Enzyme-linked immunosorbent assay using multimers of the 16th non-collagenous domain of the BP180 antigen for sensitive and specific detection of pemphigoid autoantibodies

被引:113
作者
Sitaru, Cassian
Daehnrich, Cornelia
Probst, Christian
Komorowski, Lars
Bloecker, Inga
Schmidt, Enno
Schlumberger, Wolfgang
Rose, Christian
Stoecker, Winfried
Zillikens, Detlef
机构
[1] Med Univ Lubeck, Dept Dermatol, D-23538 Lubeck, Germany
[2] Euroimmun AG, Inst Expt Immunol, Lubeck, Germany
[3] Univ Wurzburg, Dept Dermatol, D-8700 Wurzburg, Germany
关键词
autoantigen; autoimmunity; BPAG2; collagen; ELISA; tetramer;
D O I
10.1111/j.1600-0625.2007.00592.x
中图分类号
R75 [皮肤病学与性病学];
学科分类号
100206 ;
摘要
Bullous pemphigoid (BP) and pemphigoid gestationis (PG) are acquired autoimmune subepidermal blistering diseases characterized by autoantibodies against the hemidesmosomal proteins BP180/type XVII collagen and BP230. In the vast majority of BP and PG patients, these autoantibodies bind to epitopes clustered within the 16th non-collagenous domain of BP180. An ELISA system for the detection of these autoantibodies was developed and evaluated using 16th non-collagenous domain (NC16A) tetramers instead of monomers. In contrast to antigens fused to large proteins used in the past for the detection of autoantibodies against type XVII collagen, tetrameric antigen fragments bearing a small hexahistidine tag allow for high expression levels without the need to cleave off the fusion partner. Using tetrameric BP180 NC16A, positive reactions were found in 106 (89.8%) of 118 randomly selected BP sera and in all of 20 (100%) randomly selected PG sera, whereas only 2.2% of a large cohort of control subjects were positive in this assay, including patients with rheumatoid arthritis (two of 107), progressive systemic sclerosis (two of 50), systemic lupus erythematosus (one of 72), and healthy blood donors (10 of 494). Thus, the sensitivity and specificity of the new anti-tetrameric NC16A ELISA were 89.9% and 97.8% respectively. Levels of circulating autoantibodies against BP180 paralleled disease activity in the pemphigoid patients. In conclusion, the use of tetrameric NC16A in ELISA results in a sensitive and specific tool for diagnosis and monitoring of BP and PG.
引用
收藏
页码:770 / 777
页数:8
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