Interference by anti-C5a monoclonal antibodies with the binding of C5a to cellular receptors and the induction of enzyme release from granulocytes.
A sandwich ELISA based on monoclonal antibodies (MAb) for the quantification of rabbit C5a is only modestly influenced by the presence of MAb 4A3A9, which reacts with an epitope on C5a different from those of MAbs used in the sandwich ELISA. Incubation of rabbit or human granulocytes with C5a in the presence of MAb 4A3A9 results in a less pronounced decrease of the C5a concentration as compared to the concentration measured in the absence of MAb. We interpret the results to indicate an interference of MAb 4A3A9 with the binding of C5a to and/or the dissociation of C5a from its cellular receptor. MAb 4A3A9 is the least effective (25.4% inhibition) among anti-C5a MAbs in inhibiting C5a-induced beta-glucuronidase release from neutrophils. Together, these findings may indicate that a C5a-MAb complex is still able to induce a C5a effect even if MAb does interfere with the binding of ligand to the receptor.
Bergh
K
K
Department of Microbiology, Regional Hospital, University of Trondheim, Norway.
Iversen
O J
OJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001667:Binding, Competitive / Q000276:immunology; D015936:Complement C5a / Q000276:immunology*; D003571:Cytochalasin B; D004797:Enzyme-Linked Immunosorbent Assay; D005966:Glucuronidase / Q000378:metabolism*; D006098:Granulocytes / Q000201:enzymology / Q000276:immunology*; D006801:Humans; D066298:In Vitro Techniques; D011817:Rabbits; D044087:Receptor, Anaphylatoxin C5a; D011951:Receptors, Complement / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D044087:Receptor, Anaphylatoxin C5a; D011951:Receptors, Complement; D003571:Cytochalasin B; D015936:Complement C5a; D005966:Glucuronidase