Abstract
The constant interaction between intestinal epithelial cells (IECs) and intraepithelial lymphocytes (IELs) is thought to regulate mucosal barrier function and immune responses against invading pathogens. IELs represent a heterogeneous population of mostly activated and antigen-experienced T cells, but the biological function of IELs and their relationship with IECs is still poorly understood. Here, we describe a method to study T-cell-epithelial cell interactions using a recently established long-term intestinal "enteroid" culture system. This system allowed the study of peripheral T cell survival, proliferation, differentiation and behavior during long-term co-cultures with crypt-derived 3-D enteroids. Peripheral T cells activated in the presence of enteroids acquire several features of IELs, including morphology, membrane markers and movement in the epithelial layer. This co-culture system may facilitate the investigation of complex interactions between intestinal epithelial cells and immune cells, particularly allowing long term-cultures and studies targeting specific pathways in IEC or immune cell compartments.