{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,5]],"date-time":"2026-11-05T21:02:26Z","timestamp":1793912546560,"version":"4.8.0"},"reference-count":34,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2019,9,24]],"date-time":"2019-09-24T00:00:00Z","timestamp":1569283200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2019,9,24]],"date-time":"2019-09-24T00:00:00Z","timestamp":1569283200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100000060","name":"U.S. Department of Health & Human Services | NIH | National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01 AI095338"],"award-info":[{"award-number":["R01 AI095338"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"U.S. Department of Health & Human Services | NIH | National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["R25 DK103579"],"award-info":[{"award-number":["R25 DK103579"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"U.S. Department of Health & Human Services | NIH | National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["R01 DK097112"],"award-info":[{"award-number":["R01 DK097112"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000897","name":"Cystic Fibrosis Foundation","doi-asserted-by":"publisher","award":["HURLEY16G0"],"award-info":[{"award-number":["HURLEY16G0"]}],"id":[{"id":"10.13039\/100000897","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000897","name":"Cystic Fibrosis Foundation","doi-asserted-by":"publisher","award":["YONKER18Q0"],"award-info":[{"award-number":["YONKER18Q0"]}],"id":[{"id":"10.13039\/100000897","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000897","name":"Cystic Fibrosis Foundation","doi-asserted-by":"publisher","award":["MOU16G0"],"award-info":[{"award-number":["MOU16G0"]}],"id":[{"id":"10.13039\/100000897","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Pepsin represents a potential biomarker for extraesophageal reflux disease when detected in airways, however a direct role for pepsin in lung dysfunction has not been clearly established. Children experiencing gastroesophageal and extraesophageal reflux are often prescribed proton pump inhibitors (PPIs) to reduce gastric acid associated damage to esophageal and airway mucosa. The potential of pepsin and gastric fluid, from children that were either on or off PPI therapy, to cause inflammation and damage using a human\n                    <jats:italic>in vitro<\/jats:italic>\n                    co-culture model of the airway mucosa was evaluated herein. Exposure of the airway model to acidic solutions caused cellular damage and loss of viability, however, acid alone did not disrupt barrier integrity or instigate neutrophil trans-epithelial migration without pepsin. Gastric fluid from patients on PPI therapy exhibited only a slightly higher pH yet had significantly higher concentrations of pepsin and elicited more barrier disruption and neutrophil trans-epithelial migration compared to gastric fluid from patients off PPIs. Inflammatory and damaging responses observed with gastric fluid from patients on PPIs were largely driven by pepsin. These results indicate the potential for PPI usage to raise concentrations of pepsin in gastric fluid, which may enhance the pathological impact of micro-aspirations in children with extraesophageal reflux.\n                  <\/jats:p>","DOI":"10.1038\/s41598-019-50360-4","type":"journal-article","created":{"date-parts":[[2019,9,24]],"date-time":"2019-09-24T06:02:51Z","timestamp":1569304971000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":34,"title":["Pepsin Triggers Neutrophil Migration Across Acid Damaged Lung Epithelium"],"prefix":"10.1038","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6873-4762","authenticated-orcid":false,"given":"Bryan P.","family":"Hurley","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rebecca H.","family":"Jugo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryan F.","family":"Snow","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tina L.","family":"Samuels","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lael M.","family":"Yonker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongmei","family":"Mou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nikki","family":"Johnston","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachel","family":"Rosen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,9,24]]},"reference":[{"key":"50360_CR1","doi-asserted-by":"publisher","first-page":"516","DOI":"10.1097\/MPG.0000000000001889","volume":"66","author":"R Rosen","year":"2018","unstructured":"Rosen, R. et al. Pediatric Gastroesophageal Reflux Clinical Practice Guidelines: Joint Recommendations of the North American Society for Pediatric Gastroenterology, Hepatology, and Nutrition and the European Society for Pediatric Gastroenterology, Hepatology, and Nutrition. J Pediatr Gastroenterol Nutr 66, 516\u2013554 (2018).","journal-title":"J Pediatr Gastroenterol Nutr"},{"key":"50360_CR2","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1177\/000348941011900310","volume":"119","author":"TL Samuels","year":"2010","unstructured":"Samuels, T. L. & Johnston, N. Pepsin as a marker of extraesophageal reflux. Ann Otol Rhinol Laryngol 119, 203\u2013208 (2010).","journal-title":"Ann Otol Rhinol Laryngol"},{"key":"50360_CR3","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1016\/j.jpeds.2016.06.033","volume":"177","author":"F Dy","year":"2016","unstructured":"Dy, F., Amirault, J., Mitchell, P. D. & Rosen, R. Salivary Pepsin Lacks Sensitivity as a Diagnostic Tool to Evaluate Extraesophageal Reflux Disease. J Pediatr 177, 53\u201358 (2016).","journal-title":"J Pediatr"},{"key":"50360_CR4","doi-asserted-by":"publisher","first-page":"129","DOI":"10.1111\/j.1365-2982.2011.01826.x","volume":"24","author":"R Rosen","year":"2012","unstructured":"Rosen, R., Johnston, N., Hart, K., Khatwa, U. & Nurko, S. The presence of pepsin in the lung and its relationship to pathologic gastro-esophageal reflux. Neurogastroenterol Motil 24, 129\u2013133 (2012).","journal-title":"Neurogastroenterol Motil"},{"key":"50360_CR5","doi-asserted-by":"publisher","first-page":"777","DOI":"10.1002\/lary.20665","volume":"120","author":"DM Bulmer","year":"2010","unstructured":"Bulmer, D. M., Ali, M. S., Brownlee, I. A., Dettmar, P. W. & Pearson, J. P. Laryngeal mucosa: its susceptibility to damage by acid and pepsin. Laryngoscope 120, 777\u2013782 (2010).","journal-title":"Laryngoscope"},{"key":"50360_CR6","doi-asserted-by":"publisher","first-page":"e0152336","DOI":"10.1371\/journal.pone.0152336","volume":"11","author":"JH Kim","year":"2016","unstructured":"Kim, J. H. et al. Extra-Esophageal Pepsin from Stomach Refluxate Promoted Tonsil Hypertrophy. PLoS One 11, e0152336 (2016).","journal-title":"PLoS One"},{"key":"50360_CR7","doi-asserted-by":"publisher","first-page":"e230","DOI":"10.1097\/MCG.0b013e3181c74e25","volume":"44","author":"V Mertens","year":"2010","unstructured":"Mertens, V. et al. Gastric juice from patients \u201con\u201d acid suppressive therapy can still provoke a significant inflammatory reaction by human bronchial epithelial cells. J Clin Gastroenterol 44, e230\u2013235 (2010).","journal-title":"J Clin Gastroenterol"},{"key":"50360_CR8","doi-asserted-by":"publisher","first-page":"303","DOI":"10.1007\/s10620-006-3129-8","volume":"51","author":"K Nagahama","year":"2006","unstructured":"Nagahama, K., Yamato, M., Nishio, H. & Takeuchi, K. Essential role of pepsin in pathogenesis of acid reflux esophagitis in rats. Dig Dis Sci 51, 303\u2013309 (2006).","journal-title":"Dig Dis Sci"},{"key":"50360_CR9","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1111\/nyas.13080","volume":"1381","author":"N Johnston","year":"2016","unstructured":"Johnston, N. et al. Airway reflux. Ann N Y Acad Sci 1381, 5\u201313 (2016).","journal-title":"Ann N Y Acad Sci"},{"key":"50360_CR10","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.prostaglandins.2013.11.001","volume":"108","author":"SA Kubala","year":"2014","unstructured":"Kubala, S. A., Patil, S. U., Shreffler, W. G. & Hurley, B. P. Pathogen induced chemo-attractant hepoxilin A3 drives neutrophils, but not eosinophils across epithelial barriers. Prostaglandins Other Lipid Mediat 108, 1\u20138 (2014).","journal-title":"Prostaglandins Other Lipid Mediat"},{"key":"50360_CR11","doi-asserted-by":"crossref","unstructured":"Kusek, M. E., Pazos, M. A., Pirzai, W. & Hurley, B. P. In vitro coculture assay to assess pathogen induced neutrophil trans-epithelial migration. J Vis Exp, e50823 (2014).","DOI":"10.3791\/50823"},{"key":"50360_CR12","doi-asserted-by":"publisher","first-page":"1304","DOI":"10.4049\/jimmunol.1402489","volume":"194","author":"MA Pazos","year":"2015","unstructured":"Pazos, M. A. et al. Distinct cellular sources of hepoxilin A3 and leukotriene B4 are used to coordinate bacterial-induced neutrophil transepithelial migration. J Immunol 194, 1304\u20131315 (2015).","journal-title":"J