Development of a system for detection of circulating antibodies against hemidesmosomal proteins in patients with bullous pemphigoid

S. Husz, M. Kiss, Katalin Molnár, Ilona Marczinovits, János Molná, G. Tóth, A. Dobozy

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Specific antibodies directed against special hemidesmosomal proteins are involved in the pathogenesis of bullous pemphigoid (BP), and detection of these antibodies is crucial for a correct diagnosis. As the BP autoantigen primary structures are known, the question was addressed as to whether it is possible to demonstrate circulating antibodies against BP autoantigens (BPAG1 and BPAG2) by means of an ELISA system, using antigenic epitopes. With the help of the programs Peptidestructure and Plotstructure, antigenic epitopes of BP antigens were predicted, chemically synthesized and screened using serum from 43 proven BP patients. The coding sequences of the best antigenic epitopes were then chemically synthesized and inserted as monomer and homo- or hetero-oligomer forms into fusion-expression plasmids (PGEX-4T, Pharmacia) in-frame to the C-terminus of glutathione-S-transferase. Fusion products were expressed and purified from Escherichia coli cells by affinity chromatography. The recombinant proteins were used for the detection of antibodies in the serum of 43 BP patients and of 60 controls (including 30 healthy persons, 22 patients with pemphigus vulgaris and 8 patients with other bullous dermatoses). Use of the homo- and hetero-oligomers of the recombinant fusion peptides increased the sensitivity of the disease-specific antibody detection. When a mixture of the best recombinant fusion proteins was used, the sensitivity of the ELISA assays in the case of the BP patients' serum was 0.90. This system could form the basis of a rapid and simple system for the diagnosis of BP.

Original languageEnglish
Pages (from-to)217-224
Number of pages8
JournalArchives of Dermatological Research
Volume292
Issue number5
Publication statusPublished - 2000

Fingerprint

Bullous Pemphigoid
Antibodies
Proteins
Epitopes
Autoantigens
Serum
Enzyme-Linked Immunosorbent Assay
Vesiculobullous Skin Diseases
Recombinant Fusion Proteins
Pemphigus
Glutathione Transferase
Affinity Chromatography
Recombinant Proteins
Plasmids
Escherichia coli
Antigens
Peptides

Keywords

  • Antigenic epitopes
  • Bullous pemphigoid antigen 1 and 2
  • Diagnostic ELISA system
  • Recombinant bacterial proteins

ASJC Scopus subject areas

  • Dermatology

Cite this

Development of a system for detection of circulating antibodies against hemidesmosomal proteins in patients with bullous pemphigoid. / Husz, S.; Kiss, M.; Molnár, Katalin; Marczinovits, Ilona; Molná, János; Tóth, G.; Dobozy, A.

In: Archives of Dermatological Research, Vol. 292, No. 5, 2000, p. 217-224.

Research output: Contribution to journalArticle

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