{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T22:49:26Z","timestamp":1787698166548,"version":"build-2784847793"},"reference-count":47,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,3,18]],"date-time":"2025-03-18T00:00:00Z","timestamp":1742256000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["DFG GRK 2318 -318905415, Project C4"],"award-info":[{"award-number":["DFG GRK 2318 -318905415, Project C4"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Disease progression from human immunodeficiency virus (HIV) infection to acquired immunodeficiency syndrome (AIDS) is marked by chronic immune activation, partly due to increased translocation of gut-derived microbial antigens. Elevated mucosal tumour necrosis factor-\u03b1 (TNF-\u03b1) and resulting epithelial cell apoptosis may be the etiology. However, studies using carcinoma cell lines have failed to find a causal link, possibly due to cellular abnormalities related to the malignant transformation of these immortal cell lines.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We established intestinal organoid monolayers from healthy controls and HIV-infected adults and characterized their growth dynamics and cellular composition. We then examined the effects of HIV-associated cytokines on transepithelial resistance (TER), apoptosis and macromolecule translocation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Organoid monolayers from HIV-infected patients grew similarly to healthy controls, forming confluent monolayers within one to two weeks containing enterocytes, Paneth, goblet and stem cells. IFN-\u03b3 synergized with TNF-\u03b1, allowing TNF-\u03b1 to cause caspase-mediated apoptosis and TER reduction within 5 \u00b1 3 hours, reflecting patient sample heterogeneity. This led to paracellular passage of 4 kDa Dextran and transcytosis of 44 kDa horse radish peroxidase, both of which could be blocked by pan-caspase inhibitor, Q-VD-Oph.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Our study confirms that intestinal organoid monolayers from biopsies of HIV-infected individuals can be used to study apoptosis-related epithelial barrier dysfunction and macromolecular translocation. We provide direct evidence that TNF-\u03b1-induced apoptosis triggered two pathways of macromolecular translocation: paracellular passage via apoptotic leaks and transcytosis. Therapies targeting apoptosis may be useful in preventing disease progression from HIV to AIDS.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2025.1563702","type":"journal-article","created":{"date-parts":[[2025,3,18]],"date-time":"2025-03-18T07:03:41Z","timestamp":1742281421000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["Tumour necrosis factor-\u03b1 induces macromolecule translocation in HIV-derived duodenal organoids"],"prefix":"10.3389","volume":"16","author":[{"given":"Kopano Valerie","family":"Masete","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alain S.","family":"Massarani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00f6rg-Dieter","family":"Schulzke","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hans-J\u00f6rg","family":"Epple","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nina A.","family":"Hering","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,3,18]]},"reference":[{"key":"B1","volume-title":"HIV statistics, globally and by WHO region","year":"2024"},{"key":"B2","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1111\/imr.2013.254.issue-1","article-title":"HIV-associated chronic immune activation","volume":"254","author":"Paiardini","year":"2013","journal-title":"Immunol Rev"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1038\/nm1511","article-title":"Microbial translocation is a cause of systemic immune activation in chronic HIV infection","volume":"12","author":"Brenchley","year":"2006","journal-title":"Nat Med"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1086\/597476","article-title":"Plasma levels of bacterial DNA correlate with immune activation and the magnitude of immune restoration in persons with antiretroviral-treated HIV infection","volume":"199","author":"Jiang","year":"2009","journal-title":"J Infect diseases"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1097\/QAI.0b013e318271799b","article-title":"Serum (1\u2192 3)-\u03b2-D-glucan levels in HIV-infected individuals are associated with immunosuppression, inflammation, and cardiopulmonary function","volume":"61","author":"Morris","year":"2012","journal-title":"JAIDS J Acquired Immune Deficiency Syndromes"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000852","article-title":"Exposure to HIV-1 directly impairs mucosal epithelial barrier integrity allowing microbial translocation","volume":"6","author":"Nazli","year":"2010","journal-title":"PloS pathogens"},{"key":"B7","doi-asserted-by":"publisher","first-page":"6","DOI":"10.1016\/j.tim.2012.09.001","article-title":"Microbial translocation, immune activation, and HIV disease","volume":"21","author":"Klatt","year":"2013","journal-title":"Trends Microbiol"},{"key":"B8","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1111\/j.1749-6632.2012.06512.x","article-title":"HIV infection and the intestinal mucosal barrier","volume":"1258","author":"Epple","year":"2012","journal-title":"Ann New York Acad Sci"},{"key":"B9","doi-asserted-by":"publisher","first-page":"1887","DOI":"10.3390\/cells12141887","article-title":"Macromolecule Translocation across the Intestinal Mucosa of HIV-Infected Patients by Transcytosis and through Apoptotic Leaks","volume":"12","author":"Krug","year":"2023","journal-title":"Cells"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1136\/gut.2008.150425","article-title":"Impairment of the intestinal barrier is evident in untreated but absent in suppressively treated HIV-infected patients","volume":"58","author":"Epple","year":"2009","journal-title":"Gut"},{"key":"B11","doi-asserted-by":"publisher","first-page":"1289","DOI":"10.1053\/j.gastro.2010.06.065","article-title":"Acute HIV infection induces mucosal infiltration with CD4+ and CD8+ T cells, epithelial apoptosis, and a mucosal barrier defect","volume":"139","author":"Epple","year":"2010","journal-title":"Gastroenterology"},{"key":"B12","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1097\/QAD.0000000000000511","article-title":"Gut epithelial barrier and systemic inflammation during chronic HIV infection","volume":"29","author":"Somsouk","year":"2015","journal-title":"Aids"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1038\/s41435-019-0057-0","article-title":"The many faces of tumor necrosis factor signaling in the intestinal epithelium","volume":"20","author":"Delgado","year":"2019","journal-title":"Genes Immunity"},{"key":"B14","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":"B15","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.171.11.6164","article-title":"Proinflammatory cytokines disrupt epithelial barrier function by apoptosis-independent mechanisms","volume":"171","author":"Bruewer","year":"2003","journal-title":"J Immunol"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1016\/S0002-9440(10)62264-X","article-title":"Interferon-\u03b3 and tumor necrosis factor-\u03b1 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 pathology"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1111\/j.1469-7793.2001.00541.x","article-title":"Permeability of human HT-29\/B6 colonic epithelium as a function of apoptosis","volume":"535","author":"Bojarski","year":"2001","journal-title":"J Physiol"},{"key":"B18","doi-asserted-by":"publisher","first-page":"395","DOI":"10.1517\/17425255.4.4.395","article-title":"The Caco-2 cell monolayer: usefulness and limitations","volume":"4","author":"Sun","year":"2008","journal-title":"Expert Opin Drug Metab toxicology"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-319-16104-4","article-title":"The impact of food bioactives on health: in vitro and ex vivo models","author":"Verhoeckx","year":"2015"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1016\/S1359-6446(04)03354-9","article-title":"Cell culture-based models for intestinal permeability: a critique","volume":"10","author":"Balimane","year":"2005","journal-title":"Drug Discovery Today"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1152\/ajpgi.00457.2011","article-title":"Dietary 1-monoolein decreases postprandial GIP release by reducing jejunal transport of glucose and fatty acid in rodents","volume":"303","author":"Shimotoyodome","year":"2012","journal-title":"Am J Physiology-Gastrointestinal Liver Physiol"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0079643","article-title":"Improved cell line IPEC-J2, characterized as a model for porcine jejunal epithelium","volume":"8","author":"Zakrzewski","year":"2013","journal-title":"PloS One"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0257824","article-title":"Caco-2\/HT29-MTX co-cultured cells as a model for studying physiological properties and toxin-induced effects on intestinal cells","volume":"16","author":"Hoffmann","year":"2021","journal-title":"PloS One"},{"key":"B24","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1186\/s12915-024-02105-7","article-title":"Matrix-free human 2D organoids recapitulate duodenal barrier and transport properties","volume":"23","author":"Masete","year":"2025","journal-title":"BMC Biol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1038\/cddis.2014.183","article-title":"Ex vivo culture of intestinal crypt organoids as a model system for assessing cell death induction in intestinal epithelial cells and enteropathy","volume":"5","author":"Grabinger","year":"2014","journal-title":"Cell Death disease"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1007\/s00109-017-1531-7","article-title":"Stem cell-derived organoids and their application for medical research and patient treatment","volume":"95","author":"Bartfeld","year":"2017","journal-title":"J Mol Med"},{"key":"B27","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":"B28","doi-asserted-by":"publisher","first-page":"32","DOI":"10.1038\/s41421-018-0036-z","article-title":"Monolayer culture of intestinal epithelium sustains Lgr5+ intestinal stem cells","volume":"4","author":"Liu","year":"2018","journal-title":"Cell discovery"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1111\/nyas.14804","article-title":"Human duodenal organoid-derived monolayers serve as a suitable barrier model for duodenal tissue","volume":"1515","author":"Wei\u00df","year":"2022","journal-title":"Ann New York Acad Sci"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.3791\/62074","article-title":"Organoid-derived epithelial monolayer: a clinically relevant in vitro model for intestinal barrier