L-selectin ligands in rat high endothelium: multivalent sialyl Lewis x glycans are high-affinity inhibitors of lymphocyte adhesion.
Lymphocyte homing is initiated by their tethering to and rolling on the high endothelium and is followed by extravasation into the lymph nodes. We show here that glycosylated cell adhesion molecule-1 (GlyCAM-1), CD34, and sialyl Lewis x (sLex) are present on rat lymph node high endothelium analyzed by using monoclonal antibodies. alpha (1,3)fucosyltransferase VII (Fuc-TVII), the last enzyme involved in the synthesis of the sLex sequence is also expressed on the rat lymph node high endothelium. We have synthesized a family of sLex-decorated oligosaccharide structures and used them to inhibit lymphocyte binding to high endothelium in the Stamper-Woodruff assay. Monovalent sLex, branched di- and tetravalent sLex, as well as a linear tetravalent sLex significantly reduce lymphocyte binding to endothelium. The branched and linear forms of tetravalent sLex were clearly superior inhibitors of the L-selectin-dependent lymphocyte adhesion, with IC50 values in low nanomolar range. In contrast, the fucose-free analogs having the same charge and approximately the same size as the corresponding sLex glycans had no effect on lymphocyte binding and served as negative controls. Taken together, these data show the crucial importance of sLex in the endothelial ligands for L-selectin. Furthermore, we suggest that L-selectin acts as an oligomer on the lymphocyte surface as it binds multivalent sLex glycans.
Toppila
S
S
Haartman Institute, Department of Bacteriology and Immunology, University of Helsinki, Finland.
Lauronen
J
J
Mattila
P
P
Turunen
J P
JP
Penttilä
L
L
Paavonen
T
T
Renkonen
O
O
Renkonen
R
R
D000818:Animals; D018395:CA-19-9 Antigen; D002240:Carbohydrate Sequence; D002448:Cell Adhesion / Q000187:drug effects / Q000276:immunology; D004729:Endothelium, Lymphatic / Q000378:metabolism*; D005732:Gangliosides / Q000737:chemistry / Q000494:pharmacology*; D019041:L-Selectin / Q000502:physiology*; D007983:Lewis Blood Group Antigens / Q000737:chemistry / Q000494:pharmacology*; D008198:Lymph Nodes / Q000378:metabolism; D008214:Lymphocytes / Q000276:immunology* / Q000502:physiology*; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000276:immunology; D051381:Rats
D018395:CA-19-9 Antigen; D005732:Gangliosides; D007983:Lewis Blood Group Antigens; D019041:L-Selectin; C064564:sialyl Le(a) ganglioside