title: Differential effect of E-selectin antibodies on neutrophil rolling and recruitment to inflammatory sites.
abstract: The selectins are inducible adhesion molecules critically important for the inflammatory response. We investigate here the functional effects of three monoclonal antibodies (MoAbs) raised against murine E-selectin (9A9, 10E6, and 10E9.6) on neutrophil recruitment in vivo, leukocyte rolling and circulating leukocyte concentrations in vivo, and adhesion of myeloid cells to E-selectin transfectants and recombinant E-selectin-IgG fusion protein in vitro. MoAbs 9A9 and 10E6 map to the lectin and epidermal growth factor (EGF)-like domains of murine E-selectin, whereas 10E9.6 binds to the consensus repeat region. 10E9.6 blocked neutrophil recruitment in a model of thioglycollate-induced peritonitis in Balb/c mice by more than 90% but had no effect in C57BL/6 mice. 9A9 and 10E6 blocked neutrophil recruitment in this assay only when combined with a P-selectin antibody, 5H1. Neither 9A9 nor 10E9.6 alone blocked leukocyte rolling in tumor necrosis factor-alpha-treated venules of Balb/c mice, but 9A9 almost completely inhibited leukocyte rolling when combined with the function-blocking murine P-selectin MoAb, RB40.34. In contrast, 10E9.6 had no effect on leukocyte rolling in RB40.34-treated Balb/c or C57BL/6 mice. 10E9.6 did not affect adhesion of myeloid cells to E-selectin transfectants or attachment, rolling, and detachment of myeloid cells to murine E-selectin-IgG fusion protein. However, adhesion was completely blocked in the same assays by 9A9. Taken together, these results indicate that E-selectin serves a function, other than rolling, that appears to be critically important for neutrophil recruitment to inflammatory sites in Balb/c mice.
authors.affiliation: Department of Biomedical Engineering, University of Virginia Health Sciences Center, Charlottesville 22908, USA.
authors.forename: C L
authors.initials: CL
authors.lastname: Ramos
authors.forename: E J
authors.initials: EJ
authors.lastname: Kunkel
authors.forename: M B
authors.initials: MB
authors.lastname: Lawrence
authors.forename: U
authors.initials: U
authors.lastname: Jung
authors.forename: D
authors.initials: D
authors.lastname: Vestweber
authors.forename: R
authors.initials: R
authors.lastname: Bosse
authors.forename: K W
authors.initials: KW
authors.lastname: McIntyre
authors.forename: K M
authors.initials: KM
authors.lastname: Gillooly
authors.forename: C R
authors.initials: CR
authors.lastname: Norton
authors.forename: B A
authors.initials: BA
authors.lastname: Wolitzky
authors.forename: K
authors.initials: K
authors.lastname: Ley
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology*; D000918:Antibody Specificity; D019556:COS Cells; D002448:Cell Adhesion; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D019040:E-Selectin / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D000939:Epitopes / Q000276:immunology; D007249:Inflammation / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009504:Neutrophils / Q000502:physiology*; D019007:P-Selectin / Q000276:immunology; D010538:Peritonitis / Q000139:chemically induced / Q000473:pathology; D011993:Recombinant Fusion Proteins / Q000502:physiology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014699:Venules / Q000166:cytology
chemical_list: D000911:Antibodies, Monoclonal; D019040:E-Selectin; D000939:Epitopes; D019007:P-Selectin; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha