{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T23:37:51Z","timestamp":1792798671107,"version":"4.5.0"},"reference-count":52,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2020,9,10]],"date-time":"2020-09-10T00:00:00Z","timestamp":1599696000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31672534"],"award-info":[{"award-number":["31672534"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Porcine reproductive and respiratory syndrome virus (PRRSV) has disrupted the global swine industry since the 1980s. PRRSV-host interactions are largely still unknown but may involve host ISG15 protein. In this study, we developed a monoclonal antibody (Mab-3D5E6) specific for swine ISG15 (sISG15) by immunizing mice with recombinant sISG15. A sandwich enzyme-linked immunosorbent assay (ELISA) incorporating this sISG15-specific Mab was developed to detect sISG15 and provided a lower limit of sISG15 detection of 200 pg\/mL. ELISA results demonstrated that infection of porcine alveolar macrophages (PAMs) with low-virulence or attenuated PRRSV vaccine strains induced intracellular ISG15 expression that was independent of type I IFN production, while PAMs infection with a PRRSV vaccine strain promoted extracellular ISG15 secretion from infected PAMs. Conversely, the addition of recombinant sISG15 to PAMs mimicked natural extracellular ISG15 effects whereby sISG15 functioned as a cytokine by activating PAMs. Once activated, PAMs could inhibit PRRSV replication and resist infection with PRRSV vaccine strain TJM. In summary, a sandwich ELISA incorporating homemade anti-ISG15 Mab detected ISG15 secretion induced by PAMs infection with a PRRSV vaccine strain. Recombinant ISG15 added to cells exhibited cytokine-like activity that stimulated PAMs to assume an anti-viral state that enabled them to inhibit PRRSV replication and resist viral infection.<\/jats:p>","DOI":"10.3390\/v12091009","type":"journal-article","created":{"date-parts":[[2020,9,10]],"date-time":"2020-09-10T09:10:09Z","timestamp":1599729009000},"page":"1009","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["PRRSV Vaccine Strain-Induced Secretion of Extracellular ISG15 Stimulates Porcine Alveolar Macrophage Antiviral Response against PRRSV"],"prefix":"10.3390","volume":"12","author":[{"given":"Hongbin","family":"Liu","sequence":"first","affiliation":[{"name":"College of Animal Medical Sciences, Northeast Agricultural University, Harbin 150030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bingjun","family":"Shi","sequence":"additional","affiliation":[{"name":"Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&amp;F University, Yangling 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhigang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&amp;F University, Yangling 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bao","family":"Zhao","sequence":"additional","affiliation":[{"name":"Shaanxi Animal Disease Control Center, Xi\u2019an 710054, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guangming","family":"Zhao","sequence":"additional","affiliation":[{"name":"Shaanxi Animal Disease Control Center, Xi\u2019an 710054, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yijing","family":"Li","sequence":"additional","affiliation":[{"name":"College of Animal Medical Sciences, Northeast Agricultural University, Harbin 150030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4442-8004","authenticated-orcid":false,"given":"Yuchen","family":"Nan","sequence":"additional","affiliation":[{"name":"Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&amp;F University, Yangling 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,9,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"755","DOI":"10.1007\/s00705-015-2672-z","article-title":"Reorganization and expansion of the nidoviral family Arteriviridae","volume":"161","author":"Kuhn","year":"2016","journal-title":"Arch. Virol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2921","DOI":"10.1007\/s00705-016-2977-6","article-title":"Ratification vote on taxonomic proposals to the International Committee on Taxonomy of Viruses","volume":"161","author":"Adams","year":"2016","journal-title":"Arch. Virol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2131","DOI":"10.1093\/molbev\/msi208","article-title":"Divergence time of porcine reproductive and respiratory syndrome virus subtypes","volume":"22","author":"Forsberg","year":"2005","journal-title":"Mol. Biol. Evol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1136\/vr.142.19.510","article-title":"Effect on viraemia of an American and a European serotype PRRSV vaccine after challenge with European wild-type strains of the virus","volume":"142","author":"Liefkens","year":"1998","journal-title":"Vet. Rec."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1111\/tbed.12272","article-title":"Pathology and virus distribution in the lung and lymphoid tissues of pigs experimentally inoculated with three distinct type 1 PRRS virus isolates of varying pathogenicity","volume":"63","author":"Morgan","year":"2014","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1089\/jir.1998.18.485","article-title":"Interferon-alpha response to swine arterivirus (PoAV), the porcine reproductive and respiratory syndrome virus","volume":"18","author":"Albina","year":"1998","journal-title":"J. Interferon Cytokine Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/S0378-1135(96)01347-8","article-title":"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)","volume":"56","author":"Duan","year":"1997","journal-title":"Vet. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2483","DOI":"10.1007\/s007050050256","article-title":"Effects of origin and state of differentiation and activation of monocytes\/macrophages on their susceptibility to porcine reproductive and respiratory syndrome virus (PRRSV)","volume":"142","author":"Duan","year":"1997","journal-title":"Arch. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2280","DOI":"10.1128\/jcm.34.9.2280-2286.1996","article-title":"In vivo detection of porcine reproductive and respiratory syndrome virus RNA by in situ hybridization at different times postinfection","volume":"34","author":"Sur","year":"1996","journal-title":"J. Clin. Microbiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1356","DOI":"10.1099\/jgv.0.000454","article-title":"Highly pathogenic porcine reproductive and respiratory syndrome virus infection and induction of apoptosis in bone marrow cells of infected piglets","volume":"97","author":"Wang","year":"2016","journal-title":"J. Gen. Virol."},{"key":"ref_11","first-page":"1327","article-title":"Effect of cellular changes and onset of humoral immunity on the replication of porcine reproductive and respiratory syndrome virus in the lungs of pigs","volume":"81","author":"Labarque","year":"2000","journal-title":"J. Gen. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"5923","DOI":"10.1128\/JVI.78.11.5923-5933.2004","article-title":"The level of virus-specific T-cell and macrophage recruitment in porcine reproductive and respiratory syndrome virus infection in pigs is independent of virus load","volume":"78","author":"Xiao","year":"2004","journal-title":"J. Virol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1111\/j.0105-2896.2004.00204.x","article-title":"Interferons, interferon-like cytokines, and their receptors","volume":"202","author":"Pestka","year":"2004","journal-title":"Immunol. Rev."},{"key":"ref_14","first-page":"71","article-title":"The receptor of the type I interferon family","volume":"316","author":"Uze","year":"2007","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1189\/jlb.0603252","article-title":"Interferon-gamma: An overview of signals, mechanisms and functions","volume":"75","author":"Schroder","year":"2004","journal-title":"J. Leukoc. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1038\/nri888","article-title":"Viruses and interferon: A fight for supremacy","volume":"2","author":"Katze","year":"2002","journal-title":"Nat. Rev. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"912","DOI":"10.1189\/jlb.69.6.912","article-title":"Functional classification of interferon-stimulated genes identified using microarrays","volume":"69","author":"Holko","year":"2001","journal-title":"J. Leukoc. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"7806","DOI":"10.1016\/S0021-9258(18)42585-9","article-title":"The interferon-inducible 15-kDa ubiquitin homolog conjugates to intracellular proteins","volume":"267","author":"Loeb","year":"1992","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4520","DOI":"10.1016\/S0021-9258(18)68812-X","article-title":"A 15-kDa interferon-induced protein is derived by COOH-terminal processing of a 17-kDa precursor","volume":"263","author":"Knight","year":"1988","journal-title":"J. Biol. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1101\/gad.1056303","article-title":"Protein ISGylation modulates the JAK-STAT signaling pathway","volume":"17","author":"Malakhova","year":"2003","journal-title":"Genes Dev."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1073\/pnas.93.1.211","article-title":"Immunoregulatory properties of ISG15, an interferon-induced cytokine","volume":"93","author":"Knight","year":"1996","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e18","DOI":"10.1038\/emm.2013.36","article-title":"ISG15: Leading a double life as a secreted molecule","volume":"45","author":"Bogunovic","year":"2013","journal-title":"Exp. Mol. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1016\/j.bbrc.2003.08.038","article-title":"Identification of a ubiquitin family protein as a novel neutrophil chemotactic factor","volume":"309","author":"Owhashi","year":"2003","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1016\/j.molcel.2017.10.003","article-title":"Extracellular ISG15 Signals cytokine secretion through the LFA-1 integrin receptor","volume":"68","author":"Swaim","year":"2017","journal-title":"Mol. Cell"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"11045","DOI":"10.1128\/JVI.00655-10","article-title":"Porcine reproductive and respiratory syndrome virus inhibits type I interferon signaling by blocking STAT1\/STAT2 nuclear translocation","volume":"84","author":"Patel","year":"2010","journal-title":"J. Virol."},{"key":"ref_26","first-page":"315470","article-title":"Antagonizing interferon-mediated immune response by porcine reproductive and respiratory syndrome virus","volume":"2014","author":"Wang","year":"2014","journal-title":"BioMed Res. Int."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Yang, L., Wang, R., Ma, Z., Xiao, Y., Nan, Y., Wang, Y., Lin, S., and Zhang, Y.J. (2017). Porcine reproductive and respiratory syndrome virus antagonizes JAK\/STAT3 signaling via nsp5, which induces STAT3 degradation. J. Virol., 91.","DOI":"10.1128\/JVI.02087-16"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.virol.2009.11.033","article-title":"Identification of two auto-cleavage products of nonstructural protein 1 (nsp1) in porcine reproductive and respiratory syndrome virus infected cells: Nsp1 function as interferon antagonist","volume":"398","author":"Chen","year":"2010","journal-title":"Virology"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2947","DOI":"10.1099\/vir.0.025205-0","article-title":"The cysteine protease domain of porcine reproductive and respiratory syndrome virus non-structural protein 2 antagonizes interferon regulatory factor 3 activation","volume":"91","author":"Li","year":"2010","journal-title":"J. Gen. Virol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"7832","DOI":"10.1128\/JVI.00217-10","article-title":"The cysteine protease domain of porcine reproductive and respiratory syndrome virus nonstructural protein 2 possesses deubiquitinating and interferon antagonism functions","volume":"84","author":"Sun","year":"2010","journal-title":"J. Virol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"10934","DOI":"10.1128\/JVI.01396-14","article-title":"Porcine reproductive and respiratory syndrome virus nonstructural protein 4 antagonizes beta interferon expression by targeting the NF-kappaB essential modulator","volume":"88","author":"Huang","year":"2014","journal-title":"J. Virol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1568","DOI":"10.1016\/j.molimm.2011.03.004","article-title":"Endoribonuclease activities of porcine reproductive and respiratory syndrome virus nsp11 was essential for nsp11 to inhibit IFN-beta induction","volume":"48","author":"Shi","year":"2011","journal-title":"Mol. Immunol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3839","DOI":"10.1128\/JVI.06466-11","article-title":"Nonstructural protein 2 of porcine reproductive and respiratory syndrome virus inhibits the antiviral function of interferon-stimulated gene 15","volume":"86","author":"Sun","year":"2012","journal-title":"J. Virol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.antiviral.2014.07.011","article-title":"Heme oxygenase-1 acts as an antiviral factor for porcine reproductive and respiratory syndrome virus infection and over-expression inhibits virus replication in vitro","volume":"110","author":"Xiao","year":"2014","journal-title":"Antivir. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.vetmic.2006.06.006","article-title":"Suppression of porcine reproductive and respiratory syndrome virus replication by morpholino antisense oligomers","volume":"117","author":"Zhang","year":"2006","journal-title":"Vet. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Zhang, A., Zhao, L., Li, N., Duan, H., Liu, H., Pu, F., Zhang, G., Zhou, E.M., and Xiao, S. (2017). Carbon monoxide inhibits porcine reproductive and respiratory syndrome virus replication by the cyclic GMP\/Protein Kinase G and NF-kappaB signaling pathway. J. Virol., 91.","DOI":"10.1128\/JVI.01866-16"},{"key":"ref_37","unstructured":"Committee on Methods of Producing Monoclonal Antibodies, and National Research Council (1999). Monoclonal Antibody Production, National Academy Press."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.antiviral.2007.09.002","article-title":"Peptide-conjugated morpholino oligomers inhibit porcine reproductive and respiratory syndrome virus replication","volume":"77","author":"Patel","year":"2008","journal-title":"Antivir. Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1016\/j.virol.2015.05.019","article-title":"Glycoprotein 5 of porcine reproductive and respiratory syndrome virus strain SD16 inhibits viral replication and causes G2\/M cell cycle arrest, but does not induce cellular apoptosis in Marc-145 cells","volume":"484","author":"Mu","year":"2015","journal-title":"Virology"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1016\/j.virol.2012.05.015","article-title":"Induction of type I interferons by a novel porcine reproductive and respiratory syndrome virus isolate","volume":"432","author":"Nan","year":"2012","journal-title":"Virology"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1016\/j.vetmic.2013.07.022","article-title":"Variable interference with interferon signal transduction by different strains of porcine reproductive and respiratory syndrome virus","volume":"166","author":"Wang","year":"2013","journal-title":"Vet. Microbiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1758","DOI":"10.3389\/fimmu.2017.01758","article-title":"Interplay between janus kinase\/signal transducer and activator of transcription signaling activated by type I interferons and viral antagonism","volume":"8","author":"Nan","year":"2017","journal-title":"Front. Immunol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1016\/j.jhep.2017.10.027","article-title":"Safety, efficacy and pharmacodynamics of vesatolimod (GS-9620) in virally suppressed patients with chronic hepatitis B","volume":"68","author":"Janssen","year":"2018","journal-title":"J. Hepatol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"725","DOI":"10.3390\/v4050725","article-title":"Post-transcriptional control of type I interferon induction by porcine reproductive and respiratory syndrome virus in its natural host cells","volume":"4","author":"Wang","year":"2012","journal-title":"Viruses"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2453","DOI":"10.1007\/s00705-004-0377-9","article-title":"Interferon type I response in porcine reproductive and respiratory syndrome virus-infected MARC-145 cells","volume":"149","author":"Miller","year":"2004","journal-title":"Arch. Virol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1038\/nri2314","article-title":"Interferon-inducible antiviral effectors","volume":"8","author":"Sadler","year":"2008","journal-title":"Nat. Rev. Immunol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1089\/jir.2010.0110","article-title":"Interferon-stimulated gene 15 and the protein ISGylation system","volume":"31","author":"Zhang","year":"2011","journal-title":"J. Interferon Cytokine Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"13974","DOI":"10.1128\/JVI.79.22.13974-13983.2005","article-title":"Identification of interferon-stimulated gene 15 as an antiviral molecule during Sindbis virus infection in vivo","volume":"79","author":"Lenschow","year":"2005","journal-title":"J. Virol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"6338","DOI":"10.1128\/MCB.25.15.6338-6345.2005","article-title":"ISG15, an interferon-stimulated ubiquitin-like protein, is not essential for STAT1 signaling and responses against vesicular stomatitis and lymphocytic choriomeningitis virus","volume":"25","author":"Osiak","year":"2005","journal-title":"Mol. Cell. Biol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"532723","DOI":"10.1155\/2012\/532723","article-title":"Budding of enveloped viruses: Interferon-induced ISG15-antivirus mechanisms targeting the release process","volume":"2012","author":"Seo","year":"2012","journal-title":"Adv. Virol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1684","DOI":"10.1126\/science.1224026","article-title":"Mycobacterial disease and impaired IFN-gamma immunity in humans with inherited ISG15 deficiency","volume":"337","author":"Bogunovic","year":"2012","journal-title":"Science"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1635","DOI":"10.3389\/fmicb.2017.01635","article-title":"Improved vaccine against PRRSV: Current progress and future perspective","volume":"8","author":"Nan","year":"2017","journal-title":"Front. Microbiol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/12\/9\/1009\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:08:34Z","timestamp":1760177314000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/12\/9\/1009"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,9,10]]},"references-count":52,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2020,9]]}},"alternative-id":["v12091009"],"URL":"https:\/\/doi.org\/10.3390\/v12091009","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,9,10]]}}}