Document Detail


Detection of antimitochondrial autoantibodies in immunofluorescent AMA-negative patients with primary biliary cirrhosis using recombinant autoantigens.
MedLine Citation:
PMID:  11481607     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Antimitochondrial antibodies (AMA) are the serologic hallmark of primary biliary cirrhosis (PBC). However, depending on the clinical laboratory, from 5% to 17% of PBC patients are consistently AMA-negative, using native mitochondrial antigens and a variety of conventional assays including immunofluorescence (IMF) and enzyme-linked immunosorbent assay (ELISA). The major immunoreactive mitochondrial autoantigens are the E2 members of the 2-oxo-acid dehydrogenase complex family, including pyruvate dehydrogenase complex-E2 (PDC-E2), branched chain 2-oxo acid dehydrogenase complex-E2 (BCOADC-E2), and oxo-glutarate dehydrogenase complex-E2 (OGDC-E2); cDNAs of these proteins have now been cloned, sequenced, and their B-cell epitopes defined. In the present study, we cloned cDNAs encoding these proteins from human, not bovine, sources, and expressed the recombinant proteins in a newly developed ELISA that employs a unique Escherichia coli buffer, and compared the data with previous assays using both AMA-positive and -negative patients. Using this new assay and our criteria for positive as an optical density (OD) greater than 10 SD above the mean of control sera, the AMA-positive rate of 191 PBC sera was 94% (179 of 191) compared with 84% (161 of 191) by IMF. None of the 316 control sera were reactive. Using our recombinant assays, we focused attention on the 30 IMF-AMA-negative patients. Twenty-two of 30 (73%) of these patients were positive using this new ELISA. The group of 30 IMF-AMA-negative/ELISA-positive patients did not differ significantly from a comparable population of IMF-AMA-positive patients with respect to age, sex distribution, liver function tests, elevation of serum IgM, or pathologic stage.
Authors:
H Miyakawa; A Tanaka; K Kikuchi; M Matsushita; E Kitazawa; N Kawaguchi; H Fujikawa; M E Gershwin
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Hepatology (Baltimore, Md.)     Volume:  34     ISSN:  0270-9139     ISO Abbreviation:  Hepatology     Publication Date:  2001 Aug 
Date Detail:
Created Date:  2001-08-01     Completed Date:  2001-08-23     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  8302946     Medline TA:  Hepatology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  243-8     Citation Subset:  IM    
Affiliation:
Fourth Department of Internal Medicine, Teikyo University School of Medicine, Kanagawa, Japan.
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MeSH Terms
Descriptor/Qualifier:
Autoantibodies / analysis*
Autoantigens / immunology*
Blotting, Western
Enzyme-Linked Immunosorbent Assay
Epitopes
Female
Fluorescent Antibody Technique
Humans
Liver Cirrhosis, Biliary / immunology*
Male
Middle Aged
Mitochondria / immunology*
Recombinant Proteins
Sensitivity and Specificity
Grant Support
ID/Acronym/Agency:
DK 39588/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Autoantibodies; 0/Autoantigens; 0/Epitopes; 0/Recombinant Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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