Interferon Gamma Inhibits Equine Herpesvirus 1 Replication in a Cell Line-Dependent Manner.
Abstract: The sole equine herpesvirus 1 (EHV-1) immediate-early protein (IEP) is essential for viral replication by transactivating viral immediate-early (IE), early (E), and late (L) genes. Here, we report that treatment of mouse MH-S, equine NBL6, and human MRC-5 cells with 20 ng/mL of IFN-γ reduced EHV-1 yield by 1122-, 631-, and 10,000-fold, respectively. However, IFN-γ reduced virus yield by only 2-4-fold in mouse MLE12, mouse L-M, and human MeWo cells compared to those of untreated cells. In luciferase assays with the promoter of the EHV-1 early regulatory EICP0 gene, IFN-γ abrogated -activation activity of the IEP by 96% in MH-S cells, but only by 21% in L-M cells. Similar results were obtained in assays with the early regulatory UL5 and IR4 promoter reporter plasmids. IFN-γ treatment reduced IEP protein expression by greater than 99% in MH-S cells, but only by 43% in L-M cells. The expression of IEP and UL5P suppressed by IFN-γ was restored by JAK inhibitor treatment, indicating that the inhibition of EHV-1 replication is mediated by JAK/STAT1 signaling. These results suggest that IFN-γ blocks EHV-1 replication by inhibiting the production of the IEP in a cell line-dependent manner. Affymetrix microarray analyses of IFN-γ-treated MH-S and L-M cells revealed that five antiviral ISGs (MX1, SAMHD1, IFIT2, NAMPT, TREX1, and DDX60) were upregulated 3.2-18.1-fold only in MH-S cells.
Publication Date: 2021-04-16 PubMed ID: 33923733PubMed Central: PMC8073143DOI: 10.3390/pathogens10040484Google Scholar: Lookup
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Summary
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This research article investigates how the Interferon Gamma (IFN-γ) protein impacts the replication of the equine herpesvirus 1 (EHV-1) in different cell lines. The findings show that IFN-γ can significantly reduce EHV-1 replication in certain cell lines through JAK/STAT1 signaling and various antiviral genes.
Understanding the Research
- The primary focus of this study stems from understanding the role of the immediate-early protein (IEP) in EHV-1 (a type of horse herpesvirus) and its vital role in viral replication. Specifically, the researchers aimed to assess how IFN-γ treatment affects the replication of EHV-1 across different cell lines.
- Three different cell lines – mouse MH-S, equine NBL6, and human MRC-5 – were treated with 20 ng/mL of IFN-γ. The results displayed a significant decrease in EHV-1 yield in these cell lines compared to other cell lines treated with the same dosage of IFN-γ.
Role of IFN-γ and IEP
- Interferon Gamma (IFN-γ) plays a crucial role in contributing to immune responses against viral infections. This research highlights the role this protein plays in inhibiting the EHV-1 as it apparently blocks the replication process by suppressing the production of IEP, but this happened to be cell line dependent.
- The IEP, on the other hand, is essential for the replication of the EHV-1 as it facilitates the transactivation of several viral genes. Due to this, IEP protein expression and its function came under scrutiny during the IFN-γ treatment.
JAK/STAT1 Signaling and Capacity of IFN-γ Treatment
- The JAK/STAT1 pathway is integral for turning on the responses for the cell when it comes under any immune attack. This study shows that the suppression of IEP and UL5P by IFN-γ can be restored by JAK inhibitor treatment. This indicates the important role the JAK/STAT1 pathway plays in supporting the IFN-γ’s inhibition of EHV-1 replication.
- The IFN-γ treatment, however, seemingly worked differently across various cell lines. While in MH-S cells, IEP protein expression was reduced by over 99% with IFN-γ treatment, it was only reduced by 43% in L-M cells. This indicated that the effectiveness of IFN-γ treatment in inhibiting EHV-1 replication had cell-line dependencies.
Microarray Analyses and Antiviral Interferon-stimulated Genes (ISGs)
- The study’s microarray analyses identified five antiviral ISGs that were upregulated between 3.2 to 18.1-fold only in the MH-S cells. These ISGs – MX1, SAMHD1, IFIT2, NAMPT, TREX1, and DDX60 – might significantly contribute to the cell-line dependent manner of the IFN-γ’s inhibitive mechanism on the EHV-1 replication.
