title: Immunodominant epitopes in nsp2 of porcine reproductive and respiratory syndrome virus are dispensable for replication, but play an important role in modulation of the host immune response.
abstract: Non-structural protein 2 (nsp2) of porcine reproductive and respiratory syndrome virus (PRRSV) is the largest protein of this virus. In addition to its crucial role in virus replication, recent studies have indicated its involvement in modulating host immunity. In this study, each of the six identified immunodominant nsp2 B-cell epitopes (ES2-ES7) was deleted using a type I PRRSV cDNA infectious clone. Deletion of ES3, ES4 or ES7 allowed the generation of viable virus. In comparison with the parental virus, the DeltaES3 mutant showed increased cytolytic activity and more vigorous growth kinetics, whilst the DeltaES4 and DeltaES7 mutants displayed decreased cytolytic activity and slower growth kinetics in MARC-145 cells. These nsp2 mutants were characterized further in a nursery pig disease model. The results showed that the DeltaES4 and DeltaES7 mutants exhibited attenuated phenotypes, whereas the DeltaES3 mutant produced a higher peak viral load in pigs. The antibody response reached similar levels, as measured by IDEXX ELISA at 21 days post-infection, and slightly higher levels of mean virus neutralizing titres were observed from pigs infected by the DeltaES4 and DeltaES7 mutants. The expression of innate and T-helper 1 cytokines was measured in peripheral blood mononuclear cells or virus-infected macrophages. The results consistently showed that interleukin-1beta and tumour necrosis factor alpha expression levels were downregulated in cells that were stimulated (or infected) with the DeltaES3 mutant compared with parental virus and the other nsp2 deletion mutants. These results suggest that certain regions in nsp2 are non-essential for PRRSV replication but may play an important role in modulation of host immunity in vivo.
authors.affiliation: Veterinary Science Department, South Dakota State University, Brookings, SD 57007, USA.
authors.forename: Zhenhai
authors.initials: Z
authors.lastname: Chen
authors.forename: Xiaoxin
authors.initials: X
authors.lastname: Zhou
authors.forename: Joan K
authors.initials: JK
authors.lastname: Lunney
authors.forename: Steven
authors.initials: S
authors.lastname: Lawson
authors.forename: Zhi
authors.initials: Z
authors.lastname: Sun
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Brown
authors.forename: Jane
authors.initials: J
authors.lastname: Christopher-Hennings
authors.forename: David
authors.initials: D
authors.lastname: Knudsen
authors.forename: Eric
authors.initials: E
authors.lastname: Nelson
authors.forename: Ying
authors.initials: Y
authors.lastname: Fang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics; D007113:Immunity, Innate; D016056:Immunodominant Epitopes; D008969:Molecular Sequence Data; D019318:Porcine Reproductive and Respiratory Syndrome / Q000276:immunology*; D019316:Porcine respiratory and reproductive syndrome virus / Q000254:growth & development / Q000276:immunology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D017361:Viral Nonstructural Proteins / Q000276:immunology*; D014779:Virus Replication
chemical_list: D000914:Antibodies, Viral; D016207:Cytokines; D016056:Immunodominant Epitopes; D017361:Viral Nonstructural Proteins