{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,30]],"date-time":"2026-10-30T16:56:55Z","timestamp":1793379415251,"version":"4.5.0"},"reference-count":56,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,11,5]],"date-time":"2024-11-05T00:00:00Z","timestamp":1730764800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Milk allergy commonly occurs in children, mainly caused by bovine-derived casein (CAS) protein. Neutrophil-activating protein (NAP) of <jats:italic>Helicobacter pylori<\/jats:italic> plays an immunomodulatory role with potential to suppress Th2-type immune responses. <jats:italic>Bacillus subtilis<\/jats:italic> (<jats:italic>B. subtilis<\/jats:italic>) spores are commonly used as oral vectors for drug delivery.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective<\/jats:title><jats:p>To investigate whether recombinantly expressed NAP on <jats:italic>B. subtilis<\/jats:italic> spores could be an effective treatment for CAS allergy in mouse.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>After CAS sensitization, mice were orally administered <jats:italic>B. subtilis<\/jats:italic> spores expressing recombinant NAP for 6 weeks. Allergic symptoms and parameters were evaluated after CAS challenge oral gavage, including allergic inflammation, splenic cytokines, and serum-specific antibodies. Protein levels of Toll-like receptor 2 (TLR2) and c-JUN in the jejunum tissue were measured by western blot. Bone marrow-derived macrophages (BMDMs) were stimulated with inactivated NAP spores to measure the influence on cytokine profiles <jats:italic>in vitro<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>NAP recombinant spore treatment significantly reduced allergic symptoms and intestinal inflammation. Interleukin-12 and interferon-gamma levels increased, whereas serum CAS-specific IgG1 and IgE levels decreased. TLR2 and c-JUN expression levels were elevated in the jejunal tissue. Inactivated NAP spores polarized BMDMs to the M1 phenotype and enhanced cytokine expression, which were inhibited by a TLR2 neutralizing antibody.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>NAP offers a new strategy in the treatment of CAS allergy by inhibiting the Th2 response, while eliciting macrophages to promote Th1 immune responses.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2024.1428079","type":"journal-article","created":{"date-parts":[[2024,11,5]],"date-time":"2024-11-05T09:40:46Z","timestamp":1730799646000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":4,"title":["Neutrophil-activating protein in Bacillus spores inhibits casein allergy via TLR2 signaling"],"prefix":"10.3389","volume":"15","author":[{"given":"Zhuwei","family":"Liang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chao","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyu","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaiyue","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhile","family":"Xiong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bingshao","family":"Liang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jialiang","family":"Mai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaojun","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pingchang","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Damo","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenwen","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,11,5]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"291","DOI":"10.1016\/j.jaci.2013.11.020","article-title":"Food allergy: Epidemiology, pathogenesis, diagnosis, and treatment","volume":"133","author":"Sicherer","year":"2014","journal-title":"J Allergy Clin Immunol"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2012.06.029","article-title":"Overall prevalence of self-reported food allergy in Canada","volume":"130","author":"Soller","year":"2012","journal-title":"J Allergy Clin Immunol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1111\/j.1398-9995.1998.tb04979.x","article-title":"Immunochemical and molecular characterization of milk allergens","volume":"53","author":"Wal","year":"1998","journal-title":"Allergy"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2012.06.049","article-title":"Utility of casein-specific IgE levels in predicting reactivity to baked milk","volume":"131","author":"Caubet","year":"2013","journal-title":"J Allergy Clin Immunol"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2012.06.003","article-title":"Basophil reactivity, wheal size, and immunoglobulin levels distinguish degrees of cow's milk tolerance","volume":"131","author":"Ford","year":"2013","journal-title":"J Allergy Clin