Monoclonal antibody characterization of a chymotrypsin-like molecule on neutrophil membrane associated with cellular activation.
Monoclonal antibody 1-15 (Ab 1-15), is a murine anti-human neutrophil (PMN) IgG1 that inhibits PMN effector responses to N-formyl-met-leu-phe (FMLP) and phorbol myristate acetate. In this study, the effects of Ab 1-15 on PMN membrane-related functions were characterized: Ab 1-15 inhibited PMN superoxide (O-2) response to FMLP by 60% (P less than 0.005) without effect on the onset or duration of O-2 production. This inhibition of O-2 response was associated with a significant inhibition of PMN chymotrypsin-like, but not trypsin-like, protease activity. Cell fractionation studies indicated the presence of an Ab 1-15 inhibitable, chymotryptic neutral protease activity in PMN membranes. In studies of Ab 1-15 effects on membrane-related second messenger pathways, Ab 1-15 augmented both FMLP- and isoproterenol-induced intracellular cAMP accumulation, whereas alpha-chymotrypsin decreased PMN cAMP response to these stimuli. Our data suggest that the function-inhibiting, anti-PMN monoclonal Ab 1-15 defines a PMN chymotryptic enzyme on the membrane surface that is involved in regulation of two membrane-related functions, O-2 generation and cAMP generation.
D000911:Antibodies, Monoclonal; D002462:Cell Membrane / Q000201:enzymology; D002918:Chymotrypsin / Q000032:analysis*; D000242:Cyclic AMP / Q000378:metabolism; D004151:Dipeptides / Q000494:pharmacology; D006801:Humans; D008213:Lymphocyte Activation; D009504:Neutrophils / Q000201:enzymology*; D010179:Pancreas / Q000201:enzymology; D010447:Peptide Hydrolases / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D004151:Dipeptides; D013481:Superoxides; C017330:N-formylmethionylphenylalanine; D000242:Cyclic AMP; D010447:Peptide Hydrolases; D002918:Chymotrypsin; D013755:Tetradecanoylphorbol Acetate