A SV‐40 immortalized murine endothelial cell line from peripheral lymph node high endothelium expresses a new α‐L‐fucose binding protein

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Abstract
Summry— Endothelial cells from mouse peripheral lymph nodes were immortalized by cationic liposome‐mediate 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 (HECal10) 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 VII‐related antigen. Upon stimulation, they express E‐selectin which binds oligosaccharides containing the Lewisx determinant (Fucα3[Galβ4 GlcNacβ3Galβ) and the MECA 79 addressin which is characteristics for the peripheral lymph node high endothelium and is a L‐selectin. 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 cells 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 specifc neoglycoprotein.

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