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Development of a novel ELISA system for detection of anti-BP180 IgG and characterization of autoantibody profile in bullous pemphigoid patients.
Br J Dermatol. 2004 Nov; 151(5):1004-10.BJ

Abstract

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.

Authors+Show Affiliations

Molecular and Cell Biology Laboratory, Istituto Dermopatico dell'Immacolata, IDI-IRCCS, via Monti di Creta 104, 00167 Rome, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15541078

Citation

Mariotti, F, et al. "Development of a Novel ELISA System for Detection of anti-BP180 IgG and Characterization of Autoantibody Profile in Bullous Pemphigoid Patients." The British Journal of Dermatology, vol. 151, no. 5, 2004, pp. 1004-10.
Mariotti F, Grosso F, Terracina M, et al. Development of a novel ELISA system for detection of anti-BP180 IgG and characterization of autoantibody profile in bullous pemphigoid patients. Br J Dermatol. 2004;151(5):1004-10.
Mariotti, F., Grosso, F., Terracina, M., Ruffelli, M., Cordiali-Fei, P., Sera, F., Zambruno, G., Mastrogiacomo, A., & Di Zenzo, G. (2004). Development of a novel ELISA system for detection of anti-BP180 IgG and characterization of autoantibody profile in bullous pemphigoid patients. The British Journal of Dermatology, 151(5), 1004-10.
Mariotti F, et al. Development of a Novel ELISA System for Detection of anti-BP180 IgG and Characterization of Autoantibody Profile in Bullous Pemphigoid Patients. Br J Dermatol. 2004;151(5):1004-10. PubMed PMID: 15541078.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of a novel ELISA system for detection of anti-BP180 IgG and characterization of autoantibody profile in bullous pemphigoid patients. AU - Mariotti,F, AU - Grosso,F, AU - Terracina,M, AU - Ruffelli,M, AU - Cordiali-Fei,P, AU - Sera,F, AU - Zambruno,G, AU - Mastrogiacomo,A, AU - Di Zenzo,G, PY - 2004/11/16/pubmed PY - 2005/3/25/medline PY - 2004/11/16/entrez SP - 1004 EP - 10 JF - The British journal of dermatology JO - Br J Dermatol VL - 151 IS - 5 N2 - 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. SN - 0007-0963 UR - https://www.unboundmedicine.com/medline/citation/15541078/Development_of_a_novel_ELISA_system_for_detection_of_anti_BP180_IgG_and_characterization_of_autoantibody_profile_in_bullous_pemphigoid_patients_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=0007-0963&date=2004&volume=151&issue=5&spage=1004 DB - PRIME DP - Unbound Medicine ER -