Cite This Article
APA
Kim SK, Shakya AK, O'Callaghan DJ.
(2021).
Interferon Gamma Inhibits Equine Herpesvirus 1 Replication in a Cell Line-Dependent Manner.
Pathogens, 10(4), 484.
https://doi.org/10.3390/pathogens10040484 Publication
Researcher Affiliations
- Center for Molecular and Tumor Virology, Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, Shreveport, LA 71130-3932, USA.
- Center for Molecular and Tumor Virology, Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, Shreveport, LA 71130-3932, USA.
- Center for Molecular and Tumor Virology, Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, Shreveport, LA 71130-3932, USA.
Grant Funding
- P30GM110703 / NIGMS NIH HHS
- the National Research Initiative Competitive Grant 2013-67015-21311 / the USDA (NIFA) Cooperative State Research, Education and Extension Service
Conflict of Interest Statement
The authors declare no conflict of interest.
References
This article includes 63 references
- Allen GP, Bryans JT. Molecular epizootiology, pathogenesis, and prophylaxis of equine herpesvirus-1 infections.. Prog Vet Microbiol Immunol 1986;2:78-144.
- Carroll CL, Westbury HA. Isolation of equine herpesvirus 1 from the brain of a horse affected with paresis.. Aust Vet J 1985 Oct;62(10):345-6.
- Jackson T, Kendrick JW. Paralysis of horses associated with equine herpesvirus 1 infection.. J Am Vet Med Assoc 1971 Apr 15;158(8):1351-7.
- Bryans JT, Allen GP. Equine viral rhinopneumonitis.. Rev Sci Tech 1986 Dec;5(4):837-867.
- Crandell RA, Mock RE, Lock TF. Vaccination of pregnant ponies against equine rhinopneumonitis.. Am J Vet Res 1980 Jul;41(7):994-6.
- Farrar MA, Schreiber RD. The molecular cell biology of interferon-gamma and its receptor.. Annu Rev Immunol 1993;11:571-611.
- Schoenborn JR, Wilson CB. Regulation of interferon-gamma during innate and adaptive immune responses.. Adv Immunol 2007;96:41-101.
- Spellberg B, Edwards JE Jr. Type 1/Type 2 immunity in infectious diseases.. Clin Infect Dis 2001 Jan;32(1):76-102.
- Ealick SE, Cook WJ, Vijay-Kumar S, Carson M, Nagabhushan TL, Trotta PP, Bugg CE. Three-dimensional structure of recombinant human interferon-gamma.. Science 1991 May 3;252(5006):698-702.
- Platanias LC. Mechanisms of type-I- and type-II-interferon-mediated signalling.. Nat Rev Immunol 2005 May;5(5):375-86.
- Haan C, Kreis S, Margue C, Behrmann I. Jaks and cytokine receptors--an intimate relationship.. Biochem Pharmacol 2006 Nov 30;72(11):1538-46.
- Rodig SJ, Meraz MA, White JM, Lampe PA, Riley JK, Arthur CD, King KL, Sheehan KC, Yin L, Pennica D, Johnson EM Jr, Schreiber RD. Disruption of the Jak1 gene demonstrates obligatory and nonredundant roles of the Jaks in cytokine-induced biologic responses.. Cell 1998 May 1;93(3):373-83.
- Yeh TC, Pellegrini S. The Janus kinase family of protein tyrosine kinases and their role in signaling.. Cell Mol Life Sci 1999 Sep;55(12):1523-34.
- Boehm U, Klamp T, Groot M, Howard JC. Cellular responses to interferon-gamma.. Annu Rev Immunol 1997;15:749-95.
- Colle CF 3rd, Flowers CC, O'Callaghan DJ. Open reading frames encoding a protein kinase, homolog of glycoprotein gX of pseudorabies virus, and a novel glycoprotein map within the unique short segment of equine herpesvirus type 1.. Virology 1992 Jun;188(2):545-57.
- Frampton AR Jr, Smith PM, Zhang Y, Matsumura T, Osterrieder N, O'Callaghan DJ. Contribution of gene products encoded within the unique short segment of equine herpesvirus 1 to virulence in a murine model.. Virus Res 2002 Dec;90(1-2):287-301.
- Lewis JB, Thompson YG, Feng X, Holden VR, O'Callaghan D, Caughman GB. Structural and antigenic identification of the ORF12 protein (alpha TIF) of equine herpesvirus 1.. Virology 1997 Apr 14;230(2):369-75.
- Matsumura T, O'Callaghan DJ, Kondo T, Kamada M. Lack of virulence of the murine fibroblast adapted strain, Kentucky A (KyA), of equine herpesvirus type 1 (EHV-1) in young horses.. Vet Microbiol 1996 Feb;48(3-4):353-65.
- Smith PM, Kahan SM, Rorex CB, von Einem J, Osterrieder N, O'Callaghan DJ. Expression of the full-length form of gp2 of equine herpesvirus 1 (EHV-1) completely restores respiratory virulence to the attenuated EHV-1 strain KyA in CBA mice.. J Virol 2005 Apr;79(8):5105-15.
- Smith PM, Zhang Y, Grafton WD, Jennings SR, O'Callaghan DJ. Severe murine lung immunopathology elicited by the pathogenic equine herpesvirus 1 strain RacL11 correlates with early production of macrophage inflammatory proteins 1alpha, 1beta, and 2 and tumor necrosis factor alpha.. J Virol 2000 Nov;74(21):10034-40.
- Kim SK, Shakya AK, O'Callaghan DJ. Immunization with Attenuated Equine Herpesvirus 1 Strain KyA Induces Innate Immune Responses That Protect Mice from Lethal Challenge.. J Virol 2016 Sep 15;90(18):8090-104.
- Coombs DK, Patton T, Kohler AK, Soboll G, Breathnach C, Townsend HG, Lunn DP. Cytokine responses to EHV-1 infection in immune and non-immune ponies.. Vet Immunol Immunopathol 2006 May 15;111(1-2):109-16.
- Bartels T, Steinbach F, Hahn G, Ludwig H, Borchers K. In situ study on the pathogenesis and immune reaction of equine herpesvirus type 1 (EHV-1) infections in mice.. Immunology 1998 Mar;93(3):329-34.
- Kim SK, Shakya AK, O'Callaghan DJ. Intranasal treatment with CpG-B oligodeoxynucleotides protects CBA mice from lethal equine herpesvirus 1 challenge by an innate immune response.. Antiviral Res 2019 Sep;169:104546.
- Smith RH, Caughman GB, O'Callaghan DJ. Characterization of the regulatory functions of the equine herpesvirus 1 immediate-early gene product.. J Virol 1992 Feb;66(2):936-45.
- Smith RH, Holden VR, O'Callaghan DJ. Nuclear localization and transcriptional activation activities of truncated versions of the immediate-early gene product of equine herpesvirus 1.. J Virol 1995 Jun;69(6):3857-62.
- Buczynski KA, Kim SK, O'Callaghan DJ. Initial characterization of 17 viruses harboring mutant forms of the immediate-early gene of equine herpesvirus 1.. Virus Genes 2005 Oct;31(2):229-39.
- Harris N, Buller RM, Karupiah G. Gamma interferon-induced, nitric oxide-mediated inhibition of vaccinia virus replication.. J Virol 1995 Feb;69(2):910-5.
- Harty RN, O'Callaghan DJ. An early gene maps within and is 3' coterminal with the immediate-early gene of equine herpesvirus 1.. J Virol 1991 Jul;65(7):3829-38.
- Kim SK, Shakya AK, O'Callaghan DJ. Full trans-activation mediated by the immediate-early protein of equine herpesvirus 1 requires a consensus TATA box, but not its cognate binding sequence.. Virus Res 2016 Jan 4;211:222-32.
- Smith RH, Zhao Y, O'Callaghan DJ. The equine herpesvirus type 1 immediate-early gene product contains an acidic transcriptional activation domain.. Virology 1994 Aug 1;202(2):760-70.
- Caughman GB, Staczek J, O'Callaghan DJ. Equine herpesvirus type 1 infected cell polypeptides: evidence for immediate early/early/late regulation of viral gene expression.. Virology 1985 Aug;145(1):49-61.
- Gray WL, Baumann RP, Robertson AT, Caughman GB, O'Callaghan DJ, Staczek J. Regulation of equine herpesvirus type 1 gene expression: characterization of immediate early, early, and late transcription.. Virology 1987 May;158(1):79-87.
- Gray WL, Baumann RP, Robertson AT, O'Callaghan DJ, Staczek J. Characterization and mapping of equine herpesvirus type 1 immediate early, early, and late transcripts.. Virus Res 1987 Sep;8(3):233-44.
- Henry BE, Robinson RA, Dauenhauer SA, Atherton SS, Hayward GS, O'Callaghan DJ. Structure of the genome of equine herpesvirus type 1.. Virology 1981 Nov;115(1):97-114.
- Shakya AK, O'Callaghan DJ, Kim SK. Interferon Gamma Inhibits Varicella-Zoster Virus Replication in a Cell Line-Dependent Manner.. J Virol 2019 Jun 15;93(12).
- Goodwin MM, Canny S, Steed A, Virgin HW. Murine gammaherpesvirus 68 has evolved gamma interferon and stat1-repressible promoters for the lytic switch gene 50.. J Virol 2010 Apr;84(7):3711-7.
- Schoggins JW, Rice CM. Interferon-stimulated genes and their antiviral effector functions.. Curr Opin Virol 2011 Dec;1(6):519-25.
- Chen S, Bonifati S, Qin Z, St Gelais C, Kodigepalli KM, Barrett BS, Kim SH, Antonucci JM, Ladner KJ, Buzovetsky O, Knecht KM, Xiong Y, Yount JS, Guttridge DC, Santiago ML, Wu L. SAMHD1 suppresses innate immune responses to viral infections and inflammatory stimuli by inhibiting the NF-κB and interferon pathways.. Proc Natl Acad Sci U S A 2018 Apr 17;115(16):E3798-E3807.
- Chen Z, Zhu M, Pan X, Zhu Y, Yan H, Jiang T, Shen Y, Dong X, Zheng N, Lu J, Ying S, Shen Y. Inhibition of Hepatitis B virus replication by SAMHD1.. Biochem Biophys Res Commun 2014 Aug 8;450(4):1462-8.
- Sze A, Belgnaoui SM, Olagnier D, Lin R, Hiscott J, van Grevenynghe J. Host restriction factor SAMHD1 limits human T cell leukemia virus type 1 infection of monocytes via STING-mediated apoptosis.. Cell Host Microbe 2013 Oct 16;14(4):422-34.
- Wagner B, Robeson J, McCracken M, Wattrang E, Antczak DF. Horse cytokine/IgG fusion proteins--mammalian expression of biologically active cytokines and a system to verify antibody specificity to equine cytokines.. Vet Immunol Immunopathol 2005 May 1;105(1-2):1-14.
- Gutmann S, Zawatzky R, Müller M. Characterisation and quantification of equine interferon gamma.. Vet Immunol Immunopathol 2005 Mar 10;104(1-2):105-15.
- Smith PM, Zhang Y, Jennings SR, O'Callaghan DJ. Characterization of the cytolytic T-lymphocyte response to a candidate vaccine strain of equine herpesvirus 1 in CBA mice.. J Virol 1998 Jul;72(7):5366-72.
- Zhang Y, Smith PM, Jennings SR, O'Callaghan DJ. Quantitation of virus-specific classes of antibodies following immunization of mice with attenuated equine herpesvirus 1 and viral glycoprotein D.. Virology 2000 Mar 15;268(2):482-92.
- Colle CF 3rd, Tarbet EB, Grafton WD, Jennings SR, O'Callaghan DJ. Equine herpesvirus-1 strain KyA, a candidate vaccine strain, reduces viral titers in mice challenged with a pathogenic strain, RacL.. Virus Res 1996 Aug;43(2):111-24.
- Ito S, Ishii KJ, Gursel M, Shirotra H, Ihata A, Klinman DM. CpG oligodeoxynucleotides enhance neonatal resistance to Listeria infection.. J Immunol 2005 Jan 15;174(2):777-82.
- Wattrang E, Palm AK, Wagner B. Cytokine production and proliferation upon in vitro oligodeoxyribonucleotide stimulation of equine peripheral blood mononuclear cells.. Vet Immunol Immunopathol 2012 Apr 15;146(2):113-24.
- Holden VR, Yalamanchili RR, Harty RN, O'Callaghan DJ. ICP22 homolog of equine herpesvirus 1: expression from early and late promoters.. J Virol 1992 Feb;66(2):664-73.
- Kim SK, Ahn BC, Albrecht RA, O'Callaghan DJ. The unique IR2 protein of equine herpesvirus 1 negatively regulates viral gene expression.. J Virol 2006 May;80(10):5041-9.
- Zhao Y, Holden VR, Smith RH, O'Callaghan DJ. Regulatory function of the equine herpesvirus 1 ICP27 gene product.. J Virol 1995 May;69(5):2786-93.
- Roizman B. The family Herpesviridae: A brief introduction.. Fields Virol 2001;2:2381–2397.
- Nathan C. Nitric oxide as a secretory product of mammalian cells.. FASEB J 1992 Sep;6(12):3051-64.
- Nathan CF, Hibbs JB Jr. Role of nitric oxide synthesis in macrophage antimicrobial activity.. Curr Opin Immunol 1991 Feb;3(1):65-70.
- Karupiah G, Xie QW, Buller RM, Nathan C, Duarte C, MacMicking JD. Inhibition of viral replication by interferon-gamma-induced nitric oxide synthase.. Science 1993 Sep 10;261(5127):1445-8.
- Trilling M, Le VT, Zimmermann A, Ludwig H, Pfeffer K, Sutter G, Smith GL, Hengel H. Gamma interferon-induced interferon regulatory factor 1-dependent antiviral response inhibits vaccinia virus replication in mouse but not human fibroblasts.. J Virol 2009 Apr;83(8):3684-95.
- Der SD, Zhou A, Williams BR, Silverman RH. Identification of genes differentially regulated by interferon alpha, beta, or gamma using oligonucleotide arrays.. Proc Natl Acad Sci U S A 1998 Dec 22;95(26):15623-8.
- Sen GC, Peters GA. Viral stress-inducible genes.. Adv Virus Res 2007;70:233-63.
- O'Callaghan DJ, Cheevers WP, Gentry GA, Randall CC. Kinetics of cellular and viral DNA synthesis in equine abortion (herpes) virus infection of L-M cells.. Virology 1968 Sep;36(1):104-14.
- Perdue ML, Kemp MC, Randall CC, O'Callaghan DJ. Studies of the molecular anatomy of the L-M cell strain of equine herpes virus type 1: proteins of the nucleocapsid and intact virion.. Virology 1974 May;59(1):201-16.
- RECZKO E, MAYR A. [ON THE FINE STRUCTURE OF A VIRUS OF THE HERPES GROUP ISOLATED FROM HORSES (SHORT REPORT)].. Arch Gesamte Virusforsch 1963 Aug 26;13:591-3.
- Kim SK, Kim S, Dai G, Zhang Y, Ahn BC, O'Callaghan DJ. Identification of functional domains of the IR2 protein of equine herpesvirus 1 required for inhibition of viral gene expression and replication.. Virology 2011 Sep 1;417(2):430-42.
- Sambrook J., Fritsh E., Maniatis T. Molecular Cloning: A Laboratory Manual. 2nd ed. Cold Spring Harbor Press; New York, NY, USA: 1989.
Citations
This article has been cited 2 times.- Karasawa T, Sato R, Imaizumi T, Hashimoto S, Fujita M, Aizawa T, Tsugawa K, Kawaguchi S, Seya K, Terui K, Tanaka H. Glomerular endothelial expression of type I IFN-stimulated gene, DExD/H-Box helicase 60 via toll-like receptor 3 signaling: possible involvement in the pathogenesis of lupus nephritis. Ren Fail 2022 Dec;44(1):137-145.
- Frøvoll TL, Lybeck K, Lund H, Makvandi-Nejad S, Grimholt U, das Neves CG, Tryland M, Nymo IH, Klevar S. Measuring cytokines in Eurasian tundra reindeer (Rangifer tarandus tarandus) with a bovine bead-based multiplex immunoassay and real-time PCR. Acta Vet Scand 2025 Jun 18;67(1):34.
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