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. 1994 Dec 15;304(Pt 3):769–774. doi: 10.1042/bj3040769

Mobilization of intracellular Ca2+ by adenine nucleotides in human T-leukaemia cells: evidence for ADP-specific and P2y-purinergic receptors.

M Biffen 1, D R Alexander 1
PMCID: PMC1137400  PMID: 7818479

Abstract

The expression of purinergic receptors on human T-cells was investigated and the receptors were shown to be functionally coupled to intracellular signals in two out of eight T-leukaemia cell-lines. Addition of adenine nucleotides resulted in mobilization of intracellular Ca2+ in HPB-ALL cells and a cell line (CB1) recently isolated from a patient with T-acute lymphoblastic leukaemia. Of a range of nucleotides tested only ADP and ATP elevated intracellular levels of Ca2+, with ADP being the more potent agonist. Ca2+ mobilization by ATP was accompanied by increased inositol phosphate production and was blocked by the purinergic receptor antagonist, Reactive Blue 2, indicating that ATP was interacting with a P2y receptor. Intracellular Ca2+ release triggered by ADP was independent of both inositol phosphate production and protein tyrosine phosphorylation. Expression of the transmembrane phosphotyrosine phosphatase, CD45, had no effect on ADP-stimulated Ca2+ mobilization. Our results show that functional P2y receptors can be expressed on T-cells, and also identify a novel T-cell ADP receptor. Signals mediated by these purinergic receptors could play important roles in modulating T-cell function.

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Selected References

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

  1. Berridge M. J., Irvine R. F. Inositol phosphates and cell signalling. Nature. 1989 Sep 21;341(6239):197–205. doi: 10.1038/341197a0. [DOI] [PubMed] [Google Scholar]
  2. Biffen M., McMichael-Phillips D., Larson T., Venkitaraman A., Alexander D. The CD45 tyrosine phosphatase regulates specific pools of antigen receptor-associated p59fyn and CD4-associated p56lck tyrosine in human T-cells. EMBO J. 1994 Apr 15;13(8):1920–1929. doi: 10.1002/j.1460-2075.1994.tb06461.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Biffen M., Shiroo M., Alexander D. R. G-proteins are not directly involved in the CD3-antigen-mediated production of inositol phosphates in HPB-ALL T-leukaemia cells expressing phospholipase C isoforms gamma 1 and beta 3. Biochem J. 1993 Jan 15;289(Pt 2):387–394. doi: 10.1042/bj2890387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burnstock G., Warland J. J. P2-purinoceptors of two subtypes in the rabbit mesenteric artery: reactive blue 2 selectively inhibits responses mediated via the P2y-but not the P2x-purinoceptor. Br J Pharmacol. 1987 Feb;90(2):383–391. doi: 10.1111/j.1476-5381.1987.tb08968.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Charest R., Blackmore P. F., Exton J. H. Characterization of responses of isolated rat hepatocytes to ATP and ADP. J Biol Chem. 1985 Dec 15;260(29):15789–15794. [PubMed] [Google Scholar]
  6. Fisher G. J., Bakshian S., Baldassare J. J. Activation of human platelets by ADP causes a rapid rise in cytosolic free calcium without hydrolysis of phosphatidylinositol-4,5-bisphosphate. Biochem Biophys Res Commun. 1985 Jun 28;129(3):958–964. doi: 10.1016/0006-291x(85)91984-9. [DOI] [PubMed] [Google Scholar]
  7. Fishman R. F., Rubin A. L., Novogrodsky A., Stenzel K. H. Selective suppression of blastogenesis induced by different mitogens: effect of noncyclic adenosine-containing compounds. Cell Immunol. 1980 Aug 15;54(1):129–139. doi: 10.1016/0008-8749(80)90196-3. [DOI] [PubMed] [Google Scholar]
  8. Frelin C., Breittmayer J. P., Vigne P. ADP induces inositol phosphate-independent intracellular Ca2+ mobilization in brain capillary endothelial cells. J Biol Chem. 1993 Apr 25;268(12):8787–8792. [PubMed] [Google Scholar]
  9. Gordon J. L. Extracellular ATP: effects, sources and fate. Biochem J. 1986 Jan 15;233(2):309–319. doi: 10.1042/bj2330309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gregory C. D., Dive C., Henderson S., Smith C. A., Williams G. T., Gordon J., Rickinson A. B. Activation of Epstein-Barr virus latent genes protects human B cells from death by apoptosis. Nature. 1991 Feb 14;349(6310):612–614. doi: 10.1038/349612a0. [DOI] [PubMed] [Google Scholar]
  11. Holter W., Spiegel A. M., Howard B. H., Weber S., Brann M. R. Expression of GTP-binding proteins and prostaglandin E2 receptors during human T cell activation. Cell Immunol. 1991 May;134(2):287–295. doi: 10.1016/0008-8749(91)90303-s. [DOI] [PubMed] [Google Scholar]
  12. Hurley T. R., Hyman R., Sefton B. M. Differential effects of expression of the CD45 tyrosine protein phosphatase on the tyrosine phosphorylation of the lck, fyn, and c-src tyrosine protein kinases. Mol Cell Biol. 1993 Mar;13(3):1651–1656. doi: 10.1128/mcb.13.3.1651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ikehara S., Pahwa R. N., Lunzer D. G., Good R. A., Modak M. J. Adenosine-5'-triphosphate-(ATP) mediated stimulation and suppression of DNA synthesis in lymphoid cells. I. Characterization of ATP responsive cells in mouse lymphoid organs. J Immunol. 1981 Nov;127(5):1834–1838. [PubMed] [Google Scholar]
  14. Inoue K., Nakazawa K., Ohara-Imaizumi M., Obama T., Fujimori K., Takanaka A. Antagonism by reactive blue 2 but not by brilliant blue G of extracellular ATP-evoked responses in PC12 phaeochromocytoma cells. Br J Pharmacol. 1991 Apr;102(4):851–854. doi: 10.1111/j.1476-5381.1991.tb12265.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Koretzky G. A., Picus J., Schultz T., Weiss A. Tyrosine phosphatase CD45 is required for T-cell antigen receptor and CD2-mediated activation of a protein tyrosine kinase and interleukin 2 production. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2037–2041. doi: 10.1073/pnas.88.6.2037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lustig K. D., Shiau A. K., Brake A. J., Julius D. Expression cloning of an ATP receptor from mouse neuroblastoma cells. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5113–5117. doi: 10.1073/pnas.90.11.5113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Macfarlane D. E., Mills D. C. The effects of ATP on platelets: evidence against the central role of released ADP in primary aggregation. Blood. 1975 Sep;46(3):309–320. [PubMed] [Google Scholar]
  18. Park D. J., Min H. K., Rhee S. G. Inhibition of CD3-linked phospholipase C by phorbol ester and by cAMP is associated with decreased phosphotyrosine and increased phosphoserine contents of PLC-gamma 1. J Biol Chem. 1992 Jan 25;267(3):1496–1501. [PubMed] [Google Scholar]
  19. Pirotton S., Raspe E., Demolle D., Erneux C., Boeynaems J. M. Involvement of inositol 1,4,5-trisphosphate and calcium in the action of adenine nucleotides on aortic endothelial cells. J Biol Chem. 1987 Dec 25;262(36):17461–17466. [PubMed] [Google Scholar]
  20. Sage S. O., Rink T. J. Kinetic differences between thrombin-induced and ADP-induced calcium influx and release from internal stores in fura-2-loaded human platelets. Biochem Biophys Res Commun. 1986 May 14;136(3):1124–1129. doi: 10.1016/0006-291x(86)90450-x. [DOI] [PubMed] [Google Scholar]
  21. Sargeant P., Farndale R. W., Sage S. O. ADP- and thapsigargin-evoked Ca2+ entry and protein-tyrosine phosphorylation are inhibited by the tyrosine kinase inhibitors genistein and methyl-2,5-dihydroxycinnamate in fura-2-loaded human platelets. J Biol Chem. 1993 Aug 25;268(24):18151–18156. [PubMed] [Google Scholar]
  22. Schmidt A., Ortaldo J. R., Herberman R. B. Inhibition of human natural killer cell reactivity by exogenous adenosine 5'-triphosphate. J Immunol. 1984 Jan;132(1):146–150. [PubMed] [Google Scholar]
  23. Shiroo M., Goff L., Biffen M., Shivnan E., Alexander D. CD45 tyrosine phosphatase-activated p59fyn couples the T cell antigen receptor to pathways of diacylglycerol production, protein kinase C activation and calcium influx. EMBO J. 1992 Dec;11(13):4887–4897. doi: 10.1002/j.1460-2075.1992.tb05595.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Smith C. A., Williams G. T., Kingston R., Jenkinson E. J., Owen J. J. Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures. Nature. 1989 Jan 12;337(6203):181–184. doi: 10.1038/337181a0. [DOI] [PubMed] [Google Scholar]
  25. Smith S. D., McFall P., Morgan R., Link M., Hecht F., Cleary M., Sklar J. Long-term growth of malignant thymocytes in vitro. Blood. 1989 Jun;73(8):2182–2187. [PubMed] [Google Scholar]
  26. Smith S. D., Morgan R., Gemmell R., Amylon M. D., Link M. P., Linker C., Hecht B. K., Warnke R., Glader B. E., Hecht F. Clinical and biologic characterization of T-cell neoplasias with rearrangements of chromosome 7 band q34. Blood. 1988 Feb;71(2):395–402. [PubMed] [Google Scholar]
  27. Soltoff S. P., McMillian M. K., Talamo B. R. Coomassie Brilliant Blue G is a more potent antagonist of P2 purinergic responses than Reactive Blue 2 (Cibacron Blue 3GA) in rat parotid acinar cells. Biochem Biophys Res Commun. 1989 Dec 29;165(3):1279–1285. doi: 10.1016/0006-291x(89)92741-1. [DOI] [PubMed] [Google Scholar]
  28. Stewart S. J., Cunningham G. R., Strupp J. A., House F. S., Kelley L. L., Henderson G. S., Exton J. H., Bocckino S. B. Activation of phospholipase D: a signaling system set in motion by perturbation of the T lymphocyte antigen receptor/CD3 complex. Cell Regul. 1991 Oct;2(10):841–850. doi: 10.1091/mbc.2.10.841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ward S. G., Cantrell D. A. Heterogeneity of the regulation of phospholipase C by phorbol esters in T lymphocytes. J Immunol. 1990 May 1;144(9):3523–3528. [PubMed] [Google Scholar]
  30. Yule D. I., Wu D., Essington T. E., Shayman J. A., Williams J. A. Sphingosine metabolism induces Ca2+ oscillations in rat pancreatic acinar cells. J Biol Chem. 1993 Jun 15;268(17):12353–12358. [PubMed] [Google Scholar]
  31. el-Moatassim C., Dornand J., Mani J. C. Extracellular ATP increases cytosolic free calcium in thymocytes and initiates the blastogenesis of the phorbol 12-myristate 13-acetate-treated medullary population. Biochim Biophys Acta. 1987 Mar 11;927(3):437–444. doi: 10.1016/0167-4889(87)90110-8. [DOI] [PubMed] [Google Scholar]
  32. el-Moatassim C., Dubyak G. R. A novel pathway for the activation of phospholipase D by P2z purinergic receptors in BAC1.2F5 macrophages. J Biol Chem. 1992 Nov 25;267(33):23664–23673. [PubMed] [Google Scholar]

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