A SV-40 immortalized murine endothelial cell line from peripheral lymph node high endothelium expresses a new alpha-L-fucose binding protein.
Endothelial cells from mouse peripheral lymph nodes were immortalized by cationic liposome-mediated transfection using a plasmid construct containing both the gene coding for the large T antigen of simian virus 40 and a geneticin resistance gene suitable for selection. A cell line (HECa10) was isolated on the basis of its capacity to specifically bind fucoside carrying glycoconjugates; these cells present the main characteristics of endothelial cells: production of angiotensin converting enzyme and of factor VIII-related antigen. Upon stimulation, they express E-selectin which binds oligosaccharides containing the Lewisx determinant (Fuc alpha 3[Gal beta 4 GlcNAc beta 3Gal beta) and the MECA 79 addressin which is characteristic for the peripheral lymph node high endothelium and is a L-selectin ligand. HECa10 cells, as well as peripheral lymph node high endothelial cells in primary culture, express a second fucoside binding protein which differs from E-selectin. Indeed, this new fucoside-binding protein is constitutively expressed on unstimulated cells while E-selectin is not. Furthermore, HECa10 cells mediate selective lymphoid cell adhesion in a selectin/addressin-dependent mechanism, mainly inhibited by MECA 79 antibody and, in a fucose-binding lectin-dependent manner, mainly inhibited by the specific neoglycoprotein.
Bizouarne
N
N
Laboratoire de Biochimie des Glycoconjugués et Lectines Endogènes, Université d'Orléans, France.
Denis
V
V
Legrand
A
A
Monsigny
M
M
Kieda
C
C
D000818:Animals; D000954:Antigens, Surface / Q000378:metabolism; D015415:Biomarkers; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002461:Cell Line, Transformed; D019040:E-Selectin; D004729:Endothelium, Lymphatic / Q000276:immunology / Q000378:metabolism*; D005643:Fucose / Q000378:metabolism*; D037102:Lectins / Q000276:immunology / Q000378:metabolism*; D008565:Membrane Proteins; D051379:Mice; D013539:Simian virus 40; D014407:Tumor Cells, Cultured / Q000276:immunology
D000954:Antigens, Surface; D015415:Biomarkers; D015815:Cell Adhesion Molecules; D019040:E-Selectin; C069886:L-selectin counter-receptors; D037102:Lectins; D008565:Membrane Proteins; C027112:fucose-binding lectin; D005643:Fucose