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. 1984 Aug 1;160(2):584–599. doi: 10.1084/jem.160.2.584

Characterization of soluble factors that induce the cytolytic activity and the expression of T cell growth factor receptors of a T cell hybrid

PMCID: PMC2187453  PMID: 6236274

Abstract

A rat X mouse T cell hybrid (PC60) proliferates in the absence of T cell growth factor (TCGF) and its cytolytic activity can be induced by culture in mixed leukocyte culture supernatants or concanavalin A- activated rat spleen cell supernatant (CS) to lyse 51Cr-labeled tumor target cells. To characterize the factor(s) responsible for this reversible induction, serum-free CS was fractionated by reverse phase high performance liquid chromatography and by phenyl-Sepharose chromatography. A cytotoxicity-inducing activity (CIA) was separated from TCGF and macrophage-activating factor/interferon-gamma. CIA was found to be a macromolecule with an apparent molecular weight of 12,000- 18,000 and a pI of 5.0 and 6.2. Its activity on PC60 cells depended on the addition of TCGF. Thus TCGF may have other effects on T cells than the induction of entry into cell cycle. The number of TCGF surface receptors on PC60 cells was measured using purified 3H-TCGF. TCGF receptors were undetectable on noninduced cells but appeared during induction. The expression of TCGF receptors was not induced either by TCGF or by CIA-containing supernatants or fractions alone, only by a combination of both. These results show that TCGF plays a role in the regulation of the expression of its own receptors.

