{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T15:42:08Z","timestamp":1792683728520,"version":"4.5.0"},"reference-count":44,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,6,11]],"date-time":"2025-06-11T00:00:00Z","timestamp":1749600000000},"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":"\n Background<\/jats:title>\n \n Lactic acid bacteria (LAB) have been widely used as probiotics which contribute to our health. We previously reported that\n Lactobacillus delbrueckii<\/jats:italic>\n subsp.\n bulgaricus<\/jats:italic>\n 2038 and\n Streptococcus thermophilus<\/jats:italic>\n 1131, two yogurt starter strains, ameliorate the intestinal barrier dysfunction caused by tumor necrosis factor (TNF)-\u03b1 and interferon (IFN)-\u03b3 in Caco-2 cells. However, Caco-2 cells differ from living organisms in various ways. We have developed a human induced pluripotent stem cell-derived crypt-villus structural small intestine (hiPSC-SI) was established with a villus-like structure containing constituent cells of the small intestine.\n <\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n A hiPSC-SI and LAB co-culture model was established to assess the impact of LAB on barrier function and elucidate the underlying mechanisms.<\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n The medium on the luminal side for co-culturing cells and bacteria was examined and determined to use Hanks\u2019 balanced salt solution without glucose in terms of bacterial survival rate. LAB were found to ameliorate permeability and decrease the gene expression of tight junction associated proteins induced by TNF-\u03b1 and IFN-\u03b3. Regarding cell differentiation, LAB suppressed the downregulation of\n LGR5<\/jats:italic>\n ,\n VIL1<\/jats:italic>\n ,\n LYZ<\/jats:italic>\n and\n MUC2<\/jats:italic>\n by cytokines. Moreover, they ameliorated reduced mucin 2 protein production and decreased the number of mucin 2-positive cells. Finally, transcriptome analysis suggested that they ameliorated the aberration in cytokine-induced cell differentiation via an anti-inflammatory effect on intestinal stem cells.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusions<\/jats:title>\n The results indicate that LAB ameliorate the cytokine-induced dysfunction of intestinal barrier integrity and homeostasis disrupted by cytokines in a co-culture model of hiPSC-SI and LAB.<\/jats:p>\n <\/jats:sec>","DOI":"10.3389\/fimmu.2025.1585007","type":"journal-article","created":{"date-parts":[[2025,6,11]],"date-time":"2025-06-11T14:02:51Z","timestamp":1749650571000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":4,"title":["Lactobacillus delbrueckii subsp. bulgaricus 2038 and Streptococcus thermophilus 1131 ameliorate barrier dysfunction in human induced pluripotent stem cell-derived crypt-villus structural small intestine"],"prefix":"10.3389","volume":"16","author":[{"given":"Kyosuke","family":"Kobayashi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuri","family":"Imai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuka","family":"Shimizu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isamu","family":"Ogawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takaaki","family":"Nakai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuri","family":"Mizuno","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshika","family":"Kaneda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yumiko","family":"Watanabe-Yasuoka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fuka","family":"Yamazaki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junko","family":"Mochizuki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshihiro","family":"Sashihara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tamihide","family":"Matsunaga","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takahiro","family":"Iwao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,6,11]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1097\/00001574-199903000-00004","article-title":"Water and electrolyte absorption and secretion in the small intestine","volume":"15","author":"Turvill","year":"1999","journal-title":"Curr Opin Gastroenterol"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1038\/s41575-023-00766-3","article-title":"Paracellular permeability and tight junction regulation in gut health and disease","volume":"20","author":"Horowitz","year":"2023","journal-title":"Nat Rev Gastroenterol Hepatol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.19.2.315-337.2006","article-title":"The front line of enteric host defense against unwelcome intrusion of harmful microorganisms: mucins, antimicrobial peptides, and microbiota","volume":"19","author":"Lievin-Le Moal","year":"2006","journal-title":"Clin Microbiol Rev"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.3390\/nu11122998","article-title":"Lactobacillus delbrueckii subsp. bulgaricus 2038 and Streptococcus thermophilus 1131 induce the expression of the REG3 family in the small intestine of mice via the stimulation of dendritic cells and type 3 innate lymphoid cells","volume":"11","author":"Kobayashi","year":"2019","journal-title":"Nutrients"},{"key":"B5","doi-asserted-by":"publisher","first-page":"2184157","DOI":"10.1080\/21688370.2023.2184157","article-title":"Yogurt starter strains ameliorate intestinal barrier dysfunction via activating AMPK in Caco-2 cells","volume":"12","author":"Kobayashi","year":"2023","journal-title":"Tissue Barriers"},{"key":"B6","doi-asserted-by":"publisher","first-page":"4","DOI":"10.1124\/dmd.30.1.4","article-title":"Real-time quantitative polymerase chain reaction for