title: Sulfated sialyl Lewis X, the putative L-selectin ligand, detected on endothelial cells of high endothelial venules by a distinct set of anti-sialyl Lewis X antibodies.
abstract: Endothelial cells of high endothelial venules (HEV) in human peripheral lymph nodes expressed a distinct type of sialyl Lewis X antigen, which was detected preferentially with a set of anti-sialyl Lewis X antibodies, 2F3, 2H5 and HECA-452 in immunohistochemistry, while another set of anti-sialyl Lewis X antibodies, FH-6 and CSLEX-1, failed to detect it. The adhesion of cells expressing L-selectin to HEV was inhibited by members of the former set of antibodies in Stamper-Woodruff assays performed on frozen sections of human peripheral lymph nodes. Transfection of a cultured endothelial cell line with a human alpha1-->3 fucosyltransferase, Fuc-T VII, resulted in the expression of a distinct type of sialyl Lewis X antigen having the reactivity similar to that of HEV; i.e., the antigen appearing on the transfectant clone was detectable only with the set of 2F3, 2H5 and HECA-452, but not with the set of FH-6 and CSLEX-1. Treatment of transfectant cells with sodium chlorate, a metabolic inhibitor of sulfation, resulted in reactivity to the members of the latter set of antibodies, suggesting that sulfation of sialyl Lewis X moiety was the cause of the discrepancy in the reactivity of the anti-sialyl Lewis X antibodies. When tested against various authentic sulfated sialyl Lewis X determinants, 6-sulfo sialyl Lewis X and 6,6'-bis-sulfo sialyl Lewis X were found to be reactive to the antibodies, 2F3, 2H5 and HECA-452, but not with antibodies FH-6 and CSLEX-1, suggesting that the distinct type of sialyl Lewis X determinant on the HEV endothelial cells and Fuc-T VII-transfected endothelial cell clone are mainly 6-sulfo and/or 6,6'-bis-sulfo sialyl Lewis X determinants.
authors.affiliation: Program of Experimental Pathology, Aichi Cancer Center, Research Institute, Nagoya, Japan.
authors.forename: C
authors.initials: C
authors.lastname: Mitsuoka
authors.forename: N
authors.initials: N
authors.lastname: Kawakami-Kimura
authors.forename: M
authors.initials: M
authors.lastname: Kasugai-Sawada
authors.forename: N
authors.initials: N
authors.lastname: Hiraiwa
authors.forename: K
authors.initials: K
authors.lastname: Toda
authors.forename: H
authors.initials: H
authors.lastname: Ishida
authors.forename: M
authors.initials: M
authors.lastname: Kiso
authors.forename: A
authors.initials: A
authors.lastname: Hasegawa
authors.forename: R
authors.initials: R
authors.lastname: Kannagi
mesh_terms: D000818:Animals; D000937:Antigen-Antibody Reactions; D001667:Binding, Competitive / Q000276:immunology; D002448:Cell Adhesion / Q000276:immunology; D002460:Cell Line; D004729:Endothelium, Lymphatic / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D000939:Epitopes / Q000096:biosynthesis; D005647:Fucosyltransferases / Q000235:genetics; D006801:Humans; D007518:Isoantibodies / Q000737:chemistry* / Q000494:pharmacology; D015458:Leukemia, T-Cell; D016256:Lewis X Antigen / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D008198:Lymph Nodes / Q000276:immunology / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D013431:Sulfates / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D000939:Epitopes; D007518:Isoantibodies; D016256:Lewis X Antigen; D013431:Sulfates; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase