{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T20:00:36Z","timestamp":1772136036838,"version":"3.50.1"},"reference-count":56,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,7,12]],"date-time":"2021-07-12T00:00:00Z","timestamp":1626048000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,7,12]],"date-time":"2021-07-12T00:00:00Z","timestamp":1626048000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100001280","name":"Horserace Betting Levy Board","doi-asserted-by":"publisher","award":["R45502"],"award-info":[{"award-number":["R45502"]}],"id":[{"id":"10.13039\/501100001280","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>Airway inflammation is highly prevalent in horses, with the majority of non-infectious cases being defined as equine asthma. Currently, cytological analysis of airway derived samples is the principal method of assessing lower airway inflammation. Samples can be obtained by tracheal wash (TW) or by lavage of the lower respiratory tract (bronchoalveolar lavage (BAL) fluid; BALF). Although BALF cytology carries significant diagnostic advantages over TW cytology for the diagnosis of equine asthma, sample acquisition is invasive, making it prohibitive for routine and sequential screening of airway health. However, recent technological advances in sample collection and processing have made it possible to determine whether a wider range of analyses might be applied to TW samples. Considering that TW samples are relatively simple to collect, minimally invasive and readily available in the horse, it was considered appropriate to investigate whether, equine tracheal secretions represent a rich source of cells and both transcriptomic and proteomic data. Similar approaches have already been applied to a comparable sample set in humans; namely, induced sputum. Sputum represents a readily available source of airway biofluids enriched in proteins, changes in the expression of which may reveal novel mechanisms in the pathogenesis of respiratory diseases, such as asthma and chronic obstructive pulmonary disease. The aim of this study was to establish a robust protocol to isolate macrophages, protein and RNA for molecular characterization of TW samples and demonstrate the applicability of sample handling to rodent and human pediatric bronchoalveolar lavage fluid isolates. TW samples provided a good quality and yield of both RNA and protein for downstream transcriptomic\/proteomic analyses. The sample handling methodologies were successfully applicable to BALF for rodent and human research. TW samples represent a rich source of airway cells, and molecular analysis to facilitate and study airway inflammation, based on both transcriptomic and proteomic analysis. This study provides a necessary methodological platform for future transcriptomic and\/or proteomic studies on equine lower respiratory tract secretions and BALF samples from humans and mice.<\/jats:p>","DOI":"10.1038\/s41598-021-93839-9","type":"journal-article","created":{"date-parts":[[2021,7,12]],"date-time":"2021-07-12T06:03:06Z","timestamp":1626069786000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Application across species of a one health approach to liquid sample handling for respiratory based -omics analysis"],"prefix":"10.1038","volume":"11","author":[{"given":"Anna E.","family":"Karagianni","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Samantha L.","family":"Eaton","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dominic","family":"Kurian","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eugenio","family":"Cill\u00e1n-Garcia","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jonathan","family":"Twynam-Perkins","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna","family":"Raper","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas M.","family":"Wishart","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R. Scott","family":"Pirie","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,7,12]]},"reference":[{"key":"93839_CR1","doi-asserted-by":"publisher","first-page":"236","DOI":"10.2746\/0425164054530579","volume":"37","author":"JLN Wood","year":"2005","unstructured":"Wood, J. L. N., Newton, J. R., Chanter, N. & Mumford, J. A. Inflammatory airway disease, nasal discharge and respiratory infections in young British racehorses. Equine Vet. J. 37, 236\u2013242 (2005).","journal-title":"Equine Vet. J."},{"key":"93839_CR2","doi-asserted-by":"publisher","first-page":"358","DOI":"10.1111\/evj.12368","volume":"47","author":"PS Morley","year":"2015","unstructured":"Morley, P. S., Bromberek, J. L., Saulez, M. N., Hinchcliff, K. W. & Guthrie, A. J. Exercise-induced pulmonary haemorrhage impairs racing performance in thoroughbred racehorses. Equine Vet. J. 47, 358\u2013365. https:\/\/doi.org\/10.1111\/evj.12368 (2015).","journal-title":"Equine Vet. J."},{"key":"93839_CR3","doi-asserted-by":"publisher","first-page":"450","DOI":"10.3389\/fvets.2020.00450","volume":"7","author":"L Couetil","year":"2020","unstructured":"Couetil, L. et al. Equine asthma: Current understanding and future directions. Front. Vet. Sci. 7, 450. https:\/\/doi.org\/10.3389\/fvets.2020.00450 (2020).","journal-title":"Front. Vet. Sci."},{"key":"93839_CR4","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2018.00061","author":"H Rossi","year":"2018","unstructured":"Rossi, H. et al. Comparison of tracheal wash and bronchoalveolar lavage cytology in 154 horses with and without respiratory signs in a referral hospital over 2009\u20132015. Front. Vet. Sci. https:\/\/doi.org\/10.3389\/fvets.2018.00061 (2018).","journal-title":"Front. Vet. Sci."},{"key":"93839_CR5","doi-asserted-by":"publisher","first-page":"87","DOI":"10.1016\/j.cveq.2019.12.006","volume":"36","author":"LL Couetil","year":"2020","unstructured":"Couetil, L. L. & Thompson, C. A. Airway diagnostics: Bronchoalveolar lavage, tracheal wash, and pleural fluid. Vet. Clin. N. Am. Equine Pract. 36, 87\u2013103. https:\/\/doi.org\/10.1016\/j.cveq.2019.12.006 (2020).","journal-title":"Vet. Clin. N. Am. Equine Pract."},{"key":"93839_CR6","doi-asserted-by":"publisher","first-page":"743","DOI":"10.1111\/jvim.12593","volume":"29","author":"KW Hinchcliff","year":"2015","unstructured":"Hinchcliff, K. W. et al. Exercise induced pulmonary hemorrhage in horses: American College of Veterinary Internal Medicine consensus statement. J. Vet. Intern. Med. 29, 743\u2013758. https:\/\/doi.org\/10.1111\/jvim.12593 (2015).","journal-title":"J. Vet. Intern. Med."},{"key":"93839_CR7","doi-asserted-by":"publisher","first-page":"503","DOI":"10.1111\/jvim.13824","volume":"30","author":"LL Couetil","year":"2016","unstructured":"Couetil, L. L. et al. Inflammatory airway disease of horses\u2014Revised consensus statement. J. Vet. Intern. Med. 30, 503\u2013515. https:\/\/doi.org\/10.1111\/jvim.13824 (2016).","journal-title":"J. Vet. Intern. Med."},{"key":"93839_CR8","doi-asserted-by":"publisher","first-page":"330","DOI":"10.1111\/j.2042-3292.1999.tb01566.x","volume":"11","author":"AM Hoffman","year":"1999","unstructured":"Hoffman, A. M. Bronchoalveolar lavage technique and cytological diagnosis of small airway inflammatory disease. Equine Vet. Educ. 11, 330\u2013336 (1999).","journal-title":"Equine Vet. Educ."