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Viruses 2017, 9(5), 113;

Antigenic and Biological Characterization of ORF2–6 Variants at Early Times Following PRRSV Infection

Department of Animal Science, Iowa State University, Ames, IA 50011, USA
Departments of Statistics and Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50011, USA
Author to whom correspondence should be addressed.
Academic Editors: Linda Dixon and Simon Graham
Received: 8 March 2017 / Revised: 18 April 2017 / Accepted: 24 April 2017 / Published: 16 May 2017
(This article belongs to the Special Issue Porcine Viruses)
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Genetic diversity of porcine reproductive and respiratory syndrome virus (PRRSV) challenges efforts to develop effective and broadly acting vaccines. Although genetic variation in PRRSV has been extensively documented, the effects of this variation on virus phenotype are less well understood. In the present study, PRRSV open reading frame (ORF)2–6 variants predominant during the first six weeks following experimental infection were characterized for antigenic and replication phenotype. There was limited genetic variation during these early times after infection; however, distinct ORF2–6 haplotypes that differed from the NVSL97-7895 inoculum were identified in each of the five pigs examined. Chimeric viruses containing all or part of predominant ORF2–6 haplotypes were constructed and tested in virus neutralization and in vitro replication assays. In two pigs, genetic variation in ORF2–6 resulted in increased resistance to neutralization by autologous sera. Mapping studies indicated that variation in either ORF2–4 or ORF5–6 could confer increased neutralization resistance, but there was no single amino acid substitution that was predictive of neutralization phenotype. Detailed analyses of the early steps in PRRSV replication in the presence and absence of neutralizing antibody revealed both significant inhibition of virion attachment and, independently, a significant delay in the appearance of newly synthesized viral RNA. In all pigs, genetic variation in ORF2–6 also resulted in significant reduction in infectivity on MARC-145 cells, suggesting variation in ORF2–6 may also be important for virus replication in vivo. Together, these data reveal that variation appearing early after infection, though limited, alters important virus phenotypes and contributes to antigenic and biologic diversity of PRRSV. View Full-Text
Keywords: PRRSV; genetic diversity; antigenic variation; replication phenotype; neutralization PRRSV; genetic diversity; antigenic variation; replication phenotype; neutralization

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Evans, A.B.; Loyd, H.; Dunkelberger, J.R.; van Tol, S.; Bolton, M.J.; Dorman, K.S.; Dekkers, J.C.M.; Carpenter, S. Antigenic and Biological Characterization of ORF2–6 Variants at Early Times Following PRRSV Infection. Viruses 2017, 9, 113.

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