An extended C1q-binding assay using lactoperoxidase- and chloramine-T-iodinated C1q. Immediate distinction between immune-aggregate-mediated and non-immune-aggregate-mediated C1q binding.
An extension of the C1q-binding assay for the detection of immune-aggregate-mediated and non-immune-aggregate-mediated C1q binding is reported. The assay involves the use of two different C1q preparations, one radioiodinated by means of lactoperoxidase (LPO-125I-C1q) and the other by means of chloramine-T (CT-125I-C1q). The treatment with CT for 20 min at room temperature before iodination for 1 min led to abolishment of the C1q-binding capacities to complexed IgG: approximately 50% of LPO-125I-C1q but only 2% of CT-125I-C1q bound to 80 micrograms/ml of IgG forming part of tetanus toxoid/anti-tetanus toxoid complexes or to 200 micrograms/ml of heat-aggregated human gamma globulin. Similar results were obtained with staphylococcal protein-A-aggregated IgG. CT-treated C1q was haemolytically inactive. In contrast to the results with complexed IgG, CT treatment did not markedly reduce binding capacities of C1q to heparin: approximately 55% of LPO- and CT-125I-C1q were bound by 127 U/ml of commercial heparin in normal human serum. Both C1q preparations bound to a comparable extent to fibronectin, fibrinogen, and various bacterial endotoxins. When the LPO- and CT-125I-C1q-binding patterns obtained on serum samples from patients with systemic lupus erythematosus, rheumatoid arthritis, or essential mixed cryoglobulinaemia were compared with binding patterns observed using laboratory reactants, an immediate detection of non-immune-aggregate-mediated C1q binding became possible.
D000936:Antigen-Antibody Complex / Q000276:immunology; D001172:Arthritis, Rheumatoid / Q000276:immunology; D002700:Chloramines / Q000276:immunology*; D003166:Complement Activating Enzymes / Q000276:immunology*; D015922:Complement C1q; D003168:Complement Fixation Tests / Q000379:methods*; D003449:Cryoglobulinemia / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D007784:Lactoperoxidase / Q000276:immunology*; D008180:Lupus Erythematosus, Systemic / Q000276:immunology; D010544:Peroxidases / Q000276:immunology*; D014105:Tosyl Compounds
D000936:Antigen-Antibody Complex; D002700:Chloramines; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D014105:Tosyl Compounds; C016300:chloramine-T; D015922:Complement C1q; D007784:Lactoperoxidase; D010544:Peroxidases; D003166:Complement Activating Enzymes