{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T17:09:09Z","timestamp":1785431349965,"version":"3.56.0"},"posted":{"date-parts":[[2025,7,11]]},"group-title":"Immunology","reference-count":46,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,7,11]],"date-time":"2025-07-11T00:00:00Z","timestamp":1752192000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"name":"European Research Council Starting Grant","award":["WePredict project number: 949613"],"award-info":[{"award-number":["WePredict project number: 949613"]}]},{"id":[{"id":"https:\/\/ror.org\/00dkwrz68","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100013850","name":"Helmut Horten Stiftung","doi-asserted-by":"crossref","award":["2021-YIG-083"],"award-info":[{"award-number":["2021-YIG-083"]}],"id":[{"id":"10.13039\/501100013850","id-type":"DOI","asserted-by":"crossref"}]},{"award":["310030_215675"],"award-info":[{"award-number":["310030_215675"]}],"id":[{"id":"https:\/\/ror.org\/00yjd3n13","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100013362","name":"Krebsforschung Schweiz","doi-asserted-by":"crossref","award":["KFS-5691-08-2022"],"award-info":[{"award-number":["KFS-5691-08-2022"]}],"id":[{"id":"10.13039\/501100013362","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100022796","name":"Jubil\u00e4umsstiftung von Swiss Life","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100022796","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Sassella Stiftung"},{"name":"Ruth & Arthur Scherbarth Stiftung"},{"id":[{"id":"https:\/\/ror.org\/04f9t1x17","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100018234","name":"Inselspital","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100018234","id-type":"DOI","asserted-by":"crossref"}]},{"name":"French DGOS for the Rare Center of Rare Digestive Disorders"},{"id":[{"id":"https:\/\/ror.org\/02vjkv261","id-type":"ROR","asserted-by":"publisher"}]},{"id":[{"id":"https:\/\/ror.org\/004raaa70","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Association Fran\u00e7ois Aupetit"},{"name":"NIH","award":["R01DK068271"],"award-info":[{"award-number":["R01DK068271"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,7,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Inflammatory bowel diseases (IBD), including Crohn\u2019s disease (CD) and ulcerative colitis (UC), are characterized by intestinal inflammation and barrier dysfunction. While disruption of the intestinal barrier contributes to the pathogenesis of IBD, yet how colonic inflammation alters small intestinal homeostasis remains poorly defined. Here, we demonstrate that three models of colitis, including 2,4,6-Trinitrobenzenesulfonic acid (TNBS), Dextran Sulfate Sodium (DSS), and oxazolone, induce paracellular barrier dysfunction in the small intestine, with model-specific immune profiles. Th1\/Th17-skewed responses (TNBS and DSS) were associated with microbiota-dependent upregulation of myosin light chain kinase (MLCK), ROR\u03b3t\u207a CD4\u207a T cell expansion, and elevated expression of Nod2, which was not observed under Th2-dominant (oxazolone) conditions. Inhibition of MLCK restored barrier function and suppressed inflammation in a CD4\u207a T cell\u2013dependent manner. Using Nod2-deficient and 2939insC mutant mice, we showed Nod2 as a critical regulator of small intestinal permeability during colitis. Bone marrow chimeras revealed compartment-specific roles of Nod2, with non-hematopoietic Nod2 sufficient to preserve epithelial integrity, while hematopoietic Nod2 expression is required to limit cytokine-mediated inflammation. Moreover, MLCK inhibition ameliorated intestinal lesions only in mice with Nod2 deficiency in the hematopoietic compartment. Finally, microbiota transfer experiments ruled out a causal role for dysbiosis in driving small intestinal permeability defects in Nod2-deficient mice. These findings uncover a Nod2\u2013MLCK\u2013CD4\u207a T cell axis linking colonic inflammation to small intestinal barrier dysfunction and highlight distinct immune\u2013epithelial\u2013microbial mechanisms shaping intestinal homeostasis during colitis.<\/jats:p>\n                <jats:sec>\n                  <jats:title>Graphical Abstract<\/jats:title>\n                  <jats:p>Circulating ROR\u03b3t\u207a Foxp3\u207b CD4\u207a T cells from mice with TNBS- or DSS-induced colitis increase small intestinal permeability by promoting cytokine secretion that enhances MLCK activity. This effect is attenuated by Nod2 activation. Specifically, Nod2 expression in hematopoietic cells reduces cytokine secretion and permeability, while activation of Nod2 in non-hematopoietic cells alone is sufficient to prevent increased permeability by directly regulating MLCK activity, independently of cytokine influence. Moreover, the absence of Nod2 is associated with inflammatory lesions in the small intestine of TNBS-treated mice.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.07.10.663215","type":"posted-content","created":{"date-parts":[[2025,7,12]],"date-time":"2025-07-12T02:15:18Z","timestamp":1752286518000},"source":"Crossref","is-referenced-by-count":0,"title":["Colitis-primed circulating T cells drive small intestinal barrier function through Nod2, microbiota and myosin light chain kinase-dependent mechanisms"],"prefix":"10.64898","author":[{"given":"Arthur","family":"Mauduit","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Talal","family":"Nabhani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5155-1107","authenticated-orcid":false,"given":"Luisa","family":"Morelli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4181-0283","authenticated-orcid":false,"given":"Floriane","family":"Cherrier","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosemary Abdoul","family":"Ahad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nidal","family":"Nabhani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gurminder","family":"Singh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maryline","family":"Roy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gilles","family":"Dietrich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0627-9455","authenticated-orcid":false,"given":"Jerrold R.","family":"Turner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8446-6056","authenticated-orcid":false,"given":"Jean-Pierre","family":"Hugot","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew","family":"Macpherson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5425-7352","authenticated-orcid":false,"given":"Emmanuel","family":"Mas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7150-9619","authenticated-orcid":false,"given":"Fr\u00e9d\u00e9rick","family":"Barreau","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4179-9555","authenticated-orcid":false,"given":"Ziad","family":"Al Nabhani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025071407150935000_2025.07.10.663215v1.1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.180.8.5653"},{"key":"2025071407150935000_2025.07.10.663215v1.2","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1093\/ecco-jcc\/jjz196","article-title":"Nod2 Protects the Gut From Experimental Colitis Spreading to Small Intestine","volume":"14","year":"2020","journal-title":"J Crohns Colitis"},{"key":"2025071407150935000_2025.07.10.663215v1.3","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006177"},{"key":"2025071407150935000_2025.07.10.663215v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2019.02.014"},{"key":"2025071407150935000_2025.07.10.663215v1.5","doi-asserted-by":"publisher","DOI":"10.1093\/ecco-jcc\/jjw095"},{"key":"2025071407150935000_2025.07.10.663215v1.6","doi-asserted-by":"publisher","DOI":"10.1097\/MIB.0000000000001135"},{"key":"2025071407150935000_2025.07.10.663215v1.7","doi-asserted-by":"publisher","DOI":"10.1097\/MIB.0000000000000749"},{"key":"2025071407150935000_2025.07.10.663215v1.8","first-page":"721","article-title":"Mucosal function in rat jejunum and ileum is altered by induction of colitis","volume":"18","year":"2006","journal-title":"Int J Mol Med"},{"key":"2025071407150935000_2025.07.10.663215v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.lfs.2006.06.047"},{"key":"2025071407150935000_2025.07.10.663215v1.10","doi-asserted-by":"publisher","DOI":"10.1136\/gut.2008.171546"},{"key":"2025071407150935000_2025.07.10.663215v1.11","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0000523"},{"key":"2025071407150935000_2025.07.10.663215v1.12","doi-asserted-by":"publisher","DOI":"10.1038\/labinvest.3700373"},{"key":"2025071407150935000_2025.07.10.663215v1.13","doi-asserted-by":"publisher","DOI":"10.1016\/S0016-5085(99)70181-2"},{"key":"2025071407150935000_2025.07.10.663215v1.14","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1186\/s12989-023-00555-5","article-title":"Perinatal foodborne titanium dioxide exposure-mediated dysbiosis predisposes mice to develop colitis through life","volume":"20","year":"2023","journal-title":"Part Fibre Toxicol"},{"key":"2025071407150935000_2025.07.10.663215v1.15","doi-asserted-by":"publisher","DOI":"10.1126\/science.aad0314"},{"key":"2025071407150935000_2025.07.10.663215v1.16","doi-asserted-by":"publisher","DOI":"10.1172\/JCI24970"},{"key":"2025071407150935000_2025.07.10.663215v1.17","doi-asserted-by":"crossref","unstructured":"Couturier-Maillard, A. , T. Secher , A. Rehman , S. Normand , A. De Arcangelis , R. Haesler , L. Huot , T. Grandjean , A. Bressenot , A. Delanoye-Crespin , O. Gaillot , S. Schreiber , Y. Lemoine , B. Ryffel , D. Hot , G. Nunez , G. Chen , P. Rosenstiel , and M. Chamaillard . 2013. NOD2-mediated dysbiosis predisposes mice to transmissible colitis and colorectal cancer. J Clin Invest","DOI":"10.1172\/JCI62236"},{"key":"2025071407150935000_2025.07.10.663215v1.18","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.178.4.2458"},{"key":"2025071407150935000_2025.07.10.663215v1.19","doi-asserted-by":"publisher","DOI":"10.1155\/2016\/7374197"},{"key":"2025071407150935000_2025.07.10.663215v1.20","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2010.08.004"},{"key":"2025071407150935000_2025.07.10.663215v1.21","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-019-0393-7"},{"key":"2025071407150935000_2025.07.10.663215v1.22","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.2011.224568"},{"key":"2025071407150935000_2025.07.10.663215v1.23","doi-asserted-by":"crossref","unstructured":"Jin, Y. , and A.T. Blikslager . 2020. The Regulation of Intestinal Mucosal Barrier by Myosin Light Chain Kinase\/Rho Kinases. Int J Mol Sci 21:","DOI":"10.3390\/ijms21103550"},{"key":"2025071407150935000_2025.07.10.663215v1.24","doi-asserted-by":"publisher","DOI":"10.1172\/JCI58147"},{"key":"2025071407150935000_2025.07.10.663215v1.25","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(01)00227-8"},{"key":"2025071407150935000_2025.07.10.663215v1.26","doi-asserted-by":"crossref","first-page":"1196","DOI":"10.1053\/j.gastro.2016.01.031","article-title":"Variants in TRIM22 That Affect NOD2 Signaling Are Associated With Very-Early-Onset Inflammatory Bowel Disease","volume":"150","year":"2016","journal-title":"Gastroenterology"},{"key":"2025071407150935000_2025.07.10.663215v1.27","doi-asserted-by":"crossref","first-page":"e13348","DOI":"10.1111\/nmo.13348","article-title":"MLCK-mediated intestinal permeability promotes immune activation and visceral hypersensitivity in PI-IBS mice","volume":"30","year":"2018","journal-title":"Neurogastroenterol Motil"},{"key":"2025071407150935000_2025.07.10.663215v1.28","doi-asserted-by":"publisher","DOI":"10.1126\/science.1103685"},{"key":"2025071407150935000_2025.07.10.663215v1.29","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-009-0751-8"},{"key":"2025071407150935000_2025.07.10.663215v1.30","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2012.02.009"},{"key":"2025071407150935000_2025.07.10.663215v1.31","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.2100869"},{"key":"2025071407150935000_2025.07.10.663215v1.32","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2011.08.035"},{"key":"2025071407150935000_2025.07.10.663215v1.33","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20090623"},{"key":"2025071407150935000_2025.07.10.663215v1.34","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMra020831"},{"key":"2025071407150935000_2025.07.10.663215v1.35","doi-asserted-by":"publisher","DOI":"10.4161\/gmic.24373"},{"key":"2025071407150935000_2025.07.10.663215v1.36","doi-asserted-by":"publisher","DOI":"10.1053\/gast.2003.50157"},{"key":"2025071407150935000_2025.07.10.663215v1.37","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2008.02.017"},{"key":"2025071407150935000_2025.07.10.663215v1.38","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.02915"},{"key":"2025071407150935000_2025.07.10.663215v1.39","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2008.10.081"},{"key":"2025071407150935000_2025.07.10.663215v1.40","doi-asserted-by":"publisher","DOI":"10.1111\/j.1572-0241.2002.05914.x"},{"key":"2025071407150935000_2025.07.10.663215v1.41","doi-asserted-by":"publisher","DOI":"10.1002\/ibd.20636"},{"key":"2025071407150935000_2025.07.10.663215v1.42","doi-asserted-by":"publisher","DOI":"10.1038\/nri2653"},{"key":"2025071407150935000_2025.07.10.663215v1.43","doi-asserted-by":"publisher","DOI":"10.1152\/ajpgi.1990.258.4.G527"},{"key":"2025071407150935000_2025.07.10.663215v1.44","doi-asserted-by":"publisher","DOI":"10.1016\/S0002-9440(10)62264-X"},{"key":"2025071407150935000_2025.07.10.663215v1.45","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2006.08.022"},{"key":"2025071407150935000_2025.07.10.663215v1.46","doi-asserted-by":"publisher","DOI":"10.1136\/gut.2006.094375"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.07.10.663215","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T09:06:06Z","timestamp":1768467966000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.07.10.663215"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,11]]},"references-count":46,"URL":"https:\/\/doi.org\/10.1101\/2025.07.10.663215","relation":{},"subject":[],"published":{"date-parts":[[2025,7,11]]},"subtype":"preprint"}}