Quantification by ELISA of Erythrocyte-Bound C3 Fragments Expressing C3d and/or C3c Epitopes in Patients with Factor I Deficiency and with Autoimmune Diseases
Corresponding Author
Dr. Jens Møller Rasmussen
Institute of Medical Microbiology, Odense University, Odense C, Denmark
Institute of Medical Microbiology, Odense University, J.B. Winsløvsvej 19, DK–5000 Odense (Denmark)Search for more papers by this authorHans Henrik Jepsen
Institute of Medical Microbiology, Odense University, Odense C, Denmark
Search for more papers by this authorBørge Teisner
Institute of Medical Microbiology, Odense University, Odense C, Denmark
Search for more papers by this authorUffe Holmskov-Nielsen
Departements of Internal Medicine C, Odense University Hospital, Odense, Denmark
Search for more papers by this authorGlen Gorm Rasmussen
Departements of Rheumatology, Odense University Hospital, Odense, Denmark
Search for more papers by this authorSven-Erik Svehag
Institute of Medical Microbiology, Odense University, Odense C, Denmark
Search for more papers by this authorCorresponding Author
Dr. Jens Møller Rasmussen
Institute of Medical Microbiology, Odense University, Odense C, Denmark
Institute of Medical Microbiology, Odense University, J.B. Winsløvsvej 19, DK–5000 Odense (Denmark)Search for more papers by this authorHans Henrik Jepsen
Institute of Medical Microbiology, Odense University, Odense C, Denmark
Search for more papers by this authorBørge Teisner
Institute of Medical Microbiology, Odense University, Odense C, Denmark
Search for more papers by this authorUffe Holmskov-Nielsen
Departements of Internal Medicine C, Odense University Hospital, Odense, Denmark
Search for more papers by this authorGlen Gorm Rasmussen
Departements of Rheumatology, Odense University Hospital, Odense, Denmark
Search for more papers by this authorSven-Erik Svehag
Institute of Medical Microbiology, Odense University, Odense C, Denmark
Search for more papers by this authorAbstract
Abstract. A sensitive ELISA assay for quantifying erythrocyte (E) bound C3 fragments was developed. The assay employs a double-antibody sandwich technique, using polyclonal anti-C3d or anti-C3c antibodies to quantify C3 fragments, expressing C3d and/or C3c epitopes in washed, detergent-solubilized E. The assay detected 50–120 molecules of C3d per E in healthy individuals. Antigens reacting with anti-C3c antibodies were also detected on E from normal individuals, but the density of C3c-epitopes was 0.9–2.4 times lower than that of C3d-epitopes. In 2 patients with congenital factor I deficiency significantly increased density of E-bound C3c- as well as C3d-antigen was observed. Plasma infusion in one of the patients induced a loss of E-bound C3c-antigens, indicating cleavage of E-bound C3b to iC3b and further to C3c and E-bound C3d. Loss of C3c-antigens also occurred following in vitro treatment with normal human serum of E from one of the patients. Two thirds of 22 patients with systemic lupus erythematosus (SLE) and of 18 patients with rheumatoid arthritis had significantly increased density of E-bound C3d, the highest density being 490 C3d molecules/E in an SLE patient. The density of E-bound C3d correlated with the plasma-C3d concentration, indicating that the coating of E with C3d reflects the degree of complement activation.
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