Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1986 Dec 1;164(6):1988–2005. doi: 10.1084/jem.164.6.1988

Human T cell activation. III. Rapid induction of a phosphorylated 28 kD/32 kD disulfide-linked early activation antigen (EA 1) by 12-o- tetradecanoyl phorbol-13-acetate, mitogens, and antigens

PMCID: PMC2188483  PMID: 2946796

Abstract

With human T cells activated by 12-o-tetradecanoyl phorbol-13-acetate (TPA) as immunogen, an IgG2a mAb, early activation antigen 1 (EA 1), was generated against a 60-kD protein with disulfide-linked 28-kD and 32-kD subunits. Both subunits were phosphorylated. The antigen, EA 1, was readily detected on approximately 60% of isolated and cryopreserved thymocytes, as determined by indirect immunofluorescence. A low level of EA 1 expression was detectable on 6-7% of blood lymphocytes. TPA- activated T cells expressed EA 1 as early as 30 min after activation. By 1 h, 85-90% of the T cells stained with mAb EA 1. By 3-4 h, the expression of EA 1 was detected in greater than 95% of the T cells. Although the percentages of EA 1+ T cells did not change, the intensity of staining increased slightly. After 18-24 h, both the percentage of EA 1+ cells and the intensity of staining decreased gradually. TPA- induced EA 1 expression was independent of monocytes. EA 1 expression was slightly delayed in T cells that were isolated without the rosette selection and treated with TPA. Nevertheless, greater than 85% of these T cells expressed EA 1 within 1 h, and the maximal number of EA 1+ T cells was also detected at 3-4 h. In T cell populations with 1-2% monocytes, about 50-90% of the PHA- or Con A-activated T cells expressed EA 1 with a slower kinetics. EA 1 expression preceded that of IL-2-R in these activation processes. Similarly, T cells activated by soluble antigens (tetanus toxoid and PPD) and alloantigens in MLR also expressed EA 1 after a longer incubation. Approximately 20% of the T cells stained for EA 1 at day 6. EA 1 expression was not limited to activated T cells. B cells activated by TPA or anti-IgM antibody plus B cell growth factor expressed EA 1. The kinetics of EA 1 expression was markedly slower and the staining was less intense. Repeated attempts to detect EA 1 on resting and TPA-activated monocytes and granulocytes have not been successful. However, the detection of EA 1 in nonlymphoid cell lines would indicate that EA 1 may have a broader cell distribution. EA 1 expression was due to de novo synthesis, as the induction of EA 1 was blocked by cycloheximide and actinomycin D.(ABSTRACT TRUNCATED AT 400 WORDS)

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Bell R. M. Protein kinase C activation by diacylglycerol second messengers. Cell. 1986 Jun 6;45(5):631–632. doi: 10.1016/0092-8674(86)90774-9. [DOI] [PubMed] [Google Scholar]
  2. Burckhardt J. J., Anderson W. H., Kearney J. F., Cooper M. D. Human blood monocytes and platelets share a cell surface component. Blood. 1982 Sep;60(3):767–771. [PubMed] [Google Scholar]
  3. Burns G. F., Triglia T., Werkmeister J. A., Begley C. G., Boyd A. W. TLiSA1, a human T lineage-specific activation antigen involved in the differentiation of cytotoxic T lymphocytes and anomalous killer cells from their precursors. J Exp Med. 1985 May 1;161(5):1063–1078. doi: 10.1084/jem.161.5.1063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chiorazzi N., Fu S. M., Kunkel H. G. Stimulation of human B lymphocytes by antibodies to IgM and IgG: functional evidence for the expression of IgG on B-lymphocyte surface membranes. Clin Immunol Immunopathol. 1980 Mar;15(3):301–313. doi: 10.1016/0090-1229(80)90042-2. [DOI] [PubMed] [Google Scholar]
  5. Cotner T., Williams J. M., Christenson L., Shapiro H. M., Strom T. B., Strominger J. Simultaneous flow cytometric analysis of human T cell activation antigen expression and DNA content. J Exp Med. 1983 Feb 1;157(2):461–472. doi: 10.1084/jem.157.2.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dower S. K., Kronheim S. R., March C. J., Conlon P. J., Hopp T. P., Gillis S., Urdal D. L. Detection and characterization of high affinity plasma membrane receptors for human interleukin 1. J Exp Med. 1985 Aug 1;162(2):501–515. doi: 10.1084/jem.162.2.501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fox D. A., Hussey R. E., Fitzgerald K. A., Acuto O., Poole C., Palley L., Daley J. F., Schlossman S. F., Reinherz E. L. Ta1, a novel 105 KD human T cell activation antigen defined by a monoclonal antibody. J Immunol. 1984 Sep;133(3):1250–1256. [PubMed] [Google Scholar]
  8. Freedman A. S., Boyd A. W., Anderson K. C., Fisher D. C., Schlossman S. F., Nadler L. M. B5, a new B cell-restricted activation antigen. J Immunol. 1985 Apr;134(4):2228–2235. [PubMed] [Google Scholar]
  9. Fu S. M., Hurley J. N., McCune J. M., Kunkel H. G., Good R. A. Pre-B cells and other possible precursor lymphoid cell lines derived from patients with X-linked agammaglobulinemia. J Exp Med. 1980 Dec 1;152(6):1519–1526. doi: 10.1084/jem.152.6.1519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Granelli-Piperno A., Andrus L., Steinman R. M. Lymphokine and nonlymphokine mRNA levels in stimulated human T cells. Kinetics, mitogen requirements, and effects of cyclosporin A. J Exp Med. 1986 Apr 1;163(4):922–937. doi: 10.1084/jem.163.4.922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hara T., Fu S. M., Hansen J. A. Human T cell activation. II. A new activation pathway used by a major T cell population via a disulfide-bonded dimer of a 44 kilodalton polypeptide (9.3 antigen). J Exp Med. 1985 Jun 1;161(6):1513–1524. doi: 10.1084/jem.161.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hara T., Fu S. M. Human T cell activation. I. Monocyte-independent activation and proliferation induced by anti-T3 monoclonal antibodies in the presence of tumor promoter 12-o-tetradecanoyl phorbol-13 acetate. J Exp Med. 1985 Apr 1;161(4):641–656. doi: 10.1084/jem.161.4.641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Haynes B. F., Hemler M. E., Mann D. L., Eisenbarth G. S., Shelhamer J., Mostowski H. S., Thomas C. A., Strominger J. L., Fauci A. S. Characterization of a monoclonal antibody (4F2) that binds to human monocytes and to a subset of activated lymphocytes. J Immunol. 1981 Apr;126(4):1409–1414. [PubMed] [Google Scholar]
  14. Hercend T., Nadler L. M., Pesando J. M., Reinherz E. L., Schlossman S. F., Ritz J. Expression of a 26,000-dalton glycoprotein on activated human T cells. Cell Immunol. 1981 Oct;64(1):192–199. doi: 10.1016/0008-8749(81)90470-6. [DOI] [PubMed] [Google Scholar]
  15. Hercend T., Ritz J., Schlossman S. F., Reinherz E. L. Comparative expression of T9, T10, and Ia antigens on activated human T cell subsets. Hum Immunol. 1981 Nov;3(3):247–259. doi: 10.1016/0198-8859(81)90021-5. [DOI] [PubMed] [Google Scholar]
  16. Hirschhorn R., Levytska V. Alterations in isozymes of adenosine deaminase during stimulation of human peripheral blood lymphocytes. Cell Immunol. 1974 Jun;12(3):387–395. doi: 10.1016/0008-8749(74)90095-1. [DOI] [PubMed] [Google Scholar]
  17. Holter W., Fischer G. F., Majdic O., Stockinger H., Knapp W. T cell stimulation via the erythrocyte receptor. Synergism between monoclonal antibodies and phorbol myristate acetate without changes of free cytoplasmic Ca++ levels. J Exp Med. 1986 Mar 1;163(3):654–664. doi: 10.1084/jem.163.3.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Holter W., Majdic O., Stockinger H., Knapp W. Analysis of T cell activation with a non-mitogenic anti CD3 antibody and the phorbol ester TPA. Clin Exp Immunol. 1985 Dec;62(3):600–606. [PMC free article] [PubMed] [Google Scholar]
