title: Detection of autoantibodies to recombinant mitochondrial proteins in patients with primary biliary cirrhosis.
abstract: Primary biliary cirrhosis is characterized by the presence of autoantibodies to mitochondria with specific reactivity to proteins of 74 and 52 kilodaltons (kd). The 74-kd mitochondrial protein is the E2 component--dihydrolipoamide acetyltransferase--of the pyruvate dehydrogenase complex, and the 52-kd protein is the equivalent E2 component--dihydrolipoamide acyltransferase--of the branched-chain alpha-keto acid dehydrogenase complex. Current methods for the detection of antibodies to these proteins lack specificity or sensitivity, or they are time-consuming and not readily available. We therefore developed an enzyme-linked immunoassay to quantify specific antimitochondrial antibodies in patients with primary biliary cirrhosis. Recombinant polypeptides coding for both the 74-kd and the 52-kd mitochondrial autoantigens were used to analyze 217 coded serum samples, including samples from 93 patients with primary biliary cirrhosis and 124 controls, for reactivity by our immunoassay, immunoblotting, and immunofluorescence testing. Serum samples from 89 of the 93 patients with primary biliary cirrhosis reacted with either the pyruvate dehydrogenase-E2 or the branched-chain alpha-keto acid dehydrogenase protein. None of the 124 control samples from healthy volunteers (n = 86) or patients with primary sclerosing cholangitis (n = 38) had significant reactivity. Our results indicate that the use of recombinant, cloned autoantigens provides a simple, accurate, and rapid method of quantifying and monitoring the levels of specific mitochondrial autoantibodies in the serum of patients with primary biliary cirrhosis.
authors.affiliation: Division of Rheumatology/Allergy and Clinical Immunology, University of California, Davis 95616.
authors.forename: J
authors.initials: J
authors.lastname: Van de Water
authors.forename: A
authors.initials: A
authors.lastname: Cooper
authors.forename: C D
authors.initials: CD
authors.lastname: Surh
authors.forename: R
authors.initials: R
authors.lastname: Coppel
authors.forename: D
authors.initials: D
authors.lastname: Danner
authors.forename: A
authors.initials: A
authors.lastname: Ansari
authors.forename: R
authors.initials: R
authors.lastname: Dickson
authors.forename: M E
authors.initials: ME
authors.lastname: Gershwin
mesh_terms: D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide); D001323:Autoantibodies / Q000032:analysis*; D001324:Autoantigens / Q000032:analysis; D008058:Dihydrolipoamide Dehydrogenase / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007658:Ketone Oxidoreductases / Q000276:immunology*; D008105:Liver Cirrhosis, Biliary / Q000276:immunology*; D008928:Mitochondria / Q000201:enzymology / Q000276:immunology*; D009097:Multienzyme Complexes / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D007658:Ketone Oxidoreductases; D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide); D008058:Dihydrolipoamide Dehydrogenase