The cutaneous lymphocyte antigen is an essential component of the L-selectin ligand induced on human vascular endothelial cells.
L-selectin mediates leukocyte rolling on vascular endothelium during inflammation. Although vascular endothelium can be activated with inflammatory cytokines to express functional L-selectin ligands, these ligands have not been well characterized. In this study, fucosyltransferase VII cDNA (Fuc-TVII) transfection of the EA.hy926 human vascular endothelial cell line (926-FtVII) induced functional L-selectin ligand expression and expression of sialyl Lewisx (sLex), as defined by HECA-452 (cutaneous lymphocyte antigen; CLA) and CSLEX-1 mAbs. Cytokine activation of human umbilical vein endothelial cells (HUVEC) also induced functional L-selectin ligand expression, with increased CLA expression and Fuc-TVII transcription. The majority of L-selectin-dependent lymphocyte attachment to activated HUVEC and 926-FtVII cells was blocked specifically by treating the endothelial cells with the HECA-452 mAb, but not the CSLEX-1 mAb. CLA-bearing ligands on vascular endothelium also required sulfation and appropriate molecular scaffolds for functional activity, but were distinct from the L-selectin ligands previously identified by the MECA-79 mAb. These findings demonstrate that the HECA-452- defined antigen, CLA, is an essential carbohydrate component of vascular L-selectin ligands.
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Department of Immunology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Delahunty
M D
MD
Ding
H
H
Luscinskas
F W
FW
Tedder
T F
TF
D000911:Antibodies, Monoclonal / Q000276:immunology; D000945:Antigens, Differentiation, T-Lymphocyte; D000951:Antigens, Neoplasm; D002448:Cell Adhesion / Q000276:immunology; D002460:Cell Line; D004730:Endothelium, Vascular / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D005647:Fucosyltransferases / Q000235:genetics; D006801:Humans; D007075:Immunoglobulin M / Q000235:genetics / Q000276:immunology; D007249:Inflammation / Q000276:immunology; D019041:L-Selectin / Q000276:immunology*; D007962:Leukocytes / Q000276:immunology; D008024:Ligands; D008214:Lymphocytes / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000276:immunology*; D009844:Oligosaccharides / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics; D000080506:Sialyl Lewis X Antigen; D014162:Transfection / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte; D000951:Antigens, Neoplasm; C063419:CTAGE1 protein, human; D007075:Immunoglobulin M; D008024:Ligands; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D011993:Recombinant Fusion Proteins; D000080506:Sialyl Lewis X Antigen; D014409:Tumor Necrosis Factor-alpha; D019041:L-Selectin; D005647:Fucosyltransferases