title: Properdin binding to complement activating surfaces depends on initial C3b deposition.
abstract: Two functions have been assigned to properdin; stabilization of the alternative convertase, C3bBb, is well accepted, whereas the role of properdin as pattern recognition molecule is controversial. The presence of nonphysiological aggregates in purified properdin preparations and experimental models that do not allow discrimination between the initial binding of properdin and binding secondary to C3b deposition is a critical factor contributing to this controversy. In previous work, by inhibiting C3, we showed that properdin binding to zymosan and Escherichia coli is not a primary event, but rather is solely dependent on initial C3 deposition. In the present study, we found that properdin in human serum bound dose-dependently to solid-phase myeloperoxidase. This binding was dependent on C3 activation, as demonstrated by the lack of binding in human serum with the C3-inhibitor compstatin Cp40, in C3-depleted human serum, or when purified properdin is applied in buffer. Similarly, binding of properdin to the surface of human umbilical vein endothelial cells or Neisseria meningitidis after incubation with human serum was completely C3-dependent, as detected by flow cytometry. Properdin, which lacks the structural homology shared by other complement pattern recognition molecules and has its major function in stabilizing the C3bBb convertase, was found to bind both exogenous and endogenous molecular patterns in a completely C3-dependent manner. We therefore challenge the view of properdin as a pattern recognition molecule, and argue that the experimental conditions used to test this hypothesis should be carefully considered, with emphasis on controlling initial C3 activation under physiological conditions.
authors.affiliation: Department of Immunology, Oslo University Hospital, Rikshospitalet, 0424 Oslo, Norway.
authors.forename: Morten
authors.initials: M
authors.lastname: Harboe
authors.affiliation: Department of Immunology, Oslo University Hospital, Rikshospitalet, 0424 Oslo, Norway.
authors.forename: Christina
authors.initials: C
authors.lastname: Johnson
authors.affiliation: Research Laboratory, Nordland Hospital, 8092 Bodo, Norway.
authors.forename: Stig
authors.initials: S
authors.lastname: Nymo
authors.affiliation: Department of Immunology, Oslo University Hospital, Rikshospitalet, 0424 Oslo, Norway.
authors.forename: Karin
authors.initials: K
authors.lastname: Ekholt
authors.affiliation: Department of Immunology, Oslo University Hospital, Rikshospitalet, 0424 Oslo, Norway.
authors.forename: Camilla
authors.initials: C
authors.lastname: Schjalm
authors.affiliation: Department of Immunology, Oslo University Hospital, Rikshospitalet, 0424 Oslo, Norway.
authors.forename: Julie K
authors.initials: JK
authors.lastname: Lindstad
authors.affiliation: Department of Immunology, Oslo University Hospital, Rikshospitalet, 0424 Oslo, Norway.
authors.forename: Anne
authors.initials: A
authors.lastname: Pharo
authors.affiliation: Department of Immunology, Oslo University Hospital, Rikshospitalet, 0424 Oslo, Norway.
authors.forename: Bernt Christian
authors.initials: BC
authors.lastname: Hellerud
authors.affiliation: Department of Immunology, Genetics, and Pathology, Rudbeck Laboratory C5:3, Uppsala University, 751 85 Uppsala, Sweden.|Linnaeus Centre for Biomaterials Chemistry, Linnaeus University, 391 82 Kalmar, Sweden.
authors.forename: Kristina
authors.initials: K
authors.lastname: Nilsson Ekdahl
authors.affiliation: Department of Immunology, Oslo University Hospital, Rikshospitalet, 0424 Oslo, Norway.|Research Laboratory, Nordland Hospital, 8092 Bodo, Norway.|K. G. Jebsen Inflammatory Research Center, University of Oslo, 0424 Oslo, Norway.|Faculty of Health Sciences, K. G. Jebsen Thrombosis Research and Expertise Center, University of Tromsø, 9037 Tromso, Norway.|Centre of Molecular Inflammation Research, Norwegian University of Science and Technology, 7491 Trondheim, Norway.
authors.forename: Tom Eirik
authors.initials: TE
authors.lastname: Mollnes
authors.affiliation: Department of Immunology, Oslo University Hospital, Rikshospitalet, 0424 Oslo, Norway; per.nilsson@rr-research.no.|Linnaeus Centre for Biomaterials Chemistry, Linnaeus University, 391 82 Kalmar, Sweden.|K. G. Jebsen Inflammatory Research Center, University of Oslo, 0424 Oslo, Norway.
authors.forename: Per H
authors.identifier: 0000-0002-7192-5794
authors.initials: PH
authors.lastname: Nilsson
mesh_terms: D002478:Cells, Cultured; D003167:Complement Activation; D003179:Complement C3b / Q000378:metabolism*; D006098:Granulocytes / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D009345:Neisseria meningitidis; D010456:Peptides, Cyclic / Q000494:pharmacology; D009195:Peroxidase / Q000378:metabolism; D011414:Properdin / Q000378:metabolism*; D044967:Serum
chemical_list: D010456:Peptides, Cyclic; C111828:compstatin; D011414:Properdin; D003179:Complement C3b; D009195:Peroxidase
keywords: C3; Neisseria meningitidis; complement; myeloperoxidase; properdin