function","volume":"173","author":"van Dooremalen","year":"2021","journal-title":"JoVE (Journal Visualized Experiments)"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1136\/gut.45.2.191","article-title":"Synergistic effects of interferon \u03b3 and tumour necrosis factor \u03b1 on T84 cell function","volume":"45","author":"Fish","year":"1999","journal-title":"Gut"},{"key":"B32","doi-asserted-by":"publisher","first-page":"87","DOI":"10.1016\/0014-5793(92)81416-J","article-title":"Interferon-\u03b3 induces cell surface expression for both types of tumor necrosis factor receptors","volume":"312","author":"Pandita","year":"1992","journal-title":"FEBS letters"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1038\/318665a0","article-title":"Characterization of receptors for human tumour necrosis factor and their regulation by \u03b3-interferon","volume":"318","author":"Aggarwal","year":"1985","journal-title":"Nature"},{"key":"B34","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s12964-015-0102-1","article-title":"Dynasore-not just a dynamin inhibitor","volume":"13","author":"Preta","year":"2015","journal-title":"Cell Communication Signaling"},{"key":"B35","doi-asserted-by":"publisher","first-page":"103331","DOI":"10.1016\/j.autrev.2023.103331","article-title":"Cytokines and intestinal epithelial permeability: A systematic review","volume":"22","author":"Meyer","year":"2023","journal-title":"Autoimmun Rev"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2013.00498","article-title":"The significance of interferon-\u03b3 in HIV-1 pathogenesis, therapy, and prophylaxis","volume":"4","author":"Roff","year":"2014","journal-title":"Front Immunol"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1093\/ibd\/izz266","article-title":"The JAK-inhibitor tofacitinib rescues human intestinal epithelial cells and colonoids from cytokine-induced barrier dysfunction","volume":"26","author":"Sayoc-Becerra","year":"2020","journal-title":"Inflammatory Bowel Diseases"},{"key":"B38","doi-asserted-by":"publisher","first-page":"288","DOI":"10.3390\/ijms22010288","article-title":"Modeling intestinal epithelial response to interferon-\u03b3 in induced pluripotent stem cell-derived human intestinal organoids","volume":"22","author":"Workman","year":"2020","journal-title":"Int J Mol Sci"},{"key":"B39","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1089\/jir.2008.0128","article-title":"Mechanism of interferon-\u03b3\u2013induced increase in T84 intestinal epithelial tight junction","volume":"29","author":"Boivin","year":"2009","journal-title":"J Interferon Cytokine Res"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1023\/A:1024116916932","article-title":"Q-VD-OPh, a broad spectrum caspase inhibitor with potent antiapoptotic properties","volume":"8","author":"Caserta","year":"2003","journal-title":"Apoptosis"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1810017115","article-title":"Intracellular transport is accelerated in early apoptotic cells","volume":"115","author":"Li","year":"2018","journal-title":"Proc Natl Acad Sci"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1097\/QAI.0000000000002157","article-title":"Definition of immunological nonresponse to antiretroviral therapy: a systematic review","volume":"82","author":"Rb-Silva","year":"2019","journal-title":"JAIDS J Acquired Immune Deficiency Syndromes"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1136\/gut.2003.036632","article-title":"Downregulation of epithelial apoptosis and barrier repair in active Crohn\u2019s disease by tumour necrosis factor \u03b1 antibody treatment","volume":"53","author":"Zeissig","year":"2004","journal-title":"Gut"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1002\/cti2.v11.8","article-title":"An autologous colonic organoid-derived monolayer model to study immune: bacterial interactions in Crohn's disease patients","volume":"11","author":"Angus","year":"2022","journal-title":"Clin Trans Immunol"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2017.10.007","article-title":"DNA methylation and transcription patterns in intestinal epithelial cells from pediatric patients with inflammatory bowel diseases differentiate disease subtypes and associate with outcome","volume":"154","author":"Howell","year":"2018","journal-title":"Gastroenterology"},{"key":"B46","doi-asserted-by":"publisher","first-page":"1307","DOI":"10.1016\/j.cell.2018.10.008","article-title":"T helper cell cytokines modulate intestinal stem cell renewal and differentiation","volume":"175","author":"Biton","year":"2018","journal-title":"Cell"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1172\/JCI95127","article-title":"The anti-caspase inhibitor Q-VD-OPH prevents AIDS disease progression in SIV-infected rhesus macaques","volume":"128","author":"Laforge","year":"2018","journal-title":"J Clin Invest"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1563702\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,18]],"date-time":"2025-03-18T07:03:45Z","timestamp":1742281425000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1563702\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,18]]},"references-count":47,"alternative-id":["10.3389\/fimmu.2025.1563702"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2025.1563702","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,18]]},"article-number":"1563702"}}