Multiplexed ChipCytometry for immune cell profiling in colonic tissues of patients with inflammatory bowel disease

Immune cell infiltration and activation are key drivers of inflammatory bowel disease (IBD), including ulcerative colitis (UC) and Crohn’s disease (CD). Sphingosine-1-phosphate receptor 1 (S1P1) signaling regulates lymphocyte trafficking and inflammatory responses. We developed and applied a multiplex ChipCytometry platform to characterize immune cell populations and S1P1 expression in formalin-fixed paraffin-embedded colonic tissues from healthy volunteers (HV, n = 2), patients with UC ( n = 5), and patients with CD ( n = 5). A 20-marker panel was used for tissue imaging, and immune cell subsets and S1P1 expression were quantified by bivariate gating. The assay enabled high-dimensional immune profiling of colonic tissues. Compared with HV tissues, UC samples showed increased T-cell infiltration, particularly helper T cells, and a higher abundance of S1P1-positive immune cells. CD samples exhibited increased plasma cell abundance, whereas natural killer cell frequencies were reduced in both UC and CD tissues. Among S1P1-positive immune cells, T cells, especially helper T cells, constituted a larger proportion in diseased tissues than in controls. These findings demonstrate the utility of ChipCytometry for spatially resolved imaging and quantification of immune-cell populations and characterization of S1P1-associated cellular populations in IBD.