title: A single amino acid substitution alter antigenicity of Glycosylated protein 4 of HP-PRRSV.
abstract: Porcine reproductive and respiratory syndrome (PRRS) is an important pig endemic disease in pork-producing countries worldwide. The etiology, porcine reproductive and respiratory syndrome virus (PRRSV), is characterized by fast antigen variability. Glycosylated protein 4 (GP4) is a minor protein in PRRSV virion, but contributes to induce protective immune responses. However, the antigenic characterization of PRRSV GP4 and the role of the mutations in this protein in PRRSV evolution are not clear.
  
  
  
  Peptides chip scanning and peptide based ELISA was used to analyze the antigenic characterization of HP-PRRSV GP4. A total of 142 peptides printed on a chip were used to reveal the antigen reaction characteristics of the HP-PRRSV. The reactions of these peptides with HP-PRRSV-specific pig serum were scanned and quantified using the software PepSlide® Analyzer by fluorescence intensity. The active reaction regions (AR) were identified based on the scanning results and then the amino acids (aa) sequences of AR(s) is aligned among PRRSV strains for further identify the key aa site(s) impact the antigenicity of the protein. Peptide based ELISA is then reacted with PRRSV positive sera derived from pig inoculated with different PRRSV strains for further analysis the role of specific amino acid in AR.
  
  
  
  The intensity plot was used to show the reactions of the peptides with PRRSV serum and it showed that enormously different response happened to various parts of GP4. The highest reaction intensity value reached 6401.5 against one peptide with the sequence DIKTNTTAASDFVVL. An AR from S29 to G56 was identified. Sequence alignment revealed various mutations in site 43 and possibly played an important role in this AR. Peptides ELISA reaction with sera from pigs inoculated with different PRRSV strain revealed that the change of aa in site 43 reduced the reaction of the peptide with PRRSV positive sera derived from pigs inoculated with the peptide related PRRSV strains.
  
  
  
  In this study, one AR covering S29 to G56 was identified in GP4. The aa in site 43 play an important role in determining the antigenic character of GP4. The continual mutations (S → G → D → N) occurred in this site alter the antigenicity of PRRSV GP4.
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China. wxlong@nwsuaf.edu.cn.|Northwest A&F University, No. 3 Taicheng Road, 712100, Yangling, Shaanxi, China. wxlong@nwsuaf.edu.cn.
authors.forename: Xinglong
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Zhenbin
authors.initials: Z
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Hongyu
authors.initials: H
authors.lastname: Xu
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Biao
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China. nwafucrb@gmail.com.|Northwest A&F University, No. 3 Taicheng Road, 712100, Yangling, Shaanxi, China. nwafucrb@gmail.com.
authors.forename: Zengqi
authors.initials: Z
authors.lastname: Yang
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology*; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000276:immunology*; D013552:Swine
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D006023:Glycoproteins