title: Pre-neutralization of C5a-mediated effects by the monoclonal antibody 137-26 reacting with the C5a moiety of native C5 without preventing C5 cleavage.
abstract: Complement C5a is aetiologically linked to inflammatory tissue damage in conditions like septicaemia, immune complex diseases and ischaemia-reperfusion injury. We here describe a monoclonal antibody (mAb), 137-26, that binds to the C5a moiety of human C5 and neutralizes the effects of C5a without interfering with C5 cleavage and the subsequent formation of lytic C5b-9 complex. Mouse anti-human C5 mAbs were generated and the reactivity with C5 and C5a was detected by ELISA and surface plasmon resonance. The inhibition of C5a binding to C5a receptor was studied using a radioligand binding assay. The effects of the antibody on C5a functions were examined using isolated neutrophils and a novel human whole blood model of inflammation. Haemolytic assays were used to study the effect on complement-mediated lysis. mAb 137-26 reacted with both solid- and solution-phase C5 and C5a in a dose-dependent manner with high affinity. The antibody competed C5a binding to C5a receptor and inhibited C5a-mediated chemotaxis of neutrophils. Furthermore, the antibody effectively abrogated complement-dependent E. coli-induced CD11b up-regulation and oxidative burst in neutrophils of human whole blood. mAb 137-26 was more potent than a C5a receptor antagonist and a previously described anti-C5a antibody. mAb 137-26 did not inhibit complement-mediated lysis, nor did it activate complement itself. Together, mAb 137-26 binds both the C5a moiety of native C5 and free C5a, thereby effectively neutralizing the biological effects of C5a. The antibody may have therapeutic potential in inflammatory diseases where C5a inhibition combined with an operative lytic pathway of C5b-9 is particularly desired.
authors.affiliation: Tanox Inc., Houston, Texas 77025, USA. mfung@tanox.com
authors.forename: M
authors.initials: M
authors.lastname: Fung
authors.forename: M
authors.initials: M
authors.lastname: Lu
authors.forename: H
authors.initials: H
authors.lastname: Fure
authors.forename: W
authors.initials: W
authors.lastname: Sun
authors.forename: C
authors.initials: C
authors.lastname: Sun
authors.forename: N Y
authors.initials: NY
authors.lastname: Shi
authors.forename: Y
authors.initials: Y
authors.lastname: Dou
authors.forename: J
authors.initials: J
authors.lastname: Su
authors.forename: X
authors.initials: X
authors.lastname: Swanson
authors.forename: T E
authors.initials: TE
authors.lastname: Mollnes
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D001667:Binding, Competitive; D039481:CD11b Antigen / Q000097:blood; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte / Q000276:immunology; D003167:Complement Activation / Q000276:immunology; D003182:Complement C5 / Q000276:immunology; D015936:Complement C5a / Q000037:antagonists & inhibitors* / Q000276:immunology; D003169:Complement Inactivator Proteins / Q000276:immunology*; D004306:Dose-Response Relationship, Immunologic; D006461:Hemolysis / Q000276:immunology; D006801:Humans; D008297:Male; D051379:Mice; D008805:Mice, Inbred A; D009504:Neutrophils / Q000378:metabolism; D016897:Respiratory Burst / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D039481:CD11b Antigen; D003182:Complement C5; D003169:Complement Inactivator Proteins; D015936:Complement C5a