{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T07:28:07Z","timestamp":1790234887738,"version":"4.1.0"},"reference-count":37,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2019,1,24]],"date-time":"2019-01-24T00:00:00Z","timestamp":1548288000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2019,1,24]],"date-time":"2019-01-24T00:00:00Z","timestamp":1548288000000},"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 &amp; Human Services | NIH | National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["AI123661"],"award-info":[{"award-number":["AI123661"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"U.S. Department of Health &amp; Human Services | NIH | National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["DK101323"],"award-info":[{"award-number":["DK101323"]}],"id":[{"id":"10.13039\/100000062","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><jats:p>Sublingual immunization is emerging as an alternative to nasal immunization and induction of mucosal IgA responses. Using<jats:italic>Bacillus anthracis<\/jats:italic>edema toxin (EdTx) as an adjuvant, we previously showed that innate responses triggered after sublingual immunization could limit generation of IgA responses. We tested whether co-administration of a neutrophil elastase inhibitor (NEI) could rescue the ability of EdTx to induce broad antibody responses, including mucosal IgA. NEI supplementation of sublingual vaccines containing EdTx promoted antigen-specific serum IgA responses but also enhanced serum IgG1, and IgG2b responses. This enhancing effect of NEI did not extend to all antibody isotypes and IgG sublclasses, since NEI\u00a0 reduced serum IgE responses and did not affect IgG2a\/c and IgG3 responses. NEI supplementation\u00a0also promoted anti-<jats:italic>Bacillus anthracis<\/jats:italic>protective antigen (PA) neutralizing antibodies and enhanced high affinity IgG1 and IgA antibodies. In addition to serum IgA, NEI supplementation stimulated antigen-specific mucosal IgA responses in the GI tract, and enhanced antigen-specific IgG responses in vaginal washes. Analysis of CD4<jats:sup>+<\/jats:sup>T helper cell responses revealed that co-administration of NEI broadened the profile of cytokine responses, by stimulating Th1, Th2, Th17, and Tfh cytokines. We also noted that NEI had a higher stimulatory effect on IL-5, IL-10, IL-17 responses.<\/jats:p>","DOI":"10.1038\/s41598-018-36370-8","type":"journal-article","created":{"date-parts":[[2019,1,24]],"date-time":"2019-01-24T11:04:17Z","timestamp":1548327857000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["A Novel Supplementation Approach to Enhance Host Response to Sublingual Vaccination"],"prefix":"10.1038","volume":"9","author":[{"given":"John C.","family":"Rowe","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zayed","family":"Attia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4196-8280","authenticated-orcid":false,"given":"Eunsoo","family":"Kim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Estelle","family":"Cormet-Boyaka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Prosper N.","family":"Boyaka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,1,24]]},"reference":[{"key":"36370_CR1","doi-asserted-by":"publisher","first-page":"S45","DOI":"10.1038\/nm1213","volume":"11","author":"J Holmgren","year":"2005","unstructured":"Holmgren, J. & Czerkinsky, C. Mucosal immunity and vaccines. Nat Med 11, S45\u201353 (2005).","journal-title":"Nat Med"},{"key":"36370_CR2","doi-asserted-by":"publisher","first-page":"9","DOI":"10.4049\/jimmunol.1601775","volume":"199","author":"PN Boyaka","year":"2017","unstructured":"Boyaka, P. N. Inducing Mucosal IgA: A Challenge for Vaccine Adjuvants and Delivery Systems. J Immunol 199, 9\u201316 (2017).","journal-title":"J Immunol"},{"key":"36370_CR3","doi-asserted-by":"publisher","first-page":"545","DOI":"10.1038\/mi.2010.55","volume":"3","author":"C Czerkinsky","year":"2010","unstructured":"Czerkinsky, C. & Holmgren, J. Topical immunization strategies. Mucosal Immunol 3, 545\u2013555 (2010).","journal-title":"Mucosal Immunol"},{"key":"36370_CR4","doi-asserted-by":"publisher","first-page":"5943","DOI":"10.4049\/jimmunol.0902795","volume":"185","author":"A Duverger","year":"2010","unstructured":"Duverger, A. et al. Contributions of edema factor and protective antigen to the induction of protective immunity by Bacillus anthracis edema toxin as an intranasal adjuvant. J Immunol 185, 5943\u20135952 (2010).","journal-title":"J Immunol"},{"key":"36370_CR5","doi-asserted-by":"publisher","first-page":"1776","DOI":"10.4049\/jimmunol.176.3.1776","volume":"176","author":"A Duverger","year":"2006","unstructured":"Duverger, A. et al. Bacillus anthracis edema toxin acts as an adjuvant for mucosal immune responses to nasally administered vaccine antigens. J Immunol 176, 1776\u20131783 (2006).","journal-title":"J Immunol"},{"key":"36370_CR6","doi-asserted-by":"publisher","first-page":"735","DOI":"10.1038\/mi.2014.105","volume":"8","author":"J Jee","year":"2015","unstructured":"Jee, J. et al. Neutrophils negatively regulate induction of mucosal IgA responses after sublingual immunization. Mucosal Immunol 8, 735\u2013745 (2015).","journal-title":"Mucosal Immunol"},{"key":"36370_CR7","doi-asserted-by":"publisher","first-page":"1223","DOI":"10.3389\/fimmu.2017.01223","volume":"8","author":"T Ebensen","year":"2017","unstructured":"Ebensen, T. et al. Mucosal Administration of Cycle-Di-Nucleotide-Adjuvanted Virosomes Efficiently Induces Protection against Influenza H5N1 in Mice. Front Immunol 8, 1223 (2017).","journal-title":"Front Immunol"},{"key":"36370_CR8","doi-asserted-by":"publisher","first-page":"468","DOI":"10.1111\/j.1600-0463.2011.02761.x","volume":"119","author":"CF Huang","year":"2011","unstructured":"Huang, C. F. et al. Sublingual vaccination with sonicated Salmonella proteins and mucosal adjuvant induces mucosal and systemic immunity and protects mice from lethal enteritis. APMIS 119, 468\u2013478 (2011).","journal-title":"APMIS"},{"key":"36370_CR9","doi-asserted-by":"publisher","first-page":"580","DOI":"10.1016\/j.jconrel.2014.05.060","volume":"190","author":"H Kraan","year":"2014","unstructured":"Kraan, H. et al. Buccal and sublingual vaccine delivery. J Control Release 190, 580\u2013592 (2014).","journal-title":"J Control Release"},{"key":"36370_CR10","doi-asserted-by":"publisher","first-page":"2511","DOI":"10.1016\/j.vaccine.2017.02.064","volume":"35","author":"TL Martin","year":"2017","unstructured":"Martin, T. L. et al. Sublingual targeting of STING with 3\u20323\u2032-cGAMP promotes systemic and mucosal immunity against anthrax toxins. Vaccine 35, 2511\u20132519 (2017).","journal-title":"Vaccine"},{"key":"36370_CR11","doi-asserted-by":"publisher","first-page":"1644","DOI":"10.1073\/pnas.0708684105","volume":"105","author":"JH Song","year":"2008","unstructured":"Song, J. H. et al. Sublingual vaccination with influenza virus protects mice against lethal viral infection. Proc Natl Acad Sci USA 105, 1644\u20131649 (2008).","journal-title":"Proc Natl Acad Sci USA"},{"key":"36370_CR12","doi-asserted-by":"publisher","first-page":"459","DOI":"10.1146\/annurev-immunol-020711-074942","volume":"30","author":"B Amulic","year":"2012","unstructured":"Amulic, B., Cazalet, C., Hayes, G. L., Metzler, K. D. & Zychlinsky, A. Neutrophil function: from mechanisms to disease. Annu Rev Immunol 30, 459\u2013489 (2012).","journal-title":"Annu Rev Immunol"},{"key":"36370_CR13","doi-asserted-by":"publisher","first-page":"4613","DOI":"10.1182\/blood-2009-06-221630","volume":"114","author":"O Soehnlein","year":"2009","unstructured":"Soehnlein, O., Lindbom, L. & Weber, C. Mechanisms underlying neutrophil-mediated monocyte recruitment. Blood 114, 4613\u20134623 (2009).","journal-title":"Blood"},{"key":"36370_CR14","doi-asserted-by":"publisher","first-page":"S114","DOI":"10.1164\/ajrccm\/150.6_Pt_2.S114","volume":"150","author":"G Doring","year":"1994","unstructured":"Doring, G. The role of neutrophil elastase in chronic inflammation. Am J Respir Crit Care Med 150, S114\u2013117 (1994).","journal-title":"Am J Respir Crit Care Med"},{"key":"36370_CR15","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1097\/MOH.0000000000000001","volume":"21","author":"PA Henriksen","year":"2014","unstructured":"Henriksen, P. A. The potential of neutrophil elastase inhibitors as anti-inflammatory therapies. Curr Opin Hematol 21, 23\u201328 (2014).","journal-title":"Curr Opin Hematol"},{"key":"36370_CR16","doi-asserted-by":"publisher","first-page":"313","DOI":"10.1124\/jpet.111.182139","volume":"339","author":"T Stevens","year":"2011","unstructured":"Stevens, T. et al. AZD9668: pharmacological characterization of a novel oral inhibitor of neutrophil elastase. J Pharmacol Exp Ther 339, 313\u2013320 (2011).","journal-title":"J Pharmacol Exp Ther"},{"key":"36370_CR17","doi-asserted-by":"publisher","first-page":"651","DOI":"10.1073\/pnas.96.2.651","volume":"96","author":"JW Lillard Jr.","year":"1999","unstructured":"Lillard, J. W. Jr., Boyaka, P. N., Chertov, O., Oppenheim, J. J. & McGhee, J. R. Mechanisms for induction of acquired host immunity by neutrophil peptide defensins. Proc Natl Acad Sci USA 96, 651\u2013656 (1999).","journal-title":"Proc Natl Acad Sci USA"},{"key":"36370_CR18","doi-asserted-by":"publisher","first-page":"340","DOI":"10.1016\/j.it.2007.06.002","volume":"28","author":"N Borregaard","year":"2007","unstructured":"Borregaard, N., Sorensen, O. E. & Theilgaard-Monch, K. Neutrophil granules: a library of innate immunity proteins. Trends Immunol 28, 340\u2013345 (2007).","journal-title":"Trends Immunol"},{"key":"36370_CR19","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1097\/MOH.0b013e32834115d1","volume":"18","author":"U Meyer-Hoffert","year":"2011","unstructured":"Meyer-Hoffert, U. & Wiedow, O. Neutrophil serine proteases: mediators of innate immune responses. Curr Opin Hematol 18, 19\u201324 (2011).","journal-title":"Curr Opin Hematol"},{"key":"36370_CR20","doi-asserted-by":"crossref","unstructured":"Zayed A. et al. Inhibitors of elastase stimulate murine B lymphocyte differentiation into IgG- and IgA-producing cells. European Journal of Immunology 48(8), 1295\u20131301, (2018).","DOI":"10.1002\/eji.201747264"},{"key":"36370_CR21","doi-asserted-by":"publisher","first-page":"821","DOI":"10.1038\/nri3322","volume":"12","author":"O Pabst","year":"2012","unstructured":"Pabst, O. New concepts in the generation and functions of IgA. Nat Rev Immunol 12, 821\u2013832 (2012).","journal-title":"Nat Rev Immunol"},{"key":"36370_CR22","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1189\/jlb.2A0314-178R","volume":"97","author":"KA Fairfax","year":"2015","unstructured":"Fairfax, K. A., Gantier, M. P., Mackay, F., Williams, B. R. & McCoy, C. E. IL-10 regulates Aicda expression through miR-155. J Leukoc Biol 97, 71\u201378 (2015).","journal-title":"J Leukoc Biol"},{"key":"36370_CR23","doi-asserted-by":"publisher","first-page":"3051","DOI":"10.1172\/JCI44262","volume":"121","author":"SI Hammerschmidt","year":"2011","unstructured":"Hammerschmidt, S. I. et al. Retinoic acid induces homing of protective T and B cells to the gut after subcutaneous immunization in mice. J Clin Invest 121, 3051\u20133061 (2011).","journal-title":"J Clin Invest"},{"key":"36370_CR24","doi-asserted-by":"publisher","first-page":"96","DOI":"10.1038\/mi.2007.14","volume":"1","author":"JR Mora","year":"2008","unstructured":"Mora, J. R. & von Andrian, U. H. Differentiation and homing of IgA-secreting cells. Mucosal Immunol 1, 96\u2013109 (2008).","journal-title":"Mucosal Immunol"},{"key":"36370_CR25","doi-asserted-by":"publisher","first-page":"1072","DOI":"10.1038\/mi.2014.134","volume":"8","author":"AT Cao","year":"2015","unstructured":"Cao, A. T. et al. Interleukin (IL)\u221221 promotes intestinal IgA response to microbiota. Mucosal Immunol 8, 1072\u20131082 (2015).","journal-title":"Mucosal Immunol"},{"key":"36370_CR26","doi-asserted-by":"publisher","first-page":"1157","DOI":"10.1126\/science.1132742","volume":"314","author":"JR Mora","year":"2006","unstructured":"Mora, J. R. et al. Generation of gut-homing IgA-secreting B cells by intestinal dendritic cells. Science 314, 1157\u20131160 (2006).","journal-title":"Science"},{"key":"36370_CR27","doi-asserted-by":"publisher","first-page":"1189","DOI":"10.1164\/rccm.200605-632OC","volume":"174","author":"A Roghanian","year":"2006","unstructured":"Roghanian, A., Drost, E. M., MacNee, W., Howie, S. E. & Sallenave, J. M. Inflammatory lung secretions inhibit dendritic cell maturation and function via neutrophil elastase. Am J Respir Crit Care Med 174, 1189\u20131198 (2006).","journal-title":"Am J Respir Crit Care Med"},{"key":"36370_CR28","doi-asserted-by":"publisher","first-page":"458","DOI":"10.4049\/jimmunol.1700354","volume":"199","author":"AK Bird","year":"2017","unstructured":"Bird, A. K. et al. Neutrophils Slow Disease Progression in Murine Lupus via Modulation of Autoreactive Germinal Centers. J Immunol 199, 458\u2013466 (2017).","journal-title":"J Immunol"},{"key":"36370_CR29","doi-asserted-by":"publisher","first-page":"5644","DOI":"10.4049\/jimmunol.0901499","volume":"183","author":"VA Blaho","year":"2009","unstructured":"Blaho, V. A., Buczynski, M. W., Dennis, E. A. & Brown, C. R. Cyclooxygenase-1 orchestrates germinal center formation and antibody class-switch via regulation of IL-17. J Immunol 183, 5644\u20135653 (2009).","journal-title":"J Immunol"},{"key":"36370_CR30","doi-asserted-by":"publisher","first-page":"372","DOI":"10.1038\/ni.2552","volume":"14","author":"K Hirota","year":"2013","unstructured":"Hirota, K. et al. Plasticity of Th17 cells in Peyer\u2019s patches is responsible for the induction of T cell-dependent IgA responses. Nat Immunol 14, 372\u2013379 (2013).","journal-title":"Nat Immunol"},{"key":"36370_CR31","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1111\/j.1365-2567.2006.02382.x","volume":"118","author":"K Horikawa","year":"2006","unstructured":"Horikawa, K. & Takatsu, K. Interleukin-5 regulates genes involved in B-cell terminal maturation. Immunology 118, 497\u2013508 (2006).","journal-title":"Immunology"},{"key":"36370_CR32","doi-asserted-by":"crossref","first-page":"2812","DOI":"10.4049\/jimmunol.162.5.2812","volume":"162","author":"C Mizoguchi","year":"1999","unstructured":"Mizoguchi, C., Uehara, S., Akira, S. & Takatsu, K. IL-5 induces IgG1 isotype switch recombination in mouse CD38-activated sIgD-positive B lymphocytes. J Immunol 162, 2812\u20132819 (1999).","journal-title":"J Immunol"},{"key":"36370_CR33","doi-asserted-by":"publisher","first-page":"1415","DOI":"10.1084\/jem.170.4.1415","volume":"170","author":"E Sonoda","year":"1989","unstructured":"Sonoda, E. et al. Transforming growth factor beta induces IgA production and acts additively with interleukin 5 for IgA production. J Exp Med 170, 1415\u20131420 (1989).","journal-title":"J Exp Med"},{"key":"36370_CR34","doi-asserted-by":"publisher","first-page":"288","DOI":"10.5414\/CP201674","volume":"51","author":"KA Gunawardena","year":"2013","unstructured":"Gunawardena, K. A., Gullstrand, H. & Perrett, J. Pharmacokinetics and safety of AZD9668, an oral neutrophil elastase inhibitor, in healthy volunteers and patients with COPD. Int J Clin Pharmacol Ther 51, 288\u2013304 (2013).","journal-title":"Int J Clin Pharmacol Ther"},{"key":"36370_CR35","doi-asserted-by":"publisher","first-page":"257","DOI":"10.1038\/mi.2013.43","volume":"7","author":"A Bonnegarde-Bernard","year":"2014","unstructured":"Bonnegarde-Bernard, A. et al. IKKbeta in intestinal epithelial cells regulates allergen-specific IgA and allergic inflammation at distant mucosal sites. Mucosal Immunol 7, 257\u2013267 (2014).","journal-title":"Mucosal Immunol"},{"key":"36370_CR36","doi-asserted-by":"publisher","first-page":"e92307","DOI":"10.1371\/journal.pone.0092307","volume":"9","author":"A Bonnegarde-Bernard","year":"2014","unstructured":"Bonnegarde-Bernard, A. et al. Routes of allergic sensitization and myeloid cell IKKbeta differentially regulate antibody responses and allergic airway inflammation in male and female mice. PLoS One 9, e92307 (2014).","journal-title":"PLoS One"},{"key":"36370_CR37","doi-asserted-by":"publisher","first-page":"1433","DOI":"10.1128\/CVI.00056-07","volume":"14","author":"KH Chan","year":"2007","unstructured":"Chan, K. H. et al. Use of antibody avidity assays for diagnosis of severe acute respiratory syndrome coronavirus infection. Clin Vaccine Immunol 14, 1433\u20131436 (2007).","journal-title":"Clin Vaccine Immunol"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-36370-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-36370-8","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-36370-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,7,14]],"date-time":"2024-07-14T09:01:23Z","timestamp":1720947683000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-36370-8"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,1,24]]},"references-count":37,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["36370"],"URL":"https:\/\/doi.org\/10.1038\/s41598-018-36370-8","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,1,24]]},"assertion":[{"value":"29 January 2018","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 November 2018","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"24 January 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":"715"}}