Changes in neutrophil surface protein composition accompany phagocytosis.
Phagocytosis is a critical host defense mechanism of neutrophils. In this study, membrane protein changes occurring during phagocytosis were studied in human neutrophils using surface radiolabeling before or after phagocytosis of various target particles. Cells were labeled at the cell surface using lactoperoxidase-catalyzed iodination or neuraminidase-galactose oxidase-NaB3H4, galactose oxidase-NaB3H4, or periodate-NaB3H4 techniques. Such studies are complicated by the fact that these techniques identify many surface proteins on the phagocyte, and labeling after phagocytosis occurs often results in radiolabeling proteins of the target particle, thus making changes in cell-surface proteins more difficult to detect. Immunoprecipitation with monoclonal antibody AHN-1, which reacts with a carbohydrate present on several human neutrophil surface proteins and inhibits phagocytosis, eliminated interference caused by radiolabeled proteins of the target particle and simplified analysis by restricting the study to a limited number of proteins. AHN-1 immunoprecipitated less radiolabeled protein from neutrophils labeled after phagocytosis of particles opsonized with IgG or complement than from cells labeled before phagocytosis. Isolation of phagocytic vesicles containing opsonized emulsified paraffin oil demonstrated that three proteins of mol wt 105,000, 140,000, and 170,000 recognized by AHN-1 were internalized in the phagocytic vesicle during phagocytosis.
D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D000954:Antigens, Surface / Q000032:analysis*; D006801:Humans; D002479:Inclusion Bodies / Q000032:analysis; D007457:Iodine Radioisotopes; D008565:Membrane Proteins / Q000032:analysis* / Q000378:metabolism*; D009504:Neutrophils / Q000276:immunology / Q000648:ultrastructure*; D010232:Paraffin / Q000378:metabolism; D010587:Phagocytosis; D011233:Precipitin Tests; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D014316:Tritium
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D014316:Tritium; D010232:Paraffin