Download PDF
Horm Metab Res 2015; 47(12): 880-888
DOI: 10.1055/s-0035-1554662
Endocrine Care
© Georg Thieme Verlag KG Stuttgart · New York

Bridge Technology with TSH Receptor Chimera for Sensitive Direct Detection of TSH Receptor Antibodies Causing Graves’ Disease: Analytical and Clinical Evaluation

C. U. Frank   *
1   KreLo GmbH Medical Diagnostics, Ulm, Germany
,
S. Braeth   *
1   KreLo GmbH Medical Diagnostics, Ulm, Germany
,
J. W. Dietrich
2   Department for Endocrinology and Diabetology, Ruhr University of Bochum, Bochum, Germany
,
D. Wanjura
3   Gemeinschaftspraxis Radiologie & Nuklearmedizin, Ulm, Germany
,
U. Loos
1   KreLo GmbH Medical Diagnostics, Ulm, Germany
› Author Affiliations
Further Information

Publication History

received 14 January 2015

accepted 19 May 2015

Publication Date:
16 June 2015 (online)

Also available at
   Permissions and Reprints

Abstract

Graves’ disease is caused by stimulating autoantibodies against the thyrotropin receptor inducing uncontrolled overproduction of thyroid hormones. A Bridge Assay is presented for direct detection of these thyroid-stimulating immunoglobulins using thyrotropin receptor chimeras. A capture receptor, formed by replacing aa residues 261–370 of the human thyrotropin receptor with residues 261–329 from rat lutropin/choriogonadotropin receptor and fixed to microtiter plates, binds one arm of the autoantibody. The second arm bridges to the signal receptor constructed from thyrotropin receptor (aa 21–261) and secretory alkaline phosphatase (aa 1–519) inducing chemiluminescence. The working range of the assay is from 0.3 IU/l to 50 IU/l with a cutoff of 0.54 IU/l and functional sensitivity of 0.3 IU/l. Sensitivity and specificity are 99.8 and 99.1%, respectively, with a diagnostic accuracy of 0.998. The low grey zone is from 0.3–0.54 IU/l. The stimulatory character of the assayed antibodies is shown through a good correlation (r=0.7079, p<10−7) to serum T4 levels of untreated patients. In Graves’ disease, titers are increased in associated eye disease. In 3 hypothyroid patients with sera positive in the thyrotropin receptor competition assay and in the blocking bioassay, antibodies are not detected by the Bridge Assay, while the monoclonal blocking antibody K1–70 was detected. In Hashimoto disease thyrotropin receptor autoantibodies are detected in some patients, but not in goiter. This Bridge Assay delivers good diagnostic accuracy for identification of Graves’ disease patients. Its high sensitivity may facilitate early detection of onset, remission, or recurrence of Graves’ disease enabling timely adaption of the treatment.

Human genes:
TSHR, Homo sapiens, acc. no. M31774.1

Key words

graves’ disease - TSH receptor - thyroid stimulating autoantibodies - bridge assay

* equal authorship


Supporting Information

    for this article is available online at for this article is available online at http://www.thieme-connect.de/products
  • Supporting Information
 

  • References

  • 1 Weetman AP. Graves’ disease. N Engl J Med 2000; 343: 1236-1248
    CrossrefPubMedSearch in Google Scholar
  • 2 Prabhakar BS, Bahn RS, Smith TJ. Current perspective of the pathogenesis of Graves’ disease and ophthalmopathy. Endocr Rev 2003; 24: 802-835
    CrossrefPubMedSearch in Google Scholar
  • 3 Bartalena L. Diagnosis and management of Graves’ disease: a global overview. Nat Rev Endocrinol 2013; 9: 724-734
    Search in Google Scholar
  • 4 Jameson JL. Endocrinology and metabolism. In: Fauci A, Braunwald E, Kasper D, Hauser S, Longo D, Jameson J, Loscalzo J. (eds.) Harrison’s Prinicples of Internal Medicine. 17th edition New York: McGraw Hill; 2008: 2233
    Search in Google Scholar
  • 5 Abraham-Nordling M, Byström K, Törring O, Lantz M, Berg G, Calissendorff J, Nyström HF, Jansson S, Jörneskog G, Karlsson FA, Nyström E, Ohrling H, Orn T, Hallengren B, Wallin G. Incidence of hyperthyroidism in Sweden. Eur J Endocrinol 2011; 165: 899-905
    CrossrefPubMedSearch in Google Scholar
  • 6 Johns Hopkins Medical Institutions. Autoimmune Disease Research Center. Information on Graves’ disease. Available at http://autoimmune.pathology.jhmi.edu/diseases.cfm?systemID=3&diseaseID=21 Accessed 18 July 2014
    PubMedSearch in Google Scholar
  • 7 Shewring G, Smith BR. An improved radioreceptor assay for TSH receptor antibodies. Clin Endocrinol (Oxf) 1982; 17: 409-417
    CrossrefPubMedSearch in Google Scholar
  • 8 Costagliola S, Morgenthaler NG, Hoermann R, Badenhoop K, Struck J, Freitag D, Poertl S, Weglöhner W, Hollidt JM, Quadbeck B, Dumont JE, Schumm-Draeger PM, Bergmann A, Mann K, Vassart G, Usadel KH. Second generation assay for thyrotropin receptor antibodies has superior diagnostic sensitivity for Graves’ disease. J Clin Endocrinol Metab 1999; 84: 90-97
    CrossrefPubMedSearch in Google Scholar
  • 9 Gassner D, Stock W, Golla R, Roth HJ. First automated assay for thyrotropin receptor autoantibodies. Clin Chem Lab Med 2009; 47: 1091-1095
    PubMedSearch in Google Scholar
  • 10 Vitti P, Valente WA, Ambesi-Impiombato FS, Fenzi GF, Pinchera A, Kohn LD. Graves’ IgG stimulation of continuously cultured rat thyroid cells: a sensitive and potentially useful clinical assay. J Endocrinol Invest 1982; 5: 179-182
    CrossrefPubMedSearch in Google Scholar
  • 11 Giuliani C, Cerrone D, Harii N, Thornton M, Kohn LD, Dagia NM, Fiore E, Bucci I, Chamblin T, Vitti P, Monaco F, Napolitano G. A TSHr-LH/CGr chimera that measures functional TSAb in Graves’ disease. J Clin Endocrinol Metab 2012; 97: E1106-E1115
    CrossrefPubMedSearch in Google Scholar
  • 12 Lytton SD, Ponto KA, Kanitz M, Matheis N, Kohn LD, Kahaly GJ. A novel thyroid stimulating immunoglobulin bioassay is a functional indicator of activity and severity of Graves’ orbitopathy. J Clin Endocrinol Metab 2010; 95: 2123-2131
    Search in Google Scholar
  • 13 Diana T, Kanitz M, Lehmann M, Li Y, Olivo PD, Kahaly GJ. Standardization of a bioassay for thyrotropin receptor stimulating autoantibodies. Thyroid 2015; 25: 169-175
    CrossrefPubMedSearch in Google Scholar
  • 14 Araki N, Iida M, Amino N, Morito S, Ide A, Nishihara E, Ito M, Saito J, Nishikawa T, Katsuragi K, Miyauchi A. Rapid bioassay for the detection of thyroid-stimulating antibodies using cyclic adenosine monophosphate-gated calcium channel and aequorin. Eur Thyroid J 2014; 4: 14-19
    PubMedSearch in Google Scholar
  • 15 Minich WB, Loos U. Isolation of radiochemically pure 125I-labeled human thyrotropin receptor and its use for the detection of pathological autoantibodies in sera from Graves’ patients. J Endocrinol 1999; 160: 239-245
    CrossrefPubMedSearch in Google Scholar
  • 16 Minich WB, Loos U. Detection of functionally different types of pathological autoantibodies against thyrotropin receptor in Graves’ patients sera by luminescent immunoprecipitation analysis. Exp Clin Endocrinol Diabetes 2000; 108: 110-119
    Thieme ConnectPubMedSearch in Google Scholar
  • 17 Chazenbalk GD, Pichurin P, McLachlan SM, Rapoport B. A direct binding assay for thyrotropin receptor autoantibodies. Thyroid 1999; 9: 1057-1061
    CrossrefPubMedSearch in Google Scholar
