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学科主题: 临床医学
题名:
Circulating anti-glomerular basement membrane autoantibodies against a3(IV)NC1 undetectable by commercially available enzyme-linked immunosorbent assays
作者: Jia, Xiao-Yu; Qu, Zhen; Cui, Zhao; Yang, Rui; Zhao, Juan; Zhao, Ming-Hui
关键词: anti-glomerular basement membrane disease ; conformational structure ; epitope ; Goodpasture disease ; a3(IV)NC1
刊名: NEPHROLOGY
发表日期: 2012-02-01
DOI: 10.1111/j.1440-1797.2011.01511.x
卷: 17, 期:2, 页:160-166
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Urology & Nephrology
研究领域[WOS]: Urology & Nephrology
关键词[WOS]: RESONANT MIRROR BIOSENSOR ; GOODPASTURES-SYNDROME ; ALPHA-3(IV) COLLAGEN ; CRYPTIC EPITOPES ; NC1 DOMAIN ; DISEASE ; ANTIBODIES ; ANTIGEN ; GLOMERULONEPHRITIS ; AUTOANTIGEN
英文摘要:

Aim: Cases with anti-glomerular basement membrane (GBM) disease have been reported with linear deposit of immunoglobulin G (IgG) along GBM, but have undetectable anti-GBM antibodies in circulation by enzyme linked immunosorbent assays (ELISA). We speculated that the structure of the antigens recognized by these antibodies may contribute to the negative results of ELISA. Methods: Sera from four patients were collected, with typical linear deposit of IgG along GBM but no anti-GBM reactivity by commercial ELISA kits. Circulating anti-GBM antibodies were detected by indirect immunofluorescence. Antigen specificity and its conformational structure was investigated by western-blot analysis, using recombinant human alpha 1-alpha 5(IV) NC1 and chimeric proteins EA and EB as antigens. Results: The presence of circulating anti-GBM antibodies were confirmed by indirect immunofluorescence with linear deposit of IgG towards cryptic epitopes along GBM on normal kidney sections. These antibodies did not recognize recombinant human alpha 1, alpha 2, alpha 4 or alpha 5(IV)NC1, but could blot alpha 3(IV) NC1 under non-reducing non-boiling conditions on western-blot analysis, when the conformational epitope(s) on alpha 3(IV)NC1 were thought to be preserved. When alpha 3(IV) NC1 was prepared under reducing conditions with beta-mercaptoethanol and/ or boiled to destroy the disulfide bonds, the binding with the antibodies disappeared. Moreover, these antibodies recognized neither EA nor EB, indicating their distinct epitope repertoire. Conclusion: Circulating anti-GBM antibodies undetectable by ELISA could recognize cryptic and conformation-dependent epitopes restricted on alpha 3(IV) NC1, distinct from EA and EB. Indirect immunofluorescence was necessary for antibody detection and treatment monitoring under such circumstances.

语种: 英语
所属项目编号: 30725034 ; 30801020
项目资助者: National Science Foundation for Distinguished Young Scholars ; National Natural Science Fund of China
WOS记录号: WOS:000299252400008
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/53745
Appears in Collections:北京大学第一临床医学院_肾脏内科_期刊论文

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作者单位: Peking Univ, Div Renal, Dept Med,Hosp 1, Inst Nephrol,Key Lab Renal Dis,Minist Hlth China, Beijing 100034, Peoples R China

Recommended Citation:
Jia, Xiao-Yu,Qu, Zhen,Cui, Zhao,et al. Circulating anti-glomerular basement membrane autoantibodies against a3(IV)NC1 undetectable by commercially available enzyme-linked immunosorbent assays[J]. NEPHROLOGY,2012,17(2):160-166.
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