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, Volume 35, Issue 3–4, pp 238–244 | Cite as

Differential effects of the calcium ionophore A23187 and the phorbol ester PMA on lymphocyte proliferation

  • D. Planelles
  • J. Hernández-Godoy
  • A. González-Molina
Inflammation and Immunomodulation


The effects of the phorbol ester, phorbol-12-myristate-13-acetate (PMA), and of the calcium ionophore A23187 on DNA synthesis in murine quiescent and mitogen-stimulated lymphocytes have been examined. Neither PMA nor A23187 had any mitogen effect on their own on quiescent lymphocytes. However, stimulation of cells sequentially with A23187 and then PMA resulted in a proliferative response in proportion to the duration of the exposure to A23187, and the sustained simultaneous presence of both agents was necessary for maximum proliferation. On the other hand, while short incubations with A23187 potentiated mitogen-induced DNA synthesis, prolonged exposure inhibited it. Furthermore, on lymphocytes stimulated with two T cell mitogens, the effects of A23187 and PMA depended on the proliferation-inducing mitogens and the responsiveness level induced by them. Therefore, while PMA and short pretreatments with A23187 had no effect on high intensity mitogenic responses, low intensity responses were significantly enhanced. These results demonstrate differential effects of A23187 and PMA on DNA synthesis that should be useful in studies on the mechanisms of activation of lymphocyte proliferation.


