Differential effect of E-selectin antibodies on neutrophil rolling and recruitment to inflammatory sites

• Published in: Blood Vol. 89; no. 8; pp. 3009 - 3018
• Main Authors: RAMOS, C. L, KUNKEL, E. J, LEY, K, LAWRENCE, M. B, JUNG, U, VESTWEBER, D, BOSSE, R, MCINTYRE, K. W, GILLOOLY, K. M, NORTON, C. R, WOLITZKY, B. A
• Format: Journal Article
• Language: English
• Published: Washington, DC The Americain Society of Hematology 15-04-1997
• Subjects: Animals; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - pharmacology; Antibody Specificity; Biological and medical sciences; Cell Adhesion; Chemotaxis, Leukocyte - drug effects; COS Cells; E-Selectin - genetics; E-Selectin - immunology; E-Selectin - physiology; Epitopes - immunology; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Immunobiology; Inflammation - pathology; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Myeloid cells: ontogeny, maturation, markers, receptors; Neutrophils - physiology; P-Selectin - immunology; Peritonitis - chemically induced; Peritonitis - pathology; Polynuclears; Recombinant Fusion Proteins - physiology; Tumor Necrosis Factor-alpha - pharmacology; Venules - cytology; Site; Leukocyte; E Selectin; Rodentia; Granulocyte; Cell adhesion molecule; Inflammation; Monoclonal antibody; Recruitment; Vertebrata; Mammalia; Mouse; Animal; Neutrophil
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.v89.8.3009

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.