Immunol"},{"key":"50360_CR13","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-08567-w","volume":"7","author":"LM Yonker","year":"2017","unstructured":"Yonker, L. M. et al. Development of a Primary Human Co-Culture Model of Inflamed Airway Mucosa. Sci Rep 7, 8182 (2017).","journal-title":"Sci Rep"},{"key":"50360_CR14","doi-asserted-by":"publisher","first-page":"2873","DOI":"10.4049\/jimmunol.1700539","volume":"199","author":"LM Yonker","year":"2017","unstructured":"Yonker, L. M. et al. Neutrophil-Derived Cytosolic PLA2alpha Contributes to Bacterial-Induced Neutrophil Transepithelial Migration. J Immunol 199, 2873\u20132884 (2017).","journal-title":"J Immunol"},{"key":"50360_CR15","doi-asserted-by":"publisher","first-page":"5712","DOI":"10.4049\/jimmunol.173.9.5712","volume":"173","author":"BP Hurley","year":"2004","unstructured":"Hurley, B. P., Siccardi, D., Mrsny, R. J. & McCormick, B. A. Polymorphonuclear cell transmigration induced by Pseudomonas aeruginosa requires the eicosanoid hepoxilin A3. J Immunol 173, 5712\u20135720 (2004).","journal-title":"J Immunol"},{"key":"50360_CR16","doi-asserted-by":"publisher","first-page":"1036","DOI":"10.1097\/MLG.0b013e31804154c3","volume":"117","author":"N Johnston","year":"2007","unstructured":"Johnston, N., Dettmar, P. W., Bishwokarma, B., Lively, M. O. & Koufman, J. A. Activity\/stability of human pepsin: implications for reflux attributed laryngeal disease. Laryngoscope 117, 1036\u20131039 (2007).","journal-title":"Laryngoscope"},{"key":"50360_CR17","doi-asserted-by":"publisher","first-page":"547","DOI":"10.1177\/000348941011900808","volume":"119","author":"N Johnston","year":"2010","unstructured":"Johnston, N., Wells, C. W., Samuels, T. L. & Blumin, J. H. Rationale for targeting pepsin in the treatment of reflux disease. Ann Otol Rhinol Laryngol 119, 547\u2013558 (2010).","journal-title":"Ann Otol Rhinol Laryngol"},{"key":"50360_CR18","doi-asserted-by":"publisher","first-page":"2205","DOI":"10.1016\/0006-2952(80)90199-9","volume":"29","author":"DH Rich","year":"1980","unstructured":"Rich, D. H. & Sun, E. T. Mechanism of inhibition of pepsin by pepstatin. Effect of inhibitor structure on dissociation constant and time-dependent inhibition. Biochem Pharmacol 29, 2205\u20132212 (1980).","journal-title":"Biochem Pharmacol"},{"key":"50360_CR19","doi-asserted-by":"publisher","first-page":"4960","DOI":"10.4049\/jimmunol.1201922","volume":"189","author":"DL Tamang","year":"2012","unstructured":"Tamang, D. L. et al. Hepoxilin A(3) facilitates neutrophilic breach of lipoxygenase-expressing airway epithelial barriers. J Immunol 189, 4960\u20134969 (2012).","journal-title":"J Immunol"},{"key":"50360_CR20","doi-asserted-by":"publisher","first-page":"431","DOI":"10.1016\/j.jcf.2015.02.008","volume":"14","author":"LM Yonker","year":"2015","unstructured":"Yonker, L. M., Cigana, C., Hurley, B. P. & Bragonzi, A. Host-pathogen interplay in the respiratory environment of cystic fibrosis. J Cyst Fibros 14, 431\u2013439 (2015).","journal-title":"J Cyst Fibros"},{"key":"50360_CR21","doi-asserted-by":"crossref","first-page":"150","DOI":"10.4037\/ajcc2002.11.2.150","volume":"11","author":"NA Metheny","year":"2002","unstructured":"Metheny, N. A. et al. Pepsin as a marker for pulmonary aspiration. Am J Crit Care 11, 150\u2013154 (2002).","journal-title":"Am J Crit Care"},{"key":"50360_CR22","doi-asserted-by":"publisher","first-page":"217","DOI":"10.1016\/j.stem.2016.05.012","volume":"19","author":"H Mou","year":"2016","unstructured":"Mou, H. et al. Dual SMAD Signaling Inhibition Enables Long-Term Expansion of Diverse Epithelial Basal Cells. Cell Stem Cell 19, 217\u2013231 (2016).","journal-title":"Cell Stem Cell"},{"key":"50360_CR23","doi-asserted-by":"crossref","unstructured":"Levardon, H., Yonker, L. M., Hurley, B. P. & Mou, H. Expansion of Airway Basal Cells and Generation of Polarized Epithelium. Bio Protoc 8, (2018).","DOI":"10.21769\/BioProtoc.2877"},{"key":"50360_CR24","doi-asserted-by":"publisher","first-page":"1881","DOI":"10.1097\/00003246-199611000-00019","volume":"24","author":"MM Badellino","year":"1996","unstructured":"Badellino, M. M. et al. Detection of pulmonary aspiration of gastric contents in an animal model by assay of peptic activity in bronchoalveolar fluid. Crit Care Med 24, 1881\u20131885 (1996).","journal-title":"Crit Care Med"},{"key":"50360_CR25","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1097\/00000542-199502000-00023","volume":"82","author":"I Kudoh","year":"1995","unstructured":"Kudoh, I. et al. The effect of