title: Processing of C5a by human polymorphonuclear leukocytes.
abstract: Uptake of human C5a by neutrophils was monitored in vitro using both 125I-labeled and unlabeled C5a. The ligand was internalized by the cells in a dose-dependent manner and maximal binding/uptake was observed after 5 min of incubation. Neutrophils were incubated with labeled C5a and the cytosol and supernatant were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. C5a degradation products were primarily observed in the supernatant, whereas most of the protein remained intact in the cytosol even after 60 min of incubation. Cytosol from neutrophils incubated for 20 min with unlabeled C5a was examined by radioimmunoassay and found to contain antigenically intact C5a and retained the ability to induce a neutrophil (shape change) response. The functional activity of C5a recovered from the cytosol was inhibited by antibodies to either C5a or the C5a receptor (CD88). This data supports our hypothesis that although C5a is internalized it remains antigenically intact and functionally active inside the cell and is primarily degraded extracellularly.
authors.affiliation: Department of Immunology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: G
authors.initials: G
authors.lastname: Hetland
authors.forename: P H
authors.initials: PH
authors.lastname: Pfeifer
authors.forename: T E
authors.initials: TE
authors.lastname: Hugli
mesh_terms: D015936:Complement C5a / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D004306:Dose-Response Relationship, Immunologic; D006801:Humans; D018375:Neutrophil Activation; D009504:Neutrophils / Q000378:metabolism*; D011863:Radioimmunoassay
chemical_list: D015936:Complement C5a