Monoclonal antibodies that recognize lacto-N-fucopentaose III (CD15) react with the adhesion-promoting glycoprotein family (LFA-1/HMac-1/gp 150,95) and CR1 on human neutrophils.
A variety of monoclonal antibodies has been used to study the roles of surface proteins in neutrophil function. Many monoclonal antibodies that bind to human neutrophils react with the oligosaccharide lacto-N-fucopentaose III. Sequential immunoprecipitation of radiolabeled proteins from extracts of neutrophils labeled at the cell surface with 125I, and partial proteolysis peptide mapping studies were used to compare the proteins recognized by several widely used monoclonal antibodies that react with human neutrophils. The monoclonal antibodies that react with lacto-N-fucopentaose III (CD15) immunoprecipitated five distinct neutrophil surface proteins. The data indicate that CD15 monoclonal antibodies react with a subset of the LFA-1/HMac-1/gp 150,95 glycoprotein family as well as with CR1 on human neutrophils. The CD15 antibodies studied differed in their avidities for these proteins. The molecules immunoprecipitated by the CD15 antibodies tested were more resistant to proteolysis than the homologous proteins immunoprecipitated by the other monoclonal antibodies studied that react directly with the alpha M (CD11) or beta (CD18) chains of the LFA-1/HMac-1/gp 150,95 glycoprotein family. Some of the differences in antibody reactivity and protease sensitivity of the membrane proteins recognized by these antibodies may be due to differences in glycosylation. The data suggest that the antibodies studied can detect differences in post-translational modification among copies of certain surface proteins.
Skubitz
K M
KM
Snook
R W
RW
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D000954:Antigens, Surface / Q000276:immunology*; D002448:Cell Adhesion; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D016256:Lewis X Antigen / Q000276:immunology*; D016169:Lymphocyte Function-Associated Antigen-1; D008565:Membrane Proteins / Q000276:immunology*; D009504:Neutrophils / Q000276:immunology*; D009844:Oligosaccharides / Q000276:immunology*; D010449:Peptide Mapping
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D006023:Glycoproteins; D016256:Lewis X Antigen; D016169:Lymphocyte Function-Associated Antigen-1; D008565:Membrane Proteins; D009844:Oligosaccharides