Phorbol Myristate Acetate-Induced Changes in Protein Kinase C Isozymes (α, β, γ and ζ) in Human T Cell Subsets

  • Sudhir Gupta
  • William Harris
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 365)


Protein kinase C (PKC), a serine/threonine kinase, plays an important role in downstream signaling pathway of T cell activation (1,2). Activation of PKC produces varied, and at times disparate, cell specific responses. In T cells, direct activation of PKC results in an induction of interleukin-2 receptor (IL-2R) expression and IL-2 production, initiation of DNA synthesis, and regulation of cytotoxic function (3–11). Although PKC was originally thought to be a single molecule, recent molecular and biochemical studies have revealed that PKC exists as a family of different isozymes that differ in their structure, dependence on Ca++, tissue distribution and substrate specificity (12–15). They have been broadly categorized into three major subgroups; Group 1 including classical Ca++-dependent PKCs (α, β and γ); Group 2 comprised of Ca++-independent PKCs (δ, ϵ, η, θ); and Group 3, consisting of atypical PKC (ζ, λ). PKCζ isozyme is considered to be resistant to the effect of phorbol esters (12). Ways et al (16), using a variety of non-lymphoid cell lines, have failed to demonstrate translocation of PKCζ following stimulation with phorbol ester. To the best of our knowledge there is no study of PKCζ in normal human peripheral blood T cells. Further there is a controversy regarding the role of PKCβ isozyme T cell functions (17–19) and regarding the presence of PKCγ in T cells (22–26). In this study we examined the translocation of Ca++-dependent (PKCα, β, γ) and atypical (PKCζ) PKC isozymes in human peripheral blood T cells. This study demonstrates that PKC isozymes are differentially translocated among CD4+ and CD8+ T cells and PKCγ isozyme is present in T cells and translocated following PMA activation.


Phorbol Myristate Acetate Phorbol Ester Phorbol Myristate Acetate Normal Human Peripheral Blood Cell Specific Response 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    A. Weiss, J.B. Imboden, K. Hardy, B. Manger, C. Terhorst, and J.D. Stobo. The role of T3/antigen receptor complex in T cell activation. Ann. Rev. Immunol. 4:593 (1986).CrossRefGoogle Scholar
  2. 2.
    S. Gupta, M. Shimizu, K. Ohira, and B. Vayuvegula. T cell activation via the T cell receptor: A comparison between WT31 (defining α/β TcR)-induced and anti-.CD3-induced activation of human T lymphocytes. Cell. Immunol. 132: 26 (1991).PubMedCrossRefGoogle Scholar
  3. 3.
    M. Isokov and A. Altman. Human T cell activation by tumor promoters: role of protein kinase C. J. Immunol. 138: 3100 (1987).Google Scholar
  4. 4.
    A.M. Mastro and M.C. Smith. Calcium-dependent activation of lymphocytes by ionophore, A23187, and phorbol esters tumor promotor. J. Cell Physiol. 116: 51 (1993).CrossRefGoogle Scholar
  5. 5.
    A.F. Truneh, F. Albeit, P. Goldstein, and A-M. Schmitt-Verhulst. Early steps of lymphocyte activation by-passed by synergy between calcium ionophore and phorbol ester. Nature (Lond) 313: 318 (1985).CrossRefGoogle Scholar
  6. 6.
    N. Berry, K. Ase, U. Kikkawa, A. Kishimoto and Y. Nishizuka. Human T cell activation by phorbol esters and diacylglycerol. J. Immunol. 143: 1417 (1989).Google Scholar
  7. 7.
    J.M. Depper, W.J. Leonard, M. Kronke, P.D. Noguchi, R.E. Cunningham, T.A. Waldmann, and W.C. Greene. Regulation of Interleukin 2 receptor expression: effects of phorbol diester, phospholipase C and reexposure to lectin or antigen. J. Immunol. 133: 3054 (1984).PubMedGoogle Scholar
  8. 8.
    T. Hirano, K. Fujimoto, T. Teranishi, N. Nishino, K. Onoue, S. Maeda and K. Shimada. Phorbol ester increases the level of interleukin 2 mRNA in mitogen-stimulated human lymphocytes. J. Immunol. 132: 2165 (1984).PubMedGoogle Scholar
  9. 9.
    H. Schrezenmeir, R. Kurrle, and B. Fleischer. Stimulus-dependent triggering or inhibition of cytotoxicity in human cytotoxic T-lymphocytes by activators of protein kinase C. Immunology 59: 359 (1986).Google Scholar
  10. 10.
    J.A. Ledbetter, L.E. Gentry, C.H. June, P.S. Rabinovitch and A.F. Purchio. Stimulation of T cells through the CD3/T-cell receptor complex: role of cytoplasmic calcium, protein kinase C translocation and phosphorylation of pp60c-src in the activation pathway. Mol. Cell. Biol. 7: 650 (1987).PubMedGoogle Scholar
  11. 11.
    J. Abb, G.J. Bayliss, and F. Deinhardt. Lymphocyte activation by the tumor promoting agent 12-O-tetradecanoyl-phorbol-13-acetate (TPA). J. Immunol. 122: 1639 (1979).PubMedGoogle Scholar
  12. 12.
    Y. Nishizuka. The molecular heterogenity of protein kinase C and its implication for cellular regulation. Science 334: 661 (1988).Google Scholar
  13. 13.
    P.M. Blumberg. Complexities of the protein kinase C pathway. Mol. Carcinogenesis 4: 339 (1991).CrossRefGoogle Scholar
  14. 14.
    P.J. Parker, G. Kour, R. Marais, F. Mitchell, C. Pears, D. Schaap, S. Stabel and C. Webester. Protein kinase C-a family affair. Mol. Cell. Endocr. 65: 1 (1989).CrossRefGoogle Scholar
  15. 15.
    Y. Nishizuka. Intracellular signalling by hydrolysis of phospholipids and activation of protein kinase C. Science 258: 607 (1992).PubMedCrossRefGoogle Scholar
  16. 16.
    D.K. Ways, P.P. Cook, C. Webster, and P.J. Parker. Effect of phorbol esters on protein kinase C-ζ. J. Biol. Chem. 267: 4799 (1992).PubMedGoogle Scholar
  17. 17.
    G.A. Koretzky, M. Wahi, M.E. Newton and A. Weiss. Heterogeneity of protein kinase C isozyme gene expression in human T cell lines. Protein kinase C-β is not required for several T cell functions. J. Immunol. 143: 1692 (1989).PubMedGoogle Scholar
  18. 18.
    D. Kelleher and A. Long. Development and characterization of the protein kinase Cβ-isozyme-deficient T-cell line. FEBS Lett. 3:310(1992).CrossRefGoogle Scholar
  19. 19.
    S. Aggarwal, S. Lee, A. Mathur, S. Gollapudi and S. Gupta. 12-deoxyphorbol-13-O-phenylacetate 20 acetate [an agonist of protein kinase Cβ1 (PKCα1)] induces DNA synthesis, interleukin-2 (IL-2) production, IL-2 receptor α-chain (CD25) and β-chain (CD122) expression, and translocation of PKCβ isozyme in human peripheral blood lymphocytes: Evidence for a role of PKCβ1 in human T cell activation. J. Clin. Immunol 14: (1994) (in press).Google Scholar
  20. 20.
    M.S. Shearman, N. Berry, T. Oda, K. Ase, U. Kikkawa and Y. Nishizuka. Isolation of protein kinase C subspecies from a preparation of human T lymphocytes. FEBS Lett. 234:387(1988).PubMedCrossRefGoogle Scholar
  21. 21.
    S. Lucas, R. Marais, J.D. Graves, D. Alexander, P. Parker and D.A. Cantrell Heterogeneity of protein kinase C expression and regulation in T lymphocytes. FEBS Lett. 260: 53 (1990).PubMedCrossRefGoogle Scholar
  22. 22.
    A. Kvanta, M. Jondal and B.B. Freedholm. Translocation of the alpha-isoform and beta isoforms of protein kinase C following activation of human T lymphocytes. FEBS Lett. 283: 321 (1991).PubMedCrossRefGoogle Scholar
  23. 23.
    A. Altman, M.I. Mally and N. Isakov. Phorbol ester synergizes with Ca2+ ionophore in activation of protein kinase C (PKC) alpha and beta isozymes in human T cells and in induction of related cellular functions. Immunology 76: 465 (1992).PubMedGoogle Scholar
  24. 24.
    W.G. Harris and S. Gupta. Anti-CD3-mediated activation of PKC isoforms in human T cell subsets. J. Allergy Immunol. 91: 214 (1993).Google Scholar
  25. 25.
    W. Harris, S. Aggarwal, S. Gollapudi and S. Gupta. Anti-CD3 antibody-induced changes in PKC isozymes (α, β, γ,δ, ϵ and ζ) in human T cell subsets. (Submitted).Google Scholar
  26. 26.
    A. Tsutsumi, M. Kubo, H. Fujii, J. Freire-Moar, C.W. Turck and J.T. Ransom. Regulation of protein kinase C isoform proteins in phorbol ester-stimulated Jurkat T lymphoma cells. J. Immunol. 150: 1746 (1993).PubMedGoogle Scholar
  27. 27.
    M. Szamel, F. Bartels, and K. Resch. Cyclosporin A inhibits T cell receptor-induced interleukin-2 synthesis in human T lymphocytes selectively preventing a transmembrane signal transduction pathway leading to sustained activation of a protein kinase C isozyme, protein kinase C-beta. Eur. J. Immunol. 23: 3072 (1993).PubMedCrossRefGoogle Scholar
  28. 28.
    J. Terajima, A. Tsutsumi, J. Freire-Moar, H.M. Cherwinski and J.T. Ramson. Evidence of clonal heterogeneity of the expression of six protein kinase C isoforms in murine B and T lymphocytes. Cell. Immunol. 142: 197 (1992).PubMedCrossRefGoogle Scholar
  29. 29.
    T. Ono, T. Fuji, K. Ogata, U. Kikkawa, K. Igarashi and Y. Nishizuka. The structure, expression, and properties of additional members of the protein kinase C family. J. Biol. Chem. 263: 6927 (1988).PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1994

Authors and Affiliations

  • Sudhir Gupta
    • 1
  • William Harris
    • 1
  1. 1.Division of Basic and Clinical ImmunologyUniversity of CaliforniaIrvineUSA

Personalised recommendations