Halogenation and proteolysis of complement component C3 on Salmonella typhimurium during phagocytosis by human neutrophils.
We examined the fate of C component C3 on the surface of Salmonella typhimurium during ingestion by human neutrophils. Initial experiments showed that C3 fragments and C3-acceptor complexes were the major serum ligands which were surface iodinated by canine myeloperoxidase on serum-incubated rough and smooth isolates of S. typhimurium. In contrast, labeled C3 was not identified when the same organisms were ingested by neutrophils in the presence of 125I-Na, a situation previously shown to iodinate particulate targets via the neutrophil myeloperoxidase-halide-H2O2 system. Pretreatment of neutrophils before phagocytosis with the lipid-soluble protease inhibitor diisopropylfluorophosphate (DFP), but not with other protease inhibitors (p-nitrophenylguanidinobenzoate, leupeptin, pepstatin), substantially blocked proteolysis of 125I-C3 on S. typhimurium strain RG108 during ingestion by neutrophils. Purification of neutrophil phagosomes containing S. typhimurium-bearing 125I-C3 showed that DFP but no other protease inhibitors blocked proteolysis of 125I-C3 within phagosomes. Iodinated C3-acceptor complexes were identified by immunoprecipitation from the detergent-insoluble fraction of phagosomes prepared from DFP-treated cells ingesting S. typhimurium in the presence of 125I-Na. These results show that C3 fragments on the surface of S. typhimurium are the major serum ligands which are halogenated and degraded by proteolysis during phagocytosis by human neutrophils, and suggest that the majority of proteolysis on the ingested target occurs within the neutrophil phagosome.
Joiner
K A
KA
Laboratory of Parastic Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.
Schweinle
J E
JE
D000327:Adsorption; D003176:Complement C3 / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006868:Hydrolysis; D007455:Iodine / Q000378:metabolism*; D007457:Iodine Radioisotopes / Q000378:metabolism; D009504:Neutrophils / Q000378:metabolism* / Q000382:microbiology / Q000502:physiology; D010447:Peptide Hydrolases; D009195:Peroxidase; D010587:Phagocytosis; D011480:Protease Inhibitors; D012486:Salmonella typhimurium / Q000378:metabolism* / Q000502:physiology; D012977:Sodium Nitrite; D013347:Subcellular Fractions / Q000201:enzymology / Q000378:metabolism
D003176:Complement C3; D007457:Iodine Radioisotopes; D011480:Protease Inhibitors; D007455:Iodine; D009195:Peroxidase; D010447:Peptide Hydrolases; D012977:Sodium Nitrite