Ann Dermatol.  2012 Feb;24(1):45-55. 10.5021/ad.2012.24.1.45.

Usefulness of Enzyme-linked Immunosorbent Assay Using Recombinant BP180 and BP230 for Serodiagnosis and Monitoring Disease Activity of Bullous Pemphigoid

• Lee EH
• Kim YH
• Kim S
• Kim SE
• Kim SC

• KMID: 2156855
• DOI: http://doi.org/10.5021/ad.2012.24.1.45

Abstract

BACKGROUND
Bullous pemphigoid (BP) is an autoimmune subepidermal bullous disease associated with autoantibodies against BP180 and BP230. Enzyme-linked immunosorbent assay (ELISA) is a sensitive tool for the detection of immunoglobulin G (IgG) anti-BP180 and anti-BP230 autoantibodies.
OBJECTIVE
The aim of this study was to evaluate the usefulness of ELISA for diagnosing and monitoring the disease activity of BP.
METHODS
We evaluated serum IgG levels of anti-BP180 and anti-BP230 autoantibodies in 47 BP patients, 16 epidermolysis bullosa aquisita patients, and 15 healthy volunteers using ELISA. Through retrospective review of the medical records, the clinical characteristics of BP including disease activity, duration, pruritus severity and peripheral blood eosinophil counts were assessed.
RESULTS
The sensitivity of BP180 ELISA was 97.9%, BP230 ELISA 72.3%, and a combination of the two was 100%. The specificity of BP180 ELISA was 90.3%, BP230 ELISA 100%, and a combination of the two was 90.3%. BP180 ELISA scores showed strong associations with disease activity, pruritus severity, peripheral blood eosinophil counts, and disease duration, whereas BP230 ELISA scores did not.
CONCLUSION
BP180 and BP230 ELISAs are highly sensitive methods for the diagnosis of BP, and BP180 ELISA, in particular, is a sensitive tool for monitoring the disease activity of BP.

Keyword

BP180 protein; BP230 protein; Bullous pemphigoid; Enzyme-linked immunosorbent assay

MeSH Terms

Autoantibodies
Enzyme-Linked Immunosorbent Assay
Eosinophils
Epidermolysis Bullosa
Humans
Immunoglobulin G
Medical Records
Pemphigoid, Bullous
Pruritus
Retrospective Studies
Serologic Tests
Autoantibodies
Immunoglobulin G

Figure

• 

  Fig. 1 Schematic diagrams depicting the structure and regions of the recombinant proteins (marked by the blue line) of human BP180 (A) and BP230 (B) utilized in this enzymelinked immunosorbent assay study. NH2: N-terminus, NC16A: 16th noncollagenous domain, COOH: Cterminus, BP: bullous pemphigoid.

• 

  Fig. 2 Scatter plot representation of BP180 (NC16A domain, represented in black dots) and BP230 (C- and N-terminal recombinant proteins, represented in red dots) ELISA results (index value). Forty-seven BP sera and 31 control sera (16 EBA sera and 15 healthy volunteer sera) were examined. A dashed line indicates the cut-off value (BP180, 9 index; BP230, 9 index). The arithmetic means of index value are indicated by a line. Index value=(optical density [OD] of tested serum-OD of negative control)/(OD of positive control-OD of negative control)×100. Cut-off values were determined based on the manufacturer's recommendation. BP: bullous pemphigoid, EBA: epidermolysis bullosa aquisita, ELISA: enzyme-linked immunosorbent assay, NC16A: 16th non-collagenous domain.

• 

  Fig. 3 Correlation between (A) BP180 ELISA scores and disease activity scores, (B) BP230 ELISA scores and disease activity scores, (C) BP180 ELISA scores and blood eosinophil counts, (D) BP230 ELISA scores and blood eosinophil counts in BP patients. BP: bullous pemphigoid, ELISA: enzyme-linked immunosorbent assay.

• 

  Fig. 4 Comparison of ELISA scores and clinical characteristics of BP patients. (A) BP180 ELISA scores and disease duration, (B) BP230 ELISA scores and disease duration, (C) BP180 ELISA scores and disease severity grade, (D) BP230 ELISA scores and disease severity grade, (E) BP180 ELISA scores and pruritus severity grade, (F) BP230 ELISA scores and pruritus severity grade. BP: bullous pemphigoid, ELISA: enzyme-linked immunosorbent assay.

• 

  Fig. 5 The index values of BP180 ELISA and BP230 ELISA and titers of indirect immunofluorescence (IIF) in five BP patients before treatment and after remission. BP: bullous pemphigoid, ELISA: enzyme-linked immunosorbent assay.

• 

  Fig. 6 (A, B) The index values of BP180 ELISA and BP230 ELISA, titers of indirect immunofluorescence (IIF) and disease activity in one BP patient along the entire disease course. BP: bullous pemphigoid, ELISA: enzyme-linked immunosorbent assay.

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