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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 ¹

Ryo Takahashi^2,*, Yoshiko Mizukawa, Yoshimi Yamazaki, Kazuhito Hayakawa, Jun Hayakawa, Akihiko Kudo and Tetsuo Shiohara^*,

^* Division of Flow Cytometry, Department of Dermatology, and Department of Anatomy, Kyorin University School of Medicine, Tokyo, Japan

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.

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