{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T12:06:51Z","timestamp":1787314011789,"version":"build-2736575974"},"reference-count":30,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,2,5]],"date-time":"2021-02-05T00:00:00Z","timestamp":1612483200000},"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","id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>\n                    Neutrophil extracellular traps (NETs) are upregulated and promote thrombosis in Beh\u00e7et\u2019s disease (BD). However, whether NETs promote autoinflammation in BD remains unclear. This study aimed to investigate the potential role of NETs in promoting macrophage activation in BD. Firstly, we quantified NETs by measuring double-stranded DNA (dsDNA) using PicoGreen and calculating the proportion of NETosis. Then macrophages were stimulated with BD- or healthy controls (HC)-derived NETs, and IL-8 and TNF-\u03b1 production and IFN-\u03b3\n                    <jats:sup>+<\/jats:sup>\n                    CD4\n                    <jats:sup>+<\/jats:sup>\n                    T cells differentiation were measured using ELISA and flow cytometry, respectively. The protein components in NETs were analyzed by western blot. Macrophages were stimulated with Histone H4 neutralized NETs, and IL-8 and TNF-\u03b1 production were measured using ELISA. The level of 8-hydroxydeoxyguanosine (8-OHdG) DNA in NETs was measured using ELISA. The levels of reactive oxygen species (ROS) in serum and neutrophils were measured using ROS probes by a microplate reader and flow cytometry. We found that circulating NETs and neutrophil-derived NETs were significantly higher in BD than HC. BD NETs stimulated macrophages produced higher levels of IL-8 and TNF-\u03b1, and promoted IFN-\u03b3\n                    <jats:sup>+<\/jats:sup>\n                    CD4\n                    <jats:sup>+<\/jats:sup>\n                    T cells differentiation. BD NETs were enriched in Histone H4, and neutralizing Histone H4 abrogated the BD NETs-mediated IL-8 production by macrophages, but not TNF-\u03b1. Also, BD neutrophils produced more 8-OHdG DNA than HC neutrophils, and the percentage of 8-OHdG DNA in dsDNA from BD neutrophils was also higher than that of HC neutrophils. The ROS levels in serum and neutrophils were both higher in BD than HC. Our findings suggested that excessive BD NETs promoted macrophages activation and facilitated IFN-\u03b3\n                    <jats:sup>+<\/jats:sup>\n                    CD4\n                    <jats:sup>+<\/jats:sup>\n                    T cells differentiation. Higher levels of Histone H4 and oxidized DNA in BD NETs might mediate macrophages hyperactivation.\n                  <\/jats:p>","DOI":"10.3389\/fimmu.2020.590622","type":"journal-article","created":{"date-parts":[[2021,2,5]],"date-time":"2021-02-05T01:25:44Z","timestamp":1612488344000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":57,"title":["Neutrophil Extracellular Traps Promote Aberrant Macrophages Activation in Beh\u00e7et\u2019s Disease"],"prefix":"10.3389","volume":"11","author":[{"given":"Lu","family":"Li","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinjing","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhimian","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chaoran","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Shi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luxi","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fengchun","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hua","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenjie","family":"Zheng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,2,5]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"813","DOI":"10.1016\/s0140-6736(75)80031-6","article-title":"Leucocyte movement and colchicine treatment in Behcet\u2019s disease","volume":"2","author":"Matsumura","year":"1975","journal-title":"Lancet"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1092385","article-title":"Neutrophil extracellular traps kill bacteria","volume":"303","author":"Brinkmann","year":"2004","journal-title":"Science"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1136\/annrheumdis-2018-214335","article-title":"Critical role of neutrophil extracellular traps (NETs) in patients with Behcet\u2019s disease","volume":"78","author":"Le Joncour","year":"2019","journal-title":"Ann Rheum Dis"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1016\/j.jdermsci.2018.08.010","article-title":"Neutrophils contribute to vasculitis by increased release of neutrophil extracellular traps in Behcet\u2019s disease","volume":"92","author":"Safi","year":"2018","journal-title":"J Dermatol Sci"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1300436","article-title":"Macrophage clearance of neutrophil extracellular traps is a silent process","volume":"191","author":"Farrera","year":"2013","journal-title":"J Immunol"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1136\/annrheumdis-2018-213746","article-title":"Dysregulated neutrophil responses and neutrophil extracellular trap formation and degradation in PAPA syndrome","volume":"77","author":"Mistry","year":"2018","journal-title":"Ann Rheum Dis"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1136\/annrheumdis-2017-212617","article-title":"Differential ubiquitination in NETs regulates macrophage responses in systemic lupus erythematosus","volume":"77","author":"Barrera-Vargas","year":"2018","journal-title":"Ann Rheum Dis"},{"key":"B8","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1186\/s13075-018-1800-z","article-title":"Increased neutrophil extracellular traps activate NLRP3 and inflammatory macrophages in adult-onset Still\u2019s disease","volume":"21","author":"Hu","year":"2019","journal-title":"Arthritis Res Ther"},{"key":"B9","doi-asserted-by":"publisher","first-page":"e52687","DOI":"10.3791\/52687","article-title":"Simplified Human Neutrophil Extracellular Traps (NETs) Isolation and Handling","volume":"98","author":"Najmeh","year":"2015","journal-title":"J Vis Exp"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1038\/nm.4027","article-title":"Neutrophil extracellular traps