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Archives of Virology

, Volume 164, Issue 4, pp 1147–1157 | Cite as

Porcine epidemic diarrhea virus nsp4 induces pro-inflammatory cytokine and chemokine expression inhibiting viral replication in vitro

  • Linyang Yu
  • Jianguo Dong
  • Yanwu Wang
  • Pengfei Zhang
  • Yanling Liu
  • Leyi Zhang
  • Pengshuai Liang
  • Lei Wang
  • Changxu SongEmail author
Original Article
  • 112 Downloads

Abstract

Porcine epidemic diarrhea virus (PEDV) causes severe economic loss in the pig industry each year. To better understand the relationship between cytokines and PEDV replication, in this study, pro-inflammatory cytokine and chemokine expression profiles in Vero cells infected with PEDV were analyzed. Real-time quantitative PCR assay indicated that IL-1α, IL-1β, TNF-α, CCL2, CCL5 and CXCL8 expression levels were significantly upregulated. Moreover, overexpression and siRNA silencing assays showed that overexpression of IL-1α, IL-1β, TNF-α, CCL2, CCL5 and CXCL8 could significantly inhibit PEDV replication, while silencing of IL-1α, IL-1β, TNF-α, CCL2, CCL5 and CXCL8 could significantly promote PEDV replication. Finally, a dual-luciferase reporter assay showed that nsp4 contributed to the expression of IL-1α, IL-1β, TNF-α, CCL2, CCL5 and CXCL8 via the NF-κB pathway. Together, these data determined that PEDV nsp4 could upregulate pro-inflammatory cytokine and chemokine expression, inhibiting viral replication in vitro. These results provided novel insights for understanding the roles of cytokines in PEDV replication.

Abbreviations

PEDV

Porcine epidemic diarrhea virus

ORFs

Open reading frames

nsps

Nonstructural proteins

S

Spike protein

M

Membrane protein

E

Envelope protein

N

Nucleocapsid protein

IFN

Interferon

FBS

Fetal bovine serum (Thermo)

PAMs

Porcine alveolar macrophages

RT-PCR

Reverse transcription polymerase chain reaction

RT-qPCR

Real-time quantitative polymerase chain reaction

CT

Threshold cycle

OD

Optical density

SD

Standard deviations

PBL

Peripheral blood leucocytes

PCV2

Porcine circovirus type 2

PBMCs

Peripheral blood mononuclear cells

Notes

Acknowledgements

This work was supported by the National Key Research and Development Program of China (2018YFD0501102), the National Key Technologies R&D Program (2015BAD12B02-5), the Guangzhou City Project (201508020062), and the Henan Science and Technology Project (182102110037).

Author contributions

Conceived and designed the experiments: Linyang Yu, Jianguo Dong, Changxu Song. Performed the experiments: Linyang Yu, Jianguo Dong, Yanwu Wang, Pengfei Zhang, Yanling Liu, Leyi Zhang, Pengshuai Liang, Lei Wang, Changxu Song. Analyzed the data and wrote the paper: Linyang Yu, Jianguo Dong, Changxu Song.

Funding

This work was supported by the National Key Research and Development Program of China (2018YFD0501102), the National Key Technologies R&D Program (2015BAD12B02-5), the Guangzhou City Project (201508020062), and the Henan Science and Technology Project (182102110037).

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.

Ethics approval and consent to participate

All of the samples were collected according to the animal ethics regulations of the National Engineering Center for Swine Breeding Industry (NECSBI 2015-16).

Consent for publication

Not applicable.

Availability of data and materials

Genetic data presented in this paper are publicly available via GenBank.

Supplementary material

705_2019_4176_MOESM1_ESM.asp (31 kb)
Supplementary material 1 (ASP 30 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Linyang Yu
    • 1
  • Jianguo Dong
    • 2
  • Yanwu Wang
    • 1
    • 3
  • Pengfei Zhang
    • 1
  • Yanling Liu
    • 1
  • Leyi Zhang
    • 1
  • Pengshuai Liang
    • 1
  • Lei Wang
    • 1
  • Changxu Song
    • 1
    Email author
  1. 1.Animal Science College/National Engineering Center for Swine Breeding IndustrySouth China Agriculture UniversityGuangzhouChina
  2. 2.School of Animal Husbandry and Medical EngineeringXinyang Agriculture and Forestry UniversityXinyangChina
  3. 3.Guangxi State Farms Yongxin Animal Husbandry Group Co., LtdNanningChina

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