pentoxifylline on acid-induced alveolar epithelial injury. Anesthesiology 82, 531\u2013541 (1995).","journal-title":"Anesthesiology"},{"key":"50360_CR26","doi-asserted-by":"publisher","first-page":"L1103","DOI":"10.1152\/ajplung.00197.2014","volume":"309","author":"K Tetenev","year":"2015","unstructured":"Tetenev, K. et al. Synergy between acid and endotoxin in an experimental model of aspiration-related lung injury progression. Am J Physiol Lung Cell Mol Physiol 309, L1103\u20131111 (2015).","journal-title":"Am J Physiol Lung Cell Mol Physiol"},{"key":"50360_CR27","first-page":"119","volume":"121","author":"DJ Schwartz","year":"1980","unstructured":"Schwartz, D. J., Wynne, J. W., Gibbs, C. P., Hood, C. I. & Kuck, E. J. The pulmonary consequences of aspiration of gastric contents at pH values greater than 2.5. Am Rev Respir Dis 121, 119\u2013126 (1980).","journal-title":"Am Rev Respir Dis"},{"key":"50360_CR28","doi-asserted-by":"publisher","first-page":"843","DOI":"10.1016\/j.healun.2008.05.022","volume":"27","author":"R Vos","year":"2008","unstructured":"Vos, R. et al. Airway colonization and gastric aspiration after lung transplantation: do birds of a feather flock together? J Heart Lung Transplant 27, 843\u2013849 (2008).","journal-title":"J Heart Lung Transplant"},{"key":"50360_CR29","doi-asserted-by":"publisher","first-page":"161","DOI":"10.1097\/MPG.0b013e3181acae98","volume":"50","author":"K Blondeau","year":"2010","unstructured":"Blondeau, K. et al. Characteristics of gastroesophageal reflux and potential risk of gastric content aspiration in children with cystic fibrosis. J Pediatr Gastroenterol Nutr 50, 161\u2013166 (2010).","journal-title":"J Pediatr Gastroenterol Nutr"},{"key":"50360_CR30","doi-asserted-by":"publisher","first-page":"89","DOI":"10.1002\/jcph.977","volume":"58","author":"JP Franciosi","year":"2018","unstructured":"Franciosi, J. P. et al. Association Between CYP2C19*17 Alleles and pH Probe Testing Outcomes in Children With Symptomatic Gastroesophageal Reflux. J Clin Pharmacol 58, 89\u201396 (2018).","journal-title":"J Clin Pharmacol"},{"key":"50360_CR31","doi-asserted-by":"publisher","first-page":"207","DOI":"10.1016\/S0021-9673(99)87022-8","volume":"646","author":"AT Jones","year":"1993","unstructured":"Jones, A. T., Keen, J. N. & Roberts, N. B. Human pepsin 3b peptide map sequence analysis, genotype and hydrophobic nature. J Chromatogr 646, 207\u2013212 (1993).","journal-title":"J Chromatogr"},{"key":"50360_CR32","doi-asserted-by":"publisher","first-page":"1473","DOI":"10.1097\/01.mlg.0000172043.51871.d9","volume":"115","author":"J Knight","year":"2005","unstructured":"Knight, J., Lively, M. O., Johnston, N., Dettmar, P. W. & Koufman, J. A. Sensitive pepsin immunoassay for detection of laryngopharyngeal reflux. Laryngoscope 115, 1473\u20131478 (2005).","journal-title":"Laryngoscope"},{"issue":"1","key":"50360_CR33","doi-asserted-by":"publisher","first-page":"e12269","DOI":"10.14814\/phy2.12269","volume":"3","author":"Emily Foltz","year":"2015","unstructured":"Foltz, E. et al. An assessment of human gastric fluid composition as a function of PPI usage. Physiol Rep 3, (2015).","journal-title":"Physiological Reports"},{"key":"50360_CR34","doi-asserted-by":"publisher","first-page":"262","DOI":"10.1016\/j.fct.2016.11.006","volume":"98","author":"BP Hurley","year":"2016","unstructured":"Hurley, B. P., Eaton, A. D., Zimmermann, C. & Delaney, B. Polarized monolayer cultures of human intestinal epithelial cell lines exposed to intractable proteins - In vitro hazard identification studies. Food Chem Toxicol 98, 262\u2013268 (2016).","journal-title":"Food Chem Toxicol"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-50360-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-50360-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-50360-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,17]],"date-time":"2022-12-17T08:52:38Z","timestamp":1671267158000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-50360-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,9,24]]},"references-count":34,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["50360"],"URL":"https:\/\/doi.org\/10.1038\/s41598-019-50360-4","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,9,24]]},"assertion":[{"value":"19 July 2018","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 September 2019","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"24 September 2019","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"13778"}}