Characterization of the neutralization determinants of equine arteritis virus using recombinant chimeric viruses and site-specific mutagenesis of an infectious cDNA clone

Udeni B R Balasuriya, Jessika C. Dobbe, Hans W. Heidner, Victoria L. Smalley, Andrea Navarrette, Eric J. Snijder, Nigel J Maclachlan

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

We have used an infectious cDNA clone of equine arteritis virus (EAV) and reverse genetics technology to further characterize the neutralization determinants in the GP5 envelope glycoprotein of the virus. We generated a panel of 20 recombinant viruses, including 10 chimeric viruses that each contained the ORF5 (which encodes GP5) of different laboratory, field, and vaccine strains of EAV, a chimeric virus containing the N-terminal ectodomain of GP5 of a European strain of porcine reproductive and respiratory syndrome virus, and 9 mutant viruses with site-specific substitutions in their GP5 proteins. The neutralization phenotype of each recombinant chimeric/mutant strain of EAV was determined with EAV-specific monoclonal antibodies and EAV strain-specific polyclonal equine antisera and compared to that of their parental viruses from which the substituted ORF5 was derived. The data unequivocally confirm that the GP5 ectodomain contains critical determinants of EAV neutralization. Furthermore, individual neutralization sites are conformationally interactive, and the interaction of GP5 with the unglycosylated membrane protein M is likely critical to expression of individual epitopes in neutralizing conformation. Substitution of individual amino acids within the GP5 ectodomain usually resulted in differences in neutralization phenotype of the recombinant viruses, analogous to differences in the neutralization phenotype of field strains of EAV and variants generated during persistent infection of EAV carrier stallions.

Original languageEnglish (US)
Pages (from-to)235-246
Number of pages12
JournalVirology
Volume321
Issue number2
DOIs
StatePublished - Apr 10 2004

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Equine Arteritis Virus
Site-Directed Mutagenesis
Complementary DNA
Clone Cells
Viruses
Phenotype
Porcine respiratory and reproductive syndrome virus
Reverse Genetics
Amino Acid Substitution
Horses
Immune Sera
Epitopes
Glycoproteins
Membrane Proteins
Vaccines
Monoclonal Antibodies
Technology

Keywords

  • cDNA clone
  • Chimeric viruses
  • Envelope
  • Equine arteritis virus
  • Neutralization

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Characterization of the neutralization determinants of equine arteritis virus using recombinant chimeric viruses and site-specific mutagenesis of an infectious cDNA clone. / Balasuriya, Udeni B R; Dobbe, Jessika C.; Heidner, Hans W.; Smalley, Victoria L.; Navarrette, Andrea; Snijder, Eric J.; Maclachlan, Nigel J.

In: Virology, Vol. 321, No. 2, 10.04.2004, p. 235-246.

Research output: Contribution to journalArticle

Balasuriya, Udeni B R ; Dobbe, Jessika C. ; Heidner, Hans W. ; Smalley, Victoria L. ; Navarrette, Andrea ; Snijder, Eric J. ; Maclachlan, Nigel J. / Characterization of the neutralization determinants of equine arteritis virus using recombinant chimeric viruses and site-specific mutagenesis of an infectious cDNA clone. In: Virology. 2004 ; Vol. 321, No. 2. pp. 235-246.
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