Porcine reproductive and respiratory syndrome virus suppresses post-transcriptionally the protein expression of IFN-β by upregulating cellular microRNAs in porcine alveolar macrophages in vitro

Wang,Lilin; Zhou,Lei; Hu,Dongmei; Ge,Xinna; Guo,Xin; Yang,Hanchun

doi:10.3892/etm.2017.5397

Porcine reproductive and respiratory syndrome virus suppresses post-transcriptionally the protein expression of IFN-β by upregulating cellular microRNAs in porcine alveolar macrophages in vitro

Authors:
- Lilin Wang
- Lei Zhou
- Dongmei Hu
- Xinna Ge
- Xin Guo
- Hanchun Yang
View Affiliations

Affiliations: Key Laboratory of Animal Epidemiology of Ministry of Agriculture, College of Veterinary Medicine and State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing 100193, P.R. China, Veterinary Diagnostic Laboratory, China Animal Disease Control Center, Beijing 102600, P.R. China
Published online on: October 30, 2017 https://doi.org/10.3892/etm.2017.5397
Pages: 115-126
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) has been recognized to inhibit the response of type I interferon (IFN) both in vivo and in vitro. However, the post‑transcriptional mechanism by which PRRSV suppresses type Ⅰ IFN induction in virus-infected host cells remains unclear. The present study first demonstrated that PRRSV inhibited post-transcriptionally the protein induction of IFN-β in primary porcine alveolar macrophages (PAMs) during early infection, and the inhibition effect mediated by the Chinese highly pathogenic (HP)-PRRSV was stronger. Next, we analyzed the cellular microRNA (miRNA)-modulated protein expression of porcine IFN-β by dual firefly/Renilla luciferase reporter assay, transfection of miRNA mimics and inhibitor assay and polyinosinic-polycytidylic acid (poly I:C) treatment of PAMs, showing that porcine miRNAs including let-7b, miR‑26a, miR-34a and miR-145 are able to inhibit IFN-β protein expression in primary PAMs by directly targeting sequences within the porcine IFN-β 3'UTR locating at 160-181, 9-31, 27-47 and 12-32 bp, respectively. Finally, we confirmed that let-7b, miR-26a, miR-34a and miR-145, were upregulated in PRRSV‑infected PAMs early in vitro, and the expression level of these miRNAs in HP-PRRSV JXwn06‑infected PAMs were higher than those in low pathogenic PRRSV HB-1/3.9-infected PAMs. The endogenous cellular miRNA-mediated inhibition of IFN-β induction in PRRSV‑infected PAMs early could be relieved by miRNA antagonists. Taken together, our findings suggest for the first time that PRRSV can suppress post-transcriptionally protein expression of IFN-β by upregulating cellular miRNAs in PAMs in vitro, providing novel insight into mechanisms in relation to the PRRSV-mediated immunomodulation of porcine innate immunity.

View Figures

View References

1	Albina E: Epidemiology of porcine reproductive and respiratory syndrome (PRRS): An overview. Vet Microbiol. 55:309–316. 1997. View Article : Google Scholar : PubMed/NCBI
2	Rossow KD: Porcine reproductive and respiratory syndrome. Vet Pathol. 35:1–20. 1998. View Article : Google Scholar : PubMed/NCBI
