{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T08:59:19Z","timestamp":1764665959232,"version":"3.46.0"},"reference-count":98,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T00:00:00Z","timestamp":1764633600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["-398066876\/GRK 2485\/2-VIPER-GRK"],"award-info":[{"award-number":["-398066876\/GRK 2485\/2-VIPER-GRK"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"name":"German Academic Scholarship Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Viral infections in humans and animals are increasing, and retrospective studies using formalin-fixed, paraffin-embedded (FFPE) samples reveal recurring outbreaks over past decades. However, the impact of pre-analytical factors like fixation and autolysis on immunohistochemistry (IHC) remains insufficiently understood. To examine how autolysis, fixation duration (6\u201372 h) and formalin concentration (2.5\u201325%) influence histology and IHC of canine distemper virus (CDV, Morbillivirus canis), interferon-\u03b2 (IFN-\u03b2), and selected IFN-stimulated genes (ISGs), the study was conducted using an in vitro model based on persistently CDV-infected and non-infected DH82 cells (canine histiocytic sarcoma cell line). Autolysis led to a progressive loss of cell morphology, whereas formalin fixation had minimal impact. CDV nucleoprotein, ISG15, and myxovirus resistance protein (Mx) showed stable immunohistochemical signals across all fixation conditions and remained detectable after prolonged autolysis. CDV infection upregulated ISG15 and Mx. In contrast, IFN-\u03b2 and phosphorylated protein kinase R (pPKR) exhibited variable staining and did not distinguish infected from non-infected samples. Overall, autolysis had a stronger negative impact on IHC signal quality than fixation parameters. Despite the limitations of the in vitro model, the robustness of CDV, ISG15, and Mx under suboptimal conditions highlights their potential utility as virus-sensing markers in FFPE material.<\/jats:p>","DOI":"10.3390\/v17121575","type":"journal-article","created":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T08:49:22Z","timestamp":1764665362000},"page":"1575","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["An In Vitro Approach to Evaluate the Impact of Autolysis and Formalin Fixation on the Detection of Canine Distemper Virus and Innate Immune Response Antigens"],"prefix":"10.3390","volume":"17","author":[{"given":"Hannah","family":"Gerhards","sequence":"first","affiliation":[{"name":"Department of Pathology, University of Veterinary Medicine Hannover, Foundation, 30559 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karl","family":"Rohn","sequence":"additional","affiliation":[{"name":"Department of Biometry, Epidemiology and Data Management, University of Veterinary Medicine Hannover, Foundation, 30559 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2592-6868","authenticated-orcid":false,"given":"Christina","family":"Puff","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Veterinary Medicine Hannover, Foundation, 30559 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8151-5644","authenticated-orcid":false,"given":"Wolfgang","family":"Baumg\u00e4rtner","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Veterinary Medicine Hannover, Foundation, 30559 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"499","DOI":"10.20506\/rst.36.2.2669","article-title":"Viral discovery as a tool for pandemic preparedness","volume":"36","author":"Epstein","year":"2017","journal-title":"Rev. Sci. Tech."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Smits, S.L., Bodewes, R., Ruiz-Gonzalez, A., Baumg\u00e4rtner, W., Koopmans, M.P., Osterhaus, A.D., and Schurch, A.C. (2014). Assembly of viral genomes from metagenomes. Front. Microbiol., 5.","DOI":"10.3389\/fmicb.2014.00714"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.1016\/j.coviro.2013.03.007","article-title":"Virus discovery: One step beyond","volume":"3","author":"Smits","year":"2013","journal-title":"Curr. Opin. Virol."},{"key":"ref_4","first-page":"1042","article-title":"Rustrela virus as putative cause of nonsuppurative meningoencephalitis in lions","volume":"29","author":"Puff","year":"2023","journal-title":"Emerg. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Bassi, C., Guerriero, P., Pierantoni, M., Callegari, E., and Sabbioni, S. (2022). Novel virus identification through metagenomics: A systematic review. Life, 12.","DOI":"10.3390\/life12122048"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"753","DOI":"10.1369\/jhc.2009.953877","article-title":"Effects of prolonged formalin fixation on diagnostic immunohistochemistry in domestic animals","volume":"57","author":"Webster","year":"2009","journal-title":"J. Histochem. Cytochem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e377","DOI":"10.7717\/peerj.377","article-title":"Comparison of histomorphology and DNA preservation produced by fixatives in the veterinary diagnostic laboratory setting","volume":"2","author":"Craft","year":"2014","journal-title":"PeerJ"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"102496","DOI":"10.1016\/j.legalmed.2024.102496","article-title":"The influence of fixing condition on myoglobin stainability of striated muscle as a tool for forensic diagnosis","volume":"71","author":"Kobayashi","year":"2024","journal-title":"Leg. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1177\/0300985809359607","article-title":"Effects of prolonged formalin fixation on the immunohistochemical detection of infectious agents in formalin-fixed, paraffin-embedded tissues","volume":"47","author":"Webster","year":"2010","journal-title":"Vet. Pathol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"151879","DOI":"10.1016\/j.acthis.2022.151879","article-title":"The effect of prolonged formalin fixation on the expression of proteins in human brain tissues","volume":"124","author":"Wu","year":"2022","journal-title":"Acta Histochem."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Bilgin, C., Dai, D., Johnson, C., Mereuta, O.M., Kallmes, D.F., Brinjikji, W., and Kadirvel, R. (2023). Quality assessment of histopathological stainings on prolonged formalin fixed thrombus tissues retrieved by mechanical thrombectomy. Front. Neurol., 14.","DOI":"10.3389\/fneur.2023.1223947"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Chafin, D., Theiss, A., Roberts, E., Borlee, G., Otter, M., and Baird, G.S. (2013). Rapid two-temperature formalin fixation. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0054138"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"850","DOI":"10.1038\/modpathol.3800354","article-title":"Ultrasound-accelerated formalin fixation of tissue improves morphology, antigen and mRNA preservation","volume":"18","author":"Chu","year":"2005","journal-title":"Mod. Pathol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e1996","DOI":"10.7717\/peerj.1996","article-title":"Determining the utility of veterinary tissue archives for retrospective DNA analysis","volume":"4","author":"Abed","year":"2016","journal-title":"PeerJ"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.1111\/j.1464-410X.2006.06699.x","article-title":"Immunohistochemical markers in urinary bladder carcinomas from paraffin-embedded archival tissue after storage for 5\u201370 years","volume":"99","author":"Litlekalsoy","year":"2007","journal-title":"BJU Int."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/S1385-299X(97)00029-9","article-title":"Immunocytochemistry of formalin-fixed human brain tissues: Microwave irradiation of free-floating sections","volume":"2","author":"Shiurba","year":"1998","journal-title":"Brain Res. Brain Res. Protoc."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/j.vascn.2009.12.002","article-title":"Troubleshooting tissue specificity and antibody selection: Procedures in immunohistochemical studies","volume":"61","author":"Daneshtalab","year":"2010","journal-title":"J. Pharmacol. Toxicol. Methods"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.aanat.2019.03.013","article-title":"Influence of fixation method and duration of archiving on immunohistochemical staining intensity in embryonic and fetal tissues","volume":"224","author":"Tauber","year":"2019","journal-title":"Ann. Anat."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1016","DOI":"10.1097\/00000478-200007000-00014","article-title":"Effect of formalin tissue fixation and processing on immunohistochemistry","volume":"24","author":"Werner","year":"2000","journal-title":"Am. J. Surg. Pathol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1369\/jhc.7A7187.2007","article-title":"Immunohistochemical markers for quantitative studies of neurons and glia in human neocortex","volume":"56","author":"Lyck","year":"2008","journal-title":"J. Histochem. Cytochem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1111\/his.12646","article-title":"A reference image-based method for optimization of clinical immunohistochemistry","volume":"67","author":"Kap","year":"2015","journal-title":"Histopathology"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Parisi, F., Degl\u2019Innocenti, S., Ayta\u015f, \u00c7., Pirone, A., and Cantile, C. (2024). Morphological and immunohistochemical changes in progressive postmortem autolysis of the murine brain. Animals, 14.","DOI":"10.3390\/ani14243676"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.forsciint.2005.01.013","article-title":"Is there recent progress in the estimation of the postmortem interval by means of thanatochemistry?","volume":"151","author":"Madea","year":"2005","journal-title":"Forensic Sci. Int."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"897","DOI":"10.25122\/jml-2024-0260","article-title":"Postmortem interval estimation of time since death: Impact of non-histone binding proteins, immunohistochemical, and histopathological changes in vivo","volume":"17","author":"Albloshi","year":"2024","journal-title":"J. Med. Life"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/S0379-0738(99)00124-3","article-title":"Delimitation of the time of death by immunohistochemical detection of insulin in pancreatic beta-cells","volume":"105","author":"Wehner","year":"1999","journal-title":"Forensic Sci. Int."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"334","DOI":"10.1038\/labinvest.2014.139","article-title":"Preanalytical variables and phosphoepitope expression in FFPE tissue: Quantitative epitope assessment after variable cold ischemic time","volume":"95","author":"Vassilakopoulou","year":"2015","journal-title":"Lab. Invest."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Salerno, M., Cocimano, G., Roccuzzo, S., Russo, I., Piombino-Mascali, D., M\u00e1rquez-Grant, N., Zammit, C., Esposito, M., and Sessa, F. (2022). New trends in immunohistochemical methods to estimate the time since death: A review. Diagnostics, 12.","DOI":"10.3390\/diagnostics12092114"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Mauriello, S., Treglia, M., Pallocci, M., Bonfiglio, R., Giacobbi, E., Passalacqua, P., Cammarano, A., D\u2019Ovidio, C., Marsella, L.T., and Scimeca, M. (2022). Antigenicity preservation is related to tissue characteristics and the post-mortem interval: Immunohistochemical study and literature review. Healthcare, 10.","DOI":"10.3390\/healthcare10081495"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Barral-Arca, R., Pardo-Seco, J., Martin\u00f3n-Torres, F., and Salas, A. (2018). A 2-transcript host cell signature distinguishes viral from bacterial diarrhea and it is influenced by the severity of symptoms. Sci. Rep., 8.","DOI":"10.1038\/s41598-018-26239-1"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Cruz-Flores, R., Mai, H.N., Kanrar, S., Aranguren Caro, L.F., and Dhar, A.K. (2020). Genome reconstruction of white spot syndrome virus (WSSV) from archival Davidson\u2019s-fixed paraffin embedded shrimp (Penaeus vannamei) tissue. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-70435-x"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1016\/j.jcv.2010.07.015","article-title":"Use of antisera directed against dsRNA to detect viral infections in formalin-fixed paraffin-embedded tissue","volume":"49","author":"Richardson","year":"2010","journal-title":"J. Clin. Virol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1111\/apm.13300","article-title":"Immunohistochemical detection of double-stranded RNA in formalin-fixed paraffin-embedded tissue","volume":"131","author":"Thomsen","year":"2023","journal-title":"Apmis"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"9383","DOI":"10.1128\/JVI.01299-15","article-title":"Double-stranded RNA is detected by immunofluorescence analysis in RNA and DNA virus infections, including those by negative-stranded RNA viruses","volume":"89","author":"Son","year":"2015","journal-title":"J. Virol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"6062","DOI":"10.1021\/am506374t","article-title":"Comparison of three magnetic bead surface functionalities for RNA extraction and detection","volume":"7","author":"Adams","year":"2015","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"5059","DOI":"10.1128\/JVI.80.10.5059-5064.2006","article-title":"Double-stranded RNA is produced by positive-strand RNA viruses and DNA viruses but not in detectable amounts by negative-strand RNA viruses","volume":"80","author":"Weber","year":"2006","journal-title":"J. Virol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1146\/annurev-immunol-042718-041356","article-title":"Double-stranded RNA sensors and modulators in innate immunity","volume":"37","author":"Hur","year":"2019","journal-title":"Annu. Rev. Immunol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1016\/j.cytogfr.2007.06.001","article-title":"Interferon, Mx, and viral countermeasures","volume":"18","author":"Haller","year":"2007","journal-title":"Cytokine Growth Factor. Rev."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"920","DOI":"10.3390\/v3060920","article-title":"Pattern recognition receptors and the innate immune response to viral infection","volume":"3","author":"Thompson","year":"2011","journal-title":"Viruses"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1038\/nri3787","article-title":"Type I interferons in infectious disease","volume":"15","author":"McNab","year":"2015","journal-title":"Nat. Rev. Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1196\/annals.1443.020","article-title":"Toll-like receptor and RIG-I-like receptor signaling","volume":"1143","author":"Kawai","year":"2008","journal-title":"Ann. N. Y Acad. Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"886","DOI":"10.1007\/s13365-017-0583-3","article-title":"Interferon regulated gene (IRG) expression-signature in a mouse model of chikungunya virus neurovirulence","volume":"23","author":"Nair","year":"2017","journal-title":"J. Neurovirol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1007\/s13238-018-0511-1","article-title":"Identification of new type I interferon-stimulated genes and investigation of their involvement in IFN-\u03b2 activation","volume":"9","author":"Zhang","year":"2018","journal-title":"Protein Cell"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1101\/gr.20102","article-title":"Functional diversity of Mx proteins: Variations on a theme of host resistance to infection","volume":"12","author":"Lee","year":"2002","journal-title":"Genome Res."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"710","DOI":"10.1034\/j.1600-0854.2002.31003.x","article-title":"Interferon-induced mx proteins: Dynamin-like GTPases with antiviral activity","volume":"3","author":"Haller","year":"2002","journal-title":"Traffic"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1016\/j.mib.2013.04.005","article-title":"An evolutionary perspective on the broad antiviral specificity of MxA","volume":"16","author":"Mitchell","year":"2013","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Klotz, D., and Gerhauser, I. (2019). Interferon-stimulated genes-mediators of the innate immune response during canine distemper virus infection. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20071620"},{"key":"ref_47","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_48","doi-asserted-by":"crossref","first-page":"778","DOI":"10.1128\/CMR.14.4.778-809.2001","article-title":"Antiviral actions of interferons","volume":"14","author":"Samuel","year":"2001","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Huang, C.H., Laurent-Rolle, M., Grove, T.L., and Hsu, J.C. (2025). Interferon-stimulated genes and immune metabolites as broad-spectrum biomarkers for viral infections. Viruses, 17.","DOI":"10.3390\/v17010132"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"3361","DOI":"10.4049\/jimmunol.1900804","article-title":"IFN-stimulated genes in zebrafish and humans define an ancient arsenal of antiviral immunity","volume":"203","author":"Levraud","year":"2019","journal-title":"J. Immunol."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Johanson, T.M., Lew, A.M., and Chong, M.M. (2013). MicroRNA-independent roles of the RNase III enzymes Drosha and Dicer. Open Biol., 3.","DOI":"10.1098\/rsob.130144"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1146\/annurev-biophys-083012-130404","article-title":"Molecular mechanisms of RNA interference","volume":"42","author":"Wilson","year":"2013","journal-title":"Annu. Rev. Biophys."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1636","DOI":"10.1016\/j.micinf.2007.09.010","article-title":"The Mx GTPase family of interferon-induced antiviral proteins","volume":"9","author":"Haller","year":"2007","journal-title":"Microbes Infect."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/j.jviromet.2006.05.004","article-title":"Detection of canine distemper virus in dogs by real-time RT-PCR","volume":"136","author":"Elia","year":"2006","journal-title":"J. Virol. Methods"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.coviro.2016.01.019","article-title":"Mapping the evolutionary trajectories of morbilliviruses: What, where and whither","volume":"16","author":"Nambulli","year":"2016","journal-title":"Curr. Opin. Virol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.onehlt.2015.09.002","article-title":"Cross-species transmission of canine distemper virus-an update","volume":"1","author":"Beineke","year":"2015","journal-title":"One Health"},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Geiselhardt, F., Peters, M., Jo, W.K., Schadenhofer, A., Puff, C., Baumg\u00e4rtner, W., Kydyrmanov, A., Kuiken, T., Piewbang, C., and Techangamsuwan, S. (2022). Development and validation of a pan-genotypic real-time quantitative reverse transcription-PCR assay to detect canine distemper virus and phocine distemper virus in domestic animals and wildlife. J. Clin. Microbiol., 60.","DOI":"10.1128\/jcm.02505-21"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1016\/j.cvsm.2008.02.007","article-title":"Canine distemper virus","volume":"38","author":"Martella","year":"2008","journal-title":"Vet. Clin. N. Am. Small Anim. Pract."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1007\/s00401-004-0958-4","article-title":"Demyelination in canine distemper virus infection: A review","volume":"109","author":"Vandevelde","year":"2005","journal-title":"Acta Neuropathol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.vetimm.2014.11.006","article-title":"Passage-dependent morphological and phenotypical changes of a canine histiocytic sarcoma cell line (DH82 cells)","volume":"163","author":"Heinrich","year":"2015","journal-title":"Vet. Immunol. Immunopathol."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"104206","DOI":"10.1016\/j.meegid.2020.104206","article-title":"RNA sequencing analyses of gene expressions in a canine macrophages cell line DH82 infected with canine distemper virus","volume":"80","author":"Zheng","year":"2020","journal-title":"Infect. Genet. Evol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1007\/BF02623551","article-title":"A macrophage-monocyte cell line from a dog with malignant histiocytosis","volume":"24","author":"Wellman","year":"1988","journal-title":"In Vitro Cell Dev. Biol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.tvjl.2008.03.026","article-title":"Influence of persistent canine distemper virus infection on expression of RECK, matrix-metalloproteinases and their inhibitors in a canine macrophage\/monocytic tumour cell line (DH82)","volume":"182","author":"Puff","year":"2009","journal-title":"Vet. J."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1136\/jclinpath-2019-206368","article-title":"Decreasing formalin concentration improves quality of DNA extracted from formalin-fixed paraffin-embedded tissue specimens without compromising tissue morphology or immunohistochemical staining","volume":"73","author":"Cummings","year":"2020","journal-title":"J. Clin. Pathol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1177\/10406387231220649","article-title":"Effects of autolysis and prolonged formalin fixation on histomorphology and immunohistochemistry of normal canine brain tissue: An experimental study","volume":"36","author":"Koehler","year":"2024","journal-title":"J. Vet. Diagn. Invest."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1177\/10406387211021865","article-title":"Feasibility of using tissue autolysis to estimate the postmortem interval in horses","volume":"33","author":"Wenzlow","year":"2021","journal-title":"J. Vet. Diagn. Invest."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"101","DOI":"10.5603\/FM.a2021.0002","article-title":"Decay score: A guide to the immunoreactivity of human pancreatic islets in autopsy specimen","volume":"81","author":"Ravi","year":"2022","journal-title":"Folia Morphol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1097\/PAF.0000000000000408","article-title":"Usability of immunohistochemistry in forensic samples with varying decomposition","volume":"39","author":"Lesnikova","year":"2018","journal-title":"Am. J. Forensic Med. Pathol."},{"key":"ref_69","doi-asserted-by":"crossref","unstructured":"Chludzinski, E., Klemens, J., Ciurkiewicz, M., Geffers, R., P\u00f6pperl, P., Stoff, M., Shin, D.L., Herrler, G., and Beineke, A. (2022). Phenotypic and transcriptional changes of pulmonary immune responses in dogs following canine distemper virus infection. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms231710019"},{"key":"ref_70","doi-asserted-by":"crossref","unstructured":"de le Roi, M., Gerhards, H., Fayyad, A., Boelke, M., Becker, S.C., Volz, A., Gerhauser, I., Baumg\u00e4rtner, W., and Puff, C. (2025). Evaluating the potential of anti-dsRNA antibodies as an alternative viral sensing tool in encephalitides of different species. Front. Vet. Sci., 12.","DOI":"10.3389\/fvets.2025.1540437"},{"key":"ref_71","doi-asserted-by":"crossref","unstructured":"Bankhead, P., Loughrey, M.B., Fernandez, J.A., Dombrowski, Y., McArt, D.G., Dunne, P.D., McQuaid, S., Gray, R.T., Murray, L.J., and Coleman, H.G. (2017). QuPath: Open source software for digital pathology image analysis. Sci. Rep., 7.","DOI":"10.1038\/s41598-017-17204-5"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"2080","DOI":"10.1038\/s41467-025-57267-x","article-title":"Respiratory long COVID in aged hamsters features impaired lung function post-exercise with bronchiolization and fibrosis","volume":"16","author":"Heydemann","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1369\/jhc.2009.954008","article-title":"Fixative composition alters distributions of immunoreactivity for glutaminase and two markers of nociceptive neurons, Nav1.8 and TRPV1, in the rat dorsal root ganglion","volume":"58","author":"Hoffman","year":"2010","journal-title":"J. Histochem. Cytochem."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"756","DOI":"10.1053\/hupa.2002.126187","article-title":"Inadequate formalin fixation decreases reliability of p27 immunohistochemical staining: Probing optimal fixation time using high-density tissue microarrays","volume":"33","author":"Fedor","year":"2002","journal-title":"Hum. Pathol."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1177\/104063870201400611","article-title":"Effect of formalin fixation on the immunohistochemical detection of PRRS virus antigen in experimentally and naturally infected pigs","volume":"14","author":"Popielarczyk","year":"2002","journal-title":"J. Vet. Diagn. Invest."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"949","DOI":"10.1177\/03009858251349126","article-title":"Standardization of tissue handling in veterinary pathology: How fixation affects morphological and molecular examination results","volume":"62","author":"Chambers","year":"2025","journal-title":"Vet. Pathol."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1007\/s00428-023-03692-6","article-title":"Glyoxal acid-free (GAF) histological fixative is a suitable alternative to formalin: Results from an open-label comparative non-inferiority study","volume":"485","author":"Ryska","year":"2024","journal-title":"Virchows Arch."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"100198","DOI":"10.1016\/j.labinv.2023.100198","article-title":"A novel tissue preservation and transport solution as a substitute for formalin","volume":"103","author":"Vucevic","year":"2023","journal-title":"Lab. Invest."},{"key":"ref_79","first-page":"1453","article-title":"Estimation of tumor growth fractions in archival formalin-fixed, paraffin-embedded tissues using two anti-PCNA\/Cyclin monoclonal antibodies. Factors affecting reactivity","volume":"141","author":"Gelb","year":"1992","journal-title":"Am. J. Pathol."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/S0034-5288(96)90016-8","article-title":"Immunohistochemical detection of rinderpest virus: Effects of autolysis and period of fixation","volume":"60","author":"Brown","year":"1996","journal-title":"Res. Vet. Sci."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1007\/s00428-020-02848-y","article-title":"Influence of preanalytical variables on performance of delta-like protein 3 (DLL3) predictive immunohistochemistry","volume":"478","author":"Radonic","year":"2021","journal-title":"Virchows Arch."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1369\/jhc.6A7152.2007","article-title":"Robust immunohistochemical staining of several classes of proteins in tissues subjected to autolysis","volume":"55","author":"Maleszewski","year":"2007","journal-title":"J. Histochem. Cytochem."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/0046-8177(91)90156-J","article-title":"Differential antigen preservation during tissue autolysis","volume":"22","author":"Pelstring","year":"1991","journal-title":"Hum. Pathol."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"1423","DOI":"10.1007\/s00018-020-03671-z","article-title":"Viral pathogen-induced mechanisms to antagonize mammalian interferon (IFN) signaling pathway","volume":"78","author":"Rojas","year":"2021","journal-title":"Cell Mol. Life Sci."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"286","DOI":"10.1038\/s41580-021-00430-1","article-title":"Cellular origins of dsRNA, their recognition and consequences","volume":"23","author":"Chen","year":"2022","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_86","doi-asserted-by":"crossref","unstructured":"Siering, O., Sawatsky, B., and Pfaller, C.K. (2021). C protein is essential for canine distemper virus virulence and pathogenicity in ferrets. J. Virol., 95.","DOI":"10.1128\/JVI.01840-20"},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1128\/JVI.02572-13","article-title":"Measles virus C protein impairs production of defective copyback double-stranded viral RNA and activation of protein kinase R","volume":"88","author":"Pfaller","year":"2014","journal-title":"J. Virol."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1373\/clinchem.2018.292391","article-title":"Interferon-inducible myxovirus resistance proteins: Potential biomarkers for differentiating viral from bacterial infections","volume":"65","author":"Korpela","year":"2019","journal-title":"Clin. Chem."},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"981","DOI":"10.1354\/vp.43-6-981","article-title":"Immunohistochemical evaluation of mx protein expression in canine encephalitides","volume":"43","author":"Porter","year":"2006","journal-title":"Vet. Pathol."},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"612","DOI":"10.1186\/s40001-024-02221-8","article-title":"Recent updates of interferon-derived myxovirus resistance protein A as a biomarker for acute viral infection","volume":"29","author":"Abebaw","year":"2024","journal-title":"Eur. J. Med. Res."},{"key":"ref_91","doi-asserted-by":"crossref","unstructured":"Sampson, D.L., Fox, B.A., Yager, T.D., Bhide, S., Cermelli, S., McHugh, L.C., Seldon, T.A., Brandon, R.A., Sullivan, E., and Zimmerman, J.J. (2017). A four-biomarker blood signature discriminates systemic inflammation due to viral infection versus other etiologies. Sci. Rep., 7.","DOI":"10.1038\/s41598-017-02325-8"},{"key":"ref_92","doi-asserted-by":"crossref","unstructured":"Zandstra, J., Jongerius, I., and Kuijpers, T.W. (2021). Future biomarkers for infection and inflammation in febrile children. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.631308"},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"1051","DOI":"10.1016\/j.molcel.2018.07.029","article-title":"PKR senses nuclear and mitochondrial signals by interacting with endogenous double-stranded RNAs","volume":"71","author":"Kim","year":"2018","journal-title":"Mol. Cell"},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1111\/imr.13272","article-title":"Type I interferon and cancer","volume":"321","author":"Holicek","year":"2024","journal-title":"Immunol. Rev."},{"key":"ref_95","doi-asserted-by":"crossref","unstructured":"Chhipa, A.S., Boscaro, V., Gallicchio, M., and Patel, S. (2024). The curious case of type I interferon signaling in cancer. Biochim. Biophys. Acta Rev. Cancer, 1879.","DOI":"10.1016\/j.bbcan.2024.189204"},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"109185","DOI":"10.1016\/j.isci.2024.109185","article-title":"Bi-directional regulation of type I interferon signaling by heme oxygenase-1","volume":"27","author":"Wu","year":"2024","journal-title":"iScience"},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1016\/j.immuni.2013.05.007","article-title":"Cytosolic sensing of viruses","volume":"38","author":"Goubau","year":"2013","journal-title":"Immunity"},{"key":"ref_98","doi-asserted-by":"crossref","unstructured":"Nan, Y., Wu, C., and Zhang, Y.J. (2017). Interplay between janus kinase\/signal transducer and activator of transcription signaling activated by type I interferons and viral antagonism. Front. Immunol., 8.","DOI":"10.3389\/fimmu.2017.01758"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/17\/12\/1575\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T08:55:44Z","timestamp":1764665744000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/17\/12\/1575"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,2]]},"references-count":98,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["v17121575"],"URL":"https:\/\/doi.org\/10.3390\/v17121575","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,2]]}}}