},{"key":"93839_CR9","doi-asserted-by":"publisher","first-page":"3112","DOI":"10.1128\/jvi.74.7.3112-3121.2000","volume":"74","author":"SM Harrold","year":"2000","unstructured":"Harrold, S. M. et al. Tissue sites of persistent infection and active replication of equine infectious anemia virus during acute disease and asymptomatic infection in experimentally infected equids. J. Virol. 74, 3112\u20133121. https:\/\/doi.org\/10.1128\/jvi.74.7.3112-3121.2000 (2000).","journal-title":"J. Virol."},{"key":"93839_CR10","doi-asserted-by":"publisher","first-page":"282","DOI":"10.1016\/j.tvjl.2009.04.016","volume":"184","author":"EA Richard","year":"2010","unstructured":"Richard, E. A. et al. Sub-clinical diseases affecting performance in Standardbred trotters: Diagnostic methods and predictive parameters. Vet. J. 184, 282\u2013289. https:\/\/doi.org\/10.1016\/j.tvjl.2009.04.016 (2010).","journal-title":"Vet. J."},{"key":"93839_CR11","doi-asserted-by":"publisher","first-page":"99","DOI":"10.1136\/vr.148.4.99","volume":"148","author":"RM Christley","year":"2001","unstructured":"Christley, R. M. et al. Coughing in thoroughbred racehorses: Risk factors and tracheal endoscopic and cytological findings. Vet. Rec. 148, 99\u2013104 (2001).","journal-title":"Vet. Rec."},{"key":"93839_CR12","doi-asserted-by":"publisher","first-page":"322","DOI":"10.1111\/j.1939-1676.2010.0664.x","volume":"25","author":"JP Lavoie","year":"2011","unstructured":"Lavoie, J. P. et al. Bronchoalveolar lavage fluid cytology and cytokine messenger ribonucleic acid expression of racehorses with exercise intolerance and lower airway inflammation. J. Vet. Intern. Med. 25, 322\u2013329. https:\/\/doi.org\/10.1111\/j.1939-1676.2010.0664.x (2011).","journal-title":"J. Vet. Intern. Med."},{"key":"93839_CR13","doi-asserted-by":"publisher","first-page":"82","DOI":"10.1016\/j.vetimm.2010.11.018","volume":"140","author":"KJ Hughes","year":"2011","unstructured":"Hughes, K. J. et al. Evaluation of cytokine mRNA expression in bronchoalveolar lavage cells from horses with inflammatory airway disease. Vet. Immunol. Immunopathol. 140, 82\u201389. https:\/\/doi.org\/10.1016\/j.vetimm.2010.11.018 (2011).","journal-title":"Vet. Immunol. Immunopathol."},{"key":"93839_CR14","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1111\/j.1939-1676.2011.00847.x","volume":"26","author":"L Beekman","year":"2012","unstructured":"Beekman, L., Tohver, T. & L\u00e9guillette, R. Comparison of cytokine mRNA expression in the bronchoalveolar lavage fluid of horses with inflammatory airway disease and bronchoalveolar lavage mastocytosis or neutrophilia using REST software analysis. J. Vet. Intern. Med. 26, 153\u2013161. https:\/\/doi.org\/10.1111\/j.1939-1676.2011.00847.x (2012).","journal-title":"J. Vet. Intern. Med."},{"key":"93839_CR15","doi-asserted-by":"publisher","first-page":"391","DOI":"10.1080\/14789450.2017.1317246","volume":"14","author":"P Iadarola","year":"2017","unstructured":"Iadarola, P. & Viglio, S. Spit it out! How could the sputum proteome aid clinical research into pulmonary diseases? Expert Rev. Proteomics 14, 391\u2013393. https:\/\/doi.org\/10.1080\/14789450.2017.1317246 (2017).","journal-title":"Expert Rev. Proteomics"},{"key":"93839_CR16","doi-asserted-by":"publisher","first-page":"645","DOI":"10.1007\/s13167-020-00227-w","volume":"11","author":"T Zhu","year":"2020","unstructured":"Zhu, T. et al. Induced sputum metabolomic profiles and oxidative stress are associated with chronic obstructive pulmonary disease (COPD) severity: Potential use for predictive, preventive, and personalized medicine. EPMA J. 11, 645\u2013659. https:\/\/doi.org\/10.1007\/s13167-020-00227-w (2020).","journal-title":"EPMA J."