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. 1987 Jan 1;165(1):173–194. doi: 10.1084/jem.165.1.173

Cell growth cycle block of T cell hybridomas upon activation with antigen

PMCID: PMC2188264  PMID: 3491868

Abstract

Stimulation of antigen-specific T cell hybridomas with the appropriate antigen/MHC combination, at concentrations that resulted in the secretion of the lymphokine interleukin 2, resulted in a dose-dependent decrease in both [3H]thymidine incorporation and cell growth. Flow cytometric studies demonstrated that stimulation with antigen resulted in a cell cycle block that was most evident at the G1/S border, and mixing studies revealed that bystander T cells of different antigen specificities were not affected. For at least the large majority of T cells, the G1/S cell cycle block appeared to be irreversible after 24 h of exposure to antigen. This cell cycle block may be useful as a rapid and quantitative measure of T cell hybridoma activation, as a means of selecting T cell hybridomas that have functional alterations in the reception of stimulatory signals, and may serve as a model of the induction of tolerance in immature T cells.

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Selected References

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  1. Allen P. M., McKean D. J., Beck B. N., Sheffield J., Glimcher L. H. Direct evidence that a class II molecule and a simple globular protein generate multiple determinants. J Exp Med. 1985 Oct 1;162(4):1264–1274. doi: 10.1084/jem.162.4.1264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Altman A., Schreiber R. D., Katz D. H. Production of antigen-nonspecific immunoregulatory lymphokines by T cell hybridomas. Curr Top Microbiol Immunol. 1982;100:179–190. doi: 10.1007/978-3-642-68586-6_21. [DOI] [PubMed] [Google Scholar]
  3. Bijsterbosch M. K., Meade C. J., Turner G. A., Klaus G. G. B lymphocyte receptors and polyphosphoinositide degradation. Cell. 1985 Jul;41(3):999–1006. doi: 10.1016/s0092-8674(85)80080-5. [DOI] [PubMed] [Google Scholar]
  4. Boyd A. W., Schrader J. W. The regulation of growth and differentiation of a murine B cell lymphoma. II. The inhibition of WEHI 231 by anti-immunoglobulin antibodies. J Immunol. 1981 Jun;126(6):2466–2469. [PubMed] [Google Scholar]
  5. Carpenter G., Cohen S. Epidermal growth factor. Annu Rev Biochem. 1979;48:193–216. doi: 10.1146/annurev.bi.48.070179.001205. [DOI] [PubMed] [Google Scholar]
  6. Coggeshall K. M., Cambier J. C. B cell activation. VIII. Membrane immunoglobulins transduce signals via activation of phosphatidylinositol hydrolysis. J Immunol. 1984 Dec;133(6):3382–3386. [PubMed] [Google Scholar]
  7. Corradin G., Harbury H. A. Cleavage of cytochrome c with cyanogen bromide. Biochim Biophys Acta. 1970 Dec 22;221(3):489–496. doi: 10.1016/0005-2795(70)90219-9. [DOI] [PubMed] [Google Scholar]
  8. Dean P. N., Jett J. H. Mathematical analysis of DNA distributions derived from flow microfluorometry. J Cell Biol. 1974 Feb;60(2):523–527. doi: 10.1083/jcb.60.2.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Defranco A. L., Raveche E. S., Asofsky R., Paul W. E. Frequency of B lymphocytes responsive to anti-immunoglobulin. J Exp Med. 1982 May 1;155(5):1523–1536. doi: 10.1084/jem.155.5.1523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fox R. M., Kefford R. F., Tripp E. H., Taylor I. W. G1-phase arrest of cultured human leukemic T-cells induced by deoxyadenosine. Cancer Res. 1981 Dec;41(12 Pt 1):5141–5150. [PubMed] [Google Scholar]
  11. Gill G. N., Lazar C. S. Increased phosphotyrosine content and inhibition of proliferation in EGF-treated A431 cells. Nature. 1981 Sep 24;293(5830):305–307. doi: 10.1038/293305a0. [DOI] [PubMed] [Google Scholar]
  12. Glimcher L. H., Schroer J. A., Chan C., Shevach E. M. Fine specificity of cloned insulin-specific T cell hybridomas: evidence supporting a role for tertiary conformation. J Immunol. 1983 Dec;131(6):2868–2874. [PubMed] [Google Scholar]
  13. Goldsby R. A., Osborne B. A., Simpson E., Herzenberg L. A. Hybrid cell lines with T-cell characteristics. Nature. 1977 Jun 23;267(5613):707–708. doi: 10.1038/267707a0. [DOI] [PubMed] [Google Scholar]