3	Keffaber K: Reproductive failure of unknown etiology. Am Assoc Swine Prac News. 1:1–9. 1989.
4	Cho JG and Dee SA: Porcine reproductive and respiratory syndrome virus. Theriogenology. 66:655–662. 2006. View Article : Google Scholar : PubMed/NCBI
5	Tian K, Yu X, Zhao T, Feng Y, Cao Z, Wang C, Hu Y, Chen X, Hu D, Tian X, et al: Emergence of fatal PRRSV variants: Unparalleled outbreaks of atypical PRRS in China and molecular dissection of the unique hallmark. PLoS One. 2:e5262007. View Article : Google Scholar : PubMed/NCBI
6	Zhou L and Yang H: Porcine reproductive and respiratory syndrome in China. Virus Res. 154:31–37. 2010. View Article : Google Scholar : PubMed/NCBI
7	Cavanagh D: Nidovirales: A new order comprising Coronaviridae and Arteriviridae. Arch Virol. 142:629–633. 1997.PubMed/NCBI
8	Conzelmann KK, Visser N, Van Woensel P and Thiel HJ: Molecular characterization of porcine reproductive and respiratory syndrome virus, a member of the arterivirus group. Virology. 193:329–339. 1993. View Article : Google Scholar : PubMed/NCBI
9	Meulenberg JJ, Hulst MM, de Meijer EJ, Moonen PL, den Besten A, de Kluyver EP, Wensvoort G and Moormann RJ: Lelystad virus, the causative agent of porcine epidemic abortion and respiratory syndrome (PEARS), is related to LDV and EAV. Virology. 192:62–72. 1993. View Article : Google Scholar : PubMed/NCBI
10	Mardassi H, Mounir S and Dea S: Identification of major differences in the nucleocapsid protein genes of a Québec strain and European strains of porcine reproductive and respiratory syndrome virus. J Gen Virol. 75:681–685. 1994. View Article : Google Scholar : PubMed/NCBI
11	Meng XJ, Paul PS, Halbur PG and Lum MA: Phylogenetic analyses of the putative M (ORF 6) and N (ORF 7) genes of porcine reproductive and respiratory syndrome virus (PRRSV): Implication for the existence of two genotypes of PRRSV in the USA and Europe. Arch Virol. 140:745–755. 1995. View Article : Google Scholar : PubMed/NCBI
12	Nelson EA, Christopher-Hennings J, Drew T, Wensvoort G, Collins JE and Benfield DA: Differentiation of U.S. and European isolates of porcine reproductive and respiratory syndrome virus by monoclonal antibodies. J Clin Microbiol. 31:3184–3189. 1993.PubMed/NCBI
13	Wang X and Christopher-Hennings J: Post-transcriptional control of type I interferon induction by porcine reproductive and respiratory syndrome virus in its natural host cells. Viruses. 4:725–733. 2012. View Article : Google Scholar : PubMed/NCBI
14	Zhang H, Guo X, Nelson E, Christopher-Hennings J and Wang X: Porcine reproductive and respiratory syndrome virus activates the transcription of interferon alpha/beta (IFN-α/β) in monocyte-derived dendritic cells (Mo-DC). Vet Microbiol. 159:494–498. 2012. View Article : Google Scholar : PubMed/NCBI
15	Albina E, Carrat C and Charley B: Interferon-alpha response to swine arterivirus (PoAV), the porcine reproductive and respiratory syndrome virus. J Interferon Cytokine Res. 18:485–490. 1998. View Article : Google Scholar : PubMed/NCBI
16	Buddaert W, Van Reeth K and Pensaert M: In vivo and in vitro interferon (IFN) studies with the porcine reproductive and respiratory syndrome virus (PRRSV). Adv Exp Med Biol. 440:461–467. 1998. View Article : Google Scholar : PubMed/NCBI
17	Calzada-Nova G, Schnitzlein WM, Husmann RJ and Zuckermann FA: North American porcine reproductive and respiratory syndrome viruses inhibit type I interferon production by plasmacytoid dendritic cells. J Virol. 85:2703–2713. 2011. View Article : Google Scholar : PubMed/NCBI
18	Miller LC, Laegreid WW, Bono JL, Chitko-McKown CG and Fox JM: Interferon type I response in porcine reproductive and respiratory syndrome virus-infected MARC-145 cells. Arch Virol. 149:2453–2463. 2004. View Article : Google Scholar : PubMed/NCBI
