Advertisement

Virologica Sinica

, Volume 34, Issue 6, pp 631–640 | Cite as

Nsp2 and GP5-M of Porcine Reproductive and Respiratory Syndrome Virus Contribute to Targets for Neutralizing Antibodies

  • Jia Su
  • Lei Zhou
  • Bicheng He
  • Xinhui Zhang
  • Xinna Ge
  • Jun Han
  • Xin GuoEmail author
  • Hanchun Yang
Research Article
  • 223 Downloads

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) is characterized by its genetic variation and limited cross protection among heterologous strains. Even though several viral structural proteins have been regarded as inducers of neutralizing antibodies (NAs) against PRRSV, the mechanism underlying limited cross-neutralization among heterologous strains is still controversial. In the present study, examinations of NA cross reaction between a highly pathogenic PRRSV (HP-PRRSV) strain, JXwn06, and a low pathogenic PRRSV (LP-PRRSV) strain, HB-1/3.9, were conducted with viral neutralization assays in MARC-145 cells. None of the JXwn06-hyperimmuned pigs’ sera could neutralize HB-1/3.9 in vitro and vice versa. To address the genetic variation between these two viruses that are associated with limited cross-neutralization, chimeric viruses with coding regions swapped between these two strains were constructed. Viral neutralization assays indicated that variations in nonstructural protein 2 (nsp2) and structural proteins together contribute to weak cross-neutralization activity between JXwn06 and HB-1/3.9. Furthermore, we substituted the nsp2-, glycoprotein2 (GP2)-, GP3-, and GP4-coding regions together, or nsp2-, GP5-, and membrane (M) protein-coding regions simultaneously between these two viruses to construct chimeric viruses to test cross-neutralization reactivity with hyperimmunized sera induced by their parental viruses. The results indicated that the swapped nsp2 and GP5-M viruses increased the neutralization reactivity with the donor strain antisera in MARC-145 cells. Taken together, these results show that variations in nsp2 and GP5-M correlate with the limited neutralization reactivity between the heterologous strains HP-PRRSV JXwn06 and LP-PRRSV HB-1/3.9.

Keywords

Porcine reproductive and respiratory syndrome virus (PRRSV) Neutralizing antibody (NA) Non-structural protein 2 (nsp2) Structural proteins (SPs) 

Notes

Acknowledgements

This study was supported by the Major Program of National Natural Science Foundation of China (31490603, 31572549) and the National Key Technology R & D Program of China (2015BAD12B01-2).

Author Contributions

JS, XG, LZ, XNG, JH and HY contributed to the study design. JS, BH and XZ performed the experiments in the study. XG, LZ and JS analyzed the data. JS, LZ and XG wrote the manuscript. All authors read and approved the final manuscript.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Animal and Human Rights Statement

The animal experiments in this study were approved by The Laboratory Animal Ethical Committee of China Agricultural University. All institutional and national guidelines for the care and use of animals were followed.

Supplementary material

12250_2019_149_MOESM1_ESM.pdf (369 kb)
Supplementary material 1 (PDF 369 kb)