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Selected References

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  1. Cerottini J. C., Engers H. D., Macdonald H. R., Brunner T. Generation of cytotoxic T lymphocytes in vitro. I. Response of normal and immune mouse spleen cells in mixed leukocyte cultures. J Exp Med. 1974 Sep 1;140(3):703–717. doi: 10.1084/jem.140.3.703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Conzelmann A., Corthésy P., Cianfriglia M., Silva A., Nabholz M. Hybrids between rat lymphoma and mouse T cells with inducible cytolytic activity. Nature. 1982 Jul 8;298(5870):170–172. doi: 10.1038/298170a0. [DOI] [PubMed] [Google Scholar]
  3. Devos R., Plaetinck G., Cheroutre H., Simons G., Degrave W., Tavernier J., Remaut E., Fiers W. Molecular cloning of human interleukin 2 cDNA and its expression in E. coli. Nucleic Acids Res. 1983 Jul 11;11(13):4307–4323. doi: 10.1093/nar/11.13.4307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Falk W., Männel D. N., Dröge W. Activation of cytotoxic T lymphocytes requires at least two spleen cell-derived helper factors besides interleukin 2. J Immunol. 1983 May;130(5):2214–2218. [PubMed] [Google Scholar]
  5. Farrar J. J., Fuller-Farrar J., Simon P. L., Hilfiker M. L., Stadler B. M., Farrar W. L. Thymoma production of T cell growth factor (Interleukin 2). J Immunol. 1980 Dec;125(6):2555–2558. [PubMed] [Google Scholar]
  6. Finke J. H., Scott J., Gillis S., Hilfiker M. L. Generation of alloreactive cytotoxic T lymphocytes: evidence for a differentiation factor distinct from IL 2. J Immunol. 1983 Feb;130(2):763–767. [PubMed] [Google Scholar]
  7. Fontana A., McAdam K. P., Kristensen F., Weber E. Biological and biochemical characterization of an interleukin 1-like factor from rat C6 glioma cells. Eur J Immunol. 1983 Aug;13(8):685–689. doi: 10.1002/eji.1830130814. [DOI] [PubMed] [Google Scholar]
  8. Garman R. D., Fan D. P. Characterization of helper factors distinct from interleukin 2 necessary for the generation of allospecific cytolytic T lymphocytes. J Immunol. 1983 Feb;130(2):756–762. [PubMed] [Google Scholar]
  9. 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]
  10. Glasebrook A. L., Fitch F. W. T-cell lines which cooperate in generation of specific cytolytic activity. Nature. 1979 Mar 8;278(5700):171–173. doi: 10.1038/278171a0. [DOI] [PubMed] [Google Scholar]
  11. Gullberg M., Larsson E. L. Selective inhibition of antigen-induced "step one" in cytotoxic T lymphocytes by anti-Lyt-2 antibodies. Eur J Immunol. 1982 Dec;12(12):1006–1011. doi: 10.1002/eji.1830121205. [DOI] [PubMed] [Google Scholar]
  12. Hilfiker M. L., Moore R. N., Farrar J. J. Biologic properties of chromatographically separated murine thymoma-derived Interleukin 2 and colony-stimulating factor. J Immunol. 1981 Nov;127(5):1983–1987. [PubMed] [Google Scholar]
  13. Howe R. C., Russell J. H. Isolation of alloreactive CTL clones with cyclical changes in lytic activity. J Immunol. 1983 Nov;131(5):2141–2146. [PubMed] [Google Scholar]
  14. Hünig T., Loos M., Schimpl A. The role of accessory cells in polyclonal T cell activation. I. Both induction of interleukin 2 production and of interleukin 2 responsiveness by concanavalin A are accessory cell dependent. Eur J Immunol. 1983 Jan;13(1):1–6. doi: 10.1002/eji.1830130103. [DOI] [PubMed] [Google Scholar]
  15. Kanagawa O. Three different signals are required for the induction of cytolytic T lymphocytes from resting precursors. J Immunol. 1983 Aug;131(2):606–610. [PubMed] [Google Scholar]
  16. Kelso A., Glasebrook A. L., Kanagawa O., Brunner K. T. Production of macrophage-activating factor by T lymphocyte clones and correlation with other lymphokine activities. J Immunol. 1982 Aug;129(2):550–556. [PubMed] [Google Scholar]
  17. MacDonald H. R., Lees R. K. Dissociation of differentiation and proliferation in the primary induction of cytolytic T lymphocytes by alloantigens. J Immunol. 1980 Mar;124(3):1308–1313. [PubMed] [Google Scholar]
  18. Mizel S. B. Interleukin 1 and T cell activation. Immunol Rev. 1982;63:51–72. doi: 10.1111/j.1600-065x.1982.tb00411.x. [DOI] [PubMed] [Google Scholar]
  19. Mizel S. B., Oppenheim J. J., Rosenstreich D. L. Characterization of lymphocyte-activating factor (LAF) produced by the macrophage cell line, P388D1. I. Enhancement of LAF production by activated T lymphocytes. J Immunol. 1978 May;120(5):1497–1503. [PubMed] [Google Scholar]
  20. Moretta A., Colombatti M., Chapuis B. Human spleen as a source of T cell growth factor. Clin Exp Immunol. 1981 May;44(2):262–269. [PMC free article] [PubMed] [Google Scholar]
  21. Männel D. N., Falk W., Dröge W. Induction of cytotoxic T cell function requires sequential action of three different lymphokines. J Immunol. 1983 Jun;130(6):2508–2510. [PubMed] [Google Scholar]
  22. Nabholz M., Conzelmann A., Acuto O., North M., Haas W., Pohlit H., von Boehmer H., Hengartner H., Mach J. P., Engers H. Established murine cytolytic T-cell lines as tools for a somatic cell genetic analysis of T-cell functions. Immunol Rev. 1980;51:125–156. doi: 10.1111/j.1600-065x.1980.tb00319.x. [DOI] [PubMed] [Google Scholar]
  23. Nabholz M., MacDonald H. R. Cytolytic T lymphocytes. Annu Rev Immunol. 1983;1:273–306. doi: 10.1146/annurev.iy.01.040183.001421. [DOI] [PubMed] [Google Scholar]
  24. Oppenheim J. J., Stadler B. M., Siraganian R. P., Mage M., Mathieson B. Lymphokines: their role in lymphocyte responses. Properties of interleukin 1. Fed Proc. 1982 Feb;41(2):257–262. [PubMed] [Google Scholar]
  25. Perussia B., Mangoni L., Engers H. D., Trinchieri G. Interferon production by human and murine lymphocytes in response to alloantigens. J Immunol. 1980 Oct;125(4):1589–1595. [PubMed] [Google Scholar]
  26. Raulet D. H., Bevan M. J. A differentiation factor required for the expression of cytotoxic T-cell function. Nature. 1982 Apr 22;296(5859):754–757. doi: 10.1038/296754a0. [DOI] [PubMed] [Google Scholar]
  27. Robb R. J., Munck A., Smith K. A. T cell growth factor receptors. Quantitation, specificity, and biological relevance. J Exp Med. 1981 Nov 1;154(5):1455–1474. doi: 10.1084/jem.154.5.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ryser J. E., Cerottini J. C., Brunner K. T. Generation of cytolytic T lymphocytes in vitro. IX. induction of secondary CTL responses in primary long-term MLC by supernatants from secondary MLC. J Immunol. 1978 Feb;120(2):370–377. [PubMed] [Google Scholar]
  29. Sekaly R. P., MacDonald H. R., Zaech P., Nabholz M. Cell cycle regulation of cloned cytolytic T cells by T cell growth factor: analysis by flow microfluorometry. J Immunol. 1982 Oct;129(4):1407–1414. [PubMed] [Google Scholar]
  30. Silva A., MacDonald H. R., Conzelmann A., Corthésy P., Nabholz M. Rat X mouse T-cell hybrids with inducible specific cytolytic activity. Immunol Rev. 1983;76:105–129. doi: 10.1111/j.1600-065x.1983.tb01099.x. [DOI] [PubMed] [Google Scholar]
  31. Smith K. A., Favata M. F., Oroszlan S. Production and characterization of monoclonal antibodies to human interleukin 2: strategy and tactics. J Immunol. 1983 Oct;131(4):1808–1815. [PubMed] [Google Scholar]
  32. Wagner H., Hardt C., Rouse B. T., Röllinghoff M., Scheurich P., Pfizenmaier K. Dissection of the proliferative and differentiative signals controlling murine cytotoxic T lymphocyte responses. J Exp Med. 1982 Jun 1;155(6):1876–1881. doi: 10.1084/jem.155.6.1876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. von Boehmer H., Hengartner H., Nabholz M., Lernhardt W., Schreier M. H., Haas W. Fine specificity of a continuously growing killer cell clone specific for H-Y antigen. Eur J Immunol. 1979 Aug;9(8):592–597. doi: 10.1002/eji.1830090804. [DOI] [PubMed] [Google Scholar]

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