Autoantibody epitope mapping of the 21-hydroxylase antigen in autoimmune addison’s disease

Yao Hua Song, Ellen L. Connor, Andrew Muir, Jin-Xiong She, Barbara Zorovich, Debbie Derovanesian, Noel Maclaren

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Patients with idiopathic Addison’s disease are characterized by cytoplasmic adrenal autoantibodies, detectable by indirect immunofluorescence of cryocut sections of human adrenal cortex. Recently, autoantibodies that bind a 55-kilodalton protein in the μsomal fraction of adrenal gland extracts identified to be the cytochrome P450 enzyme 21-hydroxylase have been found in Addisonian patient sera. We confirm the finding and report here the autoantigenic epitopes involved in the autoantibody reactivity using recombinant DNA technology. Six cDNA fragments spanning different regions of the 21-hydroxylase gene were expressed as fusion proteins with glutathione S-transferase in Escherichia coli. Immunoblot analyses were used to evaluate the reactivity of the recombinant proteins with patients' sera to determine the autoepitopes involved. We found that a conserved region (amino acids 164-356) reacted with 25 of 30 adrenal autoantibody-positive sera tested. One serum sample reacted only with the amino portion of the 21-hydroxylase (amino acids 1-162). In addition, 4 other enzymes important to steroid hormone biosynthesis, 11 β-hydroxylase, 17 α-hydroxylase, side-chain cleavage enzyme P450, and 3 β-hydroxysteroid dehydrogenase, were expressed in E. coli, but none of them gave positive autoantibody reactions by Western blot assays, even using sera from 5 patients with type I autoimmune polyglandular syndrome. The availability of recombinant antigens has permitted structural analysis of the autoepitopes involved in the autoimmune response to 21-hydroxylase in Addison’s disease. Our findings should lead to the development of a simple and specific tool for immunodiagnosis of the disease.

Original languageEnglish (US)
Pages (from-to)1108-1112
Number of pages5
JournalJournal of Clinical Endocrinology and Metabolism
Volume78
Issue number5
DOIs
StatePublished - Jan 1 1994
Externally publishedYes

Fingerprint

Epitope Mapping
Steroid 21-Hydroxylase
Addison Disease
Autoantibodies
Autoimmune Diseases
Epitopes
Antigens
Cytochrome P-450 Enzyme System
Serum
Mixed Function Oxygenases
Escherichia coli
3-Hydroxysteroid Dehydrogenases
Steroid hormones
Autoimmune Polyendocrinopathies
Amino Acids
Genetic engineering
Immunologic Tests
Recombinant DNA
Adrenal Cortex
Biosynthesis

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Endocrinology
  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Autoantibody epitope mapping of the 21-hydroxylase antigen in autoimmune addison’s disease. / Song, Yao Hua; Connor, Ellen L.; Muir, Andrew; She, Jin-Xiong; Zorovich, Barbara; Derovanesian, Debbie; Maclaren, Noel.

In: Journal of Clinical Endocrinology and Metabolism, Vol. 78, No. 5, 01.01.1994, p. 1108-1112.

Research output: Contribution to journalArticle

Song, Yao Hua ; Connor, Ellen L. ; Muir, Andrew ; She, Jin-Xiong ; Zorovich, Barbara ; Derovanesian, Debbie ; Maclaren, Noel. / Autoantibody epitope mapping of the 21-hydroxylase antigen in autoimmune addison’s disease. In: Journal of Clinical Endocrinology and Metabolism. 1994 ; Vol. 78, No. 5. pp. 1108-1112.
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