References

  1. Collins JE, Benfield DA, Christianson WT, Harris L, Hennings JC, Shaw DP, Goyal SM, McCullough S, Morrison RB, Joo HS, Et A (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 Invest 4:117–126CrossRefGoogle Scholar
  2. Costers S, Lefebvre DJ, Van Doorsselaere J, Vanhee M, Delputte PL, Nauwynck HJ (2010a) GP4 of porcine reproductive and respiratory syndrome virus contains a neutralizing epitope that is susceptible to immunoselection in vitro. Arch Virol 155:371–378CrossRefGoogle Scholar
  3. Costers S, Vanhee M, Van Breedam W, Van Doorsselaere J, Geldhof M, Nauwynck HJ (2010b) GP4-specific neutralizing antibodies might be a driving force in PRRSV evolution. Virus Res 154:104–113CrossRefGoogle Scholar
  4. Das PB, Dinh PX, Ansari IH, de Lima M, Osorio FA, Pattnaik AK (2010) The minor envelope glycoproteins GP2a and GP4 of porcine reproductive and respiratory syndrome virus interact with the receptor CD163. J Virol 84:1731–1740CrossRefGoogle Scholar
  5. Duan X, Nauwynck HJ, Pensaert MB (1997) Virus quantification and identification of cellular targets in the lungs and lymphoid tissues of pigs at different time intervals after inoculation with porcine reproductive and respiratory syndrome virus (PRRSV). Vet Microbiol 56:9–19CrossRefGoogle Scholar
  6. Fan B, Liu X, Bai J, Zhang T, Zhang Q, Jiang P (2016) Influence of the amino acid residues at 70 in M protein of porcine reproductive and respiratory syndrome virus on viral neutralization susceptibility to the serum antibody. Virol J 13:51CrossRefGoogle Scholar
  7. Gao ZQ, Guo X, Yang HC (2004) Genomic characterization of two Chinese isolates of porcine respiratory and reproductive syndrome virus. Arch Virol 149:1341–1351CrossRefGoogle Scholar
  8. Gu W, Guo L, Yu H, Niu J, Huang M, Luo X, Li R, Tian Z, Feng L, Wang Y (2015) Involvement of CD16 in antibody-dependent enhancement of porcine reproductive and respiratory syndrome virus infection. J Gen Virol 96:1712–1722CrossRefGoogle Scholar
  9. Han J, Zhou L, Ge X, Guo X, Yang H (2017) Pathogenesis and control of the Chinese highly pathogenic porcine reproductive and respiratory syndrome virus. Vet Microbiol 209:30–47CrossRefGoogle Scholar
  10. 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 87:13456–13465CrossRefGoogle Scholar
  11. Kim WI, Kim JJ, Cha SH, Wu WH, Cooper V, Evans R, Choi EJ, Yoon KJ (2013) Significance of genetic variation of PRRSV ORF5 in virus neutralization and molecular determinants corresponding to cross neutralization among PRRS viruses. Vet Microbiol 162:10–22CrossRefGoogle Scholar
  12. Kuhn JH, Lauck M, Bailey AL, Shchetinin AM, Vishnevskaya TV, Bao Y, Ng TF, LeBreton M, Schneider BS, Gillis A, Tamoufe U, Diffo JD, Takuo JM, Kondov NO, Coffey LL, Wolfe ND, Delwart E, Clawson AN, Postnikova E, Bollinger L, Lackemeyer MG, Radoshitzky SR, Palacios G, Wada J, Shevtsova ZV, Jahrling PB, Lapin BA, Deriabin PG, Dunowska M, Alkhovsky SV, Rogers J, Friedrich TC, O’Connor DH, Goldberg TL (2016) Reorganization and expansion of the nidoviral family Arteriviridae. Arch Virol 161:755–768CrossRefGoogle Scholar
  13. Labarque GG, Nauwynck HJ, Van Reeth K, Pensaert MB (2000) Effect of cellular changes and onset of humoral immunity on the replication of porcine reproductive and respiratory syndrome virus in the lungs of pigs. J Gen Virol 81:1327–1334CrossRefGoogle Scholar
  14. Leng C, Zhang W, Zhang H, Kan Y, Yao L, Zhai H, Li M, Li Z, Liu C, An T, Peng J, Wang Q, Leng Y, Cai X, Tian Z, Tong G (2017) ORF1a of highly pathogenic PRRS attenuated vaccine virus plays a key role in neutralizing antibody induction in piglets and virus neutralization in vitro. Virol J 14:159CrossRefGoogle Scholar
  15. Li J, Murtaugh MP (2012) Dissociation of porcine reproductive and respiratory syndrome virus neutralization from antibodies specific to major envelope protein surface epitopes. Virology 433:367–376CrossRefGoogle Scholar
  16. Li X, Galliher-Beckley A, Pappan L, Trible B, Kerrigan M, Beck A, Hesse R, Blecha F, Nietfeld JC, Rowland RR, Shi J (2014) Comparison of host immune responses to homologous and heterologous type II porcine reproductive and respiratory syndrome virus (PRRSV) challenge in vaccinated and unvaccinated pigs. Biomed Res Int 2014:416727PubMedPubMedCentralGoogle Scholar
