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. 1988 Aug;85(16):6072–6076. doi: 10.1073/pnas.85.16.6072

cAMP antagonizes interleukin 2-promoted T-cell cycle progression at a discrete point in early G1.

K W Johnson 1, B H Davis 1, K A Smith 1
PMCID: PMC281907  PMID: 2842759

Abstract

T lymphocytes are stimulated to proliferate in an autocrine/paracrine manner by the lymphokine interleukin 2 (IL-2). In seeking further insight into the mechanisms by which IL-2 induces progression of T cells through the G1 phase of the cell cycle, studies were performed with agents that increase cellular adenosine 3',5'-cyclic monophosphate (cAMP), a well-known inhibitor of lymphocyte growth. The addition of dibutyryl-cAMP, cholera toxin, forskolin, or 3-isobutyl-1-methylxanthine to an IL-2-dependent murine T-cell line evoked a dose-related suppression of S-phase transition without affecting cellular viability. Moreover, elevation of cAMP levels led to an accumulation of uniformly small cells, suggesting an arrest in early G1. Consistent with these findings, dibutyryl-cAMP inhibited the incorporation of both [3H]-uridine and [3H]thymidine by IL-2-stimulated, synchronized normal human T cells. Furthermore, maximal inhibition occurred during early G1, as indicated by experiments where the addition of dibutyryl-cAMP was delayed with respect to IL-2 stimulation. Quantitative flow cytometric analysis of RNA and DNA content of IL-2-stimulated cells affirmed that increased cAMP inhibits RNA accumulation and S-phase transition. In addition, exposure of IL-2-dependent, asynchronously proliferating normal human T cells to dibutyryl-cAMP resulted in uniform growth arrest in early G1, the point at which cycling T cells accumulate when they are deprived of IL-2. These results indicate that increased cAMP inhibits G1 progression stimulated by IL-2 and provide a rationale for the use of cAMP analogues as pharmacologic probes for the dissection of molecular events occurring during IL-2 signaling and T-cell G1 transit. They also suggest the possibility of therapeutic immunosuppression by a combination of agents that act at different stages of the T-cell cycle.

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Selected References

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

  1. Appleman M. M., Thompson W. J., Russell T. R. Cyclic nucleotide phosphodiesterases. Adv Cyclic Nucleotide Res. 1973;3:65–98. [PubMed] [Google Scholar]
  2. Baker P. E., Fahey J. V., Munck A. Prostaglandin inhibition of T-cell proliferation is mediated at two levels. Cell Immunol. 1981 Jun;61(1):52–61. doi: 10.1016/0008-8749(81)90353-1. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Campisi J., Pardee A. B. Post-transcriptional control of the onset of DNA synthesis by an insulin-like growth factor. Mol Cell Biol. 1984 Sep;4(9):1807–1814. doi: 10.1128/mcb.4.9.1807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cantrell D. A., Smith K. A. The interleukin-2 T-cell system: a new cell growth model. Science. 1984 Jun 22;224(4655):1312–1316. doi: 10.1126/science.6427923. [DOI] [PubMed] [Google Scholar]
  6. Chouaib S., Welte K., Mertelsmann R., Dupont B. Prostaglandin E2 acts at two distinct pathways of T lymphocyte activation: inhibition of interleukin 2 production and down-regulation of transferrin receptor expression. J Immunol. 1985 Aug;135(2):1172–1179. [PubMed] [Google Scholar]
  7. Coffino P., Gray J. W. Regulation of S49 lymphoma cell growth by cyclic adenosine 3':5'-monophosphate. Cancer Res. 1978 Nov;38(11 Pt 2):4285–4288. [PubMed] [Google Scholar]
  8. Darzynkiewicz Z., Evenson D., Staiano-Coico L., Sharpless T., Melamed M. R. Relationship between RNA content and progression of lymphocytes through S phase of cell cycle. Proc Natl Acad Sci U S A. 1979 Jan;76(1):358–362. doi: 10.1073/pnas.76.1.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Darzynkiewicz Z., Traganos F., Staiano-Coico L. Cell and nuclear growth during G1: kinetic and clinical implications. Ann N Y Acad Sci. 1986;468:45–54. doi: 10.1111/j.1749-6632.1986.tb42027.x. [DOI] [PubMed] [Google Scholar]
  10. DosReis G. A., Nóbrega A. F., de Carvalho R. P. Purinergic modulation of T-lymphocyte activation: differential susceptibility of distinct activation steps and correlation with intracellular 3',5'-cyclic adenosine monophosphate accumulation. Cell Immunol. 1986 Aug;101(1):213–231. doi: 10.1016/0008-8749(86)90199-1. [DOI] [PubMed] [Google Scholar]
  11. Freedman M. H., Raff M. C. Induction of increased calcium uptake in mouse T lymphocytes by concanavalin A and its modulation by cyclic nucleotides. Nature. 1975 May 29;255(5507):378–382. doi: 10.1038/255378a0. [DOI] [PubMed] [Google Scholar]
  12. Gill G. N., McCune R. W. Guanosine 3',5'-monophosphate-dependent protein kinase. Curr Top Cell Regul. 1979;15:1–45. doi: 10.1016/b978-0-12-152815-7.50005-3. [DOI] [PubMed] [Google Scholar]
  13. Gullberg M., Smith K. A. Regulation of T cell autocrine growth. T4+ cells become refractory to interleukin 2. J Exp Med. 1986 Feb 1;163(2):270–284. doi: 10.1084/jem.163.2.270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Henney C. S., Lichtenstein L. M., Gillespie E., Rolley R. T. In vivo suppression of the immune response to alloantigen by cholera enterotoxin. J Clin Invest. 1973 Nov;52(11):2853–2857. doi: 10.1172/JCI107481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Herzberg V. L., Smith K. A. T cell growth without serum. J Immunol. 1987 Aug 15;139(4):998–1004. [PubMed] [Google Scholar]
  16. Hirata F., Matsuda K., Notsu Y., Hattori T., del Carmine R. Phosphorylation at a tyrosine residue of lipomodulin in mitogen-stimulated murine thymocytes. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4717–4721. doi: 10.1073/pnas.81.15.4717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Holsapple M. P., Tucker A. N., McNerney P. J., White K. L., Jr Effects of N-nitrosodimethylamine on humoral immunity. J Pharmacol Exp Ther. 1984 May;229(2):493–500. [PubMed] [Google Scholar]
  18. Insel P. A., Bourne H. R., Coffino P., Tomkins G. M. Cyclic AMP-dependent protein kinase: pivotal role in regulation of enzyme induction and growth. Science. 1975 Nov 28;190(4217):896–898. doi: 10.1126/science.171770. [DOI] [PubMed] [Google Scholar]
  19. Iwaz J., Lafont S., Revillard J. P. Elevation of cyclic 3'5' adenosine monophosphate levels by cholera toxin inhibits the generation of interleukin 2 activity. Cell Immunol. 1986 Dec;103(2):455–461. doi: 10.1016/0008-8749(86)90105-x. [DOI] [PubMed] [Google Scholar]
  20. Kelley V. E., Winkelstein A., Izui S. Effect of prostaglandin E on immune complex nephritis in NZB/W mice. Lab Invest. 1979 Dec;41(6):531–537. [PubMed] [Google Scholar]
  21. Klausner R. D., O'Shea J. J., Luong H., Ross P., Bluestone J. A., Samelson L. E. T cell receptor tyrosine phosphorylation. Variable coupling for different activating ligands. J Biol Chem. 1987 Sep 15;262(26):12654–12659. [PubMed] [Google Scholar]
  22. Leof E. B., Van Wyk J. J., O'Keefe E. J., Pledger W. J. Epidermal growth factor (EGF) is required only during the traverse of early G1 in PDGF stimulated density-arrested BALB/c-3T3 cells. Exp Cell Res. 1983 Aug;147(1):202–208. doi: 10.1016/0014-4827(83)90285-9. [DOI] [PubMed] [Google Scholar]
  23. Leof E. B., Wharton W., O'Keefe E., Pledger W. J. Elevated intracellular concentrations of cyclic AMP inhibited serum-stimulated, density-arrested BALB/c-3T3 cells in mid G1. J Cell Biochem. 1982;19(1):93–103. doi: 10.1002/jcb.240190108. [DOI] [PubMed] [Google Scholar]
  24. Leof E. B., Wharton W., van Wyk J. J., Pledger W. J. Epidermal growth factor (EGF) and somatomedin C regulate G1 progression in competent BALB/c-3T3 cells. Exp Cell Res. 1982 Sep;141(1):107–115. doi: 10.1016/0014-4827(82)90073-8. [DOI] [PubMed] [Google Scholar]
  25. Lerner A., Jacobson B., Miller R. A. Cyclic AMP concentrations modulate both calcium flux and hydrolysis of phosphatidylinositol phosphates in mouse T lymphocytes. J Immunol. 1988 Feb 1;140(3):936–940. [PubMed] [Google Scholar]
  26. Mary D., Aussel C., Ferrua B., Fehlmann M. Regulation of interleukin 2 synthesis by cAMP in human T cells. J Immunol. 1987 Aug 15;139(4):1179–1184. [PubMed] [Google Scholar]
  27. Meuer S. C., Hussey R. E., Cantrell D. A., Hodgdon J. C., Schlossman S. F., Smith K. A., Reinherz E. L. Triggering of the T3-Ti antigen-receptor complex results in clonal T-cell proliferation through an interleukin 2-dependent autocrine pathway. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1509–1513. doi: 10.1073/pnas.81.5.1509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Neckers L. M., Cossman J. Transferrin receptor induction in mitogen-stimulated human T lymphocytes is required for DNA synthesis and cell division and is regulated by interleukin 2. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3494–3498. doi: 10.1073/pnas.80.11.3494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Novogrodsky A., Patya M., Rubin A. L., Stenzel K. H. Agents that increase cellular cAMP inhibit production of interleukin-2, but not its activity. Biochem Biophys Res Commun. 1983 Jul 18;114(1):93–98. doi: 10.1016/0006-291x(83)91598-x. [DOI] [PubMed] [Google Scholar]
  30. POSTERNAK T., SUTHERLAND E. W., HENION W. F. Derivatives of cyclic 3',5'-adenosine monophosphate. Biochim Biophys Acta. 1962 Dec 17;65:558–560. doi: 10.1016/0006-3002(62)90475-4. [DOI] [PubMed] [Google Scholar]
  31. Pardee A. B. A restriction point for control of normal animal cell proliferation. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1286–1290. doi: 10.1073/pnas.71.4.1286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pastan I. H., Johnson G. S., Anderson W. B. Role of cyclic nucleotides in growth control. Annu Rev Biochem. 1975;44:491–522. doi: 10.1146/annurev.bi.44.070175.002423. [DOI] [PubMed] [Google Scholar]
  33. Prasad K. N. Butyric acid: a small fatty acid with diverse biological functions. Life Sci. 1980 Oct 13;27(15):1351–1358. doi: 10.1016/0024-3205(80)90397-5. [DOI] [PubMed] [Google Scholar]
  34. Reichard P., Ehrenberg A. Ribonucleotide reductase--a radical enzyme. Science. 1983 Aug 5;221(4610):514–519. doi: 10.1126/science.6306767. [DOI] [PubMed] [Google Scholar]
  35. Scher C. D., Shepard R. C., Antoniades H. N., Stiles C. D. Platelet-derived growth factor and the regulation of the mammalian fibroblast cell cycle. Biochim Biophys Acta. 1979 Aug 10;560(2):217–241. doi: 10.1016/0304-419x(79)90020-9. [DOI] [PubMed] [Google Scholar]
  36. Simantov R., Sachs L. Temperature sensitivity of cyclic adenosine 3':5'-monophosphate-binding proteins and the regulation of growth and differentiation in neuroblastoma cells. J Biol Chem. 1975 May 10;250(9):3236–3242. [PubMed] [Google Scholar]
  37. Smith K. A. Interleukin-2: inception, impact, and implications. Science. 1988 May 27;240(4856):1169–1176. doi: 10.1126/science.3131876. [DOI] [PubMed] [Google Scholar]
  38. Stern J. B., Smith K. A. Interleukin-2 induction of T-cell G1 progression and c-myb expression. Science. 1986 Jul 11;233(4760):203–206. doi: 10.1126/science.3523754. [DOI] [PubMed] [Google Scholar]
  39. Stewart C. C., Cramer S. F., Steward P. G. The response of human peripheral blood lymphocytes to phytohemagglutinin: determination of cell numbers. Cell Immunol. 1975 Apr;16(2):237–250. doi: 10.1016/0008-8749(75)90115-x. [DOI] [PubMed] [Google Scholar]
  40. Strom T. B., Carpenter C. B. Cyclic nucleotides in immunosuppression--neuroendocrine pharmacologic manipulation and in vivo immunoregulation of immunity acting via second messenger systems. Transplant Proc. 1980 Jun;12(2):304–310. [PubMed] [Google Scholar]
  41. Strom T. B., Carpenter C. B., Jr, Cragoe E. J., Jr, Norris S., Devlin R., Perper R. J. Suppression of in vivo and in vitro alloimmunity by prostaglandins. Transplant Proc. 1977 Mar;9(1):1075–1079. [PubMed] [Google Scholar]
  42. Stryer L., Bourne H. R. G proteins: a family of signal transducers. Annu Rev Cell Biol. 1986;2:391–419. doi: 10.1146/annurev.cb.02.110186.002135. [DOI] [PubMed] [Google Scholar]
  43. Wang H. M., Smith K. A. The interleukin 2 receptor. Functional consequences of its bimolecular structure. J Exp Med. 1987 Oct 1;166(4):1055–1069. doi: 10.1084/jem.166.4.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]

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