Conformational epitopes of C3 reflecting its mode of binding to an artificial polymer surface.
The aim of the study was to investigate the incompletely understood mechanisms of complement (C) activation and binding on artificial biomaterials. Polystyrene in the form of microtitre plates was used as target for C binding, detectable by ELISA using monoclonal anti-C3 antibodies specific for conformational epitopes expressed by bound C3 and C3 fragments. C3 binding in whole blood/plasma/serum is maximal at low dilutions and occurs predominantly by C activation. At higher dilutions, C3 binding occurs at approximately 1/3 of maximal levels and is solely an effect of adsorption. C3 adsorption in the lower serum dilution range, occurs at low but clearly detectable levels. Comparative epitope analysis between C3 fragments, actively bound to polystyrene in the presence of serum, and of iC3b bound to sheep erythrocytes, clearly indicates that C3 binding/activation on polystyrene takes place as a C3 convertase-mediated reaction, which in serum/plasma is followed by a secondary factor I-dependent degradation of the bound C3b into iC3b. The neo-epitope analysis of serum-contacting polystyrene revealed that the adsorbed C3, throughout the entire serum dilution range tested, deposits in a state closely similar to that observed for purified C3 at a high packing density. Polystyrene surfaces with adsorbed purified C3 expressing this epitope profile were found to mediate APW dependent deposition of C3b in pig serum, presumably by forming a hybrid convertase with porcine Bb. These data therefore suggest that adsorbed C3 on serum-contacting polystyrene surfaces may initiate complement activation via the APW.
Nilsson
U R
UR
Department of Clinical Immunology and Transfusion Medicine, University Hospital, Uppsala, Sweden.
Storm
K E
KE
Elwing
H
H
Nilsson
B
B
D000327:Adsorption; D000818:Animals; D003167:Complement Activation; D003176:Complement C3 / Q000276:immunology / Q000378:metabolism*; D000939:Epitopes; D005260:Female; D006801:Humans; D051379:Mice; D011137:Polystyrenes; D011487:Protein Conformation
D003176:Complement C3; D000939:Epitopes; D011137:Polystyrenes