Immunol"},{"key":"B6","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/1476-7961-10-1","article-title":"The usefulness of casein-specific IgE and IgG4 antibodies in cow's milk allergic children","volume":"10","author":"Ito","year":"2012","journal-title":"Clin Mol Allergy"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1111\/all.2017.72.issue-8","article-title":"Allergen immunotherapy for IgE-mediated food allergy: a systematic review and meta-analysis","volume":"72","author":"Nurmatov","year":"2017","journal-title":"Allergy"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2016.08.008","article-title":"Balancing tolerance or allergy to food proteins","volume":"37","author":"Bryce","year":"2016","journal-title":"Trends Immunol"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1016\/j.cyto.2013.02.010","article-title":"IFN-gamma suppresses permissive chromatin remodeling in the regulatory region of the Il4 gene","volume":"62","author":"Nishida","year":"2013","journal-title":"Cytokine"},{"key":"B10","doi-asserted-by":"publisher","first-page":"15","DOI":"10.2165\/00003495-200666010-00002","article-title":"Role of Toll-like receptors in infection and immunity: clinical implications","volume":"66","author":"Cristofaro","year":"2006","journal-title":"Drugs"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2443.2001.00458.x","article-title":"Roles of Toll-like receptors in innate immune responses","volume":"6","author":"Takeda","year":"2001","journal-title":"Genes Cells"},{"key":"B12","doi-asserted-by":"publisher","first-page":"396","DOI":"10.1016\/S1471-4892(03)00080-8","article-title":"Therapeutic targeting of Toll-like receptors for inflammatory and infectious diseases","volume":"3","author":"O'Neill","year":"2003","journal-title":"Curr Opin Pharmacol"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1093\/intimm\/13.7.933","article-title":"Discrimination of bacterial lipoproteins by Toll-like receptor 6","volume":"13","author":"Takeuchi","year":"2001","journal-title":"Int Immunol"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1038\/ni712","article-title":"Toll-like receptors control activation of adaptive immune responses","volume":"2","author":"Schnare","year":"2001","journal-title":"Nat Immunol"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1086\/590158","article-title":"Helicobacter pylori colonization is inversely associated with childhood asthma","volume":"198","author":"Chen","year":"2008","journal-title":"J Infect Dis"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0004060","article-title":"Asthma is inversely associated with Helicobacter pylori status in an urban population","volume":"3","author":"Reibman","year":"2008","journal-title":"PloS One"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1084\/jem.191.9.1467","article-title":"The neutrophil-activating protein (HP-NAP) of Helicobacter pylori is a protective antigen and a major virulence factor","volume":"191","author":"Satin","year":"2000","journal-title":"J Exp Med"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1038\/nri1001","article-title":"Interleukin-12 and the regulation of innate resistance and adaptive immunity","volume":"3","author":"Trinchieri","year":"2003","journal-title":"Nat Rev Immunol"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-5822.2008.01217.x","article-title":"The neutrophil-activating protein of Helicobacter pylori down-modulates Th2 inflammation in ovalbumin-induced allergic asthma","volume":"10","author":"Codolo","year":"2008","journal-title":"Cell Microbiol"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1016\/j.femsre.2004.12.001","article-title":"The use of bacterial spore formers as probiotics","volume":"29","author":"Hong","year":"2005","journal-title":"FEMS Microbiol Rev"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1016\/S0264-410X(03)00492-4","article-title":"Germination of the spore in the gastrointestinal tract provides a novel route for heterologous antigen delivery","volume":"21","author":"Duc le","year":"2003","journal-title":"Vaccine"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-011-4835-4","article-title":"Bacterial spores as platforms for bioanalytical and biomedical applications","volume":"400","author":"Knecht","year":"2011","journal-title":"Anal Bioanal Chem"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1128\/JB.185.21.6255-6261.2003","article-title":"Morphogenesis of bacillus spore surfaces","volume":"185","author":"Chada","year":"2003","journal-title":"J Bacteriol"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.076430-0","article-title":"Expression of Helicobacter pylori urease B on the surface of Bacillus subtilis spores","volume":"64","author":"Zhou","year":"2015","journal-title":"J Med