Inhibition of activation of the classical pathway of complement by human neutrophil defensins.
Defensins are small, cationic antimicrobial peptides that are present in the azurophilic granules of neutrophils. Earlier studies have shown that defensins may influence complement activation by specific interaction with activated C1, C1q, and C1-inhibitor. In the present study, we show that the defensin human neutrophil peptide-1 (HNP-1) is able to inhibit activation of the classical complement pathway by inhibition of C1q hemolytic activity. The binding site for HNP-1 on C1q is most likely located on the collagen-like stalks, as a clear, dose-dependent binding of HNP-1 to either intact C1q or to the collagen-like stalks of C1q was demonstrated using enzyme-linked immunosorbent assay (ELISA). Besides binding of HNP-1 to C1q, also a limited binding to C1 and to a mixture of C1r and C1s was observed, whereas no binding to C1-inhibitor was found. Because binding of HNP-1 to C1-inhibitor has been suggested in earlier studies, we also assessed the binding of HNP-1 to mixtures of C1-inhibitor with either C1r/ C1s or C1. No binding was found. Using a competition ELISA, it was found that HNP-1, but not protamine, inhibited binding of biotin-labeled HNP-1 to C1q in a dose-dependent fashion. In the fluid phase, preincubation of HNP-1 with C1q resulted in complex formation of HNP-1 and C1q and generation of stable complexes. In conclusion, HNP-1 is able to bind to C1q in the fluid phase and inhibits the classical complement pathway. This mechanism may be involved in the control of an inflammatory response in vivo.
van den Berg
R H
RH
Departments of Nephrology and Pulmonology, Leiden University Medical Center, Leiden, The Netherlands.
Faber-Krol
M C
MC
van Wetering
S
S
Hiemstra
P S
PS
Daha
M R
MR
D001665:Binding Sites; D001667:Binding, Competitive; D003174:Complement C1 Inactivator Proteins / Q000378:metabolism; D015922:Complement C1q / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D003171:Complement Pathway, Classical / Q000187:drug effects*; D023082:Defensins; D004797:Enzyme-Linked Immunosorbent Assay; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D046911:Macromolecular Substances; D009504:Neutrophils / Q000737:chemistry / Q000502:physiology*; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism / Q000494:pharmacology*; D023084:alpha-Defensins
D003174:Complement C1 Inactivator Proteins; D023082:Defensins; D046911:Macromolecular Substances; D011506:Proteins; D023084:alpha-Defensins; C406413:human neutrophil peptide 1; D015922:Complement C1q