Monoclonal antibody 91.9H raised against sulfated mucins is specific for the 3'-sulfated Lewisa tetrasaccharide sequence.
The IgG1hybridoma antibody, 91.9H, was originally raised against sulfated mucins isolated from normal human colonic mucosa. Previous studies have shown that the 91.9H antigen is expressed on normal colonic epithelial cells and the sulfomucins that they produce, but not in the normal small intestine and stomach. Tissue-specific changes occur in 91.9H antigen expression in disease: the antigen diminishes in colonic carcinomas, whereas in regions of gastric mucosa showing intestinal metaplasia and in gastric carcinomas, the antigen is expressed as a "neo-antigen." This report is concerned with elucidation, by the neoglycolipid technology, of the determinant recognized by antibody 91.9H using sulfated and sialyl oligosaccharides of Lewisa(Lea) and Lextypes, and analogs that lack sulfate, sialic acid, or fucose. Binding experiments with the lipid-linked oligosaccharides immobilized on chromatograms or on microwells, and inhibition of binding experiments with free oligosaccharides based on di-, tri- and tetrasaccharide backbones, show that the 91.9H antigenic determinant is based on a trisaccharide backbone, and consists of the 3'-sulfated Leatetrasaccharide sequence, which is a potent ligand for the E- and L-selectins. The antibody gives a relatively low signal with the 3'-sulfated non-fucosylated backbone, and has no detectable cross-reaction with the 3'-sulfated Lexisomer, nor with sialyl-Leaand -Lexanalogues. Antibody 91.9H is a valuable addition, therefore, to the repertoire of reagents for mapping details of the distribution, and determining the relative importance of sulfated and sialyl oligosaccharides as ligands for the selectins, in normal and pathological epithelia and endothelia.
Loveless
R W
RW
The Glycosciences Laboratory, Imperial College School of Medicine, Northwick Park Hospital, Harrow, Middlesex, HA1 3UJ, United Kingdom.
Yuen
C T
CT
Tsuiji
H
H
Irimura
T
T
Feizi
T
T
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism*; D001667:Binding, Competitive; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D003106:Colon / Q000276:immunology / Q000378:metabolism*; D000939:Epitopes / Q000276:immunology / Q000378:metabolism; D006017:Glycolipids / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007983:Lewis Blood Group Antigens / Q000378:metabolism*; D008969:Molecular Sequence Data; D009077:Mucins / Q000276:immunology / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism; D011485:Protein Binding
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006017:Glycolipids; D007983:Lewis Blood Group Antigens; D009077:Mucins; D009844:Oligosaccharides; C012564:sulfomucin