  14. Harwell L., Skidmore B., Marrack P., Kappler J. Concanavalin A-inducible, interleukin-2-producing T cell hybridoma. J Exp Med. 1980 Oct 1;152(4):893–904. doi: 10.1084/jem.152.4.893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hatakeyama M., Minamoto S., Uchiyama T., Hardy R. R., Yamada G., Taniguchi T. Reconstitution of functional receptor for human interleukin-2 in mouse cells. Nature. 1985 Dec 5;318(6045):467–470. doi: 10.1038/318467a0. [DOI] [PubMed] [Google Scholar]
  16. Hämmerling G. J. T lymphocyte tissue culture lines produced by cell hybridization. Eur J Immunol. 1977 Oct;7(10):743–746. doi: 10.1002/eji.1830071018. [DOI] [PubMed] [Google Scholar]
  17. Kappler J. W., Skidmore B., White J., Marrack P. Antigen-inducible, H-2-restricted, interleukin-2-producing T cell hybridomas. Lack of independent antigen and H-2 recognition. J Exp Med. 1981 May 1;153(5):1198–1214. doi: 10.1084/jem.153.5.1198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kappler J., Kubo R., Haskins K., White J., Marrack P. The mouse T cell receptor: comparison of MHC-restricted receptors on two T cell hybridomas. Cell. 1983 Oct;34(3):727–737. doi: 10.1016/0092-8674(83)90529-9. [DOI] [PubMed] [Google Scholar]
  19. Kehrl J. H., Wakefield L. M., Roberts A. B., Jakowlew S., Alvarez-Mon M., Derynck R., Sporn M. B., Fauci A. S. Production of transforming growth factor beta by human T lymphocytes and its potential role in the regulation of T cell growth. J Exp Med. 1986 May 1;163(5):1037–1050. doi: 10.1084/jem.163.5.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kishimoto T. Factors affecting B-cell growth and differentiation. Annu Rev Immunol. 1985;3:133–157. doi: 10.1146/annurev.iy.03.040185.001025. [DOI] [PubMed] [Google Scholar]
  21. Kobayashi Y., Asada M., Higuchi M., Osawa T. Human T cell hybridomas producing lymphokines. I. Establishment and characterization of human T cell hybridomas producing lymphotoxin and migration inhibitory factor. J Immunol. 1982 Jun;128(6):2714–2718. [PubMed] [Google Scholar]
  22. Krolick K. A., Isakson P. C., Uhr J. W., Vitetta E. S. BCL1, a murine model for chronic lymphocytic leukemia: use of the surface immunoglobulin idiotype for the detection and treatment of tumor. Immunol Rev. 1979;48:81–106. doi: 10.1111/j.1600-065x.1979.tb00299.x. [DOI] [PubMed] [Google Scholar]
  23. Köhler G., Lefkovits I., Elliott B., Coutinho A. Derivation of hybrids between a thymoma line and spleen cells activated in a mixed leukocyte reaction. Eur J Immunol. 1977 Nov;7(11):758–761. doi: 10.1002/eji.1830071103. [DOI] [PubMed] [Google Scholar]
  24. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  25. Köhler G., Milstein C. Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion. Eur J Immunol. 1976 Jul;6(7):511–519. doi: 10.1002/eji.1830060713. [DOI] [PubMed] [Google Scholar]
  26. Marks P. A., Rifkind R. A. Erythroleukemic differentiation. Annu Rev Biochem. 1978;47:419–448. doi: 10.1146/annurev.bi.47.070178.002223. [DOI] [PubMed] [Google Scholar]
  27. Oi V. T., Jones P. P., Goding J. W., Herzenberg L. A., Herzenberg L. A. Properties of monoclonal antibodies to mouse Ig allotypes, H-2, and Ia antigens. Curr Top Microbiol Immunol. 1978;81:115–120. doi: 10.1007/978-3-642-67448-8_18. [DOI] [PubMed] [Google Scholar]
  28. Ozato K., Epstein S. L., Henkart P., Hansen T. H., Sachs D. H. Studies on monoclonal antibodies to mouse MHC products. Transplant Proc. 1981 Mar;13(1 Pt 2):958–962. [PubMed] [Google Scholar]
  29. Ralph P. Functional subsets of murine and human B lymphocyte cell lines. Immunol Rev. 1979;48:107–121. doi: 10.1111/j.1600-065x.1979.tb00300.x. [DOI] [PubMed] [Google Scholar]
  30. Ralph P., Nakoinz I. Inhibitory effects of lectins and lymphocyte mitogens on murine lymphomas and myelomas. J Natl Cancer Inst. 1973 Sep;51(3):883–890. doi: 10.1093/jnci/51.3.883. [DOI] [PubMed] [Google Scholar]
  31. Ralph P. Retention of lymphocyte characteristics by myelomas and theta + -lymphomas: sensitivity to cortisol and phytohemagglutinin. J Immunol. 1973 Jun;110(6):1470–1475. [PubMed] [Google Scholar]
  32. Roberts A. B., Anzano M. A., Wakefield L. M., Roche N. S., Stern D. F., Sporn M. B. Type beta transforming growth factor: a bifunctional regulator of cellular growth. Proc Natl Acad Sci U S A. 1985 Jan;82(1):119–123. doi: 10.1073/pnas.82.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rock K. L., Benacerraf B. Inhibition of antigen-specific T lymphocyte activation by structurally related Ir gene-controlled polymers. Evidence of specific competition for accessory cell antigen presentation. J Exp Med. 1983 May 1;157(5):1618–1634. doi: 10.1084/jem.157.5.1618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ruddle N. H., Conta B. S. Lymphotoxin and immune (gamma) interferon production by T cell lines and hybrids. Curr Top Microbiol Immunol. 1982;100:239–248. doi: 10.1007/978-3-642-68586-6_27. [DOI] [PubMed] [Google Scholar]
  35. Samelson L. E., Germain R. N., Schwartz R. H. Monoclonal antibodies against the antigen receptor on a cloned T-cell hybrid. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6972–6976. doi: 10.1073/pnas.80.22.6972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Samelson L. E., Schwartz R. H. T cell clone-specific alloantisera that inhibit or stimulate antigen-induced T cell activation. J Immunol. 1983 Dec;131(6):2645–2650. [PubMed] [Google Scholar]
  37. Schonbrunn A., Krasnoff M., Westendorf J. M., Tashjian A. H., Jr Epidermal growth factor and thyrotropin-releasing hormone act similarly on a clonal pituitary cell strain. Modulation of hormone production and inhbition of cell proliferation. J Cell Biol. 1980 Jun;85(3):786–797. doi: 10.1083/jcb.85.3.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schrader J. W., Clark-Lewis I. The use of T cell hybridomas in the biochemical and biological characterization of multiple regulatory factors produced by T cells. Curr Top Microbiol Immunol. 1982;100:221–229. doi: 10.1007/978-3-642-68586-6_25. [DOI] [PubMed] [Google Scholar]
  39. Scott D. W., Tuttle J., Livnat D., Haynes W., Cogswell J. P., Keng P. Lymphoma models for B-cell activation and tolerance. II. Growth inhibition by anti-mu of WEHI-231 and the selection and properties of resistant mutants. Cell Immunol. 1985 Jun;93(1):124–131. doi: 10.1016/0008-8749(85)90393-4. [DOI] [PubMed] [Google Scholar]
  40. Sharpless T., Traganos F., Darzynkiewicz Z., Melamed M. R. Flow cytofluorimetry: discrimination between single cells and cell aggregates by direct size measurements. Acta Cytol. 1975 Nov-Dec;19(6):577–581. [PubMed] [Google Scholar]
  41. Shimonkevitz R., Kappler J., Marrack P., Grey H. Antigen recognition by H-2-restricted T cells. I. Cell-free antigen processing. J Exp Med. 1983 Aug 1;158(2):303–316. doi: 10.1084/jem.158.2.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sitkovsky M. V., Pasternack M. S., Lugo J. P., Klein J. R., Eisen H. N. Isolation and partial characterization of concanavalin A receptors on cloned cytotoxic T lymphocytes. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1519–1523. doi: 10.1073/pnas.81.5.1519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Smith K. A. Interleukin 2. Annu Rev Immunol. 1984;2:319–333. doi: 10.1146/annurev.iy.02.040184.001535. [DOI] [PubMed] [Google Scholar]
  44. Sugamura K., Nakai S., Fujii M., Hinuma Y. Interleukin 2 inhibits in vitro growth of human T cell lines carrying retrovirus. J Exp Med. 1985 May 1;161(5):1243–1248. doi: 10.1084/jem.161.5.1243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tucker R. F., Shipley G. D., Moses H. L., Holley R. W. Growth inhibitor from BSC-1 cells closely related to platelet type beta transforming growth factor. Science. 1984 Nov 9;226(4675):705–707. doi: 10.1126/science.6093254. [DOI] [PubMed] [Google Scholar]
  46. Zucker R. M., Whittington K., Price B. J. Differentiation of HL-60 cells: cell volume and cell cycle changes. Cytometry. 1983 May;3(6):414–418. doi: 10.1002/cyto.990030605. [DOI] [PubMed] [Google Scholar]

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