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. 1987 Mar;84(5):1384–1388. doi: 10.1073/pnas.84.5.1384

Crosslinking of surface antigens causes mobilization of intracellular ionized calcium in T lymphocytes.

J A Ledbetter, C H June, L S Grosmaire, P S Rabinovitch
PMCID: PMC304434  PMID: 3103134

Abstract

Antibodies binding to a large subset of T-cell differentiation antigens, including CD2, CD4, CD5, CD6, CD7, CD8, Tp44, and CDw18, cause an increase in the cytoplasmic calcium concentration [( Ca2+]i) after the antigens are crosslinked on the cell surface. Similar crosslinking-induced signals were seen for a subset of mouse thymocyte differentiation antigens. The various antigens on human T cells differed in the extent of crosslinking required for generating the calcium signal, as evidenced by comparisons with monoclonal versus polyclonal second-step antibody. The [Ca2+]i increase that occurs after crosslinking represents mobilization of cytoplasmic calcium since the initial component of the signal is resistant to depletion of extracellular calcium by chelation with EGTA. The [Ca2+]i increase is completely inhibited by pretreatment of cells with pertussis toxin, indicating that a substrate for pertussis toxin regulates the signal transduction. Crosslinking of antigens other than the CD3/T-cell receptor complex did not result in T-cell proliferation. Crosslinking of CD2 and Tp44, but not other antigens, resulted in expression of functional interleukin 2 receptors. Comparisons of three different anti-CD3 antibodies showed that a second calcium signal was generated by crosslinking, even when the anti-CD3 antibodies were used at optimal concentrations.

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Selected References

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  1. Abood M. E., Hurley J. B., Pappone M. C., Bourne H. R., Stryer L. Functional homology between signal-coupling proteins. Cholera toxin inactivates the GTPase activity of transducin. J Biol Chem. 1982 Sep 25;257(18):10540–10543. [PubMed] [Google Scholar]
  2. Berridge M. J., Irvine R. F. Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature. 1984 Nov 22;312(5992):315–321. doi: 10.1038/312315a0. [DOI] [PubMed] [Google Scholar]
  3. Bokoch G. M., Gilman A. G. Inhibition of receptor-mediated release of arachidonic acid by pertussis toxin. Cell. 1984 Dec;39(2 Pt 1):301–308. doi: 10.1016/0092-8674(84)90008-4. [DOI] [PubMed] [Google Scholar]
  4. Bokoch G. M., Katada T., Northup J. K., Hewlett E. L., Gilman A. G. Identification of the predominant substrate for ADP-ribosylation by islet activating protein. J Biol Chem. 1983 Feb 25;258(4):2072–2075. [PubMed] [Google Scholar]
  5. Braun J., Unanue E. R. The lymphocyte cytoskeleton and its control of surface receptor functions. Semin Hematol. 1983 Oct;20(4):322–333. [PubMed] [Google Scholar]
  6. Brottier P., Boumsell L., Gelin C., Bernard A. T cell activation via CD2 [T, gp50] molecules: accessory cells are required to trigger T cell activation via CD2-D66 plus CD2-9.6/T11(1) epitopes. J Immunol. 1985 Sep;135(3):1624–1631. [PubMed] [Google Scholar]
  7. Burn P., Rotman A., Meyer R. K., Burger M. M. Diacylglycerol in large alpha-actinin/actin complexes and in the cytoskeleton of activated platelets. Nature. 1985 Apr 4;314(6010):469–472. doi: 10.1038/314469a0. [DOI] [PubMed] [Google Scholar]
  8. Dialynas D. P., Quan Z. S., Wall K. A., Pierres A., Quintáns J., Loken M. R., Pierres M., Fitch F. W. Characterization of the murine T cell surface molecule, designated L3T4, identified by monoclonal antibody GK1.5: similarity of L3T4 to the human Leu-3/T4 molecule. J Immunol. 1983 Nov;131(5):2445–2451. [PubMed] [Google Scholar]
  9. Exton J. H. Role of calcium and phosphoinositides in the actions of certain hormones and neurotransmitters. J Clin Invest. 1985 Jun;75(6):1753–1757. doi: 10.1172/JCI111886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Florio V. A., Sternweis P. C. Reconstitution of resolved muscarinic cholinergic receptors with purified GTP-binding proteins. J Biol Chem. 1985 Mar 25;260(6):3477–3483. [PubMed] [Google Scholar]
  11. Gill D. L., Ueda T., Chueh S. H., Noel M. W. Ca2+ release from endoplasmic reticulum is mediated by a guanine nucleotide regulatory mechanism. Nature. 1986 Apr 3;320(6061):461–464. doi: 10.1038/320461a0. [DOI] [PubMed] [Google Scholar]
  12. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  13. 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]
  14. Hoessli D., Rungger-Brändle E., Jockusch B. M., Gabbiani G. Lymphocyte alpha-actinin. Relationship to cell membrane and co-capping with surface receptors. J Cell Biol. 1980 Feb;84(2):305–314. doi: 10.1083/jcb.84.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Imboden J. B., Shoback D. M., Pattison G., Stobo J. D. Cholera toxin inhibits the T-cell antigen receptor-mediated increases in inositol trisphosphate and cytoplasmic free calcium. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5673–5677. doi: 10.1073/pnas.83.15.5673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Imboden J. B., Stobo J. D. Transmembrane signalling by the T cell antigen receptor. Perturbation of the T3-antigen receptor complex generates inositol phosphates and releases calcium ions from intracellular stores. J Exp Med. 1985 Mar 1;161(3):446–456. doi: 10.1084/jem.161.3.446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Imboden J. B., Weiss A., Stobo J. D. The antigen receptor on a human T cell line initiates activation by increasing cytoplasmic free calcium. J Immunol. 1985 Feb;134(2):663–665. [PubMed] [Google Scholar]
  18. Julius M. H., Simpson E., Herzenberg L. A. A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur J Immunol. 1973 Oct;3(10):645–649. doi: 10.1002/eji.1830031011. [DOI] [PubMed] [Google Scholar]
  19. June C. H., Ledbetter J. A., Rabinovitch P. S., Martin P. J., Beatty P. G., Hansen J. A. Distinct patterns of transