Liberation of a neutrophil enzyme-releasing peptide from the surface of human alveolar macrophages.
The human alveolar macrophage product, enzyme-releasing peptide (ERP), has a molecular mass of 8,000 Da, and releases azurophilic and specific granule constituents from neutrophils. A murine monoclonal anti-ERP antibody (12E10H), previously used to show a lack of antigenic identity between ERP and C5a, interleukin 1, tumor necrosis factor, and gamma-interferon, showed no cross-reactivity with interleukin 8. 12E10H and a fluorescein-labeled second antibody were used to visualize ERP on the macrophage surface. ERP was removed from alveolar macrophages by a 3-min incubation with 5 X 10(-7) M bovine pancreatic trypsin at 37 degrees C. The washed trypsinized cells could readhere to plastic and exclude trypan blue. Dilution of the trypsin-derived ERP released myeloperoxidase from cytochalasin-B-treated neutrophils dose dependently. The enzyme-releasing ability of the trypsin-derived material was removed by immunoprecipitation using antibody 12E10H bound to Staphylococcal protein A Sepharose 4B. The estimated molecular mass of the trypsin-derived ERP (by molecular sieve chromatography on HPLC) was approximately 8,500 Da. Other proteases (plasmin, thrombin, and cathepsin G) also released ERP from the cell surface, but the ERP was not an active secretagogue for neutrophils. However, macrophages cultured with protease inhibitors did not show decreased ERP accumulation in the medium. Our data indicate that ERP exists on the surface of human alveolar macrophages and can be released by proteases found within the lung environment in some disease states.
Miller
E J
EJ
Department of Biochemistry, University of Texas Health Center, Tyler 75710.
MacArthur
C K
CK
Gray
L D
LD
Cohen
A B
AB
D000911:Antibodies, Monoclonal; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D003571:Cytochalasin B / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D008175:Lung Neoplasms / Q000473:pathology; D008264:Macrophages / Q000502:physiology*; D008970:Molecular Weight; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology*; D010455:Peptides / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology / Q000502:physiology; D014357:Trypsin
D000911:Antibodies, Monoclonal; D010455:Peptides; D003571:Cytochalasin B; D014357:Trypsin