{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T21:28:03Z","timestamp":1792013283886,"version":"4.4.0"},"reference-count":54,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,5,19]],"date-time":"2025-05-19T00:00:00Z","timestamp":1747612800000},"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>Loss of gut barrier integrity has been observed in rheumatoid arthritis (RA). While systemic inflammation in RA has been extensively investigated, intestinal-specific inflammatory processes remain poorly understood. This study is designed to identify a novel biomarker panel combining fecal cytokine profiles with gut barrier biomarkers to discriminate RA patients with varying disease progression.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Feces (Fc) and plasma (Pl) were obtained from 62 Naive RA patients (NA), 47 remission RA patients (RE), 28 difficult-to-treat RA patients(D2T), and 70 healthy controls (HC). A panel of 12 cytokines and gut barrier markers, including intestinal Fatty-Acid-Binding Protein-2 (FABP2), zonulin, Hypoxia-Inducible Factor-2\u03b1 (HIF-2\u03b1), D-lactate, LBP and fecal calprotectin (FCAL), was quantified by ELISA. Statistical integration with clinical parameters was performed using univariate and multivariate approaches.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>NA and D2T patients demonstrated marked elevations in fecal pro-inflammatory cytokines compared to RE and HC groups, including IL-6, Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF), IL-1 beta (IL-1\u03b2), Interferon-gamma (INF-\u03b3), IL-23, Tumor Necrosis Factor-Alpha (TNF-\u03b1), IL-21, IL-17A\/F, and IL-22. Fecal zonulin and plasma HIF-2\u03b1 were significantly elevated in both NA and D2T groups, whereas fecal D-lactate showed a pronounced decrease in the NA and D2T groups. These biomarkers demonstrated the strongest correlation with disease severity indices. Receiver operating characteristic (ROC) analysis revealed that fecal FABP2, zonulin and D-lactate exhibited superior discriminative capacity between the NA and RE groups. whereas fecal zonulin showed remarkable diagnostic potential for distinguishing NA from D2T groups compared to plasma counterparts. The discriminant scores (DS) model incorporating fecal zonulin and plasma HIF-2\u03b1 demonstrated superior discriminatory performance between the D2T and NA groups compared to the model utilizing the top five plasma parameters.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Our fecal profiling methodology provides novel insights into the gut mucosal cytokine microenvironment during RA progression. The dissociation between fecal and plasma inflammatory profiles underscores the critical importance of localized gut immune monitoring in RA management.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2025.1577590","type":"journal-article","created":{"date-parts":[[2025,5,19]],"date-time":"2025-05-19T05:16:13Z","timestamp":1747631773000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":4,"title":["Profiling of fecal analytes as a potential biomarker in rheumatoid arthritis"],"prefix":"10.3389","volume":"16","author":[{"given":"Zhiyi","family":"Wang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujia","family":"Shi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yachen","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bangdong","family":"Gong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianmin","family":"Xie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,5,19]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1186\/s43556-022-00076-1","article-title":"Characterization of chronic relapsing antibody mediated arthritis in mice with a mutation in Ncf1 causing reduced oxidative burst","volume":"3","author":"Liang","year":"2022","journal-title":"Mol Biomed"},{"key":"B2","doi-asserted-by":"publisher","first-page":"R142","DOI":"10.1186\/ar3875","article-title":"Investigation of the role of endosomal Toll-like receptors in murine collagen-induced arthritis reveals a potential role for TLR7 in disease maintenance","volume":"14","author":"Alzabin","year":"2012","journal-title":"Arthritis Res Ther"},{"key":"B3","doi-asserted-by":"publisher","first-page":"2386","DOI":"10.3390\/nu15102386","article-title":"Assessment of intestinal permeability and inflammation bio-markers in patients with rheumatoid arthritis","volume":"15","author":"Heidt","year":"2023","journal-title":"Nutrients"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1093\/rheumatology\/keac454","article-title":"Rheumatoid arthritis is associated with increased gut permeability and bacterial translocation which are reversed by inflammation control","volume":"62","author":"Audo","year":"2022","journal-title":"Rheumatol (Oxford)"},{"key":"B5","doi-asserted-by":"publisher","first-page":"6839356","DOI":"10.1155\/2022\/6839356","article-title":"Oral and intestinal bacterial substances associated with disease activities in patients with rheumatoid arthritis: A cross-sectional clinical study","volume":"2022","author":"Kitamura","year":"2022","journal-title":"J Immunol Res"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2023.1092118","article-title":"Gut-joint axis: Gut dysbiosis can contribute to the onset of rheumatoid arthritis via multiple pathways","volume":"13","author":"Romero-Figueroa","year":"2023","journal-title":"Front Cell Infect Microbiol"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.1007610","article-title":"The bridge of the gut-joint axis: Gut microbial metabolites in rheumatoid arthritis","volume":"13","author":"Xu","year":"2022","journal-title":"Front Immunol"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1038\/s41584-021-00585-3","article-title":"The gut-joint axis in rheumatoid arthritis","volume":"17","author":"Zaiss","year":"2021","journal-title":"Nat Rev Rheumatol"},{"key":"B9","doi-asserted-by":"publisher","first-page":"100151","DOI":"10.1016\/j.jhepr.2020.100151","article-title":"Faecal cytokine profiling as a marker of intestinal inflammation in acutely decompensated cirrhosis","volume":"2","author":"Riva","year":"2020","journal-title":"JHEP Rep"},{"key":"B10","doi-asserted-by":"publisher","first-page":"1995","DOI":"10.1038\/s41467-020-15831-7","article-title":"Targeting zonulin and intestinal epithelial barrier function to prevent onset of arthritis","volume":"11","author":"Tajik","year":"2020","journal-title":"Nat Commun"},{"key":"B11","doi-asserted-by":"publisher","first-page":"4693870","DOI":"10.1155\/2019\/4693870","article-title":"Tobacco-derived lipopolysaccharide, not microbial translocation, as a potential contributor to the pathogenesis of rheumatoid arthritis","volume":"2019","author":"Meyer","year":"2019","journal-title":"Mediators Inflamm"},{"key":"B12","doi-asserted-by":"publisher","first-page":"864","DOI":"10.1016\/j.medj.2021.04.013","article-title":"Intestinal barrier dysfunction plays an integral role in arthritis pathology and can be targeted to ameliorate disease","volume":"2","author":"Matei","year":"2021","journal-title":"Med"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1152\/ajpgi.00113.2021","article-title":"Biomarkers for assessment of intestinal permeability in clinical practice","volume":"321","author":"Seethaler","year":"2021","journal-title":"Am J Physiol Gastrointest Liver Physiol"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(17)31472-1","article-title":"Pathogenetic insights from the treatment of rheumatoid arthritis","volume":"389","author":"McInnes","year":"2017","journal-title":"Lancet"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2024.1482214","article-title":"The gut homeostasis-immune system axis: novel insights into rheumatoid arthritis pathogenesis and treatment","volume":"15","author":"Qi","year":"2024","journal-title":"Front Immunol"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1136\/ard.2010.138461","article-title":"2010 rheumatoid arthritis classification criteria: an American College of Rheumatology\/European League Against Rheumatism collaborative initiative","volume":"69","author":"Aletaha","year":"2010","journal-title":"Ann Rheum Dis"},{"key":"B17","doi-asserted-by":"publisher","first-page":"110090","DOI":"10.1016\/j.intimp.2023.110090","article-title":"IFABP2 as a new prognostic biomarker for secondary non-response in rheumatoid arthritis","volume":"119","author":"Zaragoza-Garc\u00eda","year":"2023","journal-title":"Int Immunopharmacol"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.3390\/jcm8071021","article-title":"Intestinal fatty acid binding protein, a biomarker of intestinal barrier, is associated with severity of psoriasis","volume":"8","author":"Sikora","year":"2019","journal-title":"J Clin Med"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2025.1518181","article-title":"Bifidobacterium animalis BD400 protects from collagen-induced arthritis through histidine metabolism","volume":"16","author":"Yang","year":"2025","journal-title":"Front Immunol"},{"key":"B20","doi-asserted-by":"publisher","first-page":"103001","DOI":"10.1016\/j.jaut.2023.103001","article-title":"Gut microbiota in pre-clinical rheumatoid arthritis: From pathogenesis to preventing progression","volume":"141","author":"Lin","year":"2023","journal-title":"J Autoimmun"},{"key":"B21","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1038\/nature18848","article-title":"The microbiota in adaptive immune homeostasis and