Activation of C1 by soluble IgG aggregates as detected by a novel one-step hemolytic assay that specifically measures the proenzyme form of C1s.
A new hemolytic assay is described that specifically measures the precursor form of the C1s subcomponent of the complement system. The assay employs a C1s-depleted reagent obtained by immunoadsorption of fresh human plasma on immobilized goat anti-human C1s antibodies. Linear Z plots are obtained with nanogram levels of precursor C1s, whereas C1s completely fails to induce hemolysis in the assay. Because low concentrations of C1s do not interfere with the activity of precursor C1s, the assay can be used for the stoichiometric measurement of C1 activation. The precursor C1s assay was applied to the study of C1 binding and activation by soluble aggregates of human IgG (AIgG). Incubation of purified human C1 with AIgG caused a temperature-independent consumption of whole C1 hemolytic activity, indicating binding of C1, but almost no consumption of the total (precursor + activated) C1s activity. On the other hand, activation of C1, measured as the time- and temperature-dependent consumption of precursor C1s, could greatly exceed the binding of C1. These findings can be explained by using recent findings concerning the association-dissociation equilibrium between C1q and the tetrameric complex of C1r and C1s.
D000818:Animals; D003166:Complement Activating Enzymes / Q000032:analysis* / Q000276:immunology / Q000502:physiology; D003167:Complement Activation; D003172:Complement C1 / Q000378:metabolism; D003173:Complement C1s; D003181:Complement C4 / Q000378:metabolism; D003171:Complement Pathway, Classical; D004792:Enzyme Precursors / Q000032:analysis* / Q000502:physiology; D006461:Hemolysis; D006801:Humans; D007074:Immunoglobulin G / Q000502:physiology*; D046911:Macromolecular Substances; D011817:Rabbits; D011951:Receptors, Complement / Q000032:analysis; D013696:Temperature; D013997:Time Factors
D003172:Complement C1; D003181:Complement C4; D004792:Enzyme Precursors; D007074:Immunoglobulin G; D046911:Macromolecular Substances; D011951:Receptors, Complement; D003166:Complement Activating Enzymes; D003173:Complement C1s