MDR1, MRP1, MRP2, and CYP3A-mRNA levels in Caco-2 cell lines, human duodenal enterocytes, normal colorectal tissues, and colorectal adenocarcinomas","volume":"30","author":"Nakamura","year":"2002","journal-title":"Drug Metab Dispos"},{"key":"B7","doi-asserted-by":"crossref","DOI":"10.1007\/978-3-319-16104-4_10","article-title":"Caco-2 cell line","volume-title":"The impact of food bioactives on health:","author":"Lea","year":"2015"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1038\/nature07935","article-title":"Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche","volume":"459","author":"Sato","year":"2009","journal-title":"Nature"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1038\/nature09691","article-title":"Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro","volume":"470","author":"Spence","year":"2011","journal-title":"Nature"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1152\/ajpgi.00152.2017","article-title":"A simple, cost-effective method for generating murine colonic 3D enteroids and 2D monolayers for studies of primary epithelial cell function","volume":"313","author":"Fernando","year":"2017","journal-title":"Am J Physiol Gastrointest Liver Physiol"},{"key":"B11","doi-asserted-by":"publisher","first-page":"45270","DOI":"10.1038\/srep45270","article-title":"A primary human macrophage-enteroid co-culture model to investigate mucosal gut physiology and host-pathogen interactions","volume":"7","author":"Noel","year":"2017","journal-title":"Sci Rep"},{"key":"B12","doi-asserted-by":"publisher","first-page":"44","DOI":"10.2133\/dmpk.DMPK-13-RG-005","article-title":"Differentiation of human induced pluripotent stem cells into functional enterocyte-like cells using a simple method","volume":"29","author":"Iwao","year":"2014","journal-title":"Drug Metab Pharmacokinet"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1016\/j.stemcr.2018.10.019","article-title":"Efficient generation of small intestinal epithelial-like cells from human iPSCs for drug absorption and metabolism studies","volume":"11","author":"Negoro","year":"2018","journal-title":"Stem Cell Reports"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2013.00079","article-title":"Alternative functional in vitro models of human intestinal epithelia","volume":"4","author":"Kauffman","year":"2013","journal-title":"Front Pharmacol"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1038\/nature09637","article-title":"Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts","volume":"469","author":"Sato","year":"2011","journal-title":"Nature"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1242\/dev.110.4.1001","article-title":"Stem cells: attributes, cycles, spirals, pitfalls and uncertainties","volume":"110","author":"Potten","year":"1990","journal-title":"Lessons crypt Dev"},{"key":"B17","doi-asserted-by":"publisher","first-page":"92","DOI":"10.3390\/medicina59010092","article-title":"Protective effect of Irsogladine against Aspirin-induced mucosal injury in human induced pluripotent stem cell-derived small intestine","volume":"59","author":"Kanno","year":"2022","journal-title":"Medicina (Kaunas)"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1242\/bio.061612","article-title":"Air-liquid interface culture combined with differentiation factors reproducing intestinal cell structure formation in vitro","volume":"14","author":"Ogawa","year":"2025","journal-title":"Biol Open"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1177\/2211068214561025","article-title":"TEER measurement techniques for in vitro barrier model systems","volume":"20","author":"Srinivasan","year":"2015","journal-title":"J Lab Autom"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1124\/dmd.118.080374","article-title":"Generation of intestinal organoids suitable for pharmacokinetic studies from human induced pluripotent stem cells","volume":"46","author":"Onozato","year":"2018","journal-title":"Drug Metab Dispos"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1016\/S0002-9440(10)62264-X","article-title":"Interferon-gamma and tumor necrosis factor-alpha synergize to induce intestinal epithelial barrier dysfunction by up-regulating myosin light chain kinase expression","volume":"166","author":"Wang","year":"2005","journal-title":"Am J Pathol"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2006.08.022","article-title":"IFN-gamma-induced TNFR2 expression is required for TNF-dependent intestinal epithelial barrier dysfunction","volume":"131","author":"Wang","year":"2006","journal-title":"Gastroenterology"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0506580102","article-title":"Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles","volume":"102","author":"Subramanian","year":"2005","journal-title":"Proc Natl Acad Sci U S A"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1038\/ng1180","article-title":"PGC-1alpha-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes","volume":"34","author":"Mootha","year":"2003","journal-title":"Nat Genet"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1159\/000447252","article-title":"Increased intestinal permeability and decreased barrier function: does it really influence the risk of inflammation","volume":"1","author":"Fukui","year":"2016","journal-title":"Inflammation Intest Dis"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1152\/ajpgi.00173.2003","article-title":"TNF-alpha-induced increase in intestinal epithelial tight junction permeability requires NF-kappa B activation","volume":"286","author":"Ma","year":"2004","journal-title":"Am J Physiol Gastrointest Liver