Binding of immunoglobulin G from patients with autoimmune thyroid disease to rat sodium-iodide symporter peptides: Evidence for the iodide transporter as an autoantigen

The recent cloning of the rat sodium-iodide symporter (rNIS) from FRTL-5 cells makes possible studies of the role of this thyroid-specific protein as an antigen in autoimmune diseases of the thyroid (AITD). We generated 21 synthetic peptides replicating the entire sequence of the extramembranous domains (ExMD) of rNIS. Each was synthesized by automated chemistry, purified by high-pressure liquid chromatography (HPLC), and characterized by mass spectroscopy. Immunoglobulins were purified using protein A from serum of 27 patients with Graves' disease (GD), 27 patients with autoimmune hypothyroidism (HT), and 20 normal controls. Binding of IgG from patients and controls to each of the rNIS peptides was measured by enzyme-linked immunosorbent assay (ELISA). Binding of patient IgG significantly greater than control was observed with six peptides: peptide 262-280 (representing ExMD 8 between transmembrane [TM] domains VII and WI), peptide 437-444 (ExMD 11), peptides 468-487, 483-602, and 498-517 from ExMD 12, and peptide 560-579 from the proximal portion of the carboxyl terminus (ExMD 13). 63% of GD patients and 26% of HT patients immunoglobulin G (IgG) bound peptide 498-517 compared to zero controls. Similarly, 59% of GD were positive against peptide 468-487 versus zero controls. Peptide 262-280 bound IgG from 44% of GD patients, 15% of HT patients, and none of the controls. The remaining peptides showed little or no binding of patient IgG. These data indicate that patients with GD and HT possess antibodies that recognize rNIS significantly greater than do normal individuals, suggesting that the iodide transporter represents an important autoantigen in AITD. They further suggest that the incidence of the antibodies is higher in GD than HT, and that the antigenic epitopes involve ExMD 8, 11, 12, and 13.