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  1. [1]
    M. J. Berridge and R. F. Irvine,Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature312, 315–321 (1984).PubMedGoogle Scholar
  2. [2]
    Y. Nishizuka,Turnover of inositol phospholipids and signal transduction. Science225, 1365–1370 (1984).PubMedGoogle Scholar
  3. [3]
    M. J. Berridge,Inositol trisphosphate and diacylglycerol: two interacting second messengers. Ann. Rev. Biochem.56, 159–193 (1987).PubMedGoogle Scholar
  4. [4]
    R. Y. Tsien, T. Pozzan and T. J. Rink,T-cell mitogens cause early changes in cytoplasmic free Ca 2+ and membrane potential in lymphocytes. Nature295, 68–70 (1982).PubMedGoogle Scholar
  5. [5]
    H. C. Clevers, J. M. T. Versteegen, T. Logtenberg, F. H. J. Gmelig-Meyling and R. E. Ballieux,Synergistic action of A23187 and phorbol ester on human B cell activation. J. Immunol.135, 3827–3830 (1985).PubMedGoogle Scholar
  6. [6]
    N. Kumagai, S. H. Benedict, G. B. Mills and E. W. Gelfand,Requirements for the simultaneous presence of phorbol esters and calcium ionophores in the expression of human T lymphocyte proliferation-related genes. J. Immunol.139, 1393–1399 (1987).PubMedGoogle Scholar
  7. [7]
    A. Procopio, A. Gismondi, R. Paolini, S. Morrone, R. Testi, M. Piccoli, L. Frati, R. B. Herberman and A. Santoni,Proliferative effects of 12-O-tetradecanoylphorbol-13-acetate (TPA) and calcium ionophores on human large granular lymphocytes (LGL). Cell. Immunol.113, 70–81 (1988).PubMedGoogle Scholar
  8. [8]
    T. Tanaka, T. Masuko and Y. Hashimoto,Appearance of a proliferation-associated antigen, gp 125, on rat and human lymphocytes by co-stimulation with phorbol ester and calcium ionophore. J. Biochem.103, 644–649 (1988).PubMedGoogle Scholar
  9. [9]
    A. Truneh, F. Albert, P. Golstein and A. M. Schmitt-Verhulst,Early steps of lymphocyte activation bypassed by synergy between calcium ionophores and phorbol ester. Nature313, 318–320 (1985).PubMedGoogle Scholar
  10. [10]
    J. C. Metcalfe, T. R. Hesketh, G. A. Smith, J. D. H. Morris, A. N. Corps and J. P. Moore,Early response pattern analysis of the mitogenic pathway in lymphocytes and fibroblasts. J. Cell Sci.Suppl. 3, 199–228 (1985).Google Scholar
  11. [11]
    P. S. Rabinovitch, C. H. June, A. Grossmann and J. A. Ledbetter,Heterogeneity among T cells in intracellular free calcium responses after mitogen stimulation with PHA or anti-CD3. Simultaneous use of indo-1 and immunofluorescence with flow cytometry. J. Immunol.137, 952–961 (1986).PubMedGoogle Scholar
  12. [12]
    G. Pawelec, F. Busch, U. Reinhold, A. Rehbein, I. Balko and H. J. Buhring,Division of human helper T cells into two sets on the basis of the induction of anti-tumor cytotoxicity by phorbol ester and calcium ionophore. Eur. J. Immunol.18, 1147–1150 (1988).PubMedGoogle Scholar
  13. [13]
    K. Takada, N. Amino, T. Tetsumoto and K. Miyai,Phorbol esters have a dual action through protein kinase C in regulation of proliferation of FRTL-5 cells. FEBS Lett.234, 13–16 (1988).PubMedGoogle Scholar
  14. [14]
    S. P. Soltoff and L. C. Cantley,Mitogens and ion fluxes. Ann. Rev. Physiol.50, 207–223 (1988).Google Scholar
  15. [15]
    L. J. M. Sabbe, L. De Bode and J. J. Van Rood,Analysis of variability in lymphocyte transformation tests. J. Immunol. Methods57, 21–32 (1983).PubMedGoogle Scholar
  16. [16]
    G. Ross, J. B. Dixon and A. Veevers,Variation in the lymphocyte transformation assay. Slope analysis of cell doseresponse curves. J. Immunol. Methods98, 189–193 (1987).PubMedGoogle Scholar
  17. [17]
    H. M. Hallgren, N. Bergh, K. J. Rodysill and J. J. O'Leary,Lymphocyte proliferative response to PHA and Anti-CD3/Ti monoclonal antibodies, T cell surface marker expression, and serum IL-2 receptor levels as biomarkers of age and health. Mech. Ageing Dev.43, 175–185 (1988).PubMedGoogle Scholar
  18. [18]
    C. G. Pantazis and W. T. Knicker,Assessment of blood leukocyte microbial killing by using a new fluorochrome microassay. J. Reticuloendothel. Soc.26, 155–170 (1979).PubMedGoogle Scholar
  19. [19]
    N. Papadogiannakis and S. A. Johnsen,Mitogenic action of phorbol ester TPA and calcium ionophore A23187 on human cord and maternal/adult peripheral lymphocytes: regulation by prostaglandin E2. Clin. Exp. Immunol.70, 173–181 (1987).PubMedGoogle Scholar
  20. [20]
    J. Cason, S. Chinn, C. C. Ainley, R. A. Wolstencroft and R. P. H. Thompson,Analysis of human lymphocyte transformation responses to graded doses of T cell mitogens by curve fitting. J. Immunol. Methods102, 109–117 (1987).PubMedGoogle Scholar
  21. [21]
    A. J. H. Gearing, M. Wadhwa and A. D. Perris,Interleukin 2 stimulates T cell proliferation using a calcium flux. Immunol. Lett.10, 297–302 (1985).PubMedGoogle Scholar
  22. [22]
    J. C. Allen and J. W. Mills,Two lines of MDCK epithelial cells with different volume and ion responses to calcium ionophore A23187. In Vitro Cell. Develop. Biol.24, 588–592 (1988).Google Scholar

Copyright information

© Birkhäuser Verlag 1992

Authors and Affiliations

  • D. Planelles
    • 1
  • J. Hernández-Godoy
    • 1
  • A. González-Molina
    • 1
  1. 1.Unidad de Inmunología Experimental, Centro de InvestigaciónHospital La FeValenciaSpain

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