Physiol"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1016\/j.clinre.2012.12.004","article-title":"Efficacy and safety of interferon-gamma-targeted therapy in Crohn\u2019s disease: a systematic review and meta-analysis of randomized controlled trials","volume":"37","author":"Cui","year":"2013","journal-title":"Clin Res Hepatol Gastroenterol"},{"key":"B28","doi-asserted-by":"publisher","first-page":"89","DOI":"10.1016\/0140-6736(92)90999-J","article-title":"Tumour necrosis factor alpha in stool as a marker of intestinal inflammation","volume":"339","author":"Braegger","year":"1992","journal-title":"Lancet"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1136\/gut.32.8.913","article-title":"Serum concentrations of tumour necrosis factor alpha in childhood chronic inflammatory bowel disease","volume":"32","author":"Murch","year":"1991","journal-title":"Gut"},{"key":"B30","article-title":"Plasma histamine and tumour necrosis factor-alpha levels in Crohn\u2019s disease and ulcerative colitis at various stages of disease","volume":"66","author":"Hagel","year":"2015","journal-title":"J Physiol Pharmacol"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.060665","article-title":"Claudin-2, a component of the tight junction, forms a paracellular water channel","volume":"123","author":"Rosenthal","year":"2010","journal-title":"J Cell Sci"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2019-319514","article-title":"Mitochondrial impairment drives intestinal stem cell transition into dysfunctional Paneth cells predicting Crohn\u2019s disease recurrence","volume":"69","author":"Khaloian","year":"2020","journal-title":"Gut"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1159\/000447252","article-title":"Metformin improves ileal epithelial barrier function in interleukin-10 deficient mice","volume":"11","author":"Xue","year":"2016","journal-title":"PloS One"},{"key":"B34","doi-asserted-by":"publisher","first-page":"10.1002\/mnfr.201700536","DOI":"10.1002\/mnfr.201700536","article-title":"Purple potato extract promotes intestinal epithelial differentiation and barrier function by activating AMP-activated protein kinase","volume":"62","author":"Sun","year":"2018","journal-title":"Mol Nutr Food Res"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.3390\/nu8050272","article-title":"Polyphenol-rich propolis extracts strengthen intestinal barrier function by activating AMPK and ERK signaling","volume":"8","author":"Wang","year":"2016","journal-title":"Nutrients"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1038\/cdd.2017.14","article-title":"AMPK improves gut epithelial differentiation and barrier function via regulating Cdx2 expression","volume":"24","author":"Sun","year":"2017","journal-title":"Cell Death Differ"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1002\/mnfr.201400218","article-title":"Lactobacillus delbrueckii TUA4408L and its extracellular polysaccharides attenuate enterotoxigenic Escherichia coli-induced inflammatory response in porcine intestinal epitheliocytes via Toll-like receptor-2 and 4","volume":"58","author":"Wachi","year":"2014","journal-title":"Mol Nutr Food Res"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.274.25.17406","article-title":"Peptidoglycan- and lipoteichoic acid-induced cell activation is mediated by toll-like receptor 2","volume":"274","author":"Schwandner","year":"1999","journal-title":"J Biol Chem"},{"key":"B39","doi-asserted-by":"publisher","first-page":"1","DOI":"10.4049\/jimmunol.163.1.1","article-title":"Cutting edge: recognition of Gram-positive bacterial cell wall components by the innate immune system occurs via Toll-like receptor 2","volume":"163","author":"Yoshimura","year":"1999","journal-title":"J Immunol"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1120269109","article-title":"Calcium and pH-dependent packing and release of the gel-forming MUC2 mucin","volume":"109","author":"Ambort","year":"2012","journal-title":"Proc Natl Acad Sci U S A"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1136\/gut.2007.141481","article-title":"Secreted enteric antimicrobial activity localises to the mucus surface layer","volume":"57","author":"Meyer-Hoffert","year":"2008","journal-title":"Gut"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1007\/s11357-019-00137-4","article-title":"Lipoteichoic acid from the cell wall of a heat killed Lactobacillus paracasei D3\u20135 ameliorates aging-related leaky gut, inflammation and improves physical and cognitive functions: from C","volume":"42","author":"Wang","year":"2020","journal-title":"elegans to mice Gerosci"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-011-0822-3","article-title":"Composition and functional role of the mucus layers in the intestine","volume":"68","author":"Johansson","year":"2011","journal-title":"Cell Mol Life Sci"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cws083","article-title":"Detailed O-glycomics of the Muc2 mucin from colon of wild-type, core 1- and core 3-transferase-deficient mice highlights differences compared with human MUC2","volume":"22","author":"Thomsson","year":"2012","journal-title":"Glycobiology"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1585007\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,6,11]],"date-time":"2025-06-11T14:02:54Z","timestamp":1749650574000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1585007\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,11]]},"references-count":44,"alternative-id":["10.3389\/fimmu.2025.1585007"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2025.1585007","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,11]]},"article-number":"1585007"}}