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In the PREDICT study, we enrolled treatment-na\u00efve pediatric patients with IBD, alongside disorders of gut\u2013brain interaction (DGBI) controls and healthy donors, and profiled their intestinal and blood-derived T cells using single-cell RNA sequencing (scRNA-seq). Across 107 participants, we identify a unique population of cytotoxic CD4\u207a T cells (CD4 CTL) enriched in the inflamed gut of patients with Crohn\u2019s disease (CD) and ulcerative colitis. CD4 CTLs are clonally expanded and express cytotoxic effector molecules and\n                  <jats:italic>IFNG<\/jats:italic>\n                  , consistent with antigen-driven activation. Cell\u2013cell interaction analyses implicate macrophage-derived IL-27 as the top candidate for CD4 CTL differentiation, and IL-27 blockade in a mouse model limits CD4 CTL formation. Notably, elevated CD4 CTL frequencies in gut and peripheral blood at diagnosis are associated with subsequent poor outcome of anti-TNF therapy in pediatric CD. Findings in our identification cohort are validated in an independent cohort and through reanalysis of published datasets. Importantly, we designed a simple flow cytometry panel to isolate blood CD4\n                  <jats:sup>+<\/jats:sup>\n                  CXCR6\n                  <jats:sup>+<\/jats:sup>\n                  CD27\n                  <jats:sup>-<\/jats:sup>\n                  T cells, which displayed a CD4 CTL transcriptional phenotype. Together, our results link CD4 CTLs to anti\u2013TNF nonresponse and support their potential as an early, blood-accessible biomarker for treatment stratification in pediatric CD.\n                <\/jats:p>","DOI":"10.64898\/2026.06.16.26354238","type":"posted-content","created":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T23:20:24Z","timestamp":1782516024000},"source":"Crossref","is-referenced-by-count":0,"title":["Local and circulating cytotoxic CD4\u207a T cells are early markers of disease activity in pediatric Crohn\u2019s disease"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8768-9039","authenticated-orcid":false,"given":"Veronika","family":"Niederlova","sequence":"first","affiliation":[{"name":"Division of Gastroenterology, Hepatology, and Nutrition, Boston Children\u2019s Hospital, Boston, MA 02115, USA"},{"name":"Laboratory of Adaptive Immunity, Institute of Molecular Genetics of the Czech Academy of Sciences, Prague, 142 20, Czech 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