In Vitro Differentiation from Naive to Mature E-Selectin Binding CD4 T Cells: Acquisition of Skin-Homing Properties Occurs Independently of Cutaneous Lymphocyte Antigen Expression

Abstract

We previously showed that skin-homing CD4 T cells in peripheral blood can be subdivided into three populations on the basis of the expression pattern of the cutaneous lymphocyte Ag (CLA) and fucosyltransferase VII (FucT-VII): FucT-VII+CLA−, FucT-VII+CLA+, and FucT-VII−CLA+. In view of the known late appearance of CLA during T cell differentiation, T cells programmed to attain skin-homing properties may start to generate E-selectin-binding epitopes at early stages of differentiation before induction of CLA expression. To this end, the in vitro differentiation from naive to CLA+ memory T cells was followed after activation with anti-CD3 mAb. Here we demonstrate that naive skin-homing CD4 T cell precursors undergo a linear differentiation process from the FucT-VII+CLA− phenotype to the FucT-VII+CLA+ phenotype and eventually to the FucT-VII−CLA+ phenotype. The appearance of the FucT-VII+CLA− subset coincided with or could be immediately followed by the generation of E-selectin binding epitopes, and even after E-selectin-binding epitopes were no longer detectable, CLA remained expressed for prolonged periods of time, suggesting that induction of functional E-selectin ligands depends primarily on the expression of FucT-VII, but not CLA. Immunofluorescence and confocal microscopy studies of these T cells confirm that most E-selectin ligands were found independently of CLA expression.