Endothelial leukocyte adhesion molecule 1: direct expression cloning and functional interactions.
A cDNA for endothelial leukocyte adhesion molecule 1 (ELAM-1) was isolated by transient expression in COS-7 cells of a subtracted cDNA library from cytokine-treated human umbilical vein endothelial cells (HUVECs), with selection of ELAM-1-expressing clones by adhesion of transfected cells to the human promyelocytic cell line HL-60. This cloning method requires neither antibody nor purified ligand. ELAM-1-expressing COS cells bind the promyelocytic cell line HL-60 by a Ca2(+)-dependent but temperature-independent mechanism. Although ELAM-1 is homologous to mammalian lectins, its interaction with HL-60 cells is not inhibited by simple carbohydrate structures. ELAM-1-expressing COS cells also bind human neutrophils and the human colon carcinoma cell line HT-29, but not the B-cell line Ramos. However, Ramos cells adhere to cytokine-treated HUVECs but not control HUVECs, confirming the existence of other inducible adhesion molecules. In addition, the binding of HL-60 cells or neutrophils to ELAM-1-expressing COS cells is not inhibited by a monoclonal antibody (60.3) directed to an inhibitory epitope on CD18, indicating that the ELAM-1 ligand, although uncharacterized, is not a member of the CD11/CD18 family.
Biogen, Inc., Cambridge, MA 02142.
C
C
Hession
L
L
Osborn
D
D
Goff
G
G
Chi-Rosso
C
C
Vassallo
M
M
Pasek
C
C
Pittack
R
R
Tizard
S
S
Goelz
K
K
McCarthy
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000235:genetics*; D002460:Cell Line; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D019040:E-Selectin; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology; D015870:Gene Expression; D015723:Gene Library; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000494:pharmacology; D014162:Transfection
D015815:Cell Adhesion Molecules; D019040:E-Selectin; D007375:Interleukin-1; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D004247:DNA