Sensitivity of different assays for the serological diagnosis of epidermolysis bullosa acquisita

Analysis of a cohort of 24 Italian patients

V. Calabresi, A. Sinistro, E. Cozzani, C. Cerasaro, F. Lolicato, M. Muscianese, A. Parodi, B. Didona, G. Zambruno, G. Di Zenzo

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Background: Epidermolysis bullosa acquisita (EBA) is an autoimmune blistering disease characterized by tissue-bound and circulating autoantibodies to the dermal-epidermal junction. The autoantibody target is type VII collagen (Col VII) which is involved in dermal-epidermal adhesion. Diagnosis is made by clinical and histopathological findings, linear deposition of autoantibodies at the dermal-epidermal junction detected by direct immunofluorescence, and binding to the dermal side of salt-split skin by indirect immunofluorescence (IIF). However, the detection of specific anti-Col VII reactivity has an important confirmatory value. Methods: The humoral immune response in EBA sera was analysed by (i) IIF on human skin, (ii) a commercial Col VII ELISA, and (iii) immunoblotting on Col VII produced by an epithelial cell line. Objective: The aim of this study was to compare the sensitivity of different approaches for the serological diagnosis of EBA. Results: The vast majority of EBA sera (79.2%) bound to the Col VII non-collagenous domains by a commercial ELISA, while a small proportion of patients (12.5%) exclusively reacted to the collagenous domain by immunoblotting. Of note, the autoantibodies reactivity to Col VII was more frequently detected by IB (91.7%) than by IIF (83.3%) and ELISA (79.2%). Interestingly, 2 out of 24 sera recognized Col VII epitopes undetectable in the native secreted protein but present in the context of extracellular matrix proteins, as assessed by immunomapping on Col VII-deficient skin. Conclusion: Our findings show that the use of multiple assays allows to improve diagnostic performance. An algorithm for efficient serological diagnosis of EBA is proposed.

Original languageEnglish
Pages (from-to)483-490
Number of pages8
JournalJournal of the European Academy of Dermatology and Venereology
Volume28
Issue number4
DOIs
Publication statusPublished - 2014

Fingerprint

Epidermolysis Bullosa Acquisita
Cohort Studies
Skin
Autoantibodies
Indirect Fluorescent Antibody Technique
Enzyme-Linked Immunosorbent Assay
Immunoblotting
Collagen Type VII
Serum
Direct Fluorescent Antibody Technique
Extracellular Matrix Proteins
Humoral Immunity
Autoimmune Diseases
Epitopes
Salts
Epithelial Cells
Cell Line

ASJC Scopus subject areas

  • Dermatology
  • Infectious Diseases

Cite this

Sensitivity of different assays for the serological diagnosis of epidermolysis bullosa acquisita : Analysis of a cohort of 24 Italian patients. / Calabresi, V.; Sinistro, A.; Cozzani, E.; Cerasaro, C.; Lolicato, F.; Muscianese, M.; Parodi, A.; Didona, B.; Zambruno, G.; Di Zenzo, G.

In: Journal of the European Academy of Dermatology and Venereology, Vol. 28, No. 4, 2014, p. 483-490.

Research output: Contribution to journalArticle

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abstract = "Background: Epidermolysis bullosa acquisita (EBA) is an autoimmune blistering disease characterized by tissue-bound and circulating autoantibodies to the dermal-epidermal junction. The autoantibody target is type VII collagen (Col VII) which is involved in dermal-epidermal adhesion. Diagnosis is made by clinical and histopathological findings, linear deposition of autoantibodies at the dermal-epidermal junction detected by direct immunofluorescence, and binding to the dermal side of salt-split skin by indirect immunofluorescence (IIF). However, the detection of specific anti-Col VII reactivity has an important confirmatory value. Methods: The humoral immune response in EBA sera was analysed by (i) IIF on human skin, (ii) a commercial Col VII ELISA, and (iii) immunoblotting on Col VII produced by an epithelial cell line. Objective: The aim of this study was to compare the sensitivity of different approaches for the serological diagnosis of EBA. Results: The vast majority of EBA sera (79.2{\%}) bound to the Col VII non-collagenous domains by a commercial ELISA, while a small proportion of patients (12.5{\%}) exclusively reacted to the collagenous domain by immunoblotting. Of note, the autoantibodies reactivity to Col VII was more frequently detected by IB (91.7{\%}) than by IIF (83.3{\%}) and ELISA (79.2{\%}). Interestingly, 2 out of 24 sera recognized Col VII epitopes undetectable in the native secreted protein but present in the context of extracellular matrix proteins, as assessed by immunomapping on Col VII-deficient skin. Conclusion: Our findings show that the use of multiple assays allows to improve diagnostic performance. An algorithm for efficient serological diagnosis of EBA is proposed.",
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T2 - Analysis of a cohort of 24 Italian patients