disease","volume":"535","author":"Honda","year":"2016","journal-title":"Nature"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.20630","article-title":"Fecal cytokines and markers of intestinal inflammation in international travelers with diarrhea due to Noroviruses","volume":"78","author":"Ko","year":"2006","journal-title":"J Med Virol"},{"key":"B23","doi-asserted-by":"publisher","first-page":"372","DOI":"10.1016\/0140-6736(92)91446-F","article-title":"TNF alpha in stool as marker of intestinal inflammation","volume":"340","author":"de Silva","year":"1992","journal-title":"Lancet"},{"key":"B24","doi-asserted-by":"publisher","first-page":"393","DOI":"10.1089\/jir.2019.0011","article-title":"Mechanisms by which the gut microbiota influences cytokine production and modulates host inflammatory responses","volume":"39","author":"Mendes","year":"2019","journal-title":"J Interferon Cytokine Res"},{"key":"B25","doi-asserted-by":"publisher","first-page":"488","DOI":"10.1016\/j.immuni.2015.06.024","article-title":"Growth factor FGF2 cooperates with interleukin-17 to repair intestinal epithelial damage","volume":"43","author":"Song","year":"2015","journal-title":"Immunity"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1601066","article-title":"Epithelial TNF receptor signaling promotes mucosal repair in inflammatory bowel disease","volume":"199","author":"Bradford","year":"2017","journal-title":"J Immunol"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1136\/ard-2023-224878","article-title":"Proinflammatory phenotype of B10 and B10pro cells elicited by TNF-\u03b1 in rheumatoid arthritis","volume":"83","author":"Hu","year":"2024","journal-title":"Ann Rheum Dis"},{"key":"B28","doi-asserted-by":"publisher","first-page":"21","DOI":"10.1016\/bs.ai.2024.07.002","article-title":"IL-17 family cytokines in inflammatory or autoimmune skin diseases","volume":"163","author":"Kong","year":"2024","journal-title":"Adv Immunol"},{"key":"B29","doi-asserted-by":"publisher","first-page":"892","DOI":"10.1016\/j.immuni.2019.03.021","article-title":"The IL-17 family of cytokines in health and disease","volume":"50","author":"McGeachy","year":"2019","journal-title":"Immunity"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2018.01270","article-title":"Cytokine tuning of intestinal epithelial function","volume":"9","author":"Andrews","year":"2018","journal-title":"Front Immunol"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1718902115","article-title":"A cytokine network involving IL-36\u03b3, IL-23, and IL-22 promotes antimicrobial defense and recovery from intestinal barrier damage","volume":"115","author":"Ngo","year":"2018","journal-title":"Proc Natl Acad Sci U S A"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1002\/eji.201646723","article-title":"IL-17 and IL-22 in immunity: Driving protection and pathology","volume":"47","author":"Eyerich","year":"2017","journal-title":"Eur J Immunol"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20221773","article-title":"Diet-mediated constitutive induction of novel IL-4+ ILC2 cells maintains intestinal homeostasis in mice","volume":"220","author":"Cui","year":"2023","journal-title":"J Exp Med"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1038\/nature16527","article-title":"Intestinal epithelial tuft cells initiate type 2 mucosal immunity to helminth parasites","volume":"529","author":"Gerbe","year":"2016","journal-title":"Nature"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1093\/labmed\/lmab034","article-title":"Interleukin-6 and serum\/fecal calprotectin as useful specific markers for monitoring rheumatic diseases: A pilot study","volume":"53","author":"Scapaticci","year":"2022","journal-title":"Lab Med"},{"key":"B36","doi-asserted-by":"publisher","first-page":"104808","DOI":"10.1016\/j.ebiom.2023.104808","article-title":"Intestinal barrier disruption with Plasmodium falciparum infection in pregnancy and risk of preterm birth: a cohort study","volume":"97","author":"Wright","year":"2023","journal-title":"EBioMedicine"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1126\/science.2402637","article-title":"Structure and function of lipopolysaccharide binding protein","volume":"249","author":"Schumann","year":"1990","journal-title":"Science"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1016\/j.atherosclerosis.2013.07.004","article-title":"Serum lipopolysaccharide-binding protein as a marker of atherosclerosis","volume":"230","author":"Serrano","year":"2013","journal-title":"Atherosclerosis"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00003.2008","article-title":"Zonulin and its regulation of intestinal barrier function: the biological door to inflammation, autoimmunity, and cancer","volume":"91","author":"Fasano","year":"2011","journal-title":"Physiol