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Development of a simple enzyme-linked immunosorbent assay for the detection of autoantibodies in anti-p200 pemphigoid.
Br J Dermatol. 2011 Jan; 164(1):76-82.BJ

Abstract

BACKGROUND

Anti-p200 pemphigoid is a subepidermal blistering skin disease characterized by autoantibodies against a 200-kDa protein (p200) of the dermal-epidermal junction. The laminin γ1 chain has recently been identified as target antigen in this disease and the C-terminus was described as an immunodominant region of laminin γ1. Diagnosis of anti-p200 pemphigoid requires detection of serum IgG at the dermal side of 1 mol L(-1) salt-split skin by indirect immunofluorescence microscopy and labelling of a 200-kDa protein by Western blotting of dermal extract. However, preparation of dermal extract is not widely available, limiting the possibility of diagnosing this disease to a few laboratories.

OBJECTIVES

To develop a simple, sensitive and specific diagnostic tool for anti-p200 pemphigoid.

METHODS

Sera from patients with anti-p200 pemphigoid (n = 35), bullous pemphigoid (BP, n = 101), epidermolysis bullosa acquisita (EBA, n = 10), antilaminin 332 mucous membrane pemphigoid (MMP, n = 14), pemphigus vulgaris (PV, n = 51) and healthy volunteers (HV, n = 131) were tested by a novel enzyme-linked immunosorbent assay (ELISA) that employed a recombinant monomeric C-terminal fragment of human laminin γ1 (hLAMC1-cterm) expressed in Escherichia coli.

RESULTS

Serum reactivity with hLAMC1-cterm was detected in sera from 24 of 35 (69%) patients with anti-p200 pemphigoid, two of 101 (2%) with BP, 0 of 10 with EBA, two of 14 (14%) with anti-laminin 332 MMP, 0 of 51 with PV, and 0 of 131 HV.

CONCLUSIONS

This novel ELISA will facilitate the diagnosis of anti-p200 pemphigoid.

Authors+Show Affiliations

Department of Dermatology, University of Lübeck, 23538 Lübeck, Germany.No 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

20854435

Citation

Groth, S, et al. "Development of a Simple Enzyme-linked Immunosorbent Assay for the Detection of Autoantibodies in Anti-p200 Pemphigoid." The British Journal of Dermatology, vol. 164, no. 1, 2011, pp. 76-82.
Groth S, Recke A, Vafia K, et al. Development of a simple enzyme-linked immunosorbent assay for the detection of autoantibodies in anti-p200 pemphigoid. Br J Dermatol. 2011;164(1):76-82.
Groth, S., Recke, A., Vafia, K., Ludwig, R. J., Hashimoto, T., Zillikens, D., & Schmidt, E. (2011). Development of a simple enzyme-linked immunosorbent assay for the detection of autoantibodies in anti-p200 pemphigoid. The British Journal of Dermatology, 164(1), 76-82. https://doi.org/10.1111/j.1365-2133.2010.10056.x
Groth S, et al. Development of a Simple Enzyme-linked Immunosorbent Assay for the Detection of Autoantibodies in Anti-p200 Pemphigoid. Br J Dermatol. 2011;164(1):76-82. PubMed PMID: 20854435.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of a simple enzyme-linked immunosorbent assay for the detection of autoantibodies in anti-p200 pemphigoid. AU - Groth,S, AU - Recke,A, AU - Vafia,K, AU - Ludwig,R J, AU - Hashimoto,T, AU - Zillikens,D, AU - Schmidt,E, Y1 - 2010/12/06/ PY - 2010/9/22/entrez PY - 2010/9/22/pubmed PY - 2011/4/28/medline SP - 76 EP - 82 JF - The British journal of dermatology JO - Br J Dermatol VL - 164 IS - 1 N2 - BACKGROUND: Anti-p200 pemphigoid is a subepidermal blistering skin disease characterized by autoantibodies against a 200-kDa protein (p200) of the dermal-epidermal junction. The laminin γ1 chain has recently been identified as target antigen in this disease and the C-terminus was described as an immunodominant region of laminin γ1. Diagnosis of anti-p200 pemphigoid requires detection of serum IgG at the dermal side of 1 mol L(-1) salt-split skin by indirect immunofluorescence microscopy and labelling of a 200-kDa protein by Western blotting of dermal extract. However, preparation of dermal extract is not widely available, limiting the possibility of diagnosing this disease to a few laboratories. OBJECTIVES: To develop a simple, sensitive and specific diagnostic tool for anti-p200 pemphigoid. METHODS: Sera from patients with anti-p200 pemphigoid (n = 35), bullous pemphigoid (BP, n = 101), epidermolysis bullosa acquisita (EBA, n = 10), antilaminin 332 mucous membrane pemphigoid (MMP, n = 14), pemphigus vulgaris (PV, n = 51) and healthy volunteers (HV, n = 131) were tested by a novel enzyme-linked immunosorbent assay (ELISA) that employed a recombinant monomeric C-terminal fragment of human laminin γ1 (hLAMC1-cterm) expressed in Escherichia coli. RESULTS: Serum reactivity with hLAMC1-cterm was detected in sera from 24 of 35 (69%) patients with anti-p200 pemphigoid, two of 101 (2%) with BP, 0 of 10 with EBA, two of 14 (14%) with anti-laminin 332 MMP, 0 of 51 with PV, and 0 of 131 HV. CONCLUSIONS: This novel ELISA will facilitate the diagnosis of anti-p200 pemphigoid. SN - 1365-2133 UR - https://www.unboundmedicine.com/medline/citation/20854435/Development_of_a_simple_enzyme_linked_immunosorbent_assay_for_the_detection_of_autoantibodies_in_anti_p200_pemphigoid_ L2 - https://doi.org/10.1111/j.1365-2133.2010.10056.x DB - PRIME DP - Unbound Medicine ER -