Document Detail

Development of a novel ELISA system for detection of anti-BP180 IgG and characterization of autoantibody profile in bullous pemphigoid patients.
MedLine Citation:
PMID:  15541078     Owner:  NLM     Status:  MEDLINE    
BACKGROUND: The NC16A immunodominant region of the bullous pemphigoid (BP) antigen BP180 has been used to develop several enzyme-linked immunosorbent assays (ELISAs) as diagnostic tools for BP autoantibody detection. OBJECTIVES: Because BP180 autoantibody reactivity is not restricted to NC16A, we have investigated the possibility of developing an ELISA based on selected epitopes additional to this immunodominant region. METHODS: Initially 78 BP sera were tested using an NC16A ELISA and IgG reactivity was detected in 64 BP sera (82%). The 14 NC16A-negative BP sera were then analysed by immunological screening against seven BP180-specific epitopes. Recombinant phages displaying BP180 epitopes were grown as plaques, blotted onto a nitrocellulose filter and incubated with BP sera. RESULTS: Three and five NC16A-negative BP sera reacted with epitopes AA 1080-1107 and AA 1331-1404 of the BP180 ectodomain, respectively. Thus, a novel ELISA with GST-1080 and GST-1331 (GST-1080/1331) was developed: 32 of 78 BP sera (41%) proved positive by this assay. The combined use of ELISAs with GST-NC16A and GST-1080/1331 detected IgG reactivity in 72 of 78 BP sera, increasing the sensitivity from 82% to 92%. In addition, autoreactivity against the three extracellular epitopes appeared to be related to the presence of both skin and mucosal involvement as assessed by Fisher's exact probability test. CONCLUSIONS: Our findings further characterize the autoimmune response in BP by identifying a subgroup of NC16A-negative patients who react with different BP180 extracellular epitopes. The developed ELISA system appears more sensitive than the ELISA based on NC16A alone and also informative about the epitope profile of BP patients.
F Mariotti; F Grosso; M Terracina; M Ruffelli; P Cordiali-Fei; F Sera; G Zambruno; A Mastrogiacomo; G Di Zenzo
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The British journal of dermatology     Volume:  151     ISSN:  0007-0963     ISO Abbreviation:  Br. J. Dermatol.     Publication Date:  2004 Nov 
Date Detail:
Created Date:  2004-11-15     Completed Date:  2005-03-24     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  0004041     Medline TA:  Br J Dermatol     Country:  England    
Other Details:
Languages:  eng     Pagination:  1004-10     Citation Subset:  IM    
Molecular and Cell Biology Laboratory, Istituto Dermopatico dell'Immacolata, IDI-IRCCS, via Monti di Creta 104, 00167 Rome, Italy.
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MeSH Terms
Aged, 80 and over
Autoantibodies / blood*
Autoantigens / immunology*
Enzyme-Linked Immunosorbent Assay / methods
Immunodominant Epitopes / immunology
Immunoglobulin G / blood*
Middle Aged
Mucous Membrane / immunology,  pathology
Non-Fibrillar Collagens
Pemphigoid, Bullous / diagnosis,  immunology*,  pathology
Sensitivity and Specificity
Reg. No./Substance:
0/Autoantibodies; 0/Autoantigens; 0/Immunodominant Epitopes; 0/Immunoglobulin G; 0/Non-Fibrillar Collagens; 0/collagen type XVII

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