Rev"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.3390\/nu16172949","article-title":"Can faecal zonulin and calprotectin levels be used in the diagnosis and follow-up in infants with milk protein-induced allergic proctocolitis","volume":"16","author":"Czaja-Bulsa","year":"2024","journal-title":"Nutrients"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1002\/ctm2.70136","article-title":"Intestinal oxygen utilisation and cellular adaptation during intestinal ischaemia-reperfusion injury","volume":"15","author":"Archontakis-Barakakis","year":"2025","journal-title":"Clin Transl Med"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2017-314759","article-title":"Increased human intestinal barrier permeability plasma biomarkers zonulin and FABP2 correlated with plasma LPS and altered gut microbiome in anxiety or depression","volume":"67","author":"Stevens","year":"2018","journal-title":"Gut"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms12030463","article-title":"Gut microbiota and biomarkers of intestinal barrier damage in cirrhosis","volume":"12","author":"Efremova","year":"2024","journal-title":"Microorganisms"},{"key":"B44","doi-asserted-by":"publisher","first-page":"6638","DOI":"10.1038\/s41467-023-42456-3","article-title":"Lactate dehydrogenase D is a general dehydrogenase for D-2-hydroxyacids and is associated with D-lactic acidosis","volume":"14","author":"Jin","year":"2023","journal-title":"Nat Commun"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.3390\/nu16081138","article-title":"Selection of fermentation supernatant from probiotic strains exhibiting intestinal epithelial barrier protective ability and evaluation of their effects on colitis mouse and weaned piglet models","volume":"16","author":"Abrehame","year":"2024","journal-title":"Nutrients"},{"key":"B46","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1146\/annurev-pathol-012615-044231","article-title":"Hypoxia and mucosal inflammation","volume":"11","author":"Colgan","year":"2016","journal-title":"Annu Rev Pathol"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1136\/annrheumdis-2021-222035","article-title":"Epithelial HIF2\u03b1 expression induces intestinal barrier dysfunction and exacerbation of arthritis","volume":"81","author":"Wen","year":"2022","journal-title":"Ann Rheum Dis"},{"key":"B48","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41587-024-02503-w","article-title":"Intestinal mucosal barrier repair and immune regulation with an AI-developed gut-restricted PHD inhibitor","author":"Fu","year":"2024","journal-title":"Nat Biotechnol"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2021.682614","article-title":"Claudins: beyond tight junctions in human IBD and murine models","volume":"12","author":"\u010cu\u017ei\u0107","year":"2021","journal-title":"Front Pharmacol"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.3389\/fmed.2022.1049875","article-title":"Difficult-to-treat rheumatoid arthritis: Current concept and unsolved problems","volume":"9","author":"Watanabe","year":"2022","journal-title":"Front Med (Lausanne)"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1093\/rheumatology\/keab209","article-title":"Characteristics of patients with difficult-to-treat rheumatoid arthritis in clinical practice","volume":"60","author":"Takanashi","year":"2021","journal-title":"Rheumatol (Oxford)"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1007\/s10067-019-04579-1","article-title":"Comorbidity burden and clinical characteristics of patients with difficult-to-control rheumatoid arthritis","volume":"38","author":"Batko","year":"2019","journal-title":"Clin Rheumatol"},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1093\/rheumatology\/kex349","article-title":"Difficult-to-treat rheumatoid arthritis: an area of unmet clinical need","volume":"57","author":"de Hair","year":"2018","journal-title":"Rheumatol (Oxford)"},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.1093\/rheumatology\/keab078","article-title":"Healthcare utilization and economic burden of difficult-to-treat rheumatoid arthritis: a cost-of-illness study","volume":"60","author":"Roodenrijs","year":"2021","journal-title":"Rheumatol (Oxford)"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1577590\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,5,19]],"date-time":"2025-05-19T05:16:18Z","timestamp":1747631778000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1577590\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,19]]},"references-count":54,"alternative-id":["10.3389\/fimmu.2025.1577590"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2025.1577590","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,19]]},"article-number":"1577590"}}