Skip to main content
SpringerLink
Log in

Detection of thyroid-stimulating antibodies in thyroid diseases, employing rat thyroid fragment perifusion

  • Published:
Journal of Endocrinological Investigation Aims and scope Submit manuscript

Abstract

The technique of perifusing rat thyroid fragments was used to investigate the presence of thyroid-stimulating antibodies (TSAb) in the sera of 48 patients. Response to IgG was measured by determining the mean rate of release of T4 (R) during a 30-min perifusion and the secretion peak (Imax) by means of samples taken every 5 min. Values found to be above the mean + 2 SD of the control values of R or Imax were considered to be positive. TSAb were found in all the 17 patients with untreated Graves’ disease (GD) and in the 2 treated with antithyroid drugs, but not in the 3 who had undergone surgery or131I treatment or in the 2 on corticosteroid treatment. TSAb were also found in 2 out of 3 patients with untreated nodular toxic goiter (UNTG) and in 6 out of 8 with diffuse nontoxic goiter (DNG) but at lower levels. In the untreated GD group, R and Imax correlated significantly with the corresponding IgG concentrations (from 90 to 800 µg/ml), suggesting TSAb activity which can be compared from one patient to another. TSAb activity did not correlate with thyroid function tests in any group. In all the groups it induced an early secretion peak followed by a decreasing response throughout the stimulation period, as was previously found with 65 mlU/ml TSH. The specificity of this technique was verified by five different control methods: 1) the perifusion technique was checked by using KRBG buffer alone; sera were studied from 2) a group of healthy controls, 3) a second control group with no thyroid disease but presenting “inflammatory” hypergammaglobulinemia 4) a group of patients with primary thyroid insufficiency, and 5) two patients with GD, before and after sera treatment with staphylococcal protein A. We conclude that RTFP is a rapid, sensitive and reliable technique for investigating the presence of TSAb, since thyroid-stimulating activity was present in all the patients with untreated GD; TSAb detected by this technique are thus not specific for hyperthyroid GD but they were nevertheless found to be useful humoral markers in other thyroid diseases.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Smith B.R., Munro D.S., Dorrington K.J. The distribution of the long acting thyroid stimulator among γG-immunoglobulins. Biochim. Biophys. Acta 188: 89, 1969.

    Article  PubMed  CAS  Google Scholar 

  2. Adams D.D., Purves H.D. Abnormal responses in the assay of thyrotropin. Proc. Univ. Otago Sch. Med. 34: 11, 1956.

    Google Scholar 

  3. Adams D.D., Kennedy T.H. Evidence to suggest that LATS protector stimulates the human thyroid. J. Clin. Endocrinol. Metab. 33: 47, 1971.

    Article  PubMed  CAS  Google Scholar 

  4. Onaya T., Kotani M., Yamada T., Ochi Y. New in vitro tests to detect the thyroid stimulator in sera from hyperthyroid patients by measuring colloid droplet formation and cyclic AMP in human thyroid slices. J. Clin. Endocrinol. Metab. 36: 859, 1973.

    Article  PubMed  CAS  Google Scholar 

  5. Smith B.R., Hall R. Thyroid-stimulating immunoglobulins in Graves’ disease. Lancet 2: 427, 1974.

    Article  PubMed  CAS  Google Scholar 

  6. Orgiazzi J., Williams D.E., Chopra I.J., Solomon D.H. Human thyroid adenyl-cyclase stimulating activity in immunoglobulins G of patients with Graves’ disease. J. Clin. Endocrinol. Metab. 42: 34, 1976.

    Article  Google Scholar 

  7. Zakarija M., McKenzie J.M., Banovac K. The clinical significance of the assay of thyroid stimulating antibody in Graves’ disease. Ann. Intern. Med. 93: 28, 1980.

    Article  PubMed  CAS  Google Scholar 

  8. Etienne-Decerf J., Winand R.J. A sensitive technique for determination of thyroid stimulating immunoglobulin (TSI) in unfractionated serum. Clin. Endocrinol. (Oxf.) 14: 83, 1981.

    Article  CAS  Google Scholar 

  9. Stöckle G., Wahl R., Seif F.J. Micromethod of human thyrocyte cultures for detection of thyroid stimulating antibodies and thyrotrophin. Acta Endocrinol. (Kbh.) 97: 369, 1981.