  • 18 Sanders J, Oda Y, Roberts S, Kiddie A, Richards T, Bolton J, McGrath V, Walters S, Jaskólski D, Furmaniak J, Smith BR. The interaction of TSH receptor autoantibodies with 125I-labelled TSH receptor. J Clin Endocrinol Metab 1999; 84: 3797-3802
    CrossrefPubMedSearch in Google Scholar
  • 19 Loos U, Franz C, Minich WB, Büsselmann I. Direct assay of TSH receptor autoantibodies causing Graves’ disease correlates with the clinical diagnosis closer than assays on TSH displacement. Horm Res 2007; 68 (Suppl. 03) 21-29
    PubMedSearch in Google Scholar
  • 20 Rees Smith B, Sanders J, Evans M, Tagami T, Furmaniak J. TSH receptor-autoantibody interactions. Horm Metab Res 2009; 41: 448-455
    Search in Google Scholar
  • 21 Tahara K, Ishikawa N, Yamamoto K, Hirai A, Ito K, Tamura Y, Yoshida S, Saito Y, Kohn LD. Epitopes for thyroid stimulating and blocking autoantibodies on the extracellular domain of the human thyrotropin receptor. Thyroid 1997; 7: 867-877
    CrossrefPubMedSearch in Google Scholar
  • 22 Rapoport B, Chazenbalk GD, Jaume JC, McLachlan SM. The thyrotropin (TSH) receptor: interaction with TSH and autoantibodies. Endocr Rev 1998; 19: 673-716
    PubMedSearch in Google Scholar
  • 23 Tahara K, Ban T, Minegishi T, Kohn LD. Immunoglobins from Graves’ disease patients interact with different sites on TSH receptor/LH-CG receptor chimeras then either TSH or immunoglobins from idiopathic myxedema patients. Biochem Biophys Res Commun 1991; 179: 70-77
    CrossrefPubMedSearch in Google Scholar
  • 24 McFarland KC, Sprengel R, Phillips HS, Köhler M, Rosemblit N, Nikolics K, Segaloff DL, Seeburg PH. Lutropin-choriogonadotropin receptor: an unusual member of the G protein-coupled receptor family. Science 1989; 245: 494-499
    CrossrefPubMedSearch in Google Scholar
  • 25 Loos U, Schmitt T, Frank C, Büsselmann I. Development of a novel bioassay for the detection of thyroid-stimulating autoantibodies in Graves’ disease (GD) facilitating routine measurement by reduced handling steps and high stability of cAMP monitoring reagent. Poster presentation at the ITC Paris 2010 P-0020 Available at https://b-com.mci-group.com/AbstractList/ITC2010.aspx
    PubMedSearch in Google Scholar
  • 26 Bahn RS, Burch HB, Cooper DS, Garber JR, Greenlee MC, Klein I, Laurberg P, McDougall IR, Montori VM, Rivkees SA, Ross DS, Sosa JA, Stan MN. American Thyroid Association; American Association of Clinical Endocrinologists . Hyperthyroidism and other causes of thyrotoxicosis: management guidelines of the American thyroid association and American association of clinical endocrinologists. Endocr Pract 2011; 17: e1-e65
    PubMedSearch in Google Scholar
  • 27 European Group on Graves’ Orbitopathy (EUGOGO) . Wiersinga WM, Perros P, Kahaly GJ, Mourits MP, Baldeschi L, Boboridis K, Boschi A, Dickinson AJ, Kendall-Taylor P, Krassas GE, Lane CM, Lazarus JH, Marcocci C, Marino M, Nardi M, Neoh C, Orgiazzi J, Pinchera A, Pitz S, Prummel MF, Sartini MS, Stahl M, von Arx G. Clinical assessment of patients with Graves’ orbitopathy: the EUGOGO recommendations to generalists, specialists and clinical researchers. Eur J Endocrinol 2006; 155: 387-389
    Search in Google Scholar
  • 28 NCCLS document EP5-A . Evaluation of precision performance of clinical chemistry devices; approved guideline. Wayne PA: Clinical and Laboratory Standards Institute; 1999
    Search in Google Scholar
  • 29 R development core team, 2009. R: A language and environment for statistical computing; R foundation for statistical computing; Vienna, Austria. Availabvle at http://R-project.org
    PubMed
  • 30 Morgenthaler NG, Minich WB, Willnich M, Bogusch T, Hollidt JM, Weglohner W, Lenzner C, Bergmann A. Affinity purification and diagnostic use of TSH receptor autoantibodies from human serum. Mol Cell Endocrinol 2003; 212: 73-79
    CrossrefPubMedSearch in Google Scholar
  • 31 Nakatake N, Sanders J, Richards T, Burne P, Barrett C, Dal Ora C, Presotto F, Betterle C, Furmaniak J, Rees Smith B. Estimation of serum TSH receptor autoantibody concentration and affinity. Thyroid 2006; 16: 1077-1084
    Search in Google Scholar
  • 32 Bahn R, Levy E, Wartofsky L. Graves’ disease. J Clin Endocrinol Metab 2007; 92 2p following 14A
    PubMedSearch in Google Scholar
  • 33 Eckstein A, Plicht M, Lax H, Neuhäuser M, Mann K, Lederbogen S, Heckmann C, Esser J, Morgenthaler NG. Thyrotropin receptor autoantibodies are independent risk factors for Graves’ ophtalmopathy and help to predict severity and outcome of the disease. J Clin Endocrinol Metab 2006; 91: 3464-3470
    Search in Google Scholar
  • 34 Evans M, Sanders J, Tagami T, Sanders P, Young S, Roberts E, Wilmot J, Hu X, Kabelis K, Clark J, Holl S, Richards T, Collyer A, Furmaniak J, Smith BR. Monoclonal autoantibodies to the TSH receptor, one with stimulating activity and one with blocking activity, obtained from the same blood sample. Clin Endocrinol 2010; 73: 404-412
    Search in Google Scholar
  • 35 Sanders P, Young S, Sanders J, Kabelis K, Baker S, Sullivan A, Evans M, Clark J, Wilmot J, Hu X, Roberts E, Powell M, Nunez Miguel R, Furmaniak J, Smith BR. Crystal structure of the TSH receptor (TSHR) bound to a blocking-type TSHR autoantibody. J Mol Endocrinol 2011; 46: 81-99
    Search in Google Scholar
  • 36 Li Y, Kim J, Diana T, Klasen R, Olivo PD, Kahaly GJ. A novel bioassay for anti-thyrotropin autoantibodies detects both thyroid-blocking and stimulating activity. Clin Exp Immunol 2013; 173: 390-397
    CrossrefPubMedSearch in Google Scholar
  • 37 Minich WB, Lenzner C, Bergmann A, Morgenthaler NG. A coated tube assay for the detection of blocking thyrotropin receptor autoantibodies. J Clin Endocrinol Metab 2004; 89: 352-356
    CrossrefPubMedSearch in Google Scholar
  • 38 Grasso Y, Kim MR, Faiman C, Kohn LD, Tahara K, Gupta MK. Epitope heterogeneity of thyrotropin receptor-blocking antibodies in Graves’ patients as detected with wild-type versus chimeric thyrotropin receptors. Thyroid 1999; 9: 531-537
    CrossrefPubMedSearch in Google Scholar
  • 39 Morgenthaler NG, Ho SC, Minich WB. Stimulating and Blocking Thyroid-Stimulating Hormone (TSH) Receptor Autoantibodies from patients with Graves’ disease and autoimmune hypothyroidism have very similar concentration, TSH receptor affinity, and binding sites. J Clin Endocrinol Metab 2007; 92: 1058-1065
    CrossrefPubMedSearch in Google Scholar
  • 40 Ando T, Latif R, Daniel S, Eguchi K, Davies T. Dissecting linear and conformational epitopes on the native thyrotropin receptor. Endocrinology 2004; 14: 5185-5193
    PubMedSearch in Google Scholar
  • 41 Giuliani C, Cerrone D, Harii N, Thornton M, Kohn LD, Dagia NM, Bucci I, Carpentieri M, Di Nenno B, Di Blasio A, Vitti P, Monaco F, Napolitano G. A TSHR-LH/CGR chimera that measures functional thyroid-stimulating autoantibodies (TSAb) can predict remission or recurrence in Graves’ patients undergoing antithyroid drug (ATD) treatment. J Clin Endocrinol Metab 2012; 97: E1080-E1087
    CrossrefPubMedSearch in Google Scholar
  • 42 Zöphel K, Grüning T, Roggenbuck D, Wunderlich G, Kotzerke J. On specificity of 2nd generation TSH receptor autoantibody measurements. Clin Lab 2008; 54: 243-249
    PubMedSearch in Google Scholar
  • 43 Schott M, Morgenthaler NG, Fritzen R, Feldkamp J, Willenberg HS, Scherbaum WA, Seissler J. Levels of autoantibodies against human TSH receptor predict relapse of hyperthyroidism in Graves’ disease. Horm Metab Res 2004; 36: 92-96
    Thieme ConnectPubMedSearch in Google Scholar
  • 44 Eckstein A, Mann K, Kahaly GJ, Grussendorf M, Reiners C, Feldkamp J, Quadbeck B, Bockisch A, Schott M. Role of TSH receptor autoantibodies for the diagnosis of Graves’ disease and for the prediction of the course of hyperthyroidism and ophtalmopathy. Recommendations of the Thyroid Section of the German Society of Endocrinology. Med Klin (Munich) 2009; 104: 343-348
    CrossrefPubMedSearch in Google Scholar
  • 45 Carella C, Mazziotti G, Sorvillo F, Piscopo M, Cioffi M, Pilla P, Nersita R, Iorio S, Amato G, Braverman LE, Roti E. Serum thyrotropin receptor antibodies concentrations in patients with Graves’ disease before, at the end of methimazole treatment, and after drug withdrawal: evidence that the activity of thyrotropin receptor antibody and/or thyroid response modify during the observation period. Thyroid 2006; 16: 295-302
    CrossrefPubMedSearch in Google Scholar