19	Chung HK, Lee JH, Kim SH and Chae C: Expression of interferon-alpha and Mx1 protein in pigs acutely infected with porcine reproductive and respiratory syndrome virus (PRRSV). J Comp Pathol. 130:299–305. 2004. View Article : Google Scholar : PubMed/NCBI
20	Lee SM, Schommer SK and Kleiboeker SB: Porcine reproductive and respiratory syndrome virus field isolates differ in in vitro interferon phenotypes. Vet Immunol Immunopathol. 102:217–231. 2004. View Article : Google Scholar : PubMed/NCBI
21	Sun Y, Han M, Kim C, Calvert JG and Yoo D: Interplay between interferon-mediated innate immunity and porcine reproductive and respiratory syndrome virus. Viruses. 4:424–446. 2012. View Article : Google Scholar : PubMed/NCBI
22	Lin S and Gregory RI: MicroRNA biogenesis pathways in cancer. Nat Rev Cancer. 15:321–333. 2015. View Article : Google Scholar : PubMed/NCBI
23	Forster SC, Tate MD and Hertzog PJ: MicroRNA as type I interferon-regulated transcripts and modulators of the innate immune response. Front Immunol. 6:3342015. View Article : Google Scholar : PubMed/NCBI
24	Cullen BR: MicroRNAs as mediators of viral evasion of the immune system. Nat Immunol. 14:205–210. 2013. View Article : Google Scholar : PubMed/NCBI
25	Skalsky RL and Cullen BR: Viruses, microRNAs, and host interactions. Annu Rev Microbiol. 64:123–141. 2010. View Article : Google Scholar : PubMed/NCBI
26	Cong P, Xiao S, Chen Y, Wang L, Gao J, Li M, He Z, Guo Y, Zhao G, Zhang X, et al: Integrated miRNA and mRNA transcriptomes of porcine alveolar macrophages (PAM cells) identifies strain-specific miRNA molecular signatures associated with H-PRRSV and N-PRRSV infection. Mol Biol Rep. 41:5863–5875. 2014. View Article : Google Scholar : PubMed/NCBI
27	Li J, Chen Z, Zhao J, Fang L, Fang R, Xiao J, Chen X, Zhou A, Zhang Y, Ren L, et al: Difference in microRNA expression and editing profile of lung tissues from different pig breeds related to immune responses to HP-PRRSV. Sci Rep. 5:95492015. View Article : Google Scholar : PubMed/NCBI
28	Wang C, Zhang Y, Luo J, Ding H, Liu S, Amer S, Xie L, Lyv W, Su W, Li M, et al: Identification of miRNomes reveals ssc-miR-30d-R_1 as a potential therapeutic target for PRRS viral infection. Sci Rep. 6:248542016. View Article : Google Scholar : PubMed/NCBI
29	Li L, Gao F, Jiang Y, Yu L, Zhou Y, Zheng H, Tong W, Yang S, Xia T, Qu Z, et al: Cellular miR-130b inhibits replication of porcine reproductive and respiratory syndrome virus in vitro and in vivo. Sci Rep. 5:170102015. View Article : Google Scholar : PubMed/NCBI
30	Wang D, Cao L, Xu Z, Fang L, Zhong Y, Chen Q, Luo R, Chen H, Li K and Xiao S: MiR-125b reduces porcine reproductive and respiratory syndrome virus replication by negatively regulating the NF-κB pathway. PLoS One. 8:e558382013. View Article : Google Scholar : PubMed/NCBI
31	Xiao S, Wang X, Ni H, Li N, Zhang A, Liu H, Pu F, Xu L, Gao J, Zhao Q, et al: MicroRNA miR-24-3p promotes porcine reproductive and respiratory syndrome virus replication through suppression of heme oxygenase-1 expression. J Virol. 89:4494–4503. 2015. View Article : Google Scholar : PubMed/NCBI
32	Zhang Q, Huang C, Yang Q, Gao L, Liu HC, Tang J and Feng WH: MicroRNA-30c modulates type I IFN responses to facilitate porcine reproductive and respiratory syndrome virus infection by targeting JAK1. J Immunol. 196:2272–2282. 2016. View Article : Google Scholar : PubMed/NCBI
33	Chen J, Shi X, Zhang X, Wang A, Wang L, Yang Y, Deng R and Zhang GP: MicroRNA-373 facilitated the replication of porcine reproductive and respiratory syndrome virus by its negative regulation of type I interferon Induction. J Virol. 91:e01311–e01316. 2017. View Article : Google Scholar : PubMed/NCBI
34	Jia X, Bi Y, Li J, Xie Q, Yang H and Liu W: Cellular microRNA miR-26a suppresses replication of porcine reproductive and respiratory syndrome virus by activating innate antiviral immunity. Sci Rep. 5:106512015. View Article : Google Scholar : PubMed/NCBI
35	Li L, Wei Z, Zhou Y, Gao F, Jiang Y, Yu L, Zheng H, Tong W, Yang S, Zheng H, et al: Host miR-26a suppresses replication of porcine reproductive and respiratory syndrome virus by upregulating type I interferons. Virus Res. 195:86–94. 2015. View Article : Google Scholar : PubMed/NCBI
36	He D, Overend C, Ambrogio J, Maganti RJ, Grubman MJ and Garmendia AE: Marked differences between MARC-145 cells and swine alveolar macrophages in IFNβ-induced activation of antiviral state against PRRSV. Vet Immunol Immunopathol. 139:57–60. 2011. View Article : Google Scholar : PubMed/NCBI
37	Ait-Ali T, Wilson AD, Westcott DG, Clapperton M, Waterfall M, Mellencamp MA, Drew TW, Bishop SC and Archibald AL: Innate immune responses to replication of porcine reproductive and respiratory syndrome virus in isolated Swine alveolar macrophages. Viral Immunol. 20:105–118. 2007. View Article : Google Scholar : PubMed/NCBI
38	Zhang H, Guo X, Ge X, Chen Y, Sun Q and Yang H: Changes in the cellular proteins of pulmonary alveolar macrophage infected with porcine reproductive and respiratory syndrome virus by proteomics analysis. J Proteome Res. 8:3091–3097. 2009. View Article : Google Scholar : PubMed/NCBI
39	Zhou L, Zhang J, Zeng J, Yin S, Li Y, Zheng L, Guo X, Ge X and Yang H: The 30-amino-acid deletion in the Nsp2 of highly pathogenic porcine reproductive and respiratory syndrome virus emerging in China is not related to its virulence. J Virol. 83:5156–5167. 2009. View Article : Google Scholar : PubMed/NCBI
40	John B, Enright AJ, Aravin A, Tuschl T, Sander C and Marks DS: Human MicroRNA targets. PLoS Biol. 2:e3632004. View Article : Google Scholar : PubMed/NCBI
41	Rehmsmeier M, Steffen P, Hochsmann M and Giegerich R: Fast and effective prediction of microRNA/target duplexes. RNA. 10:1507–1517. 2004. View Article : Google Scholar : PubMed/NCBI
42	Kertesz M, Iovino N, Unnerstall U, Gaul U and Segal E: The role of site accessibility in microRNA target recognition. Nat Genet. 39:1278–1284. 2007. View Article : Google Scholar : PubMed/NCBI
43	He Q, Li Y, Zhou L, Ge X, Guo X and Yang H: Both Nsp1β and Nsp11 are responsible for differential TNF-α production induced by porcine reproductive and respiratory syndrome virus strains with different pathogenicity in vitro. Virus Res. 201:32–40. 2015. View Article : Google Scholar : PubMed/NCBI
44	Guo XK, Zhang Q, Gao L, Li N, Chen XX and Feng WH: Increasing expression of microRNA 181 inhibits porcine reproductive and respiratory syndrome virus replication and has implications for controlling virus infection. J Virol. 87:1159–1171. 2013. View Article : Google Scholar : PubMed/NCBI
45	Loving CL, Brockmeier SL and Sacco RE: Differential type I interferon activation and susceptibility of dendritic cell populations to porcine arterivirus. Immunology. 120:217–229. 2007. View Article : Google Scholar : PubMed/NCBI
46	Nygard AB, Jørgensen CB, Cirera S and Fredholm M: Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR. BMC Mol Biol. 8:672007. View Article : Google Scholar : PubMed/NCBI
47	Witwer KW, Sisk JM, Gama L and Clements JE: MicroRNA regulation of IFN-beta protein expression: Rapid and sensitive modulation of the innate immune response. J Immunol. 184:2369–2376. 2010. View Article : Google Scholar : PubMed/NCBI
