{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,6]],"date-time":"2026-06-06T05:07:46Z","timestamp":1780722466217,"version":"3.54.1"},"reference-count":41,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2022,3,24]],"date-time":"2022-03-24T00:00:00Z","timestamp":1648080000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004488","name":"Croatian Science Foundation","doi-asserted-by":"publisher","award":["IP-2019-04-6048"],"award-info":[{"award-number":["IP-2019-04-6048"]}],"id":[{"id":"10.13039\/501100004488","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The low seroprevalent human adenovirus type 26 (HAdV26)-based vaccine vector was the first adenovirus-based vector to receive marketing authorization from European Commission. HAdV26-based vaccine vectors induce durable humoral and cellular immune responses and, as such, represent a highly valuable tool for fighting infectious diseases. Despite well-described immunogenicity in vivo, the basic biology of HAdV26 still needs some refinement. The aim of this study was to determine the pro-inflammatory cytokine profile of epithelial cells infected with HAdV26 and then investigate the underlying molecular mechanism. The expression of studied genes and proteins was assessed by quantitative polymerase chain reaction, western blot, and enzyme-linked immunosorbent assay. Confocal microscopy was used to visualize HAdV26 cell uptake. We found that HAdV26 infection in human epithelial cells triggers the expression of pro-inflammatory cytokines and chemokines, namely IL-6, IL-8, IL-1\u03b2, and TNF-\u03b1, with the most pronounced difference shown for IL-6. We investigated the underlying molecular mechanism and observed that HAdV26-induced IL-6 gene expression is \u03b1v\u03b23 integrin dependent and NF-\u03baB mediated. Our findings provide new data regarding pro-inflammatory cytokine and chemokine expression in HAdV26-infected epithelial cells, as well as details concerning HAdV26-induced host signaling pathways. Information obtained within this research increases our current knowledge of HAdV26 basic biology and, as such, can contribute to further development of HAdV26-based vaccine vectors.<\/jats:p>","DOI":"10.3390\/v14040672","type":"journal-article","created":{"date-parts":[[2022,3,24]],"date-time":"2022-03-24T23:31:43Z","timestamp":1648164703000},"page":"672","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Human Adenovirus Type 26 Induced IL-6 Gene Expression in an \u03b1v\u03b23 Integrin- and NF-\u03baB-Dependent Manner"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5518-5650","authenticated-orcid":false,"given":"Davor","family":"Nesti\u0107","sequence":"first","affiliation":[{"name":"Laboratory for Cell Biology and Signalling, Division of Molecular Biology, Ru\u0111er Bo\u0161kovi\u0107 Institute, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6840-6689","authenticated-orcid":false,"given":"Ksenija","family":"Bo\u017einovi\u0107","sequence":"additional","affiliation":[{"name":"Laboratory for Cell Biology and Signalling, Division of Molecular Biology, Ru\u0111er Bo\u0161kovi\u0107 Institute, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1905-0115","authenticated-orcid":false,"given":"Isabela","family":"Dra\u0161kovi\u0107","sequence":"additional","affiliation":[{"name":"Laboratory for Cell Biology and Signalling, Division of Molecular Biology, Ru\u0111er Bo\u0161kovi\u0107 Institute, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alen","family":"Kova\u010devi\u0107","sequence":"additional","affiliation":[{"name":"Laboratory for Cell Biology and Signalling, Division of Molecular Biology, Ru\u0111er Bo\u0161kovi\u0107 Institute, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jolien","family":"van den Bosch","sequence":"additional","affiliation":[{"name":"Laboratory for Cell Biology and Signalling, Division of Molecular Biology, Ru\u0111er Bo\u0161kovi\u0107 Institute, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3152-3563","authenticated-orcid":false,"given":"Jelena","family":"Kne\u017eevi\u0107","sequence":"additional","affiliation":[{"name":"Laboratory for Advanced Genomics, Division of Molecular Medicine, Ru\u0111er Bo\u0161kovi\u0107 Institute, 10000 Zagreb, Croatia"},{"name":"Faculty for Dental Medicine and Health, University of Osijek, 