J Leukoc Biol. 2026 Jun 8;118(6):qiag075. doi: 10.1093/jleuko/qiag075.
ABSTRACT
Autologous cell therapy is a patient-embraced approach to inflammatory disease. Murine models demonstrate that alternatively activated (or regulatory) macrophages can reduce the severity of disease, and human macrophages (M0) educated with interleukin-4 (M(IL4)) limit murine colitis. M0 and M(IL4) from healthy donors and individuals with Crohn’s disease (active or remission) were assessed by qPCR, characterized by a time-of-flight mass cytometry (CyTOF) comparison of 16 myeloid marker molecules expression, and subtypes of M(IL4)s tested in the murine dinitrobenzene sulfonic acid (DNBS) model of colitis and in vitro co-culture with T cells. Most macrophage preparations responded to IL-4 by increasing CD206, CCL18 and RAMP1 mRNA expression and produced mediators that promoted epithelial repair in a wound assay with the human CaCo2 epithelial line. Single-cell clustering by FlowSOM defined 8 distinct subpopulations (meta-clusters) of macrophages, the proportions of which were unaffected by biological sex, age or cryopreservation: meta-cluster 4 (CD206highPD-L1highHLA-DRhigh) represented 20 to 40% of M0s, with a higher proportion in controls compared with active Crohn’s disease. IL-4 treatment significantly expanded meta-cluster 4 in all M0s. Transfer of M(IL4)-CD206high (ie, predominantly meta-cluster 4) into rag1-/- mice significantly reduced DNBS-induced colitis, and to a greater extent than M(IL4)-CD206low from the same individual. IL-10 production was increased in M(IL4)-CD206high-T-cell co-cultures. Having confirmed the prohealing effect of the human M(IL4), the predominant meta-cluster M(IL4)-CD206high was found to have superior anticolitic effect. Reduced numbers of M0-CD206high and M(IL4)-CD206high cells in some individuals with Crohn’s disease suggest that their absence may contribute to more severe inflammation or reduced healing capacity.
PMID:42367148 | DOI:10.1093/jleuko/qiag075