  17. Li Y, Zhou L, Zhang J, Ge X, Zhou R, Zheng H, Geng G, Guo X, Yang H (2014) Nsp9 and Nsp10 contribute to the fatal virulence of highly pathogenic porcine reproductive and respiratory syndrome virus emerging in China. PLoS Pathog 10:e1004216CrossRefGoogle Scholar
  18. Li Y, Tas A, Sun Z, Snijder EJ, Fang Y (2015) Proteolytic processing of the porcine reproductive and respiratory syndrome virus replicase. Virus Res 202:48–59CrossRefGoogle Scholar
  19. Liu D, Zhou R, Zhang J, Zhou L, Jiang Q, Guo X, Ge X, Yang H (2011) Recombination analyses between two strains of porcine reproductive and respiratory syndrome virus in vivo. Virus Res 155:473–486CrossRefGoogle Scholar
  20. Lopez OJ, Osorio FA (2004) Role of neutralizing antibodies in PRRSV protective immunity. Vet Immunol Immunopathol 102:155–163CrossRefGoogle Scholar
  21. Lopez OJ, Oliveira MF, Garcia EA, Kwon BJ, Doster A, Osorio FA (2007) Protection against porcine reproductive and respiratory syndrome virus (PRRSV) infection through passive transfer of PRRSV-neutralizing antibodies is dose dependent. Clin Vaccine Immunol 14:269–275CrossRefGoogle Scholar
  22. 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 4:129–154CrossRefGoogle Scholar
  23. Ostrowski M, Galeota JA, Jar AM, Platt KB, Osorio FA, Lopez OJ (2002) Identification of neutralizing and nonneutralizing epitopes in the porcine reproductive and respiratory syndrome virus GP5 ectodomain. J Virol 76:4241–4250CrossRefGoogle Scholar
  24. Shanmukhappa K, Kim JK, Kapil S (2007) Role of CD151, A tetraspanin, in porcine reproductive and respiratory syndrome virus infection. Virol J 4:62CrossRefGoogle Scholar
  25. Shi C, Liu Y, Ding Y, Zhang Y, Zhang J (2015) PRRSV receptors and their roles in virus infection. Arch Microbiol 197:503–512CrossRefGoogle Scholar
  26. Tian D, Cao D, Lynn Heffron C, Yugo DM, Rogers AJ, Overend C, Matzinger SR, Subramaniam S, Opriessnig T, LeRoith T, Meng XJ (2017) Enhancing heterologous protection in pigs vaccinated with chimeric porcine reproductive and respiratory syndrome virus containing the full-length sequences of shuffled structural genes of multiple heterologous strains. Vaccine 35:2427–2434CrossRefGoogle Scholar
  27. Vanhee M, Costers S, Van Breedam W, Geldhof MF, Van Doorsselaere J, Nauwynck HJ (2010) A variable region in GP4 of European-type porcine reproductive and respiratory syndrome virus induces neutralizing antibodies against homologous but not heterologous virus strains. Viral Immunol 23:403–413CrossRefGoogle Scholar
  28. Vanhee M, Van Breedam W, Costers S, Geldhof M, Noppe Y, Nauwynck H (2011) Characterization of antigenic regions in the porcine reproductive and respiratory syndrome virus by the use of peptide-specific serum antibodies. Vaccine 29:4794–4804CrossRefGoogle Scholar
  29. Wang W, Zhang L, Ma X, Gao J, Xiao Y, Zhou E (2011) The role of vimentin during PRRSV infection of MARC-145 cells. Bing Du Xue Bao 27:456–461 (in Chinese) PubMedGoogle Scholar
  30. Wensvoort G, Terpstra C, Pol JM, ter Laak EA, Bloemraad M, de Kluyver EP, Kragten C, van Buiten L, den Besten A, Wagenaar F, Et A (1991) Mystery swine disease in The Netherlands: the isolation of Lelystad virus. Vet Q 13:121–130CrossRefGoogle Scholar
  31. Wissink EH, Kroese MV, van Wijk HA, Rijsewijk FA, Meulenberg JJ, Rottier PJ (2005) Envelope protein requirements for the assembly of infectious virions of porcine reproductive and respiratory syndrome virus. J Virol 79:12495–12506CrossRefGoogle Scholar
  32. Wu WH, Fang Y, Farwell R, Steffen-Bien M, Rowland RR, Christopher-Hennings J, Nelson EA (2001) A 10-kDa structural protein of porcine reproductive and respiratory syndrome virus encoded by ORF2b. Virology 287:183–191CrossRefGoogle Scholar
  33. Zhou L, Ni YY, Pineyro P, Sanford BJ, Cossaboom CM, Dryman BA, Huang YW, Cao DJ, Meng XJ (2012) DNA shuffling of the GP3 genes of porcine reproductive and respiratory syndrome virus (PRRSV) produces a chimeric virus with an improved cross-neutralizing ability against a heterologous PRRSV strain. Virology 434:96–109CrossRefGoogle Scholar

Copyright information

© Wuhan Institute of Virology, CAS 2019

Authors and Affiliations

  1. 1.Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine and State Key Laboratory of AgrobiotechnologyChina Agricultural UniversityBeijingChina

Personalised recommendations