{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T14:49:07Z","timestamp":1786978147722,"version":"build-2736575974"},"reference-count":29,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2020,5,23]],"date-time":"2020-05-23T00:00:00Z","timestamp":1590192000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000055","name":"National Institute on Deafness and Other Communication Disorders","doi-asserted-by":"publisher","award":["R01DC012377"],"award-info":[{"award-number":["R01DC012377"]}],"id":[{"id":"10.13039\/100000055","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Laryngoscope Investig Oto"],"published-print":{"date-parts":[[2020,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Chronic otitis media (COM) is characterized by middle ear fluid predominantly containing cytokines,\n                      <jats:italic>Nontypeable haemophilus influenzae<\/jats:italic>\n                      (NTHi), the mucin MUC5B, and neutrophil extracellular traps (NETs). NETs consist of extracellular DNA coated with antibacterial proteins such as myeloperoxidase (MPO) and citrullinated histone 3 (CitH3). NETs can damage tissues and sustain inflammation. Our study aimed to develop an in vitro model of NETosis, testing COM inductors.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>NETosis was evaluated in fresh blood human neutrophils attached to collagen\u2010coated plates and in suspension exposed to phorbol myristate acetate (PMA) as a control, and COM relevant mediators. Confocal microscopy, DNA fluorescence assay and flow cytometry were used to quantify NETosis.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      PMA exposure induced DNA, MPO, and CitH3 by immunofluorescence (IF) most significantly at 3 hours (3.8\u2010fold for DAPI, 7.6\u2010fold for MPO, and 6.9\u2010fold for CitH3, all\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u2009.05). IL\u20108 and TNF\u2010\u03b1 cytokines showed milder increases of DAPI, MPO, and CitH3 positive cells. NTHi had no effect on these NETs markers. Purified salivary MUC5B (10 to 40\u2009\u03bcg\/mL) produced potent increases, comparable to PMA. A composite NET score summing the fold\u2010increases for DAPI, MPO, and CitH3 demonstrated PMA at 13.6 to 19 relative to control set at 1; and MUC5B at 8.6 to 16.3 (all\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u2009.05). IL\u20108 and TNF\u2010\u03b1 showed scores of 5.4 and 3, respectively, but these were not statistically significant.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>We developed a reliable in vitro assay for NETosis which demonstrated that salivary MUC5B is a potent inductor of NETs whereas IL\u20108, TNF\u2010\u03b1, live and lyzed NTHi demonstrated minimal to no NETosis.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Level of evidence<\/jats:title>\n                    <jats:p>NA.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/lio2.396","type":"journal-article","created":{"date-parts":[[2020,5,23]],"date-time":"2020-05-23T04:40:35Z","timestamp":1590208835000},"page":"536-545","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["<scp>MUC5B<\/scp>\n                    induces in vitro neutrophil extracellular trap formation: Implication in otitis media"],"prefix":"10.1002","volume":"5","author":[{"given":"St\u00e9phanie","family":"Val","sequence":"first","affiliation":[{"name":"Sheikh Zayed Institute for Pediatric Surgical Innovation Children's National Health System  Washington District of Columbia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Krueger","sequence":"additional","affiliation":[{"name":"Sheikh Zayed Institute for Pediatric Surgical Innovation Children's National Health System  Washington District of Columbia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arman","family":"Hussain","sequence":"additional","affiliation":[{"name":"Sheikh Zayed Institute for Pediatric Surgical Innovation Children's National Health System  Washington District of Columbia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amarel","family":"Tomney","sequence":"additional","affiliation":[{"name":"Sheikh Zayed Institute for Pediatric Surgical Innovation Children's National Health System  Washington District of Columbia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yajun","family":"Chen","sequence":"additional","affiliation":[{"name":"Sheikh Zayed Institute for Pediatric Surgical Innovation Children's National Health System  Washington District of Columbia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher","family":"Lazarski","sequence":"additional","affiliation":[{"name":"Center for Cancer and Immunology Research Children's National Health System  Washington District of Columbia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0075-8279","authenticated-orcid":false,"given":"Diego","family":"Preciado","sequence":"additional","affiliation":[{"name":"Sheikh Zayed Institute for Pediatric Surgical Innovation Children's National Health System  Washington District of Columbia USA"},{"name":"Division of Pediatric Otolaryngology Children's National Medical Center  Washington District of Columbia 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