Skip to main content
Log in

A monoclonal antibody recognizing an epitope shared by receptors for growth hormone, prolactin, interleukin 2 and interleukin 6

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Monoclonal antibody (MAb) termed R7B4 was generated throughout the idiotypic-antiidiotypic network from mice immunized with human and bovine growth hormones (GH). The Ab was selected on the basis that it did not recognize human GH (hGH) neither insolubilized nor in solution but inhibited 125I-hGH binding to receptors from rat and rabbit liver and from Nb2-cell membranes.

Since it inhibited Nb2-cell mitogenesis stimulated by hGH, prolactins or placental lactogens, MAb R7B4 behaved as an antagonist of lactogenic hormones. Furthermore, the Ab impaired proliferative activity of interleukin 2 (IL-2) on Nb2 cells as well as growth of 7TD1 cells, an interleukin 6 (IL-6) dependent hybridoma not expressing GH receptors.

Biotin-labeled MAb R7B4 specifically bound to rat liver microsomes, and the Ab was able to recognize Nb2 and 7TD1-cell membranes as shown by flow cytometry experiments. However, MAb binding was not hampered by hGH, indicating that the Ab did not mimic GH binding site to receptors. Immunoblot assays indicated that rat and rabbit liver as well as Nb2-cells membrane antigens recognized by MAb R7B4 were similar to those revealed by a MAb directed to prolactin receptors. In addition, MAb R7B4 was able to detect two bands probably corresponding to the somatogenic receptor in rabbit liver microsomes as well as three different proteins in 7TD1-cells showing molecular weights similar to those of the IL-6 receptor complex.

Results suggest that MAb R7B4 is directed to an epitope shared by receptors for lactogenic and somatogenic hormones, IL-2 and IL-6. To our knowledge, these data are the first experimental evidence of the existence of structural similarity between some of the receptors grouped in the cytokine receptor superfamily.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Bazan JF: Structural design and molecular evolution of a cytokine receptor superfamily. Proc Natl Acad Sci USA 87: 6934–6938, 1990

    PubMed  Google Scholar 

  2. Ihle JN, Witthunhn BA, Quelle FW, Yamamoto K, Silvennoinen O: Signaling through the hematopoietic cytokine receptors. Annu Rev Immunol 13: 369–398, 1995

    PubMed  Google Scholar 

  3. Heim MH: The Jak-STAT pathway: Specific signal transduction from the cell membrane to the nucleus. Eur J Clin Inv 26: 1–12, 1996

    Google Scholar 

  4. Hill BL, Erlanger BF: Monoclonal antibodies to the thyrotropin receptor raised by an autoantiidiotypic protocol and their relationship to monoclonal antibodies from Graves’ patients. Endocrinology 122: 2840–2850, 1988

    PubMed  Google Scholar 

  5. Dellacha JM, Sonenberg M: Purification of bovine growth hormone. J Biol Chem 239: 1515–1520, 1964

    PubMed  Google Scholar 

  6. Roth J: Methods for assessing inununologic and biological properties of iodinated peptide hormones. In: SO Colowick, NO Kaplan (eds). Methods in Enzymology, vol. 37, part B. Peptide Hormones, Academic Press, New York, 1975, pp 223–231

    Google Scholar 

  7. Balbis A, Dellacha JM, Calandra RS, Bartke A, Turyn D: Down regulation of masked and unmasked insulin receptors in the liver of transgenic mice expressing bovine growth hormone gene. Life Sciences 51: 771–778, 1992

    PubMed  Google Scholar 

  8. Mazza MM, Retegui LA: The antigenic topography of human growth hormone. Mol Immunol 26: 231–240, 1989

    PubMed  Google Scholar 

  9. Galfre G, Milstein C: Preparation of monoclonal antibodies: strategies and procedures. In: JJ Langone, H Van Vunakis (eds). Methods in Enzymology, vol. 73, part B, Immunochemical Techniques, Academic Press, New York, 1981, pp 1–46

    Google Scholar 

  10. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurements with the folin phenol reagent. J Biol Chem 193: 265–275, 1951

    PubMed  Google Scholar 

  11. Tanaka T, Shiu RPC, Gout PW, Beer CT, Noble RL, Friesen HG: A new sensitive and specific bioassay for lactogenic hormones: measurement of prolactin and growth hormone in human serum. J Clin Endocrinol Metab 51: 1058–1063, 1980

    PubMed  Google Scholar 

  12. Landegren U: Measurement of cell numbers by means of the endogenous enzyme hexosaminidase. Applications to detection of lymphokines and cell surface antigens J Immunol Methods 67: 379–388, 1984

    PubMed  Google Scholar 

  13. Van Snick J, Cayphas S, Vink A, Uyttenhove C, Coulie PG, Rubira MR, Sirnpson RJ: Purification and NH2-terminal amino acid sequence of a t-cell-derived lymphokine with growth factor activity for B-cell hybridomas. Proc Natl Acad Sci USA 83: 9679–9683, 1986

    PubMed  Google Scholar 

  14. Ey PL, Prowse SJ, Jenkin CR: Isolation of pure IgG2a and IgG2b immunoglobulins from mouse serum using protein A-Sepharose. Immunochemistry 15: 429–436, 1978

    PubMed  Google Scholar 

  15. Laemmli UK: leavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970

    PubMed  Google Scholar 

  16. Paladini AC, Peña C, Poskus E: Molecular biology of growth hormone. In: JGR Hurrell (ed.) CRC Press, Cleveland, Ohio, 215: 25–56, 1983

    Google Scholar 

  17. Aguilar RC, Retegui LA, Postel-Vinay M-C, Roguin LP: Allosteric effects of monoclonal antibodies on human growth hormone. Mol Cell Biochem 36: 35–42, 1994

    Google Scholar 

  18. Coulic PG, Vanheeke A, Van damme J, Cayphas S, Poupart P, De Wit L, Content J: High-affinity binding sites for human 26-kDa protein (interleukin 6, B cell stimulatory factor-2, human hybridoma plasmocytoma growth factor, interferon-β2), different from those of type I interferon (α, β), on lymphoblastoid cells. Eur J Immunol 17: 1435–1440, 1987

    PubMed  Google Scholar 

  19. Robetto EJ, Caaman–o CA, Fernández HN, Dellacha JM: Proteins associated with somatogenic and lactogenic receptors in microsomal membranes and intact rat hepatocytes. Biochim Biophys Acta 1013: 223–230, 1989

    PubMed  Google Scholar 

  20. Gaulton GN, Greene MI: Idiotypic mimicry of biological receptors. Annu Rev Immunol 4: 253–280, 1986

    PubMed  Google Scholar 

  21. Jerne NK: Towards a network theory of the immune response. Annals of Inununology (Paris) 125C: 373–389, 1974

    Google Scholar 

  22. Taga T, Kishimoto T: Cytokine receptor and signal transduction. FASEB J 7: 3387–3396, 1993

    Google Scholar 

  23. Okamura H, Zachwieja J, Raguet S, Kelly PA: Characterization and applications of monoclonal antibodies to the prolactin receptor. Endocrinology 124: 2499–2508, 1989

    PubMed  Google Scholar 

  24. Elberg G, Kelly, PA, Djiane J, Binder L, Gertler A: Mitogenic and binding properties of monoclonal antibodies to the prolactin receptor in Nb2 rat lymphoma cells. J Biol Chem 265: 14770–14776, 1990

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Longhi, S.A., Miranda, M.E., Gobet, M.G. et al. A monoclonal antibody recognizing an epitope shared by receptors for growth hormone, prolactin, interleukin 2 and interleukin 6. Mol Cell Biochem 195, 235–243 (1999). https://doi.org/10.1023/A:1006984506267

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006984506267

Navigation