},{"key":"93839_CR17","doi-asserted-by":"publisher","first-page":"1176","DOI":"10.1016\/j.jaci.2011.07.053","volume":"128","author":"SA Gharib","year":"2011","unstructured":"Gharib, S. A. et al. Induced sputum proteome in healthy subjects and asthmatic patients. J. Allergy Clin. Immunol. 128, 1176\u20131184. https:\/\/doi.org\/10.1016\/j.jaci.2011.07.053 (2011).","journal-title":"J. Allergy Clin. Immunol."},{"key":"93839_CR18","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1016\/j.molimm.2014.12.005","volume":"66","author":"M Bullone","year":"2015","unstructured":"Bullone, M. & Lavoie, J. P. Asthma, \u201cof horses and men\u201d\u2013how can equine heaves help us better understand human asthma immunopathology and its functional consequences? Mol. Immunol. 66, 97\u2013105. https:\/\/doi.org\/10.1016\/j.molimm.2014.12.005 (2015).","journal-title":"Mol. Immunol."},{"key":"93839_CR19","doi-asserted-by":"publisher","DOI":"10.3390\/genes11101143","author":"MF Hulliger","year":"2020","unstructured":"Hulliger, M. F. et al. An Integrative miRNA-mRNA expression analysis reveals striking transcriptomic similarities between severe equine asthma and specific asthma endotypes in humans. Genes (Basel). https:\/\/doi.org\/10.3390\/genes11101143 (2020).","journal-title":"Genes (Basel)."},{"key":"93839_CR20","doi-asserted-by":"publisher","first-page":"465","DOI":"10.1016\/s0165-6147(97)01142-5","volume":"18","author":"CG Persson","year":"1997","unstructured":"Persson, C. G., Erjef\u00e4lt, J. S., Korsgren, M. & Sundler, F. The mouse trap. Trends Pharmacol. Sci. 18, 465\u2013467. https:\/\/doi.org\/10.1016\/s0165-6147(97)01142-5 (1997).","journal-title":"Trends Pharmacol. Sci."},{"key":"93839_CR21","doi-asserted-by":"publisher","first-page":"10","DOI":"10.4168\/aair.2009.1.1.10","volume":"1","author":"YS Shin","year":"2009","unstructured":"Shin, Y. S., Takeda, K. & Gelfand, E. W. Understanding asthma using animal models. Allergy Asthma Immunol. Res. 1, 10\u201318. https:\/\/doi.org\/10.4168\/aair.2009.1.1.10 (2009).","journal-title":"Allergy Asthma Immunol. Res."},{"key":"93839_CR22","doi-asserted-by":"publisher","DOI":"10.1111\/evj.12584","author":"AE Karagianni","year":"2016","unstructured":"Karagianni, A. E. et al. Comparative transcriptome analysis of equine alveolar macrophages. Equine Vet. J. https:\/\/doi.org\/10.1111\/evj.12584 (2016).","journal-title":"Equine Vet. J."},{"key":"93839_CR23","doi-asserted-by":"publisher","first-page":"219","DOI":"10.1016\/j.vetimm.2013.07.003","volume":"155","author":"AE Karagianni","year":"2013","unstructured":"Karagianni, A. E., Kapetanovic, R., McGorum, B. C., Hume, D. A. & Pirie, S. R. The equine alveolar macrophage: Functional and phenotypic comparisons with peritoneal macrophages. Vet. Immunol. Immunopathol. 155, 219\u2013228. https:\/\/doi.org\/10.1016\/j.vetimm.2013.07.003 (2013).","journal-title":"Vet. Immunol. Immunopathol."},{"key":"93839_CR24","doi-asserted-by":"publisher","first-page":"27","DOI":"10.4049\/immunohorizons.1700073","volume":"2","author":"R Young","year":"2018","unstructured":"Young, R. et al. Species-specific transcriptional regulation of genes involved in nitric oxide production and arginine metabolism in macrophages. ImmunoHorizons 2, 27\u201337. https:\/\/doi.org\/10.4049\/immunohorizons.1700073 (2018).","journal-title":"ImmunoHorizons"},{"key":"93839_CR25","doi-asserted-by":"publisher","DOI":"10.1111\/evj.13341","author":"AE Karagianni","year":"2020","unstructured":"Karagianni, A. E., Lisowski, Z. M., Hume, D. A. & Scott Pirie, R. The equine mononuclear phagocyte system: The relevance of the horse as a model for understanding human innate immunity. Equine Vet. J. https:\/\/doi.org\/10.1111\/evj.13341 (2020).","journal-title":"Equine Vet. J."