Skip to main content

Advertisement

SpringerLink
Log in

Identification of novel epitopes targeting non-structural protein 2 of PRRSV using monoclonal antibodies

  • Applied genetics and molecular biotechnology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) is leading to huge losses in the swine industry worldwide. Its nonstructural protein 2 (Nsp2), with a cysteine protease domain (PL2), is crucial for virus replication and as a trigger to host innate immune regulation. In this study, three monoclonal antibodies (mAbs) to Nsp2, designated 4A12, 4G8, and 8H11, were generated. Subsequently, a sequence of recombinant peptides with partial overlap was utilized to determine the epitopes using these mAbs. We found three novel minimal linear Nsp2 B cell epitopes, 188ELSDDSNRPV197, 42HLKRYSPPAE51, and 54CGWHCISA61, which were identified by the antibodies 4A12, 4G8, and 8H11, respectively. Structure analysis indicates that 42HLKRYSPPAE51 and 188ELSDDSNRPV197 are located separately in hypervariable region 1 and hypervariable region 2 of Nsp2. Interestingly, 54CGWHCISA61 is located in the PL2 region, which is highly conserved in all arteriviruses, particularly at the expected conserved catalytic site at Cys54. Importantly, 54CGWHCISA61 is located in the inner region of the expected 3D structure of Nsp2, which reveals that the epitope is cryptic. These findings not only provide valuable insight for vaccine design and hold diagnostic potential for the identified epitopes, but also reveal a protective mechanism against variation under selective pressure in an important epitope.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Albina E (1997) Epidemiology of porcine reproductive and respiratory syndrome (PRRS): an overview. Vet Microbiol 55:309–316

    Article  CAS  PubMed  Google Scholar 

  • Albina E, Leforban Y, Baron T, Plana Duran J (1992) Blue-eared pig disease in Brittany: a new test. Vet Rec 130:83–84

    Article  CAS  PubMed  Google Scholar 

  • Cavanagh D (1997) Nidovirales: a new order comprising Coronaviridae and Arteriviridae. Arch Virol 142:629–633

    CAS  PubMed  Google Scholar 

  • Chen Z, Zhou X, Lunney JK, Lawson S, Sun Z, Brown E, Christopher-Hennings J, Knudsen D, Nelson E, Fang Y (2010) 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. J Gen Virol 91:1047–1057

    Article  CAS  PubMed  Google Scholar 

  • Chen N, Cao Z, Yu X, Deng X, Zhao T, Wang L, Liu Q, Li X, Tian K (2011) Emergence of novel European genotype porcine reproductive and respiratory syndrome virus in mainland China. J Gen Virol 92:880–892

    Article  CAS  PubMed  Google Scholar 

  • Collins JE, Benfield DA, Christianson WT, Harris L, Hennings JC, Shaw DP, Goyal SM, McCullough S, Morrison RB, Joo HS, 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. J Vet Diagn Investig 4:117–126

    Article  CAS  Google Scholar 

  • de Lima M, Pattnaik AK, Flores EF, Osorio FA (2006) Serologic marker candidates identified among B-cell linear epitopes of Nsp2 and structural proteins of a north American strain of porcine reproductive and respiratory syndrome virus. Virology 353:410–421

    Article  CAS  PubMed  Google Scholar 

  • Fang Y, Kim DY, Ropp S, Steen P, Christopher-Hennings J, Nelson EA, Rowland RR (2004) Heterogeneity in Nsp2 of European-like porcine reproductive and respiratory syndrome viruses isolated in the United States. Virus Res 100:229–235

    Article  CAS  PubMed  Google Scholar 

  • Fang Y, Schneider P, Zhang WP, Faaberg KS, Nelson EA, Rowland RR (2007) Diversity and evolution of a newly emerged north American type 1 porcine arterivirus: analysis of isolates collected between 1999 and 2004. Arch Virol 152:1009–1017

    Article  CAS  PubMed  Google Scholar 

  • Han J, Wang Y, Faaberg KS (2006) Complete genome analysis of RFLP 184 isolates of porcine reproductive and respiratory syndrome virus. Virus Res 122:175–182

    Article  CAS  PubMed  Google Scholar 

  • Han J, Liu G, Wang Y, Faaberg KS (2007) Identification of nonessential regions of the nsp2 replicase protein of porcine reproductive and respiratory syndrome virus strain VR-2332 for replication in cell culture. J Virol 81:9878–9890

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Han J, Rutherford MS, Faaberg KS (2009) The porcine reproductive and respiratory syndrome virus nsp2 cysteine protease domain possesses both trans- and cis-cleavage activities. J Virol 83:9449–9463

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kappes MA, Miller CL, Faaberg KS (2013) Highly divergent strains of porcine reproductive and respiratory syndrome virus incorporate multiple isoforms of nonstructural protein 2 into virions. J Virol 87:13456–13465

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li Y, Wang X, Bo K, Wang X, Tang B, Yang B, Jiang W, Jiang P (2007) Emergence of a highly pathogenic porcine reproductive and respiratory syndrome virus in the mid-eastern region of China. Vet J 174:577–584

    Article  CAS  PubMed  Google Scholar 

  • Lunney JK, Fang Y, Ladinig A, Chen N, Li Y, Rowland B, Renukaradhya GJ (2016) Porcine reproductive and respiratory syndrome virus (PRRSV): pathogenesis and interaction with the immune system. Annu Rev Anim Biosci 4:129–154

    Article  CAS  PubMed  Google Scholar 

  • Morin M, Robinson Y (1991) Porcine reproductive and respiratory syndrome in Quebec. Vet Rec 129:367–368

    Article  CAS  PubMed  Google Scholar 

  • Mortimer GM, Minchin RF (2016) Cryptic epitopes and functional diversity in extracellular proteins. Int J Biochem Cell Biol 81:112–120

    Article  CAS  PubMed  Google Scholar 

  • Oleksiewicz MB, Botner A, Toft P, Normann P, Storgaard T (2001) Epitope mapping porcine reproductive and respiratory syndrome virus by phage display: the nsp2 fragment of the replicase polyprotein contains a cluster of B-cell epitopes. J Virol 75:3277–3290

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pejsak Z, Stadejek T, Markowska-Daniel I (1997) Clinical signs and economic losses caused by porcine reproductive and respiratory syndrome virus in a large breeding farm. Vet Microbiol 55:317–322

    Article  CAS  PubMed  Google Scholar 

  • Ramirez E, Moreno V, Diaz N, Osorio F, Ruiz A, Neira V, Quezada M (2008) Evaluation of the pathogenicity and transmissibility of a chilean isolate of porcine reproductive and respiratory syndrome virus. Transbound Emerg Dis 55:115–124

    Article  CAS  PubMed  Google Scholar 

  • Rascon-Castelo E, Burgara-Estrella A, Mateu E, Hernandez J (2015) Immunological features of the non-structural proteins of porcine reproductive and respiratory syndrome virus. Viruses 7:873–886

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rizzuto CD, Wyatt R, Hernandez-Ramos N, Sun Y, Kwong PD, Hendrickson WA, Sodroski J (1998) A conserved HIV gp120 glycoprotein structure involved in chemokine receptor binding. Science 280:1949–1953

    Article  CAS  PubMed  Google Scholar 

  • Schmitt S, Kuhn D, Klebe G (2002) A new method to detect related function among proteins independent of sequence and fold homology. J Mol Biol 323:387–406

    Article  CAS  PubMed  Google Scholar 

  • Sullivan N, Sun Y, Sattentau Q, Thali M, Wu D, Denisova G, Gershoni J, Robinson J, Moore J, Sodroski J (1998) CD4-induced conformational changes in the human immunodeficiency virus type 1 gp120 glycoprotein: consequences for virus entry and neutralization. J Virol 72:4694–4703

    CAS  PubMed  PubMed Central  Google Scholar 

  • Tian K, Yu X, Zhao T, Feng Y, Cao Z, Wang C, Hu Y, Chen X, Hu D, Tian X, Liu D, Zhang S, Deng X, Ding Y, Yang L, Zhang Y, Xiao H, Qiao M, Wang B, Hou L, Wang X, Yang X, Kang L, Sun M, Jin P, Wang S, Kitamura Y, Yan J, Gao GF (2007) Emergence of fatal PRRSV variants: unparalleled outbreaks of atypical PRRS in China and molecular dissection of the unique hallmark. PLoS One 2:e526

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang X, Yang X, Zhou R, Zhou L, Ge X, Guo X, Yang H (2016) Genomic characterization and pathogenicity of a strain of type 1 porcine reproductive and respiratory syndrome virus. Virus Res 225:40–49

    Article  CAS  PubMed  Google Scholar 

  • Wang F, Yang Y, Liu X, He M, Liu Y, Sun N, Zhu H, Ren J, Wu H, Wen Y (2017) Development of monoclonal antibody for differentiating porcine reproductive and respiratory syndrome virus and identification of a novel non-structural protein 2 epitope peptide. Virusdisease 28:408–415

    Article  PubMed  PubMed Central  Google Scholar 

  • Wensvoort G, Terpstra C, Pol JM, ter Laak EA, Bloemraad M, de Kluyver EP, Kragten C, van Buiten L, den Besten A, Wagenaar F, Broekhuijsen JM, Moonen PL, Zetstra T, de Boer EA, Tibben HJ, de Jong MF, van Veld P, Greenland GJ, van Gennep JA, Voets MT, Verheijden JH, Braamskamp J (1991) Mystery swine disease in the Netherlands: the isolation of Lelystad virus. Vet Q 13:121–130

    Article  CAS  PubMed  Google Scholar 

  • Wu L, Gerard NP, Wyatt R, Choe H, Parolin C, Ruffing N, Borsetti A, Cardoso AA, Desjardin E, Newman W, Gerard C, Sodroski J (1996) CD4-induced interaction of primary HIV-1 gp120 glycoproteins with the chemokine receptor CCR-5. Nature 384:179–183

    Article  CAS  Google Scholar 

  • Wyatt R, Kwong PD, Desjardins E, Sweet RW, Robinson J, Hendrickson WA, Sodroski JG (1998) The antigenic structure of the HIV gp120 envelope glycoprotein. Nature 393:705–711

    Article  CAS  PubMed  Google Scholar 

  • Yan Y, Guo X, Ge X, Chen Y, Cha Z, Yang H (2007) Monoclonal antibody and porcine antisera recognized B-cell epitopes of Nsp2 protein of a Chinese strain of porcine reproductive and respiratory syndrome virus. Virus Res 126:207–215

    Article  CAS  PubMed  Google Scholar 

  • Zhou YJ, Hao XF, Tian ZJ, Tong GZ, Yoo D, An TQ, Zhou T, Li GX, Qiu HJ, Wei TC, Yuan XF (2008) Highly virulent porcine reproductive and respiratory syndrome virus emerged in China. Transbound Emerg Dis 55:152–164

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

This study was supported by National Key R&D Program of China (2017YFD0501600), the Chinese Academy of Agricultural Sciences Foundation (1610302017010 and Y2016JC23), the National Natural Science Foundation of China (31640083 and 31872448), and the State Key Laboratory of Veterinary Biotechnological Foundation (SKLVBP2018002).

Author information

Author notes

  1. Caihong Bi and Zengyu Shao contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, 150069, Heilongjiang, China

    Caihong Bi, Zengyu Shao, Jiangnan Li & Changjiang Weng

Authors
  1. Caihong Bi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zengyu Shao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jiangnan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Changjiang Weng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Changjiang Weng.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests.

Ethics statement

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bi, C., Shao, Z., Li, J. et al. Identification of novel epitopes targeting non-structural protein 2 of PRRSV using monoclonal antibodies. Appl Microbiol Biotechnol 103, 2689–2699 (2019). https://doi.org/10.1007/s00253-019-09665-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-019-09665-7

Keywords

Search

Navigation