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These cytokines include heterodimers sharing distinct \u03b1 (IL-12A, IL-23A, and IL-27A) with two \u03b2 (IL-12B and Epstein-Barr virus induced gene 3 [EBI3]) chains, respectively, IL-12 (IL-12B plus IL-12A) and IL-23 (IL-12B plus IL-23A) sharing IL-12B, IL-27 (EBI3 plus IL-27A), IL-35 (EBI3 plus IL-12A), and IL-39 (EBI3 plus IL-23A) sharing EBI3. In this context, we have recently reported that highly pure neutrophils incubated with TLR8 agonists produce functional IL-23. Previously, we showed that neutrophils incubated with LPS plus IFN\u03b3 for 20 h produce IL-12. Herein, we investigated whether highly pure, TLR8-activated, neutrophils produce EBI3, and in turn IL-27, IL-35, and IL-39, the IL-12 members containing it. We report that neutrophils incubated with TLR8 ligands, TNF\u03b1 and, to a lesser extent, LPS, produce and release remarkable amounts of EBI3, but not IL-27A, consequently excluding the possibility for an IL-27 production. We also report a series of unsuccessful experiments performed to investigate whether neutrophil-derived EBI3 associates with IL-23A to form IL-39. Furthermore, we show that neutrophils incubated with IFN\u03b3 in combination with either TLR8 or TLR4 ligands express\/produce neither IL-12, nor IL-35, due to the inability of IFN\u03b3, contrary to previous findings, to activate IL12A transcription. Even IL-27 was undetectable in supernatants harvested from IFN\u03b3 plus R848-treated neutrophils, although they were found to accumulate IL27A transcripts. Finally, by immunohistochemistry experiments, EBI3-positive neutrophils were found in discrete pathologies only, including diverticulitis, cholecystitis, Gorham disease, and Bartonella Henselae infection, implying a specific role of neutrophil-derived EBI3 in vivo.<\/jats:p>","DOI":"10.1002\/jlb.3ma0520-054r","type":"journal-article","created":{"date-parts":[[2020,6,1]],"date-time":"2020-06-01T21:09:11Z","timestamp":1591045751000},"page":"1515-1526","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Human neutrophils activated by TLR8 agonists, with or without IFN\u03b3, synthesize and release EBI3, but not IL-12, IL-27, IL-35, or IL-39"],"prefix":"10.1093","volume":"108","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7824-1806","authenticated-orcid":true,"given":"Marco A","family":"Cassatella","sequence":"first","affiliation":[{"name":"Department of Medicine, Section of General Pathology, University of Verona , Verona , Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisa","family":"Gardiman","sequence":"additional","affiliation":[{"name":"Department of Medicine, Section of General Pathology, University of Verona , Verona , Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabio","family":"Arruda-Silva","sequence":"additional","affiliation":[{"name":"Department of Medicine, Section of General Pathology, University of Verona , Verona , Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francisco","family":"Bianchetto-Aguilera","sequence":"additional","affiliation":[{"name":"Department of Medicine, Section of General Pathology, University of Verona , Verona , Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Gasperini","sequence":"additional","affiliation":[{"name":"Department of Medicine, Section of General Pathology, University of Verona , Verona , Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mattia","family":"Bugatti","sequence":"additional","affiliation":[{"name":"Department of Molecular and Translational Medicine, Section of Pathology, University of Brescia , Brescia , Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William","family":"Vermi","sequence":"additional","affiliation":[{"name":"Department of Molecular and Translational Medicine, Section of Pathology, University of Brescia , Brescia , Italy"},{"name":"Department of Pathology and Immunology, Washington University School of Medicine , Saint Louis, Missouri , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frederique","family":"Larousserie","sequence":"additional","affiliation":[{"name":"Sorbonne Universit\u00e9, INSERM, CNRS, Centre d\u2019Immunologie et des Maladies Infectieuses , Paris , France"},{"name":"D\u00e9partement de Pathologie, H\u00f4pital Cochin, AP-HP, Universit\u00e9 de Paris , Paris , France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Odile","family":"Devergne","sequence":"additional","affiliation":[{"name":"Sorbonne Universit\u00e9, INSERM, CNRS, Centre d\u2019Immunologie et des Maladies Infectieuses , Paris , France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicola","family":"Tamassia","sequence":"additional","affiliation":[{"name":"Department of Medicine, Section of General Pathology, University of Verona , Verona , Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2020,6,1]]},"reference":[{"key":"2023032108064595300_jlb10690-cit-0001","first-page":"e12952","article-title":"Cytokine production by human neutrophils: revisiting the \u201cdark side of the moon\u201d","volume":"2018","author":"Tamassia","journal-title":"Eur J Clin 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