Autoantigenicity of DFS70 is restricted to the conformational epitope of C-terminal alpha-helical domain

Yasushi Ogawa, Kazumitsu Sugiura, Akihiro Watanabe, Mitoshi Kunimatsu, Masaki Mishima, Yasushi Tomita, Yoshinao Muro

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Autoantibodies against DFS70 (Dense Fine Speckles 70) are found in 30% of Japanese atopic dermatitis patients, and less frequently in patients with other diseases. We have recently reported that they are also seen in 11% of hospital workers, but in only ∼2% of patients with systemic rheumatic disease. In this study, in order to investigate the possible pathological role of anti-DFS70 antibodies, fine epitope mapping was carried out using 93 anti-DFS70 autoantibody-positive sera. Immunoblotting using overlapping peptides failed to reveal major linear epitopes. Western blotting using various truncated proteins showed a strikingly uniform epitope distribution on a suspected tertiary structure expressed by DFS70349-435. Some sera showed reactivity only in an immunoprecipitation assay using an in vitro translated DFS70. Circular dichroism analysis revealed that DFS349-435 contains an approximately 40% alpha-helical conformation, while an overlapping, non-antigenic peptide is composed of random coiled structures. The skewed single major epitope enabled us to establish a highly quantitative ELISA for the epitope region. Antibody titers showed no significant differences between the diseased group and healthy individuals. We propose that anti-DFS70 antibody may be a natural autoantibody, which might modify or reflect the inflammatory process of various disorders.

Original languageEnglish
Pages (from-to)221-231
Number of pages11
JournalJournal of Autoimmunity
Volume23
Issue number3
DOIs
Publication statusPublished - 01-01-2004
Externally publishedYes

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Epitopes
Autoantibodies
Antibodies
Epitope Mapping
Peptides
Atopic Dermatitis
Circular Dichroism
Rheumatic Diseases
Serum
Immunoprecipitation
Immunoblotting
Western Blotting
Enzyme-Linked Immunosorbent Assay
Proteins

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

Ogawa, Yasushi ; Sugiura, Kazumitsu ; Watanabe, Akihiro ; Kunimatsu, Mitoshi ; Mishima, Masaki ; Tomita, Yasushi ; Muro, Yoshinao. / Autoantigenicity of DFS70 is restricted to the conformational epitope of C-terminal alpha-helical domain. In: Journal of Autoimmunity. 2004 ; Vol. 23, No. 3. pp. 221-231.
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Autoantigenicity of DFS70 is restricted to the conformational epitope of C-terminal alpha-helical domain. / Ogawa, Yasushi; Sugiura, Kazumitsu; Watanabe, Akihiro; Kunimatsu, Mitoshi; Mishima, Masaki; Tomita, Yasushi; Muro, Yoshinao.

In: Journal of Autoimmunity, Vol. 23, No. 3, 01.01.2004, p. 221-231.

Research output: Contribution to journalArticle

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