M., Raamsman M. J. B., Horzinek M. C., Rottier P. J. M. 1995; The two major envelope proteins of equine arteritis virus associate into disulphide-linked heterodimers. Journal of Virology 69:4668–4674
     [Google Scholar]
  9. Drew T. W., Meulenberg J. J. M., Sands J. J., Paton D. J. 1995; Production, characterization and reactivity of monoclonal antibodies to porcine reproductive and respiratory syndrome virus. Journal of General Virology 76:1361–1369
     [Google Scholar]
  10. Goyal S. M. 1993; Porcine reproductive and respiratory syndrome. Journal Veterinary Diagnostic Investigation 5:656–664
     [Google Scholar]
  11. Kim H. S., Kwang J., Yoon J. J., Joo H. S., Frey M. L. 1993; Enhanced replication of porcine reproductive and respiratory syndrome (PRRS) virus in a homogeneous subpopulation of MA-104 cell line. Archives of Virology 133:477–483
     [Google Scholar]
  12. Loemba H.D, 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]
  13. Mardassi H., Mounir S., Dea S. 1994; Identification of major differences in the nucleocapsid protein genes of a Quebec strain and European strains of porcine reproductive and respiratory syndrome virus. Journal of General Virology 75:681–685
     [Google Scholar]
  14. Mardassi H., Mounir S., Dea S. 1995; Molecular analysis of the ORFs 3 to 7 genes of porcine reproductive and respiratory syndrome virus, Quebec reference strain. Archives of Virology 140:1405–1418
     [Google Scholar]
  15. Mardassi H., Massie B., Dea S. 1996; Intracellular synthesis, processing and transport of proteins encoded by ORFs 5 to 7 of porcine reproductive and respiratory syndrome virus. Virology 221:98–112
     [Google Scholar]
  16. Meng X. J., Paul P. S., Halbur P. J., Lum M. A. 1995; Phylogenetic analyses of the putative M (ORF 6) and N (ORF 7) genes of porcine reproductive and respiratory syndrome virus (PRRSV): implication for the existence of two genotypes of PRRSV in the U.S.A. and Europe. Archives of Virology 140:745–755
     [Google Scholar]
  17. Meulenberg J. J. M., den Besten A. 1996; Identification and characterization of a sixth structural protein of Lelystad virus: the glycoprotein GP encoded by ORF 2 is incorporated in virus particles. Virology 225:44–51
     [Google Scholar]
  18. Meulenberg J. J. M., Hulst M., de Meijer E. J., Moonen P. L. J. M., den Besten A., de Kluyver E. P., Wensvoort G., Moormann R. J. M. 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]
  19. Meulenberg J. J. M., den Besten A., de Kluyer E. P., Moormann R. J. M., Schaaper V. M. M., Wensvoort G. 1995; Characterization of proteins encoded by ORFs 2 to 7 of Lelystad virus. Virology 206:155–163
     [Google Scholar]
  20. Nelson E. A., Christopher-Hennings J., Drew T., Wensvoort G., Collins J. E., Benfield D. A. 1993; Differentiation of US and European isolates of porcine reproductive and respiratory syndrome virus by monoclonal antibodies. Journal of Clinical Microbiology 31:3184–3189
     [Google Scholar]
  21. Nelson E. A., Christopher-Hennings J. T., Benfield D. A. 1994; Serum immune responses to the proteins of porcine reproductive and respiratory syndrome (PRRS) virus. Journal of Veterinary Diagnostic Investigation 6: 410–415
     [Google Scholar]
  22. Plagemann P. G. W., Moennig V. 1992; Lactate dehydrogenase- elevating virus, equine arteritis virus, and simian hemorrhagic fever virus : a new group of positive-strand RNA viruses. Advances in Virus Research 41:99–192
     [Google Scholar]
  23. van Nieuwstadt A. A. P., Meulenberg J. J. M., van Essen-Zendbergen A., den Besten A., Bende R. J., Moormann R. J. M., Wenswoort G. 1996; Proteins encoded by open reading frames 3 and 4 of the genome of Lelystad virus (Arteriviridae) are structural proteins of the virion. Journal of Virology 70:4767–4772
     [Google Scholar]
  24. Wensvoort G., Terpstra C., Pol J. M. A., Terlaak E. A., Bloemraad M. D. E., Kluyver E. P., Kragten C., van Buiten L., den Besten A., Wagenaar F., Broekhuijsen J. M., Moonen P. L. J. M., Zetstra T., de Boer E. A., Tibben H. J., de Jong M. F., van’t Veld P., Groenlandg J. R., van Gennep J. A., Voets M. T., Verheijden J. H. M., Braamskamp J. 1991; Mystery swine disease in The Netherlands: the isolation of Lelystad virus. Veterinary Quarterly 13:121–130
     [Google Scholar]
  25. Wensvoort G., Kluyver E. P., Luijtze E. A., den Besten A., Harris L., Collins J. E., Christianson W. T., Chladek D. 1992; Antigenic comparison of Lelystad virus and swine infertility and respiratory syndrome (SIRS) virus. Journal of Veterinary Diagnostic Investigation 4:134–138
     [Google Scholar]
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Monoclonal antibodies to the ORF5 product of porcine reproductive and respiratory syndrome virus define linear neutralizing determinants
J. Gen. Virol. 78, 1867 (1997); https://doi.org/10.1099/0022-1317-78-8-1867
/content/journal/jgv/10.1099/0022-1317-78-8-1867
/content/journal/jgv/10.1099/0022-1317-78-8-1867
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