Resident memory T cells (TRMs) help control local immune homeostasis and contribute to tissue-protective immune responses. The local cues that guide their differentiation and localization are poorly defined. We demonstrate that mucosal vascular addressin cell adhesion molecule 1, a ligand for the gut-homing receptor α4β7integrin, in the presence of retinoic acid and transforming growth factor-β provides a co-stimulatory signal that induces blood cluster of differentiation (CD)8+T cells to adopt a TRM-like phenotype. These cells express CD103 (integrin αE) and CD69, the two major TRMcell-surface markers, along with CD101. They also express C-C motif chemokine receptors 5, C-C motif chemokine receptors 9, and α4β7, three receptors associated with gut homing. A subset also expresses E-cadherin, a ligand for αEβ7. Fluorescent lifetime imaging indicated an αEβ7and E-cadherin cis interaction on the plasma membrane. This report advances our understanding of the signals that drive the differentiation of CD8+T cells into resident memory T cells and provides a means to expand these cells in vitro, thereby affording an avenue to generate more effective tissue-specific immunotherapies.