48	Biron CA: Role of early cytokines, including alpha and beta interferons (IFN-alpha/beta), in innate and adaptive immune responses to viral infections. Semin Immunol. 10:383–390. 1998. View Article : Google Scholar : PubMed/NCBI
49	Theofilopoulos AN, Baccala R, Beutler B and Kono DH: Type I interferons (alpha/beta) in immunity and autoimmunity. Annu Rev Immunol. 23:307–336. 2005. View Article : Google Scholar : PubMed/NCBI
50	Baumann A, Mateu E, Murtaugh MP and Summerfield A: Impact of genotype 1 and 2 of porcine reproductive and respiratory syndrome viruses on interferon-α responses by plasmacytoid dendritic cells. Vet Res (Faisalabad). 44:332013. View Article : Google Scholar
51	Rose KM and Weiss SR: Murine coronavirus cell type dependent interaction with the type I interferon response. Viruses. 1:689–712. 2009. View Article : Google Scholar : PubMed/NCBI
52	Li Y, Zhou L, Zhang J, Ge X, Zhou R, Zheng H, Geng G, Guo X and Yang H: Nsp9 and Nsp10 contribute to the fatal virulence of highly pathogenic porcine reproductive and respiratory syndrome virus emerging in China. PLoS Pathog. 10:e10042162014. View Article : Google Scholar : PubMed/NCBI
53	Carrigan DR and Knox KK: Identification of interferon-resistant subpopulations in several strains of measles virus: Positive selection by growth of the virus in brain tissue. J Virol. 64:1606–1615. 1990.PubMed/NCBI
54	Marcus PI, Rojek JM and Sekellick MJ: Interferon induction and/or production and its suppression by influenza A viruses. J Virol. 79:2880–2890. 2005. View Article : Google Scholar : PubMed/NCBI
55	Khabar KS and Young HA: Post-transcriptional control of the interferon system. Biochimie. 89:761–769. 2007. View Article : Google Scholar : PubMed/NCBI
56	Sedger LM: microRNA control of interferons and interferon induced anti-viral activity. Mol Immunol. 56:781–793. 2013. View Article : Google Scholar : PubMed/NCBI
57	Hertzog PJ and Williams BR: Fine tuning type I interferon responses. Cytokine Growth Factor Rev. 24:217–225. 2013. View Article : Google Scholar : PubMed/NCBI
58	Huang C, Zhang Q and Feng WH: Regulation and evasion of antiviral immune responses by porcine reproductive and respiratory syndrome virus. Virus Res. 202:101–111. 2015. View Article : Google Scholar : PubMed/NCBI
59	Overend C, Mitchell R, He D, Rompato G, Grubman MJ and Garmendia AE: Recombinant swine beta interferon protects swine alveolar macrophages and MARC-145 cells from infection with Porcine reproductive and respiratory syndrome virus. J Gen Virol. 88:925–931. 2007. View Article : Google Scholar : PubMed/NCBI
60	A Hicks J, Yoo D and Liu HC: Characterization of the microRNAome in porcine reproductive and respiratory syndrome virus infected macrophages. PLoS One. 8:e820542013. View Article : Google Scholar : PubMed/NCBI
61	Zhang Q, Guo XK, Gao L, Huang C, Li N, Jia X, Liu W and Feng WH: MicroRNA-23 inhibits PRRSV replication by directly targeting PRRSV RNA and possibly by upregulating type I interferons. Virology 450–451. 1–195. 2014.
62	Chen Y, Chen J, Wang H, Shi J, Wu K, Liu S, Liu Y and Wu J: HCV-induced miR-21 contributes to evasion of host immune system by targeting MyD88 and IRAK1. PLoS Pathog. 9:e10032482013. View Article : Google Scholar : PubMed/NCBI
63	Ho BC, Yu IS, Lu LF, Rudensky A, Chen HY, Tsai CW, Chang YL, Wu CT, Chang LY, Shih SR, et al: Inhibition of miR-146a prevents enterovirus-induced death by restoring the production of type I interferon. Nat Commun. 5:33442014. View Article : Google Scholar : PubMed/NCBI
64	Thornburg NJ, Hayward SL and Crowe JE Jr: Respiratory syncytial virus regulates human microRNAs by using mechanisms involving beta interferon and NF-κB. MBio. 3:e0022012. View Article : Google Scholar