Microbiol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2008.02.015","article-title":"Oral administration of a Bacillus subtilis spore-based vaccine expressing Clonorchis sinensis tegumental protein 22.3 kDa confers protection against Clonorchis sinensis","volume":"26","author":"Zhou","year":"2008","journal-title":"Vaccine"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.000404","article-title":"Recombinant Bacillus subtilis spores expressing cholera toxin B subunit and Helicobacter pylori urease B confer protection against H. pylori in mice","volume":"66","author":"Zhou","year":"2017","journal-title":"J Med Microbiol"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-017-8370-x","article-title":"The recombinant fusion protein of cholera toxin B and neutrophil-activating protein expressed on Bacillus subtilis spore surface suppresses allergic inflammation in mice","volume":"101","author":"Dong","year":"2017","journal-title":"Appl Microbiol Biotechnol"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1111\/cea.v49.12","article-title":"Oral administration of recombinant Bacillus subtilis spores expressing Helicobacter pylori neutrophil-activating protein suppresses peanut allergy via up-regulation of Tregs","volume":"49","author":"Mai","year":"2019","journal-title":"Clin Exp Allergy"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jafc.9b02132","article-title":"Comparison of glycated ovalbumin-monosaccharides in the attenuation of ovalbumin-induced allergic response in a BALB\/C mouse model","volume":"67","author":"Rupa","year":"2019","journal-title":"J Agric Food Chem"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2012.10.026","article-title":"A microbiota signature associated with experimental food allergy promotes allergic sensitization and anaphylaxis","volume":"131","author":"Noval","year":"2013","journal-title":"J Allergy Clin Immunol"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1016\/j.dld.2006.06.012","article-title":"Intestinal permeability in patients with adverse reactions to food","volume":"38","author":"Ventura","year":"2006","journal-title":"Dig Liver Dis"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1128\/JB.188.7.2692-2700.2006","article-title":"The intestinal life cycle of Bacillus subtilis and close relatives","volume":"188","author":"Tam","year":"2006","journal-title":"J Bacteriol"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1007\/s12016-018-8710-3","article-title":"IgE-mediated food allergy","volume":"57","author":"Anvari","year":"2019","journal-title":"Clin Rev Allergy Immunol"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1067\/mai.2002.122503","article-title":"Human milk-specific mucosal lymphocytes of the gastrointestinal tract display a TH2 cytokine profile","volume":"109","author":"Beyer","year":"2002","journal-title":"J Allergy Clin Immunol"},{"key":"B35","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1186\/s40413-015-0063-2","article-title":"Mechanisms of allergen-specific immunotherapy and immune tolerance to allergens","volume":"8","author":"Akdis","year":"2015","journal-title":"World Allergy Organ J"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1067\/mai.2003.1463","article-title":"Genetic susceptibility to food allergy is linked to differential TH2-TH1 responses in C3H\/HeJ and BALB\/c mice","volume":"111","author":"Morafo","year":"2003","journal-title":"J Allergy Clin Immunol"},{"key":"B37","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1034\/j.1398-9995.2000.00802.x","article-title":"Allergic rhinitis: not purely a histamine-related disease","author":"Howarth","year":"2000","journal-title":"Allergy"},{"key":"B38","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1016\/j.biopha.2006.11.003","article-title":"Initiating mechanisms of food allergy: Oral tolerance versus allergic sensitization","volume":"61","author":"van Wijk","year":"2007","journal-title":"BioMed Pharmacother"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2010.09.013","article-title":"Food allergy herbal formula 2 protection against peanut anaphylactic reaction is via inhibition of mast cells and basophils","volume":"126","author":"Song","year":"2010","journal-title":"J Allergy Clin Immunol"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajpath.2011.12.036","article-title":"Intestinal mast cell levels control severity of oral antigen-induced anaphylaxis in mice","volume":"180","author":"Ahrens","year":"2012","journal-title":"Am J Pathol"},{"key":"B41","doi-asserted-by":"publisher","first-page":"491","DOI":"10.1016\/S0002-9440(10)65592-7","article-title":"Histochemical and ultrastructural modification of mucosal mast cell granules in parasitized mice lacking the beta-chymase, mouse mast cell protease-1","volume":"153","author":"Wastling","year":"1998","journal-title":"Am J Pathol"},{"key":"B42","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1016\/j.clim.2009.05.014","article-title":"Selective IgA deficiency in early life: association to infections and allergic diseases during childhood","volume":"133","author":"Janzi","year":"2009","journal-title":"Clin Immunol"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2013.00294","article-title":"Regulatory T cells control immune responses through their non-redundant tissue specific features","volume":"4","author":"Lehtimaki","year":"2013","journal-title":"Front Immunol"},{"key":"B44","doi-asserted-by":"publisher","first-page":"295","DOI":"10.1038\/nri.2016.36","article-title":"Development and maintenance of intestinal regulatory T cells","volume":"16","author":"Tanoue","year":"2016","journal-title":"Nat Rev Immunol"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.3389\/fped.2017.00117","article-title":"Regulatory T cells in allergy and asthma","volume":"5","author":"Martin-Orozco","year":"2017","journal-title":"Front Pediatr"},{"key":"B46","doi-asserted-by":"publisher","first-page":"125","DOI":"10.1186\/1479-5876-12-125","article-title":"Subsets of regulatory T cells and their roles in allergy","volume":"12","author":"Zhang","year":"2014","journal-title":"J Transl Med"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2015.00493","article-title":"Helicobacter pylori neutrophil-activating protein: a potential Treg modulator suppressing allergic asthma","volume":"6","author":"Sehrawat","year":"2015","journal-title":"Front Microbiol"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1097\/MPG.0b013e3180ca8960","article-title":"Increased interleukin-10 in Helicobacter pylori infection could be involved in the mechanism protecting from allergy","volume":"45","author":"Oderda","year":"2007","journal-title":"J Pediatr Gastroenterol Nutr"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.3892\/ol.2018.9182","article-title":"Toll-like receptor agonist rMBP-NAP enhances antitumor cytokines production and CTL activity of peripheral blood mononuclear cells from patients with lung cancer","volume":"16","author":"Ding","year":"2018","journal-title":"Oncol Lett"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.1139\/cjpp-2017-0186","article-title":"Recombinant protein rMBP-NAP restricts tumor progression by triggering antitumor immunity in mouse metastatic lung cancer","volume":"96","author":"Wang","year":"2018","journal-title":"Can J Physiol Pharmacol"},{"key":"B51","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1007\/s00262-011-1087-2","article-title":"HP-NAP inhibits the growth of bladder cancer in mice by activating a cytotoxic Th1 response","volume":"61","author":"Codolo","year":"2012","journal-title":"Cancer Immunol Immunother"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2012.4","article-title":"Expression of immunomodulatory neutrophil-activating protein of Helicobacter pylori enhances the antitumor activity of oncolytic measles virus","volume":"20","author":"Iankov","year":"2012","journal-title":"Mol Ther"},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2013.153","article-title":"An infection-enhanced oncolytic adenovirus secreting H. pylori neutrophil-activating protein with therapeutic effects on neuroendocrine tumors","volume":"21","author":"Ramachandran","year":"2013","journal-title":"Mol Ther"},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.1038\/nri1630","article-title":"Negative regulation of toll-like receptor-mediated immune responses","volume":"5","author":"Liew","year":"2005","journal-title":"Nat Rev Immunol"},{"key":"B55","doi-asserted-by":"publisher","DOI":"10.1038\/sj.onc.1204389","article-title":"AP-1 in mouse development and tumorigenesis","volume":"20","author":"Jochum","year":"2001","journal-title":"Oncogene"},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2004.09.015","article-title":"The chemokine system in diverse forms of macrophage activation and polarization","volume":"25","author":"Mantovani","year":"2004","journal-title":"Trends Immunol"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2024.1428079\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,11,5]],"date-time":"2024-11-05T09:40:49Z","timestamp":1730799649000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2024.1428079\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11,5]]},"references-count":56,"alternative-id":["10.3389\/fimmu.2024.1428079"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2024.1428079","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,11,5]]},"article-number":"1428079"}}