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. 1983 Jul;80(13):3986–3990. doi: 10.1073/pnas.80.13.3986

Receptor aggregation induced by antilutropin receptor antibody and biological response in rat testis Leydig cells.

E J Podestá, A R Solano, R Attar, M L Sánchez, L Molina y Vedia
PMCID: PMC394184  PMID: 6306657

Abstract

Antibodies against the lutropin receptor have been obtained by the monoclonal antibody technique. Mice were immunized with luteal membrane from ovaries from pseudopregnant rats, containing high lutropin receptor concentration. Hybridoma cells were obtained by fusing mouse myeloma cells with spleen cells from the immunized animal. Five clones were produced that secreted monoclonal antibodies that specifically inhibited lutropin binding to its receptor in a competitive fashion. Antibodies from three clones were capable of blocking biological response to lutropin (e.g., testosterone production by isolated rat Leydig cells). Antibodies secreted by two other clones, however, were capable of acting as Leydig cell stimulators. Immunofluorescence studies demonstrated the presence of receptor capping which may be associated with receptor-mediated testosterone production. Antagonist antibodies could be transformed into agonist by the addition of a second crosslinking anti-mouse IgG. The discovery of agonist antibodies against the receptor molecule proves that the biological information of the lutropin-receptor complex resides in the receptor and not in the hormone.

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Selected References

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

  1. Best-Belpomme M., Dessen P. Inhibition compètitive. Relations linèaires nouvellcs et gènèrales. Biochimie. 1973;55(1):11–16. doi: 10.1016/s0300-9084(73)80231-7. [DOI] [PubMed] [Google Scholar]
  2. Cuatrecasas P. Epidermal growth factor: uptake and fate. Ciba Found Symp. 1982;(92):96–108. doi: 10.1002/9780470720745.ch6. [DOI] [PubMed] [Google Scholar]
  3. Dufau M. L., Podesta E. J., Catt K. J. Physical characteristics of the gonadotropin receptor-hormone complexes formed in vivo and in vitro. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1272–1275. doi: 10.1073/pnas.72.4.1272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Galfre G., Howe S. C., Milstein C., Butcher G. W., Howard J. C. Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature. 1977 Apr 7;266(5602):550–552. doi: 10.1038/266550a0. [DOI] [PubMed] [Google Scholar]
  5. Hawkes R., Niday E., Matus A. Monoclonal antibodies identify novel neural antigens. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2410–2414. doi: 10.1073/pnas.79.7.2410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Heinrich J., Pilch P. F., Czech M. P. Purification of the adipocyte insulin receptor by immunoaffinity chromatography. J Biol Chem. 1980 Feb 25;255(4):1732–1737. [PubMed] [Google Scholar]
  7. Jacobs S., Hazum E., Cuatrecasas P. The subunit structure of rat liver insulin receptor. Antibodies directed against the insulin-binding subunit. J Biol Chem. 1980 Jul 25;255(14):6937–6940. [PubMed] [Google Scholar]
  8. Kahn C. R., Baird K., Filier J. S., Jarrett D. B. Effects of autoantibodies to the insulin receptor on isolated adipocytes. Studies of insulin binding and insulin action. J Clin Invest. 1977 Nov;60(5):1094–1106. doi: 10.1172/JCI108861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. King B. W. Components of a motivation system: communication. J Emerg Nurs. 1981 Mar-Apr;7(2):77–78. [PubMed] [Google Scholar]
  10. Lang U., Kahn C. R., Harrison L. C. Subunit structure of the insulin receptor of the human lymphocyte. Biochemistry. 1980 Jan 8;19(1):64–70. doi: 10.1021/bi00542a010. [DOI] [PubMed] [Google Scholar]
  11. Lernhardt W., Andersson J., Coutinho A., Melchers F. Cloning of murine transformed cell lines in suspension culture with efficiencies near 100%. Exp Cell Res. 1978 Feb;111(2):309–316. doi: 10.1016/0014-4827(78)90175-1. [DOI] [PubMed] [Google Scholar]
  12. Mendelson C., Dufau M., Catt K. Gonadotropin binding and stimulation of cyclic adenosine 3':5'-monophosphate and testosterone production in isolated Leydig cells. J Biol Chem. 1975 Nov 25;250(22):8818–8823. [PubMed] [Google Scholar]
  13. Podesta E. J., Dufau M. L., Solano A. R., Catt K. J. Hormonal activation of protein kinase in isolated Leydig cells. Electrophoretic analysis of cyclic AMP receptors. J Biol Chem. 1978 Dec 25;253(24):8994–9001. [PubMed] [Google Scholar]
  14. Schechter Y., Hernaez L., Schlessinger J., Cuatrecasas P. Local aggregation of hormone-receptor complexes is required for activation by epidermal growth factor. Nature. 1979 Apr 26;278(5707):835–838. doi: 10.1038/278835a0. [DOI] [PubMed] [Google Scholar]
  15. Shechter Y., Chang K. J., Jacobs S., Cuatrecasas P. Modulation of binding and bioactivity of insulin by anti-insulin antibody: relation to possible role of receptor self-aggregation in hormone action. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2720–2724. doi: 10.1073/pnas.76.6.2720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Shechter Y., Hernaez L., Cuatrecasas P. Epidermal growth factor: biological activity requires persistent occupation of high-affinity cell surface receptors. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5788–5791. doi: 10.1073/pnas.75.12.5788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Smith B. R., Hall R. Binding of thyroid stimulators to thyroid membranes. FEBS Lett. 1974 Jun 15;42(3):301–304. doi: 10.1016/0014-5793(74)80751-9. [DOI] [PubMed] [Google Scholar]
  18. Thorell J. I., Johansson B. G. Enzymatic iodination of polypeptides with 125I to high specific activity. Biochim Biophys Acta. 1971 Dec 28;251(3):363–369. doi: 10.1016/0005-2795(71)90123-1. [DOI] [PubMed] [Google Scholar]
  19. Valente W. A., Vitti P., Yavin Z., Yavin E., Rotella C. M., Grollman E. F., Toccafondi R. S., Kohn L. D. Monoclonal antibodies to the thyrotropin receptor: stimulating and blocking antibodies derived from the lymphocytes of patients with Graves disease. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6680–6684. doi: 10.1073/pnas.79.21.6680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Yavin E., Yavin Z., Schneider M. D., Kohn L. D. Monoclonal antibodies to the thyrotropin receptor: implications for receptor structure and the action of autoantibodies in Graves disease. Proc Natl Acad Sci U S A. 1981 May;78(5):3180–3184. doi: 10.1073/pnas.78.5.3180. [DOI] [PMC free article] [PubMed] [Google Scholar]

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