title: Partial characterization of a C5a-inhibitor in peritoneal fluid.
abstract: We have recently described a 40-kDa protein in peritoneal fluid that neutralized the chemotactic activity of the C fraction C5a. It was deficient in peritoneal fluids of patients suffering from familial Mediterranean fever. Further characterization of the inhibitor with the use of 125I-rC5a binding to dibutyryl cAMP-induced U937 cells revealed dependence on the peritoneal fluid concentration, on the time of incubation and on temperature and pH. Fractionation of 125I-C5a on Sephadex G-50 column, before and after incubation with peritoneal fluid, revealed similar fractionation patterns despite loss of biologic activity of the treated C5a (but not its binding to polyclonal anti-C5a antibody). Analysis of rC5a by SDS-PAGE before and after treatment with partially purified C5a inhibitor, revealed slight modification of the inhibitor-treated C5a. Using various protease inhibitors (i.e., PMSF) suggested that the C5a inhibitor is a serine protease. It neutralized C5a by means of limited proteolysis which did not change C5a immunologic properties and changed only slightly its m.w. but abolished its receptor binding and chemotactic functions. It is suggested that the C5a inhibitor plays a role in the regulation of inflammation in serosal tissues and that its deficiency in familial Mediterranean fever may explain the attacks of sterile inflammation characteristic of this disease.
authors.affiliation: Hematology Unit, Hadassah University Hospital Mount Scopus, Jerusalem, Israel.
authors.forename: S K
authors.initials: SK
authors.lastname: Ayesh
authors.forename: M
authors.initials: M
authors.lastname: Ferne
authors.forename: I
authors.initials: I
authors.lastname: Flechner
authors.forename: B M
authors.initials: BM
authors.lastname: Babior
authors.forename: Y
authors.initials: Y
authors.lastname: Matzner
mesh_terms: D001202:Ascitic Fluid / Q000032:analysis*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D015936:Complement C5a / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D012697:Serine Endopeptidases / Q000302:isolation & purification*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013696:Temperature
chemical_list: D015842:Serine Proteinase Inhibitors; D015936:Complement C5a; D012697:Serine Endopeptidases