Characterization of Autoantibodies in Pemphigus Using Antigen-Specific Enzyme-Linked Immunosorbent Assays with Baculovirus-Expressed Recombinant Desmogleins

Ken Ishii, Masayuki Amagai, Russell P. Hall, Takashi Hashimoto, Atsushi Takayanagi, Shinobu Gamou, Nobuyoshi Shimizu, Takeji Nishikawa

Research output: Contribution to journalArticle

414 Citations (Scopus)

Abstract

Pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are autoimmune skin diseases caused by autoantibodies against desmoglein (Dsg) 3 and Dsg1, respectively. Routine immunofluorescence testing of skin and serum from patients cannot distinguish between these two severe diseases since both have IgG Abs directed against keratinocyte cell surfaces. In this study, recombinant Dsg3 and Dsg1, produced as secreted proteins by baculovirus expression, have been utilized to develop ELISAs for the specific characterization of their autoantibodies. Of 49 PV sera, 46 were positive in the Dsg3 ELlSA and 44 of 46 PF sera were positive in the Dsg1 ELISA, compared with only 3 of 23 sera of bullous pemphigoid, and none of 53 normal control sera in both ELISAs. Both the Dsg3 and Dsg1 ELISAs were more specific and sensitive than conventional immunofluorescence staining. These Ag-specific ELISAs revealed that more than one-half of PV sera (26 of 49) had anti-Dsg1 Abs in addition to anti-Dsg3 Abs. PV patients who had not only oral mucous lesions but also significant skin involvement tended to have higher titers of anti-Dsg1 Abs. Furthermore, the ELISA reactivity correlated well with clinical disease activity in 5 of 6 PV and 5 of 5 PF patients. This ELISA provides a sensitive and highly specific assay for the diagnosis of patients with PV and PF, the correlation of disease activity with serum Ab levels, and a novel tool for investigating the immunopathogenesis of pemphigus.

Original languageEnglish
Pages (from-to)2010-2017
Number of pages8
JournalJournal of Immunology
Volume159
Issue number4
Publication statusPublished - 1997 Aug 15

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Desmogleins
Pemphigus
Baculoviridae
Autoantibodies
Enzyme-Linked Immunosorbent Assay
Antigens
Serum
Fluorescent Antibody Technique
Desmoglein 3
Bullous Pemphigoid
Skin
Keratinocytes
Skin Diseases
Autoimmune Diseases

ASJC Scopus subject areas

  • Immunology

Cite this

Characterization of Autoantibodies in Pemphigus Using Antigen-Specific Enzyme-Linked Immunosorbent Assays with Baculovirus-Expressed Recombinant Desmogleins. / Ishii, Ken; Amagai, Masayuki; Hall, Russell P.; Hashimoto, Takashi; Takayanagi, Atsushi; Gamou, Shinobu; Shimizu, Nobuyoshi; Nishikawa, Takeji.

In: Journal of Immunology, Vol. 159, No. 4, 15.08.1997, p. 2010-2017.

Research output: Contribution to journalArticle

Ishii, K, Amagai, M, Hall, RP, Hashimoto, T, Takayanagi, A, Gamou, S, Shimizu, N & Nishikawa, T 1997, 'Characterization of Autoantibodies in Pemphigus Using Antigen-Specific Enzyme-Linked Immunosorbent Assays with Baculovirus-Expressed Recombinant Desmogleins', Journal of Immunology, vol. 159, no. 4, pp. 2010-2017.
Ishii, Ken ; Amagai, Masayuki ; Hall, Russell P. ; Hashimoto, Takashi ; Takayanagi, Atsushi ; Gamou, Shinobu ; Shimizu, Nobuyoshi ; Nishikawa, Takeji. / Characterization of Autoantibodies in Pemphigus Using Antigen-Specific Enzyme-Linked Immunosorbent Assays with Baculovirus-Expressed Recombinant Desmogleins. In: Journal of Immunology. 1997 ; Vol. 159, No. 4. pp. 2010-2017.
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abstract = "Pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are autoimmune skin diseases caused by autoantibodies against desmoglein (Dsg) 3 and Dsg1, respectively. Routine immunofluorescence testing of skin and serum from patients cannot distinguish between these two severe diseases since both have IgG Abs directed against keratinocyte cell surfaces. In this study, recombinant Dsg3 and Dsg1, produced as secreted proteins by baculovirus expression, have been utilized to develop ELISAs for the specific characterization of their autoantibodies. Of 49 PV sera, 46 were positive in the Dsg3 ELlSA and 44 of 46 PF sera were positive in the Dsg1 ELISA, compared with only 3 of 23 sera of bullous pemphigoid, and none of 53 normal control sera in both ELISAs. Both the Dsg3 and Dsg1 ELISAs were more specific and sensitive than conventional immunofluorescence staining. These Ag-specific ELISAs revealed that more than one-half of PV sera (26 of 49) had anti-Dsg1 Abs in addition to anti-Dsg3 Abs. PV patients who had not only oral mucous lesions but also significant skin involvement tended to have higher titers of anti-Dsg1 Abs. Furthermore, the ELISA reactivity correlated well with clinical disease activity in 5 of 6 PV and 5 of 5 PF patients. This ELISA provides a sensitive and highly specific assay for the diagnosis of patients with PV and PF, the correlation of disease activity with serum Ab levels, and a novel tool for investigating the immunopathogenesis of pemphigus.",
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