A monoclonal antibody to the human leukocyte adhesion molecule intercellular adhesion molecule-2. Cellular distribution and molecular characterization of the antigen.

Abstract

The DNA of the human leukocyte adhesion molecule intercellular adhesion molecule-2 (ICAM-2) was synthesized by the polymerase chain reaction, and a protein A-ICAM-2 fusion protein was expressed in Escherichia coli. The fusion protein was used as immunogen to obtain mAb to ICAM-2. The 6D5 antibody was selected by its reactivity in immunofluorescence with the endothelial cell line Eahy926. The antibody precipitated a 55,000 m.w. glycoprotein from radioactivity surface labeled cells and also reacted in Western blotting. As measured by immunofluorescence flow cytometry, the antibody reacted with lymphoblastoid B cells of normal origin, some Burkitt lymphoma cell lines, vascular endothelial cells, the endothelial cell line Eahy926, 4 and a subpopulation of Con A-stimulated blood mononuclear cells. The two Ig-like domains of ICAM-2 were separately expressed in E. coli, and the antibody was shown to react with the NH2-terminal domain. The antibody inhibited the CD11/CD18-dependent binding of HL-60 promyelocytic leukemia cells to transfected COS-1 cells.