},{"key":"93839_CR26","doi-asserted-by":"publisher","first-page":"1004","DOI":"10.1164\/rccm.201202-0320ST","volume":"185","author":"KC Meyer","year":"2012","unstructured":"Meyer, K. C. et al. An official American Thoracic Society clinical practice guideline: the clinical utility of bronchoalveolar lavage cellular analysis in interstitial lung disease. Am. J. Respir. Crit. Care Med. 185, 1004\u20131014. https:\/\/doi.org\/10.1164\/rccm.201202-0320ST (2012).","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"93839_CR27","doi-asserted-by":"publisher","first-page":"105405","DOI":"10.1016\/j.tvjl.2019.105405","volume":"254","author":"KJ Allen","year":"2019","unstructured":"Allen, K. J., Tennant, K. V. & Franklin, S. H. Effect of inclusion or exclusion of epithelial cells in equine respiratory cytology analysis. Vet. J. 254, 105405. https:\/\/doi.org\/10.1016\/j.tvjl.2019.105405 (2019).","journal-title":"Vet. J."},{"key":"93839_CR28","doi-asserted-by":"publisher","first-page":"667","DOI":"10.1136\/thorax.57.8.667","volume":"57","author":"IS Woolhouse","year":"2002","unstructured":"Woolhouse, I. S., Bayley, D. L. & Stockley, R. A. Effect of sputum processing with dithiothreitol on the detection of inflammatory mediators in chronic bronchitis and bronchiectasis. Thorax 57, 667\u2013671. https:\/\/doi.org\/10.1136\/thorax.57.8.667 (2002).","journal-title":"Thorax"},{"key":"93839_CR29","doi-asserted-by":"publisher","first-page":"44","DOI":"10.1038\/nprot.2008.211","volume":"4","author":"W da Huang","year":"2009","unstructured":"da Huang, W., Sherman, B. T. & Lempicki, R. A. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat. Protoc. 4, 44\u201357. https:\/\/doi.org\/10.1038\/nprot.2008.211 (2009).","journal-title":"Nat. Protoc."},{"key":"93839_CR30","doi-asserted-by":"publisher","first-page":"523","DOI":"10.1093\/bioinformatics\/btt703","volume":"30","author":"A Kramer","year":"2014","unstructured":"Kramer, A., Green, J., Pollard, J. Jr. & Tugendreich, S. Causal analysis approaches in ingenuity pathway analysis. Bioinformatics 30, 523\u2013530. https:\/\/doi.org\/10.1093\/bioinformatics\/btt703 (2014).","journal-title":"Bioinformatics"},{"key":"93839_CR31","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1186\/1471-2199-12-5","volume":"12","author":"L Beekman","year":"2011","unstructured":"Beekman, L., Tohver, T., Dardari, R. & L\u00e9guillette, R. Evaluation of suitable reference genes for gene expression studies in bronchoalveolar lavage cells from horses with inflammatory airway disease. BMC Mol. Biol. 12, 5. https:\/\/doi.org\/10.1186\/1471-2199-12-5 (2011).","journal-title":"BMC Mol. Biol."},{"key":"93839_CR32","doi-asserted-by":"publisher","first-page":"43","DOI":"10.2147\/vmrr.s194427","volume":"10","author":"LA Bright","year":"2019","unstructured":"Bright, L. A. et al. Modeling the pasture-associated severe equine asthma bronchoalveolar lavage fluid proteome identifies molecular events mediating neutrophilic airway inflammation. Vet. Med. (Auckl.) 10, 43\u201363. https:\/\/doi.org\/10.2147\/vmrr.s194427 (2019).","journal-title":"Vet. Med. (Auckl.)"},{"key":"93839_CR33","doi-asserted-by":"publisher","first-page":"558","DOI":"10.1136\/thx.2006.073544","volume":"62","author":"LI Sikkeland","year":"2007","unstructured":"Sikkeland, L. I., Kongerud, J., Stangeland, A. M., Haug, T. & Alexis, N. E. Macrophage enrichment from induced sputum. Thorax 62, 558\u2013559. https:\/\/doi.org\/10.1136\/thx.2006.073544 (2007).","journal-title":"Thorax"},{"key":"93839_CR34","doi-asserted-by":"publisher","first-page":"e86912","DOI":"10.1371\/journal.pone.0086912","volume":"9","author":"Y Kumar","year":"2014","unstructured":"Kumar, Y. et al. Molecular analysis of serum and bronchoalveolar lavage in a mouse model of influenza reveals markers of disease severity that can be clinically useful in humans. PLoS ONE 9, e86912. https:\/\/doi.org\/10.1371\/journal.pone.0086912 (2014).","journal-title":"PLoS ONE"},{"key":"93839_CR35","doi-asserted-by":"publisher","first-page":"155","DOI":"10.3390\/cancers9110155","volume":"9","author":"TN Zamay","year":"2017","unstructured":"Zamay, T. N. et al. Current and prospective protein biomarkers of lung cancer. Cancers (Basel) 9, 155. https:\/\/doi.org\/10.3390\/cancers9110155 (2017).","journal-title":"Cancers (Basel)"},{"key":"93839_CR36","doi-asserted-by":"publisher","first-page":"1036","DOI":"10.1038\/s41598-018-37662-9","volume":"9","author":"SC Bishwal","year":"2019","unstructured":"Bishwal, S. C. et al. Sputum proteomics reveals a shift in vitamin D-binding protein and antimicrobial protein axis in tuberculosis patients. Sci. Rep. 9, 1036\u20131036. https:\/\/doi.org\/10.1038\/s41598-018-37662-9 (2019).","journal-title":"Sci. Rep."},{"key":"93839_CR37","doi-asserted-by":"publisher","first-page":"529","DOI":"10.1111\/j.2042-3306.2006.tb05599.x","volume":"38","author":"KJ Allen","year":"2006","unstructured":"Allen, K. J., Tremaine, W. H. & Franklin, S. H. Prevalence of inflammatory airway disease in national hunt horses referred for investigation of poor athletic performance. Equine Vet. J. Suppl. 38, 529\u2013534. https:\/\/doi.org\/10.1111\/j.2042-3306.2006.tb05599.x (2006).","journal-title":"Equine Vet. J. Suppl."},{"key":"93839_CR38","doi-asserted-by":"publisher","first-page":"1396","DOI":"10.1152\/ajplung.00444.2006","volume":"292","author":"K Rousseau","year":"2007","unstructured":"Rousseau, K. et al. Muc5b and Muc5ac are the major oligomeric mucins in equine airway mucus. Am. J. Physiol. Lung Cell Mol. Physiol. 292, 1396\u20131404. https:\/\/doi.org\/10.1152\/ajplung.00444.2006 (2007).","journal-title":"Am. J. Physiol. Lung Cell Mol. Physiol."},{"issue":"Pt 3","key":"93839_CR39","doi-asserted-by":"publisher","first-page":"685","DOI":"10.1042\/bj3340685","volume":"334","author":"C Wickstr\u00f6m","year":"1998","unstructured":"Wickstr\u00f6m, C., Davies, J. R., Eriksen, G. V., Veerman, E. C. & Carlstedt, I. MUC5B is a major gel-forming, oligomeric mucin from human salivary gland, respiratory tract and endocervix: Identification of glycoforms and C-terminal cleavage. Biochem. J. 334(Pt 3), 685\u2013693. https:\/\/doi.org\/10.1042\/bj3340685 (1998).","journal-title":"Biochem. J."},{"issue":"Pt 1","key":"93839_CR40","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1042\/bj3180319","volume":"318","author":"HW Hovenberg","year":"1996","unstructured":"Hovenberg, H. W., Davies, J. R. & Carlstedt, I. Different mucins are produced by the surface epithelium and the submucosa in human trachea: Identification of MUC5AC as a major mucin from the goblet cells. Biochem. J. 318(Pt 1), 319\u2013324. https:\/\/doi.org\/10.1042\/bj3180319 (1996).","journal-title":"Biochem. J."},{"key":"93839_CR41","doi-asserted-by":"publisher","first-page":"1044","DOI":"10.1002\/prca.200900032","volume":"3","author":"P Govender","year":"2009","unstructured":"Govender, P., Dunn, M. J. & Donnelly, S. C. Proteomics and the lung: Analysis of bronchoalveolar lavage fluid. Proteomics Clin. Appl. 3, 1044\u20131051. https:\/\/doi.org\/10.1002\/prca.200900032 (2009).","journal-title":"Proteomics Clin. Appl."},{"key":"93839_CR42","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1002\/prca.201400099","volume":"9","author":"R Terracciano","year":"2015","unstructured":"Terracciano, R., Pelaia, G., Preian\u00f2, M. & Savino, R. Asthma and COPD proteomics: Current approaches and future directions. Proteomics Clin. Appl. 9, 203\u2013220. https:\/\/doi.org\/10.1002\/prca.201400099 (2015).","journal-title":"Proteomics Clin. Appl."},{"key":"93839_CR43","doi-asserted-by":"publisher","first-page":"620","DOI":"10.1136\/thoraxjnl-2015-207358","volume":"72","author":"A Kerrin","year":"2017","unstructured":"Kerrin, A. et al. Differential lower airway dendritic cell patterns may reveal distinct endotypes of RSV bronchiolitis. Thorax 72, 620\u2013627. https:\/\/doi.org\/10.1136\/thoraxjnl-2015-207358 (2017).","journal-title":"Thorax"},{"key":"93839_CR44","doi-asserted-by":"publisher","first-page":"e1000412","DOI":"10.1371\/journal.pbio.1000412","volume":"8","author":"C Kilkenny","year":"2010","unstructured":"Kilkenny, C., Browne, W. J., Cuthill, I. C., Emerson, M. & Altman, D. G. Improving bioscience research reporting: The ARRIVE guidelines for reporting animal research. PLoS Biol. 8, e1000412. https:\/\/doi.org\/10.1371\/journal.pbio.1000412 (2010).","journal-title":"PLoS Biol."},{"key":"93839_CR45","doi-asserted-by":"publisher","first-page":"351","DOI":"10.1111\/cei.13042","volume":"190","author":"JI Kast","year":"2017","unstructured":"Kast, J. I. et al. Respiratory syncytial virus infection influences tight junction integrity. Clin. Exp. Immunol. 190, 351\u2013359. https:\/\/doi.org\/10.1111\/cei.13042 (2017).","journal-title":"Clin. Exp. Immunol."},{"key":"93839_CR46","doi-asserted-by":"publisher","first-page":"150","DOI":"10.1016\/j.tvjl.2013.10.013","volume":"199","author":"M Depecker","year":"2014","unstructured":"Depecker, M. et al. Bronchoalveolar lavage fluid in Standardbred racehorses: Influence of unilateral\/bilateral profiles and cut-off values on lower airway disease diagnosis. Vet. J. 199, 150\u2013156. https:\/\/doi.org\/10.1016\/j.tvjl.2013.10.013 (2014).","journal-title":"Vet. J."},{"key":"93839_CR47","doi-asserted-by":"publisher","first-page":"217","DOI":"10.1183\/09031936.00.15121700","volume":"15","author":"J de Blic","year":"2000","unstructured":"de Blic, J. et al. Bronchoalveolar lavage in children. ERS Task Force on bronchoalveolar lavage in children. European Respiratory Society. Eur. Respir. J. 15, 217\u2013231. https:\/\/doi.org\/10.1183\/09031936.00.15121700 (2000).","journal-title":"Eur. Respir. J."},{"key":"93839_CR48","doi-asserted-by":"publisher","first-page":"4665","DOI":"10.4049\/jimmunol.170.9.4665","volume":"170","author":"M Miyamoto","year":"2003","unstructured":"Miyamoto, M. et al. Endogenous IL-17 as a mediator of neutrophil recruitment caused by endotoxin exposure in mouse airways. J. Immunol. 170, 4665\u20134672 (2003).","journal-title":"J. Immunol."},{"key":"93839_CR49","doi-asserted-by":"publisher","first-page":"21","DOI":"10.1006\/clim.2000.4911","volume":"97","author":"N Alexis","year":"2000","unstructured":"Alexis, N., Soukup, J., Ghio, A. & Becker, S. Sputum phagocytes from healthy individuals are functional and activated: A flow cytometric comparison with cells in bronchoalveolar lavage and peripheral blood. Clin. Immunol. 97, 21\u201332. https:\/\/doi.org\/10.1006\/clim.2000.4911 (2000).","journal-title":"Clin. Immunol."},{"key":"93839_CR50","doi-asserted-by":"publisher","first-page":"e72457","DOI":"10.1371\/journal.pone.0072457","volume":"8","author":"SL Eaton","year":"2013","unstructured":"Eaton, S. L. et al. Total protein analysis as a reliable loading control for quantitative fluorescent Western blotting. PLoS ONE 8, e72457. https:\/\/doi.org\/10.1371\/journal.pone.0072457 (2013).","journal-title":"PLoS ONE"},{"key":"93839_CR51","doi-asserted-by":"publisher","first-page":"1896","DOI":"10.1038\/nprot.2007.261","volume":"2","author":"J Rappsilber","year":"2007","unstructured":"Rappsilber, J., Mann, M. & Ishihama, Y. Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips. Nat. Protoc. 2, 1896\u20131906. https:\/\/doi.org\/10.1038\/nprot.2007.261 (2007).","journal-title":"Nat. Protoc."},{"key":"93839_CR52","doi-asserted-by":"publisher","first-page":"99","DOI":"10.1007\/s10930-009-9227-7","volume":"29","author":"G Maccarrone","year":"2010","unstructured":"Maccarrone, G., Turck, C. W. & Martins-de-Souza, D. Shotgun mass spectrometry workflow combining IEF and LC-MALDI-TOF\/TOF. Protein J. 29, 99\u2013102. https:\/\/doi.org\/10.1007\/s10930-009-9227-7 (2010).","journal-title":"Protein J."},{"key":"93839_CR53","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2020.11.018","author":"BJ Dwyer","year":"2020","unstructured":"Dwyer, B. J. et al. TWEAK\/Fn14 signalling promotes cholangiocarcinoma niche formation and progression. J. Hepatol. https:\/\/doi.org\/10.1016\/j.jhep.2020.11.018 (2020).","journal-title":"J. Hepatol."},{"key":"93839_CR54","doi-asserted-by":"publisher","first-page":"e00876","DOI":"10.1128\/JVI.00876-19","volume":"93","author":"AE Karagianni","year":"2019","unstructured":"Karagianni, A. E. et al. Transcriptional response of ovine lung to infection with jaagsiekte sheep retrovirus. J. Virol. 93, e00876. https:\/\/doi.org\/10.1128\/JVI.00876-19 (2019).","journal-title":"J. Virol."},{"key":"93839_CR55","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1038\/75556","volume":"25","author":"M Ashburner","year":"2000","unstructured":"Ashburner, M. et al. Gene ontology: Tool for the unification of biology. The Gene Ontology Consortium. Nat. Genet. 25, 25\u201329. https:\/\/doi.org\/10.1038\/75556 (2000).","journal-title":"Nat. Genet."},{"key":"93839_CR56","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1093\/nar\/27.1.29","volume":"27","author":"H Ogata","year":"1999","unstructured":"Ogata, H. et al. KEGG: Kyoto encyclopedia of genes and genomes. Nucleic Acids Res. 27, 29\u201334. https:\/\/doi.org\/10.1093\/nar\/27.1.29 (1999).","journal-title":"Nucleic Acids Res."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-93839-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-93839-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-93839-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,3]],"date-time":"2022-12-03T06:30:33Z","timestamp":1670049033000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-93839-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,12]]},"references-count":56,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["93839"],"URL":"https:\/\/doi.org\/10.1038\/s41598-021-93839-9","relation":{"has-preprint":[{"id-type":"doi","id":"10.21203\/rs.3.rs-279312\/v1","asserted-by":"object"}]},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,12]]},"assertion":[{"value":"26 February 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"25 June 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"12 July 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"One of the authors of this manuscript is an academic editor for Scientific Reports. Dr T M Wishart, Reader in Molecular Anatomy, The Roslin Institute, Royal (Dick) School of Veterinary Studies, College of Medicine and Veterinary Medicine, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, SCOTLAND, UK, T.M.Wishart@Roslin.ed.ac.uk. Research explorer:\n                      \n                      . Academic Lead\u2014Proteomics and Metabolomics Facility; Roslin Institute\n                      \n                      . Co-Head\u2014Translational Biomarker Development; Centre for Dementia Prevention; University of Edinburgh\n                      \n                      . The other authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"14292"}}