    Google Scholar 

  10. Davies T.F., Platzer M., Schwartz A., Friedman E. Functionality of thyroid-stimulating antibodies assessed by cryopreserved human thyroid cell bioassay. J. Clin. Endocrinol. Metab. 57: 1021, 1983.

    Article  PubMed  CAS  Google Scholar 

  11. Bliddal H., Bech K., Petersen P.H., Stersboek-Nielsen K., Friis T. Evidence of a correlation between thyrotrophin receptor binding inhibition and thyroid adenylate cyclase activation by immunoglobulins in Graves’ disease before and during long term antithyroid treatment. Acta Endocrinol. (Kbh.) 101: 35, 1982.

    CAS  Google Scholar 

  12. Sugenoya A., Kidd A., Row V.V., Volpé R. Correlation between thyrotropin displacing activity and human thyroid stimulating activity by immunoglobulins from patients with Graves’ disease and other thyroid disorders. J. Clin. Endocrinol. Metab. 48: 398, 1979.

    Article  PubMed  CAS  Google Scholar 

  13. Macchia E., Fenzi G.F., Monzani F., Lippi F., Vitti P., Grasso L., Bartalena L., Baschieri L., Pinchera A. Comparison between thyroid-stimulating and TSH binding inhibiting immunoglobulins of Graves’ disease. Clin. Endocrinol. (Oxf.) 15: 175, 1981.

    Article  CAS  Google Scholar 

  14. Laurberg P., Weeke J. T3 release from thyroid slices as an assay for thyroid stimulators. Scand. J. Clin. Lab. Invest. 35: 723, 1975.

    Article  PubMed  CAS  Google Scholar 

  15. Takata I., Suzuki Y., Saida K., Sato T. Human thyroid stimulating activity and clinical state in antithyroid treatment of juvenile Graves’ disease. Acta Endocrinol. (Kbh.) 94: 46, 1980.

    CAS  Google Scholar 

  16. Smyth P.P.A., Neylan K., O’Donovan D.K. The prevalence of thyroid stimulating antibodies in goitrous disease assessed by cytochemical section bioassay. J. Clin. Endocrinol. Metab. 54: 357, 1982.

    Article  PubMed  CAS  Google Scholar 

  17. Smyth P.P.A., Neylan D., O’Donovan D.K. Association of thyroid stimulating immunoglobulins and thyrotropin releasing hormone responsiveness in women with euthyroid goiter. J. Clin. Endocrinol. Metab. 51: 1001, 1983.

    Article  Google Scholar 

  18. Attali J.R., Darnis D., Valensi P., Weisselberg C., Sebaoun J. Use of the perifusion technique on rat thyroid fragments in the study of thyroid hormone secretion: short-term effects of thyrotropin, theophylline and glucagon. J. Endocrinol. 102: 43, 1984.

    Article  PubMed  CAS  Google Scholar 

  19. Mashita K., Kawamura S., Kishino B., Kimura H., Nonawa K., Tarui S. Effects of iodide and propylthiouracil on the release of 3, 5, 3′—triidothyronine and of cyclic adenosine 3′, 5′-monophosphate from perifused rat thyroids. Endocrinology 110: 1023, 1982.

    Article  PubMed  CAS  Google Scholar 

  20. Attali J.R., Valensi P., Darnis D., Perret G., Sebaoun J. Evidence of direct thyroid-stimulating action of thyrotropin-releasing hormone in perifusion of rat thyroid fragments. Endocrinology 116: 561, 1985.

    Article  PubMed  CAS  Google Scholar 

  21. Attali J.R., Valensi P., Darnis D., Weisselberg C., Sebaoun J. Glucagon induced thyroid stimulating action in perifused rat thyroid fragments (RTFP) 7th International Congress of Endocrinology, Sao Paulo, Excerpta Medica 264 (Abstract), 1985.

    Google Scholar 

  22. Karlsson F.A., Dahlberg P.A., Walinder O. Activation of membrane-bound adenylcyclase by thyroid stimulating antibodies. Acta Endocrinol. (Kbh.) 97: 60, 1981.

