Preparation and characterization of monoclonal antithyrotropin receptor antibodies obtained from peripheral lymphocytes of hypothyroid patients with primary myxedema

J Clin Endocrinol Metab. 1994 Dec;79(6):1600-4. doi: 10.1210/jcem.79.6.7989462.

Abstract

Anti-TSH receptor antibodies (TSH-R Ab), which have been detected in the serum of some patients with primary myxedema, are themselves considered to induce hypothyroidism. These are termed blocking-type TSH-R Ab (TSH-R BAb), because they inhibit adenylate cyclase stimulation by TSH on thyrocytes or nonthyroidal cells transfected with TSH-R complementary DNA. We prepared monoclonal TSH-R BAb and characterized them. Peripheral lymphocytes from three patients with primary hypothyroidism and potent TSH-R BAb were transformed by Epstein-Barr virus, and the culture supernatants were screened by TSH binding inhibitor immunoglobulin (TBII) assay. Twenty positive and 7 negative lymphocyte clones were obtained; their monoclonality was confirmed by Southern blot analysis, using an immunoglobulin (Ig) JH probe. These monoclonal antibodies were then tested for TSH-R BAb activity. TSH-R BAb activity ranged from 24.1-58.5% (normal range, < 24%) in all 20 TBII-positive clones and in 2 of 7 TBII-negative clones. An enzyme-linked immunosorbent assay showed that the Ig isotypes of these clones with TBII and/or TSH-R BAb activity were IgG in 8 and IgM in 14. Another enzyme-linked immunosorbent assay and Southern blot analysis of the light chains revealed that 13 clones had kappa-chains, whereas the light chains could not be determined in the other 9 clones. To summarize, 1) we obtained 22 clones that produced monoclonal TSH-R BAb, including 8 IgG-type clones. 2) The clones exhibited dominant usage of the kappa-chain. 3) Although all TBII clones had TSH-R BAb activity, their TBII and TSH-R BAb activities were not significantly correlated, and two TSH-R BAb clones did not show TBII activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / blood*
  • Antibodies, Monoclonal / genetics
  • Autoantibodies / blood*
  • Autoantibodies / genetics
  • Blotting, Southern
  • CHO Cells
  • Clone Cells
  • Cricetinae
  • DNA / analysis
  • Gene Rearrangement, B-Lymphocyte, Heavy Chain
  • Gene Rearrangement, B-Lymphocyte, Light Chain
  • Humans
  • Hypothyroidism / immunology*
  • Immunoglobulin G / blood
  • Immunoglobulin M / blood
  • Myxedema / immunology*
  • Receptors, Thyrotropin / genetics
  • Receptors, Thyrotropin / immunology*
  • T-Lymphocytes / immunology*
  • Transfection

Substances

  • Antibodies, Monoclonal
  • Autoantibodies
  • Immunoglobulin G
  • Immunoglobulin M
  • Receptors, Thyrotropin
  • DNA