Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1981 Nov 1;154(5):1455–1474. doi: 10.1084/jem.154.5.1455

T cell growth factor receptors. Quantitation, specificity, and biological relevance

RJ Robb, A Munck, KA Smith
PMCID: PMC2186509  PMID: 6975347

Abstract

To examine directly the hypothesis that T cell growth factor (TCGF) interacts with target cells in a fashion similar to polypeptide hormones, the binding of radiolabeled TCGF to various cell populations was investigated. The results indicate that TCGF interacts with activated T cells via a receptor through which it initiates the T cell proliferative response. Internally radiolabeled TCGF, prepared from a human T leukemia cell line and purified by gel filtration and isoelectric focusing, retained biological activity and was uniform with respect to size and charge. Binding of radiolabeled TCGF to TCGF-dependent cytolytic T cells occurred rapidly (within 15 rain at 37 degrees C) and was both saturable and largely reversible. In addition, at 37 degrees C, a receptor- and lysosome-dependent degradation of TCGF occurred. Radiolabeled TCGF binding was specific for activated, TCGF-responsive T cells. Whereas unstimulated lymphocytes of human or murine origin and lipopolysaccharide-activated B cell blasts expressed few if any detectable binding sites, lectin- or alloantigen-activated cells had easily detectable binding sites. Moreover, compared with lectin- or alloantigen-activated T cells, long-term TCGF-dependent cytolytic and helper T cell lines and TCGF-dependent neo-plastic T cell lines bound TCGF with a similar affinity (dissociation constant of 5-25 pM) and expressed a similar number of receptor sites per cell (5,000-15,000). In contrast, a number of TCGF-independent cell lines of T cell, B cell, or myeloid origin did not bind detectable quantities of radiolabeled TCGF. Binding of radiolabeled TCGF to TCGF-responsive cells was specific, in that among several growth factors and polypeptide hormones tested, only TCGF competed for binding. Finally, the relative magnitude of T cell proliferation induced by a given concentration of TCGF closely paralleled the fraction of occupied receptor sites. As the extent of T cell clonal expansion depends on TCGF and on the TCGF receptor, the dissection of the molecular events surrounding the interaction of TCGF and its receptor that these studies permit, should provide new insight into the hormonelike regulation of the immune response by this lymphokine.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  1. Andersson J., Grönvik K. O., Larsson E. L., Coutinho A. Studies on T lymphocyte activation. I. Requirements for the mitogen-dependent production of T cell growth factors. Eur J Immunol. 1979 Aug;9(8):581–587. doi: 10.1002/eji.1830090802. [DOI] [PubMed] [Google Scholar]
  2. Baker P. E., Gillis S., Smith K. A. Monoclonal cytolytic T-cell lines. J Exp Med. 1979 Jan 1;149(1):273–278. doi: 10.1084/jem.149.1.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bonnard G. D., Yasaka K., Jacobson D. Ligand-activated T cell growth factor-induced proliferation: absorption of T cell growth factor by activated T cells. J Immunol. 1979 Dec;123(6):2704–2708. [PubMed] [Google Scholar]
  4. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Carpenter G., Cohen S. 125I-labeled human epidermal growth factor. Binding, internalization, and degradation in human fibroblasts. J Cell Biol. 1976 Oct;71(1):159–171. doi: 10.1083/jcb.71.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Coutinho A., Larsson E. L., Grönvik K. O., Andersson J. Studies on T lymphocyte activation II. The target cells for concanavalin A-induced growth factors. Eur J Immunol. 1979 Aug;9(8):587–592. doi: 10.1002/eji.1830090803. [DOI] [PubMed] [Google Scholar]
  8. Gillis S., Crabtree G. R., Smith K. A. Glucocorticoid-induced inhibition of T cell growth factor production. I. The effect on mitogen-induced lymphocyte proliferation. J Immunol. 1979 Oct;123(4):1624–1631. [PubMed] [Google Scholar]
  9. Gillis S., Crabtree G. R., Smith K. A. Glucocorticoid-induced inhibition of T cell growth factor production. II. The effect on the in vitro generation of cytolytic T cells. J Immunol. 1979 Oct;123(4):1632–1638. [PubMed] [Google Scholar]
  10. Gillis S., Ferm M. M., Ou W., Smith K. A. T cell growth factor: parameters of production and a quantitative microassay for activity. J Immunol. 1978 Jun;120(6):2027–2032. [PubMed] [Google Scholar]
  11. Gillis S., Smith K. A. Long term culture of tumour-specific cytotoxic T cells. Nature. 1977 Jul 14;268(5616):154–156. doi: 10.1038/268154a0. [DOI] [PubMed] [Google Scholar]
  12. Gillis S., Smith K. A., Watson J. Biochemical characterization of lymphocyte regulatory molecules. II. Purification of a class of rat and human lymphokines. J Immunol. 1980 Apr;124(4):1954–1962. [PubMed] [Google Scholar]
  13. Gillis S., Watson J. Biochemical and biological characterization of lymphocyte regulatory molecules. V. Identification of an interleukin 2-producing human leukemia T cell line. J Exp Med. 1980 Dec 1;152(6):1709–1719. doi: 10.1084/jem.152.6.1709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gootenberg J. E., Ruscetti F. W., Mier J. W., Gazdar A., Gallo R. C. Human cutaneous T cell lymphoma and leukemia cell lines produce and respond to T cell growth factor. J Exp Med. 1981 Nov 1;154(5):1403–1418. doi: 10.1084/jem.154.5.1403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gorden P., Carpentier J. L., Cohen S., Orci L. Epidermal growth factor: morphological demonstration of binding, internalization, and lysosomal association in human fibroblasts. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5025–5029. doi: 10.1073/pnas.75.10.5025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hizuka N., Gorden P., Lesniak M. A., Van Obberghen E., Carpentier J. L., Orci L. Polypeptide hormone degradation and receptor regulation are coupled to ligand internalization. A direct biochemical and morphologic demonstration. J Biol Chem. 1981 May 10;256(9):4591–4597. [PubMed] [Google Scholar]
  17. Iscove N. N., Melchers F. Complete replacement of serum by albumin, transferrin, and soybean lipid in cultures of lipopolysaccharide-reactive B lymphocytes. J Exp Med. 1978 Mar 1;147(3):923–933. doi: 10.1084/jem.147.3.923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kaplan J., Tilton J., Peterson W. D., Jr Identification of T cell lymphoma tumor antigens on human T cell lines. Am J Hematol. 1976;1(2):219–223. doi: 10.1002/ajh.2830010206. [DOI] [PubMed] [Google Scholar]
  19. Krupp M., Lane M. D. On the mechanism of ligand-induced down-regulation of insulin receptor level in the liver cell. J Biol Chem. 1981 Feb 25;256(4):1689–1694. [PubMed] [Google Scholar]
  20. Kurnick J. T., Ostberg L., Stegagno M., Kimura A. K., Orn A., Sjöberg O. A rapid method for the separation of functional lymphoid cell populations of human and animal origin on PVP-silica (Percoll) density gradients. Scand J Immunol. 1979;10(6):563–573. doi: 10.1111/j.1365-3083.1979.tb01391.x. [DOI] [PubMed] [Google Scholar]
  21. Lachman L. B., Hacker M. P., Blyden G. T., Handschumacher R. E. Preparation of lymphocyte-activating factor from continuous murine macrophage cell lines. Cell Immunol. 1977 Dec;34(2):416–419. doi: 10.1016/0008-8749(77)90263-5. [DOI] [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Larsson E. L., Coutinho A. The role of mitogenic lectins in T-cell triggering. Nature. 1979 Jul 19;280(5719):239–241. doi: 10.1038/280239a0. [DOI] [PubMed] [Google Scholar]
  24. Larsson E. L., Iscove N. N., Coutinho A. Two distinct factors are required for induction of T-cell growth. Nature. 1980 Feb 14;283(5748):664–666. doi: 10.1038/283664a0. [DOI] [PubMed] [Google Scholar]
  25. Larsson E. L. Mechanism of T cell activation. II. Antigen- and lectin-dependent acquisition of responsiveness to TCGF is a nonmitogenic, active response of resting T cells. J Immunol. 1981 Apr;126(4):1323–1326. [PubMed] [Google Scholar]
  26. Lesniak M. A., Roth J. Regulation of receptor concentration by homologous hormone. Effect of human growth hormone on its receptor in IM-9 lymphocytes. J Biol Chem. 1976 Jun 25;251(12):3720–3729. [PubMed] [Google Scholar]
  27. Morgan D. A., Ruscetti F. W., Gallo R. Selective in vitro growth of T lymphocytes from normal human bone marrows. Science. 1976 Sep 10;193(4257):1007–1008. doi: 10.1126/science.181845. [DOI] [PubMed] [Google Scholar]
  28. Ruscetti F. W., Morgan D. A., Gallo R. C. Functional and morphologic characterization of human T cells continuously grown in vitro. J Immunol. 1977 Jul;119(1):131–138. [PubMed] [Google Scholar]
  29. Schreier M. H., Iscove N. N., Tees R., Aarden L., von Boehmer H. Clones of killer and helper T cells: growth requirements, specificity and retention of function in long-term culture. Immunol Rev. 1980;51:315–336. doi: 10.1111/j.1600-065x.1980.tb00326.x. [DOI] [PubMed] [Google Scholar]
  30. Seglen P. O. Protein degradation in isolated rat hepatocytes is inhibited by ammonia. Biochem Biophys Res Commun. 1975 Sep 2;66(1):44–52. doi: 10.1016/s0006-291x(75)80292-0. [DOI] [PubMed] [Google Scholar]
  31. Smith K. A., Baker P. E., Gillis S., Ruscetti F. W. Functional and molecular characteristics of T-cell growth factor. Mol Immunol. 1980 May;17(5):579–589. doi: 10.1016/0161-5890(80)90156-x. [DOI] [PubMed] [Google Scholar]
  32. Smith K. A., Gilbride K. J., Favata M. F. Lymphocyte activating factor promotes T-cell growth factor production by cloned murine lymphoma cells. Nature. 1980 Oct 30;287(5785):853–855. doi: 10.1038/287853a0. [DOI] [PubMed] [Google Scholar]
  33. Smith K. A., Gillis S., Baker P. E., McKenzie D., Ruscetti F. W. T-cell growth factor-mediated T-cell proliferation. Ann N Y Acad Sci. 1979;332:423–432. doi: 10.1111/j.1749-6632.1979.tb47136.x. [DOI] [PubMed] [Google Scholar]
  34. Smith K. A., Lachman L. B., Oppenheim J. J., Favata M. F. The functional relationship of the interleukins. J Exp Med. 1980 Jun 1;151(6):1551–1556. doi: 10.1084/jem.151.6.1551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Smith K. A., Ruscetti F. W. T-cell growth factor and the culture of cloned functional T cells. Adv Immunol. 1981;31:137–175. doi: 10.1016/s0065-2776(08)60920-7. [DOI] [PubMed] [Google Scholar]
  36. Smith K. A. T-cell growth factor. Immunol Rev. 1980;51:337–357. doi: 10.1111/j.1600-065x.1980.tb00327.x. [DOI] [PubMed] [Google Scholar]
  37. Spits H., de Vries J. E., Terhorst C. A permanent human cytotoxic T-cell line with high killing capacity against a lymphoblastoid B-cell line shows preference for HLA A, B target antigens and lacks spontaneous cytotoxic activity. Cell Immunol. 1981 Apr;59(2):435–447. doi: 10.1016/0008-8749(81)90423-8. [DOI] [PubMed] [Google Scholar]
  38. Swain S. L., Dennert G., Warner J. F., Dutton R. W. Culture supernatants of a stimulated T-cell line have helper activity that acts synergistically with interleukin 2 in the response of B cells to antigen. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2517–2521. doi: 10.1073/pnas.78.4.2517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wagner H., Röllinghoff M. T-T-cell interactions during the vitro cytotoxic allograft responses. I. Soluble products from activated Lyl+ T cells trigger autonomously antigen-primed Ly23+ T cells to cell proliferation and cytolytic activity. J Exp Med. 1978 Dec 1;148(6):1523–1538. doi: 10.1084/jem.148.6.1523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Watson J., Mochizuki D. Interleukin 2: a class of T cell growth factors. Immunol Rev. 1980;51:257–278. doi: 10.1111/j.1600-065x.1980.tb00324.x. [DOI] [PubMed] [Google Scholar]
  41. Ziegler A., Harrison S. C., Leberman R. The minor proteins in tomato bushy stunt and turnip crinkle viruses. Virology. 1974 Jun;59(2):509–515. doi: 10.1016/0042-6822(74)90460-7. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES