title: Structural and functional studies of the endothelial activation antigen endothelial leucocyte adhesion molecule-1 using a panel of monoclonal antibodies.
abstract: We have produced a panel of mAb to the endothelial activation Ag endothelial leucocyte adhesion molecule-1 (ELAM-1), using both a conventional immunization protocol and one involving immunosuppression. By constructing ELAM-1 mutants we have demonstrated that seven of these antibodies recognize epitopes within the lectin domain of ELAM-1 and that one binds within the complement regulatory protein domains. These studies also suggest that the EGF-like domain is important in maintaining the conformation of the neighbouring lectin domain. In functional studies, U937 cells bound to Cos cells expressing either ELAM-1 or ELAM-1 with the complement regulatory protein domains deleted. No adhesion was observed to Cos cells expressing ELAM-1 mutants lacking either the lectin or EGF-like domains. The fact that antibodies directed against the lectin domain can inhibit adhesion suggest that this domain is directly involved in cell binding.
authors.affiliation: British Biotechnology, Cowley, Oxford, U.K.
authors.forename: R
authors.initials: R
authors.lastname: Pigott
authors.forename: L A
authors.initials: LA
authors.lastname: Needham
authors.forename: R M
authors.initials: RM
authors.lastname: Edwards
authors.forename: C
authors.initials: C
authors.lastname: Walker
authors.forename: C
authors.initials: C
authors.lastname: Power
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D001667:Binding, Competitive; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D019040:E-Selectin; D004730:Endothelium, Vascular / Q000276:immunology*; D000939:Epitopes; D006801:Humans; D011994:Recombinant Proteins / Q000276:immunology; D013329:Structure-Activity Relationship
chemical_list: D000911:Antibodies, Monoclonal; D015815:Cell Adhesion Molecules; D019040:E-Selectin; D000939:Epitopes; D011994:Recombinant Proteins