Characterization of murine E-selectin expression in vitro using novel anti-mouse E-selectin monoclonal antibodies.
A recombinant phage containing the structural exons for mouse E-selectin has been isolated and characterized. Utilizing PCR techniques the lectin and egf domains were fused to form an artificial cDNA for expression in eukaryotic cells. Transient expression in COS cells demonstrated the lectin and egf domains were sufficient to mediate the binding of mouse and human neutrophils as well as HL60 cells. Recombinant soluble mouse E-selectin was purified and used to immunize rats to generate mAbs specific to mouse E-selectin. A panel of mAbs directed against mouse E-selectin was characterized including five that inhibit the adhesion of HL60 cells or mouse neutrophils to COS cells expressing the mouse lectin/egf domains. These mAbs have been used to characterize the expression and function of E-selectin on cytokine stimulated eEnd.2 murine endothelial cells.
Norton
C R
CR
Department of Inflammation/Autoimmune Diseases, Hoffmann-La Roche Inc., Nutley, N.J. 07110.
Rumberger
J M
JM
Burns
D K
DK
Wolitzky
B A
BA
D000818:Animals; D000911:Antibodies, Monoclonal; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology; D002460:Cell Line; D004247:DNA / Q000235:genetics; D019040:E-Selectin; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism; D005091:Exons; D015870:Gene Expression; D006801:Humans; D018799:Intercellular Adhesion Molecule-1; D007375:Interleukin-1 / Q000494:pharmacology; D015473:Leukemia, Promyelocytic, Acute; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D019010:Vascular Cell Adhesion Molecule-1
D000911:Antibodies, Monoclonal; D015815:Cell Adhesion Molecules; D019040:E-Selectin; D007375:Interleukin-1; D008070:Lipopolysaccharides; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D004247:DNA