enriched in oxidized mitochondrial DNA are interferogenic and contribute to lupus-like disease","volume":"22","author":"Lood","year":"2016","journal-title":"Nat Med"},{"key":"B11","doi-asserted-by":"publisher","first-page":"235","DOI":"10.1186\/s13075-017-1443-5","article-title":"Soluble CD40L is associated with increased oxidative burst and neutrophil extracellular trap release in Behcet\u2019s disease","volume":"19","author":"Perazzio","year":"2017","journal-title":"Arthritis Res Ther"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1016\/j.intimp.2013.07.017","article-title":"Possible mechanisms of neutrophil activation in Behcet\u2019s disease","volume":"17","author":"Neves","year":"2013","journal-title":"Int Immunopharmacol"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2004-08-3193","article-title":"The CXC chemokine MIP-2 stimulates neutrophil mobilization from the rat bone marrow in a CD49d-dependent manner","volume":"105","author":"Burdon","year":"2005","journal-title":"Blood"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-4632.2004.02184.x","article-title":"Serum interleukin-8 as a serologic marker of activity in Behcet\u2019s disease","volume":"44","author":"Gur-Toy","year":"2005","journal-title":"Int J Dermatol"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1080\/03009740600905380","article-title":"Expression of cytokines, chemokines, and chemokine receptors in oral ulcers of patients with Behcet\u2019s disease (BD) and recurrent aphthous stomatitis is Th1-associated, although Th2-association is also observed in patients with BD","volume":"35","author":"Dalghous","year":"2006","journal-title":"Scand J Rheumatol"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1093\/rheumatology\/keq366","article-title":"A single infliximab infusion vs corticosteroids for acute panuveitis attacks in Behcet\u2019s disease: a comparative 4-week study","volume":"50","author":"Markomichelakis","year":"2011","journal-title":"Rheumatol (Oxford)"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.2169\/internalmedicine.52.0589","article-title":"Efficacy of infliximab in patients with intestinal Behcet\u2019s disease refractory to conventional medication","volume":"52","author":"Kinoshita","year":"2013","journal-title":"Intern Med"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1002\/jcp.27576","article-title":"Behcet\u2019s disease: An immunogenetic perspective","volume":"234","author":"Salmaninejad","year":"2019","journal-title":"J Cell Physiol"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2018.01554","article-title":"Thrombin and Plasmin Alter the Proteome of Neutrophil Extracellular Traps","volume":"9","author":"Lim","year":"2018","journal-title":"Front Immunol"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000639","article-title":"Neutrophil extracellular traps contain calprotectin, a cytosolic protein complex involved in host defense against Candida albicans","volume":"5","author":"Urban","year":"2009","journal-title":"PloS Pathog"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0075141","article-title":"Global substrate profiling of proteases in human neutrophil extracellular traps reveals consensus motif predominantly contributed by elastase","volume":"8","author":"O\u2019Donoghue","year":"2013","journal-title":"PloS One"},{"key":"B22","doi-asserted-by":"publisher","first-page":"e1370","DOI":"10.1038\/cddis.2014.337","article-title":"Release and activity of histone in diseases","volume":"5","author":"Chen","year":"2014","journal-title":"Cell Death Dis"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1016\/s0014-5793(03)00482-4","article-title":"Neutrophil elastase up-regulates interleukin-8 via toll-like receptor 4","volume":"544","author":"Devaney","year":"2003","journal-title":"FEBS Lett"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1038\/nm1638","article-title":"Mrp8 and Mrp14 are endogenous activators of Toll-like receptor 4, promoting lethal, endotoxin-induced shock","volume":"13","author":"Vogl","year":"2007","journal-title":"Nat Med"},{"key":"B25","doi-asserted-by":"publisher","first-page":"999","DOI":"10.1002\/hep.24501","article-title":"Endogenous histones function as alarmins in sterile inflammatory liver injury through Toll-like receptor 9 in mice","volume":"54","author":"Huang","year":"2011","journal-title":"Hepatology"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1096\/fj.13-236380","article-title":"Extracellular histones are essential effectors of C5aR- and C5L2-mediated tissue damage and inflammation in acute lung injury","volume":"27","author":"Bosmann","year":"2013","journal-title":"FASEB J"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2053","article-title":"Extracellular histones are major mediators of death in sepsis","volume":"15","author":"Xu","year":"2009","journal-title":"Nat Med"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1003930","article-title":"Extracellular histones are mediators of death through TLR2 and TLR4 in mouse fatal liver injury","volume":"187","author":"Xu","year":"2011","journal-title":"J\u00a0Immunol"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1155\/2019\/5953685","article-title":"Circulating Oxidative Stress Biomarkers in Clinical Studies on Type 2 Diabetes and Its Complications","volume":"2019","author":"Bigagli","year":"2019","journal-title":"Oxid Med Cell LONGEV"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2013.08.004","article-title":"Oxidative damage of DNA confers resistance to cytosolic nuclease TREX1 degradation and potentiates STING-dependent immune sensing","volume":"39","author":"Gehrke","year":"2013","journal-title":"Immunity"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2020.590622\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,2,5]],"date-time":"2021-02-05T01:25:46Z","timestamp":1612488346000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2020.590622\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,2,5]]},"references-count":30,"alternative-id":["10.3389\/fimmu.2020.590622"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2020.590622","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,2,5]]},"article-number":"590622"}}