title: Post challenge inhibition of C3 and CD14 attenuates Escherichia coli-induced inflammation in human whole blood.
abstract: Combined inhibition of CD14 and complement, two main inducers of the inflammatory response, have proved particularly effective in attenuating Gram-negative bacteria-induced inflammation. Approaching possible clinical relevance, we investigated the effect of such inhibition in a post-challenge setting. Human whole blood was anti-coagulated with lepirudin. Anti-CD14, compstatin (C3 inhibitor) and the combination thereof were added 5 min prior to or 5, 15 or 30 min after adding Escherichia coli. Total incubation time with Escherichia coli was 120 min. Cytokines, myeloperoxidase (MPO) and the terminal complement complex (TCC) were measured using multiplex technology and ELISA. Delayed combined inhibition significantly attenuated the inflammatory response. IL-1β, IL-8 and TNF-α were significantly inhibited in the range of 20-40%, even when adding the inhibitors with up to 30 min delay. IL-6 was significantly inhibited with 15 min delay, and MIP-1α and MPO with 5 min delay. Complement activation (TCC) was blocked completely at each time point compstatin was added, whereas the cytokines and MPO increased steadily between the time points. The combined regimen was significantly more effective than single inhibition in the pre-challenge setting. The attenuation of Escherichia coli-induced inflammation in a post-challenge setting suggests a potential therapeutic window for this treatment in sepsis.
authors.affiliation: 1Department of Immunology, Oslo University Hospital Rikshospitalet and University of Oslo, Norway.
authors.forename: Kjetil H
authors.initials: KH
authors.lastname: Egge
authors.forename: Ebbe B
authors.initials: EB
authors.lastname: Thorgersen
authors.forename: Julie K
authors.initials: JK
authors.lastname: Lindstad
authors.forename: Anne
authors.initials: A
authors.lastname: Pharo
authors.forename: John D
authors.initials: JD
authors.lastname: Lambris
authors.forename: Andreas
authors.initials: A
authors.lastname: Barratt-Due
authors.forename: Tom E
authors.initials: TE
authors.lastname: Mollnes
mesh_terms: D019138:Antibodies, Blocking / Q000494:pharmacology; D001769:Blood / Q000187:drug effects / Q000276:immunology*; D003167:Complement Activation / Q000187:drug effects; D003176:Complement C3 / Q000037:antagonists & inhibitors / Q000276:immunology*; D015938:Complement Membrane Attack Complex / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D004338:Drug Combinations; D004357:Drug Synergism; D004926:Escherichia coli / Q000276:immunology* / Q000378:metabolism; D006358:Hot Temperature; D006801:Humans; D007113:Immunity, Innate / Q000187:drug effects; D007114:Immunization; D007167:Immunotherapy; D018836:Inflammation Mediators / Q000378:metabolism; D018950:Lipopolysaccharide Receptors / Q000276:immunology / Q000378:metabolism*; D010456:Peptides, Cyclic / Q000494:pharmacology; D009195:Peroxidase / Q000378:metabolism; D018805:Sepsis / Q000276:immunology* / Q000628:therapy
chemical_list: D019138:Antibodies, Blocking; D003176:Complement C3; D015938:Complement Membrane Attack Complex; D016207:Cytokines; D004338:Drug Combinations; D018836:Inflammation Mediators; D018950:Lipopolysaccharide Receptors; D010456:Peptides, Cyclic; C111828:compstatin; D009195:Peroxidase
keywords: CD14; Escherichia coli; c3; complement; cytokines; human; post-challenge