title: Detection of binding and blocking autoantibodies to the human sodium-iodide symporter in patients with autoimmune thyroid disease.
abstract: The sodium iodide symporter (NIS) is a novel autoantigen in autoimmune thyroid disease (ATD). A recent study has described the development of a bioassay for human (h) NIS antibody detection, but this will not detect antibodies that bind the symporter without modulating its activity. Therefore, the establishment of a binding assay is of importance to determine the overall prevalence of hNIS antibodies in ATD patients. An in vitro transcription and translation system was used to produce [35S]-labeled hNIS. The radiolabeled ligand reacted specifically in immunoprecipitation experiments with rabbit antiserum raised against a peptide fragment of hNIS. Subsequently, the reactivity of control and ATD sera to translated [35S]hNIS was determined using RIAs. A significant difference in the frequency of hNIS antibody-positive sera was found when patients with either Graves' disease (GD) or autoimmune hypothyroidism (AH) were compared with normal controls (P = 0.01 and P = 0.03, respectively). Of 49 GD and 29 AH sera tested, 11 (22%) and 7 (24%), respectively, were found to contain hNIS antibodies. Differences were also significant when the antibody-binding indices of the control group of sera were compared with those of both the GD and the AH patient sera (P < 0.0001 and P = 0.001, respectively). In contrast, sera from 10 patients with Addison's disease and 10 patients with vitiligo (without associated ATD) were all negative for antibody reactivity to the symporter. No differences were detected when the antibody binding indices of either the Addison's disease or the vitiligo sera were compared with those of the normal sera group (P = 0.9 and P = 0.6, respectively). Eight of the 11 (73%) GD and 3 of the 7 (43%) AH sera, which were positive for hNIS antibodies in the immunoprecipitation assay, were also found to inhibit iodide uptake in hNIS-transfected CHO-K1 cells, suggesting the existence of antibodies in some serum samples that bind to the symporter without modulating its function. Overall, a significant correlation was found between the iodide uptake inhibition and the binding assays for hNIS antibody detection (r = 0.49, P < 0.0001). In summary, we have developed a specific and quantitative assay for the detection of hNIS binding antibodies in sera of patients with ATD. This system offers the advantage of studying antibody reactivity against conformational epitopes and will be useful in understanding the role of NIS autoreactivity in the pathogenesis of ATD.
authors.affiliation: Division of Clinical Sciences, Northern General Hospital, University of Sheffield, United Kingdom. Ramzi@Ajjan.Freeserve.co.uk
authors.forename: R A
authors.initials: RA
authors.lastname: Ajjan
authors.forename: E H
authors.initials: EH
authors.lastname: Kemp
authors.forename: E A
authors.initials: EA
authors.lastname: Waterman
authors.forename: P F
authors.initials: PF
authors.lastname: Watson
authors.forename: T
authors.initials: T
authors.lastname: Endo
authors.forename: T
authors.initials: T
authors.lastname: Onaya
authors.forename: A P
authors.initials: AP
authors.lastname: Weetman
mesh_terms: D000224:Addison Disease / Q000097:blood / Q000276:immunology; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001323:Autoantibodies / Q000097:blood*; D016466:CHO Cells; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006224:Cricetinae; D005260:Female; D006111:Graves Disease / Q000097:blood / Q000276:immunology*; D006801:Humans; D007037:Hypothyroidism / Q000097:blood / Q000276:immunology; D007454:Iodides / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D008875:Middle Aged; D011487:Protein Conformation; D011817:Rabbits; D011863:Radioimmunoassay; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D027981:Symporters; D013967:Thyroiditis, Autoimmune / Q000097:blood / Q000276:immunology*; D014162:Transfection; D000099209:Sodium-Iodide Symporters
chemical_list: D001323:Autoantibodies; D002352:Carrier Proteins; D007454:Iodides; D008565:Membrane Proteins; D011994:Recombinant Proteins; D027981:Symporters; D000099209:Sodium-Iodide Symporters