PREPARATION AND CHARACTERIZATION OF MONOCLONAL ANTITHYROTROPIN RECEPTOR ANTIBODIES OBTAINED FROM PERIPHERAL LYMPHOCYTES OF HYPOTHYROID PATIENTS WITH PRIMARY MYXEDEMA

Anti-TSH receptor antibodies (TSH-R Ab), which have been detected in t he serum of some patients with primary myxedema, are themselves consid ered to induce hypothyroidism. These are termed blocking-type TSH-R Ab (TSH-R BAb), because they inhibit adenylate cyclase stimulation by TS H on thyrocytes or nonthyroidal cells transfected with TSH-R complemen tary DNA. We prepared monoclonal TSH-R BAb and characterized them. Per ipheral lymphocytes from three patients with primary hypothyroidism an d potent TSH-R BAb were transformed by Epstein-Barr virus, and the cul ture supernatants were screened by TSH binding inhibitor immunoglobuli n (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 we re 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 show ed that the Ig isotypes of these clones with TBII and/or TSH-R BAb act ivity 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 determ ined in the other 9 clones. To summarize, 1) we obtained 22 clones tha t produced monoclonal TSH-R BAb, including 8 IgG-type clones. 2) The c lones exhibited dominant usage of the kappa-chain. 3) Although all TBI I clones had TSH-R BAb activity, their TBII and TSH-R BAb activities w ere not significantly correlated and two TSH-R BAb clones did not shaw TBII activity.