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. 1986 Jun;64(3):544–554.

Characterization of monoclonal antibodies directed towards the microsomal/microvillar thyroid autoantigen recognized by Hashimoto autoantibodies.

J P Banga, R Mirakian, L Hammond, G Pryce, S Bidey, F Bottazzo, A P Weetman, A M McGregor, I M Roitt
PMCID: PMC1542454  PMID: 3791690

Abstract

Monoclonal antibodies reacting with thyroid microsomes have been described. In this report, we present evidence that some of these monoclonal antibodies (MoAb) MAH C3 and perhaps MAH C6 are directed towards the thyroid microsomal antigen towards which microsomal autoantibodies from Hashimoto patients are directed. Immunofluorescence on cryostat sections with MAH C3 and MAH C6 on thyroid gland gave cytoplasmic staining patterns that were comparable to those obtained with autoantibodies. By western blotting, reactivity of MAH C3 was shown to be directed towards a 105,000 mol. wt component, comparable to that recognized by autoantibodies by immunoprecipitation. Double immunofluorescence with MAH C3 and autoantibodies on cultured, viable thyroid monolayers revealed a large number of comparable binding sites; in contrast, binding sites for asioloagalactothyroglobulin were different of binding of MAH C3 by the autoantibodies suggesting that the epitopes recognized were different. This was confirmed with the use of a rat thyroid cell line FRTL-5 and fluorescence activated cell sorter analysis which showed binding to the rat microsomal component with autoantibodies but negligible binding with MAH C3 and MAH C6. It therefore appears that the monoclonal antibodies recognize species specific determinants while the autoantibodies recognize species cross reactive determinants. These monoclonal antibodies will be useful for purification and immunochemical analysis of this autoantigen in human thyroid disease.

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Selected References

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

  1. Ambesi-Impiombato F. S., Parks L. A., Coon H. G. Culture of hormone-dependent functional epithelial cells from rat thyroids. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3455–3459. doi: 10.1073/pnas.77.6.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Banga J. P., Guarnotta G., Harte A., Pryce G., Campbell M. A., Quartey-Papafio R., Lydyard P. M., Roitt I. M. A common epitope identified by a monoclonal antibody, MID 2, present on all leucocytes and associated with a group of high molecular weight glycopeptides. Scand J Immunol. 1984 Jan;19(1):11–21. doi: 10.1111/j.1365-3083.1984.tb00895.x. [DOI] [PubMed] [Google Scholar]
  3. Banga J. P., Pryce G., Hammond L., Roitt I. M. Structural features of the autoantigens involved in thyroid autoimmune disease: the thyroid microsomal/microvillar antigen. Mol Immunol. 1985 Jun;22(6):629–642. doi: 10.1016/0161-5890(85)90092-6. [DOI] [PubMed] [Google Scholar]
  4. Bidey S. P., Ryder K., Gaines-Das R., Marshall N. J., Ekins R. P. A comparison of the bioactivity of human and bovine thyrotrophin preparations, as determined by intracellular cyclic AMP responses of cultured FRTL-5 cells and human thyroid cell monolayers. Acta Endocrinol (Copenh) 1984 Aug;106(4):482–489. doi: 10.1530/acta.0.1060482. [DOI] [PubMed] [Google Scholar]
  5. Bottazzo G. F., Drexhage H. A., Khoury E. L. Thyroid antibodies in thyroid diseases. Ciba Found Symp. 1982;(90):153–177. doi: 10.1002/9780470720721.ch10. [DOI] [PubMed] [Google Scholar]
  6. Chiovato L., Vitti P., Lombardi A., Kohn L. D., Pinchera A. Expression of the microsomal antigen on the surface of continuously cultured rat thyroid cells is modulated by thyrotropin. J Clin Endocrinol Metab. 1985 Jul;61(1):12–16. doi: 10.1210/jcem-61-1-12. [DOI] [PubMed] [Google Scholar]
  7. Haspel M. V., Onodera T., Prabhakar B. S., McClintock P. R., Essani K., Ray U. R., Yagihashi S., Notkins A. L. Multiple organ-reactive monoclonal autoantibodies. Nature. 1983 Jul 7;304(5921):73–76. doi: 10.1038/304073a0. [DOI] [PubMed] [Google Scholar]
  8. Khoury E. L., Hammond L., Bottazzo G. F., Doniach D. Presence of the organ-specific 'microsomal' autoantigen on the surface of human thyroid cells in culture: its involvement in complement-mediated cytotoxicity. Clin Exp Immunol. 1981 Aug;45(2):316–328. [PMC free article] [PubMed] [Google Scholar]
  9. Knight A., Knight J., Laing P., Adams D. Coexisting thyroid and gastric autoimmune diseases are not due to cross-reactive autoantibodies. J Clin Lab Immunol. 1984 Jul;14(3):141–144. [PubMed] [Google Scholar]
  10. Knight J. G., Knight A. Multiple-organ autoimmunity. Nature. 1984 Mar 22;308(5957):318–318. doi: 10.1038/308318a0. [DOI] [PubMed] [Google Scholar]
  11. Prabhakar B. S., Saegusa J., Onodera T., Notkins A. L. Lymphocytes capable of making monoclonal autoantibodies that react with multiple organs are a common feature of the normal B cell repertoire. J Immunol. 1984 Dec;133(6):2815–2817. [PubMed] [Google Scholar]
  12. Roitt I. M., Pujol-Borrell R., Hanafusa T., Delves P. J., Bottazzo G. F., Kohn L. D. Asialoagalactothyroglobulin binds to the surface of human thyroid cells at a site distinct from the 'microsomal' autoantigen. Clin Exp Immunol. 1984 Apr;56(1):129–134. [PMC free article] [PubMed] [Google Scholar]
  13. Satoh J., Prabhakar B. S., Haspel M. V., Ginsberg-Fellner F., Notkins A. L. Human monoclonal autoantibodies that react with multiple endocrine organs. N Engl J Med. 1983 Jul 28;309(4):217–220. doi: 10.1056/NEJM198307283090405. [DOI] [PubMed] [Google Scholar]
  14. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Weetman A. P., Gunn C. A., Rennie D. P., Hall R., McGregor A. M. The production and characterization of monoclonal antibodies to the human thyroid microsome. J Endocrinol. 1985 Apr;105(1):47–52. doi: 10.1677/joe.0.1050047. [DOI] [PubMed] [Google Scholar]
  16. Weetman A. P., McGregor A. M. Autoimmune thyroid disease: developments in our understanding. Endocr Rev. 1984 Spring;5(2):309–355. doi: 10.1210/edrv-5-2-309. [DOI] [PubMed] [Google Scholar]
  17. Weetman A. P., Rennie D. P., Hassman R., Hall R., McGregor A. M. Enzyme-linked immunoassay of monoclonal and serum microsomal autoantibodies. Clin Chim Acta. 1984 Apr 27;138(3):237–244. doi: 10.1016/0009-8981(84)90130-x. [DOI] [PubMed] [Google Scholar]

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