Crosstalk with infant derived Th17 cells promotes maturation of intestinal epithelial cells in an enteroid model through IL-22 signaling

The use of 3D human intestinal organoid (HIO) models is growing in biomedical research due to their ability to mimic human tissue for studying diseases, developing biomarkers, and testing therapies. However, current models of the infant gut are limited, focusing mostly on necrotizing enterocolitis, leaving gaps in our understanding of healthy infant gut development. Additionally, achieving full differentiation and maturation of secretory cells, especially Paneth cells, remains challenging. Our studies suggest IL-22 can enhance maturation, though its effects on infant enteroids remain unexplored. In this study, infant-derived ileal enteroids were cultured with Th17 supernatant or IL-22, and their development was assessed via immunofluorescence staining and RNA sequencing. Our results showed that neonatal Th17 cell-derived IL-22 supported the growth and maturation of infant enteroids, including Paneth cell differentiation. IL-22 induced gene expression changes related to cellular identity, maturation, immune response, and barrier function, as revealed by RNAseq analysis. Human infant-derived ileal enteroids were cultured with recombinant IL-22. Morphological and functional changes were assessed using immunofluorescence staining and RNA sequencing, respectively, to characterize the developmental and functional profiles of intestinal epithelial cells (IECs) and to investigate the impact of IL-22 co-culture on IEC differentiation and maturation.