31000 Osijek, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jerome","family":"Custers","sequence":"additional","affiliation":[{"name":"Janssen Vaccines and Preventions BV, 2333 CA Leiden, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7784-2466","authenticated-orcid":false,"given":"Andreja","family":"Ambriovi\u0107-Ristov","sequence":"additional","affiliation":[{"name":"Laboratory for Cell Biology and Signalling, Division of Molecular Biology, Ru\u0111er Bo\u0161kovi\u0107 Institute, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0385-0900","authenticated-orcid":false,"given":"Dragomira","family":"Majhen","sequence":"additional","affiliation":[{"name":"Laboratory for Cell Biology and Signalling, Division of Molecular Biology, Ru\u0111er Bo\u0161kovi\u0107 Institute, 10000 Zagreb, Croatia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1089\/hum.2013.235","article-title":"Adenovirus-Based Vaccines for Fighting Infectious Diseases and Cancer: Progress in the Field","volume":"25","author":"Majhen","year":"2014","journal-title":"Hum. Gene Ther."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"15265","DOI":"10.1128\/JVI.79.24.15265-15276.2005","article-title":"Phylogenetic Analysis of the Main Neutralization and Hemagglutination Determinants of All Human Adenovirus Prototypes as a Basis for Molecular Classification and Taxonomy","volume":"79","author":"Madisch","year":"2005","journal-title":"J. Virol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3482","DOI":"10.1128\/JCM.00156-11","article-title":"Computational Analysis of Two Species C Human Adenoviruses Provides Evidence of a Novel Virus","volume":"49","author":"Walsh","year":"2011","journal-title":"J. Clin. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.virol.2010.10.020","article-title":"Computational analysis and identification of an emergent human adenovirus pathogen implicated in a respiratory fatality","volume":"409","author":"Robinson","year":"2011","journal-title":"Virology"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Gallardo, J., P\u00e9rez-Illana, M., Mart\u00edn-Gonz\u00e1lez, N., and Mart\u00edn, C.S. (2021). Adenovirus Structure: What Is New?. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22105240"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"11241","DOI":"10.1128\/JVI.00721-06","article-title":"Interaction of Adenovirus Type 5 Fiber with the Coxsackievirus and Adenovirus Receptor Activates Inflammatory Response in Human Respiratory Cells","volume":"80","author":"Tamanini","year":"2006","journal-title":"J. Virol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1111\/cmi.12053","article-title":"Adenovirus signalling in entry","volume":"15","author":"Wolfrum","year":"2012","journal-title":"Cell. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2875","DOI":"10.4161\/hv.29594","article-title":"Pre-existing immunity against Ad vectors: Humoral, cellular, and innate response, what\u2019s important?","volume":"10","author":"Kobinger","year":"2014","journal-title":"Hum. Vaccin. Immunother."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"3081","DOI":"10.1016\/j.vaccine.2020.09.018","article-title":"Vaccines based on replication incompetent Ad26 viral vectors: Standardized template with key considerations for a risk\/benefit assessment","volume":"39","author":"Custers","year":"2021","journal-title":"Vaccine"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"313","DOI":"10.7326\/M15-0880","article-title":"Assessment of the Safety and Immunogenicity of 2 Novel Vaccine Platforms for HIV-1 Prevention: A Randomized Trial","volume":"164","author":"Baden","year":"2016","journal-title":"Ann. Intern. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1093\/infdis\/jiz070","article-title":"Safety and Immunogenicity of a 2-Dose Heterologous Vaccination Regimen with Ad26.ZEBOV and MVA-BN-Filo Ebola Vaccines: 12-Month Data From a Phase 1 Randomized Clinical Trial in Uganda and Tanzania","volume":"220","author":"Anywaine","year":"2019","journal-title":"J. Infect. Dis."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1093\/infdis\/jis671","article-title":"Characterization of Humoral and Cellular Immune Responses Elicited by a Recombinant Adenovirus Serotype 26 HIV-1 Env Vaccine in Healthy Adults (IPCAVD 001)","volume":"207","author":"Barouch","year":"2012","journal-title":"J. Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1038\/s41541-020-0200-y","article-title":"Adenovector 26 Encoded Prefusion Conformation Stabilized RSV-F Protein Induces Long-Lasting Th1-Biased Immunity in Neonatal Mice","volume":"5","author":"Bolder","year":"2020","journal-title":"npj Vaccines"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"9590","DOI":"10.1128\/JVI.00740-12","article-title":"Vaccination with Adenovirus Serotypes 35, 26, and 48 Elicits Higher Levels of Innate Cytokine Responses than Adenovirus Serotype 5 in Rhesus Monkeys","volume":"86","author":"Teigler","year":"2012","journal-title":"J. Virol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1128\/JVI.76.1.427-431.2002","article-title":"Interleukin-1\u03b1 Released from Epithelial Cells after Adenovirus Type 37 Infection Activates Intercellular Adhesion Molecule 1 Expression on Human Vascular Endothelial Cells","volume":"76","author":"Chang","year":"2002","journal-title":"J. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4654","DOI":"10.1128\/JVI.02696-06","article-title":"Comparative Seroprevalence and Immunogenicity of Six Rare Serotype Recombinant Adenovirus Vaccine Vectors from Subgroups B and D","volume":"81","author":"Abbink","year":"2007","journal-title":"J. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"8263","DOI":"10.1128\/JVI.77.15.8263-8271.2003","article-title":"Replication-Deficient Human Adenovirus Type 35 Vectors for Gene Transfer and Vaccination: Efficient Human Cell Infection and Bypass of Preexisting Adenovirus Immunity","volume":"77","author":"Vogels","year":"2003","journal-title":"J. Virol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e01474-18","DOI":"10.1128\/JVI.01474-18","article-title":"Av\u03b23 Integrin Is Required for Efficient Infection of Epithelial Cells with Human Adenovirus Type 26","volume":"93","author":"Uil","year":"2019","journal-title":"J. Virol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2212\u0394\u0394CT method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1089\/hum.2014.001","article-title":"Innate Immunity to Adenovirus","volume":"25","author":"Hendrickx","year":"2014","journal-title":"Hum. Gene Ther."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"19792","DOI":"10.1073\/pnas.1212597109","article-title":"\u03b1v\u03b23-integrin is a major sensor and activator of innate immunity to herpes simplex virus-1","volume":"109","author":"Gianni","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4243","DOI":"10.1128\/JVI.03175-15","article-title":"\u03b1v\u03b23 Integrin Boosts the Innate Immune Response Elicited in Epithelial Cells through Plasma Membrane and Endosomal Toll-Like Receptors","volume":"90","author":"Casiraghi","year":"2016","journal-title":"J. Virol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"935","DOI":"10.1038\/sj.gt.3302036","article-title":"Molecular basis of the inflammatory response to adenovirus vectors","volume":"10","author":"Liu","year":"2003","journal-title":"Gene Ther."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"17023","DOI":"10.1038\/sigtrans.2017.23","article-title":"NF-\u039aB Signaling in Inflammation","volume":"2","author":"Liu","year":"2017","journal-title":"Signal Transduct. Target Ther."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"416036","DOI":"10.1155\/2012\/416036","article-title":"BAY 11-7082 Is a Broad-Spectrum Inhibitor with Anti-Inflammatory Activity against Multiple Targets","volume":"2012","author":"Lee","year":"2012","journal-title":"Mediat. Inflamm."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2899","DOI":"10.1128\/JVI.76.6.2899-2911.2002","article-title":"Distinct Roles of Adenovirus Vector-Transduced Dendritic Cells, Myoblasts, and Endothelial Cells in Mediating an Immune Response against a Transgene Product","volume":"76","author":"Mercier","year":"2002","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.imlet.2010.09.003","article-title":"Adenoviral vectors stimulate innate immune responses in macrophages through cross-talk with epithelial cells","volume":"134","author":"Lee","year":"2010","journal-title":"Immunol. Lett."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"909","DOI":"10.3389\/fimmu.2020.00909","article-title":"Factors Which Contribute to the Immunogenicity of Non-replicating Adenoviral Vectored Vaccines","volume":"11","author":"Coughlan","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1002\/ppul.20574","article-title":"Cytokine induction by respiratory syncytial virus and adenovirus in bronchial epithelial cells","volume":"42","author":"Yoon","year":"2007","journal-title":"Pediatr. Pulmonol."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Lam, E., Ramke, M., Warnecke, G., Schrepfer, S., Kopfnagel, V., Dobner, T., and Heim, A. (2015). Effective Apical Infection of Differentiated Human Bronchial Epithelial Cells and Induction of Proinflammatory Chemokines by the Highly Pneumotropic Human Adenovirus Type 14p1. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0131201"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1165\/ajrcmb.21.4.3677","article-title":"Type-Specific Induction of Interleukin-8 by Adenovirus","volume":"21","author":"Booth","year":"1999","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"6450","DOI":"10.1128\/JVI.75.14.6450-6459.2001","article-title":"Adenovirus Type 7 Induces Interleukin-8 Production via Activation of Extracellular Regulated Kinase 1\/2","volume":"75","author":"Alcorn","year":"2001","journal-title":"J. Virol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"6234","DOI":"10.4049\/jimmunol.170.12.6234","article-title":"Corneal IL-8 Expression Following Adenovirus Infection Is Mediated by c-Src Activation in Human Corneal Fibroblasts","volume":"170","author":"Natarajan","year":"2003","journal-title":"J. Immunol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1186\/1743-422X-5-17","article-title":"Human adenovirus type 19 infection of corneal cells induces p38 MAPK-dependent interleukin-8 expression","volume":"5","author":"Rajaiya","year":"2008","journal-title":"Virol. J."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1016\/j.immuni.2009.04.015","article-title":"Virus Binding to a Plasma Membrane Receptor Triggers Interleukin-1\u03b1-Mediated Proinflammatory Macrophage Response In Vivo","volume":"31","author":"Miao","year":"2009","journal-title":"Immunity"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2811","DOI":"10.1158\/0008-5472.CAN-14-3761","article-title":"Pharmacological Inhibition of \u03b23 Integrin Reduces the Inflammatory Toxicities Caused by Oncolytic Adenovirus without Compromising Anticancer Activity","volume":"75","author":"Browne","year":"2015","journal-title":"Cancer Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"551413","DOI":"10.3389\/fimmu.2020.551413","article-title":"Adenovirus 7 Induces Interlukin-6 Expression in Human Airway Epithelial Cells via P38\/NF-\u039aB Signaling Pathway","volume":"11","author":"Qi","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"14507","DOI":"10.1128\/JVI.79.23.14507-14515.2005","article-title":"Akt\/Protein Kinase B Activation by Adenovirus Vectors Contributes to NFkappaB-Dependent CXCL10 Expression","volume":"79","author":"Liu","year":"2005","journal-title":"J. Virol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"19922","DOI":"10.1038\/srep19922","article-title":"NF-\u039aB Promotes Leaky Expression of Adenovirus Genes in a Replication-Incompetent Adenovirus Vector","volume":"6","author":"Machitani","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"33363","DOI":"10.18632\/oncotarget.9221","article-title":"Kaposi\u2019s Sarcoma Herpesvirus (KSHV) MicroRNA K12-1 Functions as an Oncogene by Activating NF-\u039aB\/IL-6\/STAT3 Signaling","volume":"7","author":"Chen","year":"2016","journal-title":"Oncotarget"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Morgan, E.L., and Macdonald, A. (2019). Autocrine STAT3 activation in HPV positive cervical cancer through a virus-driven Rac1-NF\u03baB-IL-6 signalling axis. PLoS Pathog., 15.","DOI":"10.1371\/journal.ppat.1007835"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/4\/672\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:42:45Z","timestamp":1760136165000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/4\/672"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,24]]},"references-count":41,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2022,4]]}},"alternative-id":["v14040672"],"URL":"https:\/\/doi.org\/10.3390\/v14040672","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,24]]}}}