Expression of Cutaneous Lymphocyte-Associated Antigen Regulated by a Set of Glycosyltransferases in Human T Cells: Involvement of α1,3-Fucosyltransferase VII and β1,4-Galactosyltransferase I

Abstract

Cutaneous lymphocyte-associated antigen (CLA), which plays a key part in skin homing of human CD4+ memory T cells via CLA/E-selectin binding, is upregulated by IL-12 and downregulated by IL-4. Although alpha1,3-fucosyltransferase VII is essential for synthesis of the CLA carbohydrate epitope, little is known about how the CLA expression is regulated by a number of glycosyltransferases. A 6 wk long-term culture for the in vitro differentiation of naïve Th cells to memory Th1 cells was employed. By repeated activation in the presence of IL-12, naïve T cells differentiated into memory Th1 cells, resulting in the upregulation of CLA expression. The switching of cytokine from IL-12 to IL-4 at three cycles resulted in a marked downregulation of CLA. The transcript levels of 16 glycosyltransferases and P-selectin glycoprotein ligand-1, all considered to be potentially involved in CLA synthesis, were determined after each cycle. The level of CLA expression was well correlated with the amounts of alpha1,3-fucosyltransferase VII and beta1,4-galactosyltransferase I. Both were upregulated by IL-12 and downregulated by IL-4. In particular, alpha1,3-fucosyltransferase VII levels decreased markedly in the presence of IL-4. P-selectin glycoprotein ligand-1 and Core 2 beta1, 6-N-acetylglucosaminyltransferase were progressively up-regulated by repeated IL-12 stimulation, but they were not downregulated by IL-4. The transcript levels of some genes examined were constitutive without any correlation to CLA expression. These results suggest that the level of CLA expression is determined by alpha1, 3-fucosyltransferase VII and beta1,4-galactosyltransferase I, the other enzymes merely participating in the synthesis of CLA. In peripheral blood mononuclear cells, IL-12 and IL-4 profoundly upregulated and downregulated the alpha1,3-fucosyltransferase VII transcripts, respectively, but not the beta1,4-galactosyltransferase I ones, within only 2 h of in vitro culture. This suggested that alpha1,3-fucosyltransferase VII is transcriptionally regulated directly by IL-12 and IL-4.

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

“…It remains unknown, however, where, when, and how the commitment of T cells to the skin-homing phenotype is determined; we cannot be sure whether the induction of CLA expression on these T cells occurred in temporal correlation with the acquisition of skin-homing properties or whether the induction of CLA appeared afterward. In this regard, recent kinetic studies have demonstrated that optimal CLA induction on T cells is a relatively slow process even in serum-free medium highly permissive for CLA expression (18,19), and we have also seen a similar requirement for 5 days or more to generate CLA-positive T cells from naive T cells in vitro (20,21). Nevertheless, the acquisition of skin-homing properties has been thought to occur rapidly upon activation.…”

Activation of Murine CD4+ and CD8+ T Lymphocytes Leads to Dramatic Remodeling ofN-Linked Glycans

“…An inherent problem with microarray data is that some probe sets "do not work". Two genes in particular, FucT-VII and core 2 GlcNAc transferase, known to be involved in Sialyl-Lewis X selectin ligand formation on O-linked glycans of CD4 Th1 T cells (11)(12)(13)(14)(15)(16)68), were not detected (see supplemental data). However, our recent experience is that redesign of the probe sets can largely remedy this problem and should lead to increasingly powerful data sets.…”

“…The additional increased expression of fucosyl-and sialyltransferases in Th1 cells promotes synthesis of the selectin ligand sialyl-Lewis X (NeuAc␣2-3Gal␤1-4[Fuca1-3]GlcNAc), while Th2 helper cells lack sialyl-Lewis X sequences because a key fucosyltransferase, Fuc-T VII, is not expressed. Such differential glycosylation accounts for the selectin-mediated recruitment of CD4 Th1 cells to sites of inflammation (11)(12)(13)(14)(15)(16)(17).…”

Immunohistochemical Detection of Skin-Homing T Cells Expressing Fucosyltransferase VII (Fuc-TVII) In Vitro and In Situ

“…Therefore, expression of Fuc-TVII is an attractive candidate for identifying skin "homing" T cells. However, analyses of Fuc-TVII expression in human T cells have been performed only at the mRNA level (Nakayama et al, 2000) because of the lack of mAb. In this study Fuc-TVII was for the first time visualized in individual cells by using a novel mAb that we developed.…”