Foot-and-mouth disease virus (FMDV) is the most contagious pathogen in cloven-hoofed (two-toed) animals. Due to the rapid replication and spread of FMDV, novel therapeutic strategies are greatly needed to reduce or block FMDV shedding in cases of disease outbreak. Here, we generated an IRES-Mx1 construct in which the internal ribosome entry site (IRES) of FMDV was inserted between the promoter and open reading frame (ORF) of porcine myxovirus resistance protein 1 (poMx1). This construct provides more powerful protection against FMDV infection than the IRES-IFN construct that was previously generated by our group. The results indicate that this IRES-Mx1 construct was able to express poMx1 12 h after transfection and induce a robust immune response. In contrast to the control, the proliferation of virus in transfected cells was significantly inhibited, as evaluated by morphology monitoring, real-time RT-PCR, virus titration and Western blot. In addition, we also found that the antiviral activity in cells transfected with pc-IRES-Mx1 was abolished when the JAK/STAT pathway was repressed, which indicates that the antiviral mechanism of poMx1 is JAK/STAT pathway dependent. Taken together, our data suggest that the antiviral activity of poMx1 is possibly produced by affecting the host cells themselves, instead of interacting with the virus directly. The new construct reported here could be used as a novel effective therapy against FMDV infection.
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We gratefully acknowledge Prof. Qingzhen Liu (College of Life Science, Wuhan University, China) for critical review of the manuscript. We also thank Dr. Maoqing Wu (Research fellow at Harvard Medical School) of Sander International Translation LLC for manuscript polishing service. This study was supported by a grant from the National Natural Science Foundation of China (No. 31370185) and National Infrastructure of Natural Resources for Science and Technology Program (No. 2011-572).
YB and SC conceived and designed the experiments. YB and FH performed the experiments. YB, SC and ZCY wrote the paper. All authors have read and agreed to the final version of the manuscript.
Moraes MP, de Los Santos T, Koster M, Turecek T, Wang H et al (2007) Enhanced antiviral activity against foot-and-mouth disease virus by a combination of type I and II porcine interferons. J Virol 81:7124–7135PubMedCentralPubMedCrossRefGoogle Scholar
Xiong Y, Lin M, Yuan B, Yuan T, Zheng C (2009) Expression of exogenous IFN-alpha by bypassing the translation block protects cells against FMDV infection. Antivir Res 84:60–66PubMedCrossRefGoogle Scholar
Devaney MA, Vakharia VN, Lloyd RE, Ehrenfeld E, Grubman MJ (1988) Leader protein of foot-and-mouth disease virus is required for cleavage of the p220 component of the cap-binding protein complex. J Virol 62:4407–4409PubMedCentralPubMedGoogle Scholar
Kirchweger R, Ziegler E, Lamphear BJ, Waters D, Liebig HD et al (1994) Foot-and-mouth disease virus leader proteinase: purification of the Lb form and determination of its cleavage site on eIF-4 gamma. J Virol 68:5677–5684PubMedCentralPubMedGoogle Scholar
de Los Santos T, Diaz-San Segundo F, Grubman MJ (2007) Degradation of nuclear factor kappa B during foot-and-mouth disease virus infection. J Virol 81:12803–12815CrossRefGoogle Scholar
Wang D, Fang L, Luo R, Ye R, Fang Y et al (2010) Foot-and-mouth disease virus leader proteinase inhibits dsRNA-induced type I interferon transcription by decreasing interferon regulatory factor 3/7 in protein levels. Biochem Biophys Res Commun 399:72–78PubMedCrossRefGoogle Scholar
Wang D, Fang L, Liu L, Zhong H, Chen Q et al (2011) Foot-and-mouth disease virus (FMDV) leader proteinase negatively regulates the porcine interferon-lambda1 pathway. Mol Immunol 49:407–412PubMedCrossRefGoogle Scholar
Haller O, Gao S, von der Malsburg A, Daumke O, Kochs G (2010) Dynamin-like MxA GTPase: structural insights into oligomerization and implications for antiviral activity. J Biol Chem 285:28419–28424PubMedCentralPubMedCrossRefGoogle Scholar
