title: The role of properdin in zymosan- and Escherichia coli-induced complement activation.
abstract: Properdin is well known as an enhancer of the alternative complement amplification loop when C3 is activated, whereas its role as a recognition molecule of exogenous pathogen-associated molecular patterns and initiator of complement activation is less understood. We therefore studied the role of properdin in activation of complement in normal human serum by zymosan and various Escherichia coli strains. In ELISA, microtiter plates coated with zymosan induced efficient complement activation with deposition of C4b and terminal complement complex on the solid phase. Virtually no deposition of C4b or terminal complement complex was observed with mannose-binding lectin (MBL)-deficient serum. Reconstitution with purified MBL showed distinct activation in both readouts. In ELISA, normal human serum-induced deposition of properdin by zymosan was abolished by the C3-inhibiting peptide compstatin. Flow cytometry was used to further explore whether properdin acts as an initial recognition molecule reacting directly with zymosan and three E. coli strains. Experiments reported by other authors were made with EGTA Mg²⁺ buffer, permitting autoactivation of C3. We found inhibition by compstatin on these substrates, indicating that properdin deposition depended on initial C3b deposition followed by properdin in a second step. Properdin released from human polymorphonuclear cells stimulated with PMA did not bind to zymosan or E. coli, but when incubated in properdin-depleted serum this form of properdin bound efficiently to both substrates in a strictly C3-dependent manner, as the binding was abolished by compstatin. Collectively, these data indicate that properdin in serum as well as polymorphonuclear-released properdin is unable to bind and initiate direct alternative pathway activation on these substrates.
authors.affiliation: Institute of Immunology, University of Oslo and Oslo University Hospital, Rikshospitalet, NO-0027 Oslo, Norway.
authors.forename: Morten
authors.initials: M
authors.lastname: Harboe
authors.forename: Peter
authors.initials: P
authors.lastname: Garred
authors.forename: Julie K
authors.initials: JK
authors.lastname: Lindstad
authors.forename: Anne
authors.initials: A
authors.lastname: Pharo
authors.forename: Fredrik
authors.initials: F
authors.lastname: Müller
authors.forename: Gregory L
authors.initials: GL
authors.lastname: Stahl
authors.forename: John D
authors.initials: JD
authors.lastname: Lambris
authors.forename: Tom E
authors.initials: TE
authors.lastname: Mollnes
mesh_terms: D000328:Adult; D003170:Complement Pathway, Alternative / Q000276:immunology*; D004926:Escherichia coli / Q000276:immunology* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008297:Male; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D011414:Properdin / Q000378:metabolism / Q000502:physiology*; D011485:Protein Binding / Q000276:immunology; D013379:Substrate Specificity / Q000276:immunology; D015054:Zymosan / Q000502:physiology*
chemical_list: D029968:Escherichia coli Proteins; D011414:Properdin; D015054:Zymosan