{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,3]],"date-time":"2026-11-03T22:44:52Z","timestamp":1793745892094,"version":"4.6.1"},"reference-count":54,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T00:00:00Z","timestamp":1773878400000},"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:p>\n                    Necroptosis is an inflammatory programmed cell death pathway linked to diverse physiological and pathological disorders, yet its role in Enterotoxigenic\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    (ETEC)\u2212induced intestinal inflammation and mucosal injury remains poorly understood. This study aimed to elucidate the contribution of intestinal epithelial cell necroptosis to the development of intestinal inflammation and injury induced by ETEC infection in piglets. Following ETEC challenge in piglets, key proteins involved in necroptosis, including phosphorylated receptor-interacting protein kinase 3 (p-RIP3) and high-mobility group box 1 (HMGB1), were upregulated in jejunal crypt epithelial cells, which are primarily composed of Paneth cells and stem cells, in a time-dependent manner. In addition, ETEC challenge triggered time\u2212dependent pyroptosis in jejunal lamina propria lymphocytes, a population that includes macrophages, as demonstrated by elevated levels of NLRP3, Caspase\u22121, GSDMD\u2212N, and Cleaved -IL\u22121\u03b2 (p17) proteins in lamina propria lymphocytes. Necroptosis of jejunal crypt epithelial cells occurred prior to pyroptosis of lamina propria lymphocytes, indicating that epithelial cell necroptosis may contribute to the induction of pyroptosis in lamina propria lymphocytes. ETEC challenge induced progressive TNF\u2212\u03b1 and IL\u22121\u03b2 upregulation in plasma, jejunal crypt epithelial cells, and lamina propria lymphocytes of piglets. These changes coincided with intestinal injury and barrier loss, which were indicated by increased plasma i\u2212FABP and decreased jejunal ZO\u22121 and Occludin. Notably, Nec\u22121 pretreatment mitigates ETEC\u2212induced intestinal inflammation and tissue damage in piglets by inhibition of crypt epithelial cells necroptosis and the ensuing pyroptosis of lymphocytes. These results indicate that targeting upstream epithelial-cell necroptosis is an important strategy to attenuate inflammation and preserve barrier integrity.\n                  <\/jats:p>","DOI":"10.3389\/fimmu.2026.1778258","type":"journal-article","created":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T05:23:19Z","timestamp":1773897799000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Enterotoxigenic Escherichia coli\u2013induced intestinal epithelial necroptosis drives lamina propria immune cell pyroptosis and mucosal injury in piglets"],"prefix":"10.3389","volume":"17","author":[{"given":"Xiaoyu","family":"Wu","sequence":"first","affiliation":[{"name":"Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Huazhong Agricultural University","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujiao","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Huazhong Agricultural University","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaofeng","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Huazhong Agricultural University","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongkui","family":"Wei","sequence":"additional","affiliation":[{"name":"Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Huazhong Agricultural University","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Peng","sequence":"additional","affiliation":[{"name":"Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Huazhong Agricultural University","place":["Wuhan, China"]},{"name":"The Cooperative Innovation Center for Sustainable Pig Production","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,3,19]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1016\/j.aninu.2017.10.001","article-title":"Intestinal challenge with enterotoxigenic Escherichia coli in pigs, and nutritional intervention to prevent postweaning diarrhea","volume":"3","author":"Sun","year":"2017","journal-title":"Anim Nutr"},{"key":"B2","first-page":"73","article-title":"Presentation of postweaning Escherichia coli diarrhea in southern Ontario, prevalence of hemolytic E. coli serogroups involved, and their antimicrobial resistance patterns","volume":"66","author":"Amezcua","year":"2002","journal-title":"Can J Vet Res"},{"key":"B3","doi-asserted-by":"publisher","first-page":"851","DOI":"10.1038\/s41419-023-06370-2","article-title":"Mechanisms of PANoptosis and relevant small-molecule compounds for fighting diseases","volume":"14","author":"Wang","year":"2023","journal-title":"Cell Death Dis"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1038\/s41580-020-0270-8","article-title":"Emerging connectivity of programmed cell death pathways and its physiological implications","volume":"21","author":"Bedoui","year":"2020","journal-title":"Nat Rev Mol Cell Biol"},{"key":"B5","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1038\/s41390-021-01457-y","article-title":"Toll-like receptor 4-mediated necroptosis in the development of necrotizing enterocolitis","volume":"91","author":"Liu","year":"2022","journal-title":"Pediatr Res"},{"key":"B6","doi-asserted-by":"publisher","first-page":"921","DOI":"10.1038\/s41419-018-0973-3","article-title":"TNFR2 unlocks a RIPK1 kinase activity-dependent mode of proinflammatory TNFR1 signaling","volume":"9","author":"Siegmund","year":"2018","journal-title":"Cell Death Dis"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1016\/j.cyto.2020.155046","article-title":"PI3K mediates tumor necrosis factor induced-necroptosis through initiating RIP1-RIP3-MLKL signaling pathway activation","volume":"129","author":"Hu","year":"2020","journal-title":"Cytokine"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1038\/s41418-022-01061-5","article-title":"MLKL post-translational modifications: road signs to infection, inflammation and unknown destinations","volume":"30","author":"Liccardi","year":"2023","journal-title":"Cell Death Differ"},{"key":"B9","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1186\/s40104-022-00816-x","article-title":"Protocatechuic acid and quercetin attenuate ETEC-caused IPEC-1 cell inflammation and injury associated with inhibition of necroptosis and pyroptosis signaling pathways","volume":"14","author":"Xiao","year":"2023","journal-title":"J Anim Sci Biotechnol"},{"key":"B10","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1017\/S0007114521005213","article-title":"Long chain PUFA ameliorate ETEC-induced intestinal inflammation and cell injury by modulating pyroptosis and necroptosis signaling pathways in IPEC-1 cells","volume":"128","author":"Xiao","year":"2022","journal-title":"Br J Nutr"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-immunol-081022-021207","article-title":"Structural mechanisms of NLRP3 inflammasome assembly and activation","volume":"41","author":"Fu","year":"2023","journal-title":"Annu Rev Immunol"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.3389\/fnagi.2022.879021","article-title":"The NLRP3 inflammasome pathway: a review of mechanisms and inhibitors for the treatment of inflammatory diseases","volume":"14","author":"Blevins","year":"2022","journal-title":"Front Aging Neurosci"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2017.03.007","article-title":"Disruption of the epithelial barrier during intestinal inflammation: Quest for new molecules and mechanisms","volume":"1864","author":"Lechuga","year":"2017","journal-title":"Biochim Biophys Acta (BBA) Mol Cell Res"},{"key":"B14","doi-asserted-by":"publisher","first-page":"232","DOI":"10.1007\/s10238-024-01496-9","article-title":"Intestinal permeability disturbances: Causes, diseases and therapy","volume":"24","author":"Macura","year":"2024","journal-title":"Clin Exp Med"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1080\/21688370.2021.1972760","article-title":"Dysfunction of epithelial permeability barrier induced by HMGB1 in 2.5 D cultures of human epithelial cells","volume":"10","author":"Kojima","year":"2022","journal-title":"Tissue Barriers"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1016\/j.lfs.2019.117120","article-title":"HMGB1 was negatively regulated by HSF1 and mediated the TLR4\/MyD88\/NF-\u03baB signal pathway in asthma","volume":"241","author":"Shang","year":"2020","journal-title":"Life Sci"},{"key":"B17","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/s11427-024-2728-2","article-title":"Bacterial infection in weaned piglets promotes diarrhea by inducing the NLRP3 inflammasome-pyroptosis pathway","volume":"68","author":"Fu","year":"2025","journal-title":"Sci China Life Sci"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1111\/jam.13936","article-title":"Dietary supplementation with Clostridium butyricum helps to improve the intestinal barrier function of weaned piglets challenged with enterotoxigenic Escherichia coli K88","volume":"125","author":"Li","year":"2018","journal-title":"J Appl Microbiol"},{"key":"B19","volume-title":"Toxicity Test and Therapeutic Effect of Diweng Keli on Piglet white Scour","author":"Xia","year":"2015"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.3390\/vetsci13020120","article-title":"Robust goat-derived enterococcus isolates with broad-spectrum antipathogenic activity as next-generation probiotic candidates","volume":"13","author":"Essa","year":"2026","journal-title":"Vet Sci"},{"key":"B21","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1038\/s41419-020-03365-1","article-title":"Necroptosis is active and contributes to intestinal injury in a piglet model with lipopolysaccharide challenge","volume":"12","author":"Liu","year":"2021","journal-title":"Cell Death Dis"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2- \u0394\u0394CT method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2008.73","article-title":"Analyzing real-time PCR data by the comparative CT method","volume":"3","author":"Schmittgen","year":"2008","journal-title":"Nat Protoc"},{"key":"B24","doi-asserted-by":"publisher","first-page":"338","DOI":"10.5713\/ab.21.0010","article-title":"Understanding intestinal health in nursery pigs and the relevant nutritional strategies","volume":"34","author":"Kim","year":"2021","journal-title":"Anim Biosci"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2021.683863","article-title":"Clostridium butyricum helps to alleviate inflammation in weaned piglets challenged with enterotoxigenic Escherichia coli K88","volume":"8","author":"Li","year":"2021","journal-title":"Front Vet Sci"},{"key":"B26","doi-asserted-by":"publisher","first-page":"2060","DOI":"10.1038\/s41598-024-52586-3","article-title":"Effect of dietary tall oil fatty acids and hydrolysed yeast in SNP2-positive and SNP2-negative piglets challenged with F4 enterotoxigenic Escherichia coli","volume":"14","author":"Middelkoop","year":"2024","journal-title":"Sci Rep"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1002\/jobm.201400901","article-title":"F4+ enterotoxigenic Escherichia coli (ETEC) adhesion mediated by the major fimbrial subunit FaeG","volume":"55","author":"Xia","year":"2015","journal-title":"J Basic Microbiol"},{"key":"B28","doi-asserted-by":"publisher","first-page":"226","DOI":"10.1186\/s12866-016-0847-8","article-title":"Differential protection by cell wall components of Lactobacillus amylovorus DSM 16698Tagainst alterations of membrane barrier and NF-kB activation induced by enterotoxigenic F4+ Escherichia coli on intestinal cells","volume":"16","author":"Roselli","year":"2016","journal-title":"BMC Microbiol"},{"key":"B29","doi-asserted-by":"publisher","first-page":"112","DOI":"10.1186\/s40104-024-01070-z","article-title":"Role of immunomodulatory probiotics in alleviating bacterial diarrhea in piglets: a systematic review","volume":"15","author":"Saha","year":"2024","journal-title":"J Anim Sci Biotechnol"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1186\/1297-9716-43-70","article-title":"The search for the gene mutations underlying enterotoxigenic Escherichia coli F4ab\/ac susceptibility in pigs: a review","volume":"43","author":"Schroyen","year":"2012","journal-title":"Vet Res"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.3390\/toxins12120760","article-title":"Impact of the Escherichia coli heat-stable enterotoxin b (STb) on gut health and function","volume":"12","author":"Butt","year":"2020","journal-title":"Toxins (Basel)"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2180-10-193","article-title":"Common origin of plasmid encoded alpha-hemolysin genes in Escherichia coli","volume":"10","author":"Burgos","year":"2010","journal-title":"BMC Microbiol"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1810819115","article-title":"Intestinal barrier dysfunction orchestrates the onset of inflammatory host\u2013microbiome cross-talk in a human gut inflammation-on-a-chip","volume":"115","author":"Shin","year":"2018","journal-title":"Proc Natl Acad Sci"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1108289","article-title":"Understanding disruption of the gut barrier during inflammation: Should we abandon traditional epithelial cell lines and switch to intestinal organoids","volume":"14","author":"Lechuga","year":"2023","journal-title":"Front Immunol"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jafc.0c01414","article-title":"A combination of formic acid and monolaurin attenuates enterotoxigenic Escherichia coli induced intestinal inflammation in piglets by inhibiting the NF-\u03baB\/MAPK pathways with modulation of gut microbiota","volume":"68","author":"Ren","year":"2020","journal-title":"J Agric Food Chem"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(00)80038-2","article-title":"Impaired on\/off regulation of TNF biosynthesis in mice lacking TNF AU-rich elements: implications for joint and gut-associated immunopathologies","volume":"10","author":"Kontoyiannis","year":"1999","journal-title":"Immunity"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1007\/s12026-017-8894-2","article-title":"Glycyrrhizin ameliorates experimental colitis through attenuating interleukin-17-producing T cell responses via regulating antigen-presenting cells","volume":"65","author":"Chen","year":"2017","journal-title":"Immunol Res"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.3390\/ijms222312729","article-title":"Investigation of H2S donor treatment on neutrophil extracellular traps in experimental colitis","volume":"22","author":"T\u00f6r\u00f6k","year":"2021","journal-title":"Int J Mol Sci"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.3892\/mmr_00000056","article-title":"Roles of high-mobility group box 1 in murine experimental colitis","volume":"2","author":"Yamasaki","year":"2009","journal-title":"Mol Med Rep"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1177\/15593258211003798","article-title":"Anti-oxidative and immuno-protective effect of camel milk on radiation-induced intestinal injury in C57BL\/6 J mice","volume":"19","author":"Chen","year":"2021","journal-title":"Dose-Response"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.3390\/molecules27217311","article-title":"Therapeutic potential of targeting the HMGB1\/RAGE axis in inflammatory diseases","volume":"27","author":"Singh","year":"2022","journal-title":"Molecules"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.0900390","article-title":"HMGB1 enhances the proinflammatory activity of lipopolysaccharide by promoting the phosphorylation of MAPK p38 through receptor for advanced glycation end products","volume":"183","author":"Qin","year":"2009","journal-title":"J Immunol"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1016\/j.fct.2023.113683","article-title":"NLRP3 Inflammasome: A key contributor to the inflammation formation","volume":"174","author":"Zhang","year":"2023","journal-title":"Food Chem Toxicol"},{"key":"B44","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1016\/j.immuni.2017.06.020","article-title":"The P2X7 receptor in infection and inflammation","volume":"47","author":"Di Virgilio","year":"2017","journal-title":"Immunity"},{"key":"B45","doi-asserted-by":"publisher","first-page":"681","DOI":"10.1016\/j.it.2021.06.001","article-title":"The \u2018cytokine storm\u2019: molecular mechanisms and therapeutic prospects","volume":"42","author":"Karki","year":"2021","journal-title":"Trends Immunol"},{"key":"B46","doi-asserted-by":"crossref","DOI":"10.1038\/nature10400","article-title":"Caspase-8 regulates TNF-\u03b1-induced epithelial necroptosis and terminal ileitis","volume":"477","author":"G\u00fcnther","year":"2011","journal-title":"Nature"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1122258","article-title":"From birth to death: The hardworking life of Paneth cell in the small intestine","volume":"14","author":"Cui","year":"2023","journal-title":"Front Immunol"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.00587","article-title":"The Paneth cell: the curator and defender of the immature small intestine","volume":"11","author":"Lueschow","year":"2020","journal-title":"Front Immunol"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1016\/j.micpath.2018.09.032","article-title":"Enterotoxigenic Escherichia coli infection promotes apoptosis in piglets","volume":"125","author":"Xia","year":"2018","journal-title":"Microb Pathogen"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcmgh.2021.06.006","article-title":"Secretory sorcery: paneth cell control of intestinal repair and homeostasis","volume":"12","author":"Cray","year":"2021","journal-title":"Cell Mol Gastroenterol Hepatol"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1002\/stem.3324","article-title":"Heat-stable enterotoxin inhibits intestinal stem cell expansion to disrupt the intestinal integrity by downregulating the Wnt\/\u03b2-catenin pathway","volume":"39","author":"Zhou","year":"2021","journal-title":"Stem Cells"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1016\/j.tcb.2018.08.001","article-title":"The intestinal stem cell niche: homeostasis and adaptations","volume":"28","author":"Santos","year":"2018","journal-title":"Trends Cell Biol"},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrep.2024.101665","article-title":"Escherichia coli K88 activates NLRP3 inflammasome-mediated pyroptosis in vitro and in vivo","volume":"38","author":"Cheng","year":"2024","journal-title":"Biochem Biophys Rep"},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2024.1394857","article-title":"Necroptosis in bacterial infections","volume":"15","author":"Yu","year":"2024","journal-title":"Front Immunol"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2026.1778258\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T05:23:21Z","timestamp":1773897801000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2026.1778258\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,19]]},"references-count":54,"alternative-id":["10.3389\/fimmu.2026.1778258"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2026.1778258","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,19]]},"article-number":"1778258"}}