Haller O, Kochs G (2011) Human MxA protein: an interferon-induced dynamin-like GTPase with broad antiviral activity. J Interferon Cytokine Res 31:79–87PubMedCrossRefGoogle Scholar
Palm M, Garigliany MM, Cornet F, Desmecht D (2010) Interferon-induced Sus scrofa Mx1 blocks endocytic traffic of incoming influenza A virus particles. Vet Res 41:29PubMedCentralPubMedCrossRefGoogle Scholar
Zhang XM, He DN, Zhou B, Pang R, Liu K et al (2013) In vitro inhibition of vesicular stomatitis virus replication by purified porcine Mx1 protein fused to HIV-1 Tat protein transduction domain (PTD). Antivir Res 99:149–157PubMedCrossRefGoogle Scholar
He DN, Zhang XM, Liu K, Pang R, Zhao J et al (2014) In vitro inhibition of the replication of classical swine fever virus by porcine Mx1 protein. Antivir Res 104:128–135PubMedCrossRefGoogle Scholar
Diaz-San Segundo F, Moraes MP, de Los Santos T, Dias CC, Grubman MJ (2010) Interferon-induced protection against foot-and-mouth disease virus infection correlates with enhanced tissue-specific innate immune cell infiltration and interferon-stimulated gene expression. J Virol 84:2063–2077PubMedCentralPubMedCrossRefGoogle Scholar
Dash P, Barnett PV, Denyer MS, Jackson T, Stirling CM et al (2010) Foot-and-mouth disease virus replicates only transiently in well-differentiated porcine nasal epithelial cells. J Virol 84:9149–9160PubMedCentralPubMedCrossRefGoogle Scholar
Rodriguez-Pulido M, Borrego B, Sobrino F, Saiz M (2011) RNA structural domains in noncoding regions of the foot-and-mouth disease virus genome trigger innate immunity in porcine cells and mice. J Virol 85:6492–6501PubMedCentralPubMedCrossRefGoogle Scholar
Borrego B, Rodriguez-Pulido M, Mateos F, de la Losa N, Sobrino F et al (2013) Delivery of synthetic RNA can enhance the immunogenicity of vaccines against foot-and-mouth disease virus (FMDV) in mice. Vaccine 31:4375–4381PubMedCrossRefGoogle Scholar
Zhang X, Shin J, Molitor TW, Schook LB, Rutherford MS (1999) Molecular responses of macrophages to porcine reproductive and respiratory syndrome virus infection. Virology 262:152–162PubMedCrossRefGoogle Scholar
Zhang Q, Gong R, Qu J, Zhou Y, Liu W et al (2012) Activation of the Ras/Raf/MEK pathway facilitates hepatitis C virus replication via attenuation of the interferon-JAK-STAT pathway. J Virol 86:1544–1554PubMedCentralPubMedCrossRefGoogle Scholar
Gu C, Zheng C, Shi L, Zhang Q, Li Y et al (2007) Plus- and minus-stranded foot-and-mouth disease virus RNA quantified simultaneously using a novel real-time RT-PCR. Virus Genes 34:289–298PubMedCrossRefGoogle Scholar
Sabara MI, Larence JE (2003) Plaque assay for avian metapneumovirus using a Japanese quail fibrosarcoma cell line (QT-35). J Virol Methods 107:9–14PubMedCrossRefGoogle Scholar
Goodbourn S, Didcock L, Randall RE (2000) Interferons: cell signalling, immune modulation, antiviral response and virus countermeasures. J Gen Virol 81:2341–2364PubMedGoogle Scholar
Valentine AD, Meyers CA, Kling MA, Richelson E, Hauser P (1998) Mood and cognitive side effects of interferon-alpha therapy. Semin Oncol 25:39–47PubMedGoogle Scholar
Billiau A (2006) Interferon: the pathways of discovery I. Molecular and cellular aspects. Cytokine Growth Factor Rev 17:381–409PubMedCrossRefGoogle Scholar
Saito T, Owen DM, Jiang F, Marcotrigiano J, Gale M Jr (2008) Innate immunity induced by composition-dependent RIG-I recognition of hepatitis C virus RNA. Nature 454:523–527PubMedCentralPubMedCrossRefGoogle Scholar
Norder H, De Palma AM, Selisko B, Costenaro L, Papageorgiou N et al (2011) Picornavirus non-structural proteins as targets for new anti-virals with broad activity. Antivir Res 89:204–218PubMedCrossRefGoogle Scholar
Haller O, Staeheli P, Kochs G (2007) Interferon-induced Mx proteins in antiviral host defense. Biochimie 89:812–818PubMedCrossRefGoogle Scholar
Cilloniz C, Pantin-Jackwood MJ, Ni C, Carter VS, Korth MJ et al (2012) Molecular signatures associated with Mx1-mediated resistance to highly pathogenic influenza virus infection: mechanisms of survival. J Virol 86:2437–2446PubMedCentralPubMedCrossRefGoogle Scholar