Epigenetically controlled ZEB2 expression promotes the cytotoxic potential of CMV-specific CD8+ T cells

Zinc finger E-box binding protein 2 (ZEB2) is a key factor in the differentiation of naive CD8+ T cells into effector and memory T cells. However, the precise regulatory role of ZEB2 in cytotoxic CD8+ T cells remains unknown. Our recent DNA methylation analysis of cytomegalovirus (CMV)-specific human CD8+ T cells revealed two differentially methylated regions (DMRs) within the ZEB2 locus. We showed that these ZEB2 DMRs undergo pronounced demethylation during T cell differentiation. In particular, terminally differentiated CD8+ T cells and cytotoxic CD4+ T cells show almost complete demethylation. A ZEB2 knockout approach in CD8+ effector T cells followed by RNA-seq analysis revealed an altered gene expression profile, affecting genes related to cell-cell adhesion and impairing cytotoxicity in CMV-specific killing assays. Our data show that ZEB2 expression contributes to the differentiation of naive CD8+ T cells into effector and memory T cells and regulates the functional properties of virus-specific cytotoxic CD8+ T cells.