Moteki S, Amaki S, Nakano E
Department of Clinical Pathology, Nihon University School of Medicine, Tokyo.
Rinsho Byori. 1998 Apr;46(4):317-23.
A serological feature of primary biliary cirrhosis (PBC) is the presence of high-titer antimitochondrial autoantibodies (AMA) in patient sera. Five different target mitochondrial autoantigens recognized by sera from PBC patients have been identified as subunits of the 2-oxoacid dehydrogenase complex (2-OADC), of which pyruvate dehydrogenase complex-E2 (PDH-E2) is the most prominent antigenic component. Extensive molecular and immunological studies in PBC such as cloning of mitochondrial autoantigens, mapping of both T and B cell epitopes and immunohistochemical studies have provided valuable reagents in the understanding of immunopathogenesis of PBC. We mapped the epitope recognized by AMA specific to 2-oxoglutarate dehydrogenase complex-E2 (OGDC-E2) in patients with PBC using full-length rat OGDC-E2 cDNA and a series of expression clones spanning the entire molecule. It appears that the epitope is dependent on conformation and includes the lipoic acid-binding region. Furthermore we have taken advantage of the antigenic mapping studies of PDC-E2, OGDC-E2 and branched chain 2-oxoacid dehydrogenase complex-E2 (BCOADC-E2) subunits and designed a hybrid clone, pML-MIT3, that expressed three different immunodominant epitopes. Our results indicate that an immunoassay using recombinant, cloned autoantigen is a powerful and very specific method for detecting AMA in PBC.
原发性胆汁性肝硬化(PBC)的一个血清学特征是患者血清中存在高滴度抗线粒体自身抗体(AMA)。已确定PBC患者血清识别的五种不同靶线粒体自身抗原为2-氧代酸脱氢酶复合体(2-OADC)的亚基,其中丙酮酸脱氢酶复合体-E2(PDH-E2)是最主要的抗原成分。PBC中广泛的分子和免疫学研究,如线粒体自身抗原的克隆、T和B细胞表位的定位以及免疫组织化学研究,为理解PBC的免疫发病机制提供了有价值的试剂。我们使用全长大鼠OGDC-E2 cDNA和一系列跨越整个分子的表达克隆,对PBC患者中AMA识别的2-氧代戊二酸脱氢酶复合体-E2(OGDC-E2)特异性表位进行了定位。该表位似乎依赖于构象,包括硫辛酸结合区域。此外,我们利用了PDC-E2、OGDC-E2和支链2-氧代酸脱氢酶复合体-E2(BCOADC-E2)亚基的抗原定位研究,设计了一个表达三种不同免疫显性表位的杂交克隆pML-MIT3。我们的结果表明,使用重组克隆自身抗原的免疫测定法是检测PBC中AMA的一种强大且非常特异的方法。