AU - Calabresi, V.

AU - Sinistro, A.

AU - Cozzani, E.

AU - Cerasaro, C.

AU - Lolicato, F.

AU - Muscianese, M.

AU - Parodi, A.

AU - Didona, B.

AU - Zambruno, G.

AU - Di Zenzo, G.

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N2 - Background: Epidermolysis bullosa acquisita (EBA) is an autoimmune blistering disease characterized by tissue-bound and circulating autoantibodies to the dermal-epidermal junction. The autoantibody target is type VII collagen (Col VII) which is involved in dermal-epidermal adhesion. Diagnosis is made by clinical and histopathological findings, linear deposition of autoantibodies at the dermal-epidermal junction detected by direct immunofluorescence, and binding to the dermal side of salt-split skin by indirect immunofluorescence (IIF). However, the detection of specific anti-Col VII reactivity has an important confirmatory value. Methods: The humoral immune response in EBA sera was analysed by (i) IIF on human skin, (ii) a commercial Col VII ELISA, and (iii) immunoblotting on Col VII produced by an epithelial cell line. Objective: The aim of this study was to compare the sensitivity of different approaches for the serological diagnosis of EBA. Results: The vast majority of EBA sera (79.2%) bound to the Col VII non-collagenous domains by a commercial ELISA, while a small proportion of patients (12.5%) exclusively reacted to the collagenous domain by immunoblotting. Of note, the autoantibodies reactivity to Col VII was more frequently detected by IB (91.7%) than by IIF (83.3%) and ELISA (79.2%). Interestingly, 2 out of 24 sera recognized Col VII epitopes undetectable in the native secreted protein but present in the context of extracellular matrix proteins, as assessed by immunomapping on Col VII-deficient skin. Conclusion: Our findings show that the use of multiple assays allows to improve diagnostic performance. An algorithm for efficient serological diagnosis of EBA is proposed.

AB - Background: Epidermolysis bullosa acquisita (EBA) is an autoimmune blistering disease characterized by tissue-bound and circulating autoantibodies to the dermal-epidermal junction. The autoantibody target is type VII collagen (Col VII) which is involved in dermal-epidermal adhesion. Diagnosis is made by clinical and histopathological findings, linear deposition of autoantibodies at the dermal-epidermal junction detected by direct immunofluorescence, and binding to the dermal side of salt-split skin by indirect immunofluorescence (IIF). However, the detection of specific anti-Col VII reactivity has an important confirmatory value. Methods: The humoral immune response in EBA sera was analysed by (i) IIF on human skin, (ii) a commercial Col VII ELISA, and (iii) immunoblotting on Col VII produced by an epithelial cell line. Objective: The aim of this study was to compare the sensitivity of different approaches for the serological diagnosis of EBA. Results: The vast majority of EBA sera (79.2%) bound to the Col VII non-collagenous domains by a commercial ELISA, while a small proportion of patients (12.5%) exclusively reacted to the collagenous domain by immunoblotting. Of note, the autoantibodies reactivity to Col VII was more frequently detected by IB (91.7%) than by IIF (83.3%) and ELISA (79.2%). Interestingly, 2 out of 24 sera recognized Col VII epitopes undetectable in the native secreted protein but present in the context of extracellular matrix proteins, as assessed by immunomapping on Col VII-deficient skin. Conclusion: Our findings show that the use of multiple assays allows to improve diagnostic performance. An algorithm for efficient serological diagnosis of EBA is proposed.

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