Analysis of autoantibody reactivity in patients with graves' disease using recombinant extracellular domain of the human thyrotropin receptor and synthetic peptides

Ji Lao Fan, Gattadahalli Seetharamaiah, Rajesh K. Desai, John S. Dallas, Neelam M. Wagle, Bellur S. Prabhakar

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Graves' disease is characterized by hyperthyroidism leading to enhanced production of thyroid hormones. Hyperthyroidism is primarily mediated by the binding of autoantibodies to the thyrotropin receptor (TSHr). In the past, either thyroid cells or thyroid membranes were used as a source of TSHr to detect anti-TSHr antibodies. Recently, we expressed the extracellular domain of the human TSHr (ETSHr) using the baculovirus expression system. In this study, we used ETSHr protein in an ELISA to detect anti-TSHr antibodies. Our data show that this assay can be used to analyze and quantitate isotype specific antibodies against the TSHr. To map immunogenic epitopes on the TSHr, we tested patients sera against synthetic peptides derived from two highly immunogenic regions (amino acid, AA 12-46 and 316-397) of the receptor. Although sera from patients with Graves' disease reacted with several peptides, they showed particularly strong reactivity against peptides from a relatively narrow region (i.e. AA 352-394) of the TSHr. The present study demonstrates the usefulness of the recombinant ETSHr to detect and characterize anti-TSHr antibodies in a simple and sensitive ELISA, and has lead to the identification of some of the immuno-reactive epitopes on the TSHr.

Original languageEnglish (US)
Pages (from-to)285-291
Number of pages7
JournalAutoimmunity
Volume15
Issue number4
DOIs
StatePublished - 1993
Externally publishedYes

Fingerprint

Thyrotropin Receptors
Graves Disease
Autoantibodies
Anti-Idiotypic Antibodies
Hyperthyroidism
Peptides
Epitopes
Thyroid Gland
Enzyme-Linked Immunosorbent Assay
Baculoviridae
Serum
Thyroid Hormones
Amino Acids
Membranes
Antibodies
Proteins

Keywords

  • Autoantibody
  • Autoimmunity
  • Graves' disease
  • Peptides
  • Thyrotropin receptor

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Analysis of autoantibody reactivity in patients with graves' disease using recombinant extracellular domain of the human thyrotropin receptor and synthetic peptides. / Fan, Ji Lao; Seetharamaiah, Gattadahalli; Desai, Rajesh K.; Dallas, John S.; Wagle, Neelam M.; Prabhakar, Bellur S.

In: Autoimmunity, Vol. 15, No. 4, 1993, p. 285-291.

Research output: Contribution to journalArticle

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