Eosinophil Specialization Is Regulated by Exposure to the Esophageal Epithelial Microenvironment

Distinct subsets of eosinophils are reported in inflammatory and healthy tissues, yet the functions of uniquely specialized eosinophils and the signals that elicit them, particularly in eosinophilic esophagitis (EoE), are not well understood. Herein, we report an ex-vivo system wherein freshly isolated human eosinophils were cocultured with esophageal epithelial cells and disease-relevant pro-inflammatory (IL-13) or pro-fibrotic (TGF-β) cytokines. Our results demonstrate that disease-relevant pro-inflammatory and pro-fibrotic signals present in the esophagus of EoE patients cause distinct profiles of eosinophil activation and gene expression. Immediately after isolation or following 96h of coculture with esophageal epithelial cells with- and without IL-13 or TGFb, eosinophils were lysed in TriPure and RNA was extracted with phenol-chloroform followed by Zymogen Quick RNA Microprep Kit. RNAseq libraries were generated as described in PMID 28041958 and 32362324.