  19. Jacobsen D. W., Montejano Y. D., Vitols K. S., Huennekens F. M. Adherence of L1210 murine leukemia cells to sephacryl-aminopropylcobalamin beads treated with transcobalamin-II. Blood. 1980 Jan;55(1):160–163. [PubMed] [Google Scholar]
  20. Jung L. K., Fu S. M. Selective inhibition of growth factor-dependent human B cell proliferation by monoclonal antibody AB1 to an antigen expressed by activated B cells. J Exp Med. 1984 Dec 1;160(6):1919–1924. doi: 10.1084/jem.160.6.1919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jung L. K., Hara T., Fu S. M. Detection and functional studies of p60-65 (Tac antigen) on activated human B cells. J Exp Med. 1984 Nov 1;160(5):1597–1602. doi: 10.1084/jem.160.5.1597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kawamoto S., Hidaka H. 1-(5-Isoquinolinesulfonyl)-2-methylpiperazine (H-7) is a selective inhibitor of protein kinase C in rabbit platelets. Biochem Biophys Res Commun. 1984 Nov 30;125(1):258–264. doi: 10.1016/s0006-291x(84)80362-9. [DOI] [PubMed] [Google Scholar]
  23. Ko H. S., Fu S. M., Winchester R. J., Yu D. T., Kunkel H. G. Ia determinants on stimulated human T lymphocytes. Occurrence on mitogen- and antigen-activated T cells. J Exp Med. 1979 Aug 1;150(2):246–255. doi: 10.1084/jem.150.2.246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Krönke M., Leonard W. J., Depper J. M., Greene W. C. Sequential expression of genes involved in human T lymphocyte growth and differentiation. J Exp Med. 1985 Jun 1;161(6):1593–1598. doi: 10.1084/jem.161.6.1593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lamont J. T., Perrotto J. L., Weiser M. M., Isselbacher K. J. Cell surface galactosyltransferase and lectin agglutination of thymus and spleen lymphocytes. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3726–3730. doi: 10.1073/pnas.71.9.3726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Marth J. D., Peet R., Krebs E. G., Perlmutter R. M. A lymphocyte-specific protein-tyrosine kinase gene is rearranged and overexpressed in the murine T cell lymphoma LSTRA. Cell. 1985 Dec;43(2 Pt 1):393–404. doi: 10.1016/0092-8674(85)90169-2. [DOI] [PubMed] [Google Scholar]
  27. Moretta A., Pantaleo G., Lopez-Botet M., Moretta L. Involvement of T44 molecules in an antigen-independent pathway of T cell activation. Analysis of the correlations to the T cell antigen-receptor complex. J Exp Med. 1985 Sep 1;162(3):823–838. doi: 10.1084/jem.162.3.823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. doi: 10.1038/308693a0. [DOI] [PubMed] [Google Scholar]
  29. Novogrodsky A., Tate S. S., Meister A. gamma-Glutamyl transpeptidase, a lymphoid cell-surface marker: relationship to blastogenesis, differentiation, and neoplasia. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2414–2418. doi: 10.1073/pnas.73.7.2414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Painter R. G., White A. Effect of concanavalin A on expression of cell surface sialyltransferase activity of mouse thymocytes. Proc Natl Acad Sci U S A. 1976 Mar;73(3):837–841. doi: 10.1073/pnas.73.3.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Palacios R. Mechanisms by which accessory cells contribute in growth of resting T lymphocytes initiated by OKT3 antibody. Eur J Immunol. 1985 Jul;15(7):645–651. doi: 10.1002/eji.1830150702. [DOI] [PubMed] [Google Scholar]
  32. Peters P. M., Kamarck M. E., Hemler M. E., Strominger J. L., Ruddle F. H. Genetic and biochemical characterization of human lymphocyte cell surface antigens. The A-1A5 and A-3A4 determinants. J Exp Med. 1984 May 1;159(5):1441–1454. doi: 10.1084/jem.159.5.1441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Posnett D. N., Wang C. Y., Chiorazzi N., Crow M. K., Kunkel H. G. An antigen characteristic of hairy cell leukemia cells is expressed on certain activated B cells. J Immunol. 1984 Sep;133(3):1635–1640. [PubMed] [Google Scholar]
  34. Slovin S. F., Frisman D. M., Tsoukas C. D., Royston I., Baird S. M., Wormsley S. B., Carson D. A., Vaughan J. H. Membrane antigen on Epstein--Barr virus-infected human B cells recognized by a monoclonal antibody. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2649–2653. doi: 10.1073/pnas.79.8.2649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sutherland R., Delia D., Schneider C., Newman R., Kemshead J., Greaves M. Ubiquitous cell-surface glycoprotein on tumor cells is proliferation-associated receptor for transferrin. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4515–4519. doi: 10.1073/pnas.78.7.4515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Takahashi K., Tavassoli M., Jacobsen D. W. Receptor binding and internalization of immobilized transcobalamin II by mouse leukaemia cells. Nature. 1980 Dec 25;288(5792):713–715. doi: 10.1038/288713a0. [DOI] [PubMed] [Google Scholar]
  37. Thorley-Lawson D. A., Schooley R. T., Bhan A. K., Nadler L. M. Epstein-Barr virus superinduces a new human B cell differentiation antigen (B-LAST 1) expressed on transformed lymphoblasts. Cell. 1982 Sep;30(2):415–425. doi: 10.1016/0092-8674(82)90239-2. [DOI] [PubMed] [Google Scholar]
  38. Trowbridge I. S., Omary M. B. Human cell surface glycoprotein related to cell proliferation is the receptor for transferrin. Proc Natl Acad Sci U S A. 1981 May;78(5):3039–3043. doi: 10.1073/pnas.78.5.3039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Udey M. C., Jendrisak M. D., Parker C. W. Properties of a surface antigen expressed on activated human thymus-derived lymphocytes. J Immunol. 1982 Apr;128(4):1870–1875. [PubMed] [Google Scholar]
  40. Udey M. C., Parker C. W. Membrane protein synthesis in mitogen-stimulated human T lymphocytes. J Immunol. 1981 Mar;126(3):1106–1113. [PubMed] [Google Scholar]
  41. Wahl L. M., Katona I. M., Wilder R. L., Winter C. C., Haraoui B., Scher I., Wahl S. M. Isolation of human mononuclear cell subsets by counterflow centrifugal elutriation (CCE). I. Characterization of B-lymphocyte-, T-lymphocyte-, and monocyte-enriched fractions by flow cytometric analysis. Cell Immunol. 1984 May;85(2):373–383. doi: 10.1016/0008-8749(84)90251-x. [DOI] [PubMed] [Google Scholar]
  42. Waldmann T. A. The structure, function, and expression of interleukin-2 receptors on normal and malignant lymphocytes. Science. 1986 May 9;232(4751):727–732. doi: 10.1126/science.3008337. [DOI] [PubMed] [Google Scholar]
  43. Weiss A., Imboden J., Hardy K., Manger B., Terhorst C., Stobo J. The role of the T3/antigen receptor complex in T-cell activation. Annu Rev Immunol. 1986;4:593–619. doi: 10.1146/annurev.iy.04.040186.003113. [DOI] [PubMed] [Google Scholar]
  44. Wright S. D., Silverstein S. C. Tumor-promoting phorbol esters stimulate C3b and C3b' receptor-mediated phagocytosis in cultured human monocytes. J Exp Med. 1982 Oct 1;156(4):1149–1164. doi: 10.1084/jem.156.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Yokochi T., Holly R. D., Clark E. A. B lymphoblast antigen (BB-1) expressed on Epstein-Barr virus-activated B cell blasts, B lymphoblastoid cell lines, and Burkitt's lymphomas. J Immunol. 1982 Feb;128(2):823–827. [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES