Skip to main content
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1980 Jun;40(3):532–539.

Thyroid-stimulating antibody (TSAb) detected in sera of Graves' patients using human thyroid cell cultures

R S Toccafondi, S Aterini, Maria Alice Medici, C M Rotella, Annalisa Tanini, R Zonefrati
PMCID: PMC1538941  PMID: 6251989

Abstract

As a homologous system is required to evaluate the effect of thyroid-stimulating antibody (TSAb) present in the serum of Graves' patients, primary cultures obtained from normal human thyroid gland have been used and the stimulatory effect measured as an increase of cAMP intracellular levels.

Monolayer cell cultures were stimulated by IgG purified from sera of Graves' patients or control subjects and compared to the effect of bovine TSH. Bovine TSH produced a dose-dependent increase in cAMP intracellular levels between 0·05 mU and 2·5 mU/ml, reaching a maximal value after 30 min with higher doses. While normal IgG had no effect, IgG prepared from untreated patients with frank Graves' disease elicited a significant increase in cAMP accumulation at a concentration between 0·05 and 0·5 mg/ml within 60 min in thirteen out of fourteen patients. A longer incubation period showed no further increase in cAMP values, even if in one case a higher concentration (5·0 mg/ml) of Graves' IgG had a delayed response. When the cAMP intracellular level modifications produced by Graves' IgG preparations in thyroid cell cultures were compared to those evoked in thyroid slices, an identical percentage (93%) of positive cases was obtained, without a coincidence of negative cases. Using thyroid slices the cAMP intracellular increase above basal levels was higher, if considered as a percentage, but in cultured cells a very low IgG concentration was sufficient to detect the presence of TSAb. No correlation between the two assays was found.

In conclusion, normal human cultured thyroid cells appeared to be a more suitable substrate when compared to human thyroid slices for detecting the presence of TSAb in Graves' disease and for studying its effect on thyroid cells. However, a 100% TSAb positivity was present in our Graves' patient series only when both assays were used.

Full text

PDF
532

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ADAMS D. D. The presence of an abnormal thyroid-stimulating hormone in the serum of some thyrotoxic patients. J Clin Endocrinol Metab. 1958 Jul;18(7):699–712. doi: 10.1210/jcem-18-7-699. [DOI] [PubMed] [Google Scholar]
  2. BAUMSTARK J. S., LAFFIN R. J., BARDAWIL W. A. A PREPARATIVE METHOD FOR THE SEPARATION OF 7S GAMMA GLOBULIN FROM HUMAN SERUM. Arch Biochem Biophys. 1964 Dec;108:514–522. doi: 10.1016/0003-9861(64)90436-9. [DOI] [PubMed] [Google Scholar]
  3. Brown B. L., Albano J. D., Ekins R. P., Sgherzi A. M. A simple and sensitive saturation assay method for the measurement of adenosine 3':5'-cyclic monophosphate. Biochem J. 1971 Feb;121(3):561–562. doi: 10.1042/bj1210561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. KISSANE J. M., ROBINS E. The fluorometric measurement of deoxyribonucleic acid in animal tissues with special reference to the central nervous system. J Biol Chem. 1958 Jul;233(1):184–188. [PubMed] [Google Scholar]
  5. KRISS J. P., PLESHAKOV V., CHIEN J. R. ISOLATION AND IDENTIFICATION OF THE LONG-ACTING THYROID STIMULATOR AND ITS RELATION TO HYPERTHYROIDISM AND CIRCUMSCRIBED PRETIBIAL MYXEDEMA. J Clin Endocrinol Metab. 1964 Oct;24:1005–1028. doi: 10.1210/jcem-24-10-1005. [DOI] [PubMed] [Google Scholar]
  6. McKENZIE J. M. Delayed thyroid response to serum from thyrotoxic patients. Endocrinology. 1958 Jun;62(6):865–868. doi: 10.1210/endo-62-6-865. [DOI] [PubMed] [Google Scholar]
  7. McKenzie J. M., Zakarija M. A reconsideration of a thyroid-stimulating immunoglobulin as the cause of hyperthyroidism in Graves' disease. J Clin Endocrinol Metab. 1976 Apr;42(4):778–781. doi: 10.1210/jcem-42-4-778. [DOI] [PubMed] [Google Scholar]
  8. Orgiazzi J., Williams D. E., Chopra I. J., Solomon D. H. Human thyroid adenyl cyclase-stimulating activity in immunoglobulin G of patients with Graves' disease. J Clin Endocrinol Metab. 1976 Feb;42(2):341–354. doi: 10.1210/jcem-42-2-341. [DOI] [PubMed] [Google Scholar]
  9. PULVERTAFT R. J., DONIACH D., ROITT I. M., HUDSON R. V. Cytotoxic effects of Hashimoto serum on human thyroid cells in tissue culture. Lancet. 1959 Aug 29;2(7096):214–216. doi: 10.1016/s0140-6736(59)90161-8. [DOI] [PubMed] [Google Scholar]
  10. Wadeleux P. A., Etienne-Decerf J., Winand R. J., Kohn L. D. Effects of thyrotropin on iodine metabolism of dog thyroid cells in tissue culture. Endocrinology. 1978 Mar;102(3):889–902. doi: 10.1210/endo-102-3-889. [DOI] [PubMed] [Google Scholar]

Articles from Clinical and Experimental Immunology are provided here courtesy of British Society for Immunology

RESOURCES