    CAS  Google Scholar 

  23. Zakarija M., McKenzie J.M. Zoological specificity of human thyroid stimulating antibody. J. Clin. Endocrinol. Metab. 47: 249, 1978.

    Article  PubMed  CAS  Google Scholar 

  24. Rapoport B., Takai N.A., Filetti S. Evidence for species specificity in the interaction between thyrotropin and thyroid stimulating immunoglobulin and their receptor in thyroid tissue. J. Clin. Endocrinol. Metab. 54: 1059, 1982.

    Article  PubMed  CAS  Google Scholar 

  25. Atkinson S., Kendall-Taylor P. The stimulation of thyroid hormone secretion in vitro by thyroid stimulating antibodies. J. Clin. Endocrinol. Metab. 53: 1263, 1981.

    Article  PubMed  CAS  Google Scholar 

  26. Vitti P., Valente W.A., Ambesiimpiombato F.S., Fenzi G.F., Pinchera A., Kohn L.D. Graves’ IgG stimulation of continuously cultured rat thyroid cells: a sensitive potentially useful clinical assay. J. Endocrinol. Invest. 5: 179, 1982.

    Article  PubMed  CAS  Google Scholar 

  27. Karlsson F.A., Dahlberg P.A. Thyroid stimulating antibodies (TSAb) in patients with Graves’ disease undergoing antithyroid drug treatment: indicators of activity of disease. Clin. Endocrinol. (Oxf.) 14: 579, 1981.

    Article  CAS  Google Scholar 

  28. Brown R.S., Jackson I.M.D., Pohl S.L., Reichlin S. Do thyroid stimulating immunoglobulins cause nontoxic multinodular goiter? Lance 1: 904, 1978.

    Article  CAS  Google Scholar 

  29. Bidey S.P., Marshall N.J., Ekins R.P. Bioassay of thyroid stimulating immunoglobulins using human thyroid cell cultures: optimization and clinical assessment. Clin. Endocrinol. (Oxf.) 18: 193, 1983.

    Article  Google Scholar 

  30. Drexhage H.A., Bottazzo G.F., Doniach D., Bietensky L., Chahen J. Evidence for thyroid growth stimulating immunoglobulins in some goitrous thyroid diseases. Lancet 2: 287, 1980.

    Article  CAS  Google Scholar 

  31. Chiovato L., Hammond L.J., Bottazzo G.F. Detection of thyroid growth stimulating immunoglobulins (TGi) by 3H-thymidine incorporation in reconstituted rat thyroid follicles. Ann. Endocrinol. (Paris) 43: 60 A, 1982.

    Google Scholar 

  32. Madec A.M., Laurent M.C., Rostagnat Stefanutti A., Allanic H., Orgiazzi J., Mornex R. Détection des immunoglobulins thyreostimulantes sur serum entier. Intérêt de la culture de cellules thyroïdiennes humaines. Nouv. Presse Med. 19: 1849, 1983.

    Google Scholar 

  33. Ikeda H., Chin S.C., Kuzuya N., Uchimura H., Nagataki S. Thyroid hormone secretion is more sensitive than thyroid cyclic AMP accumulation to stimulation with LATS in mice in vitro and in vivo. J. Endocrinol. Invest. 7: 1, 1984.

    Article  PubMed  CAS  Google Scholar 

  34. Yashamita K., Field J.B. Effects of long-acting thyroid stimulator on thyrotropin stimulation of adenyl cyclase activity in thyroid plasma membranes. J. Clin. Invest. 51: 463, 1972.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire d’Endocrinologie Clinique, Hôpital Avicenne, Uer Paris XIII, 93009, Bobigny, France

    J. R. Attali, P. Valensi, D. Darnis, C. Weisselberg & J. Sebaoun

Authors
  1. J. R. Attali
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P. Valensi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. D. Darnis
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C. Weisselberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J. Sebaoun
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This study was presented in part at the 1st International Symposium Thyroid and Auto-immunity at Amsterdam, March, 1986.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Attali, J.R., Valensi, P., Darnis, D. et al. Detection of thyroid-stimulating antibodies in thyroid diseases, employing rat thyroid fragment perifusion. J Endocrinol Invest 10, 171–177 (1987). https://doi.org/10.1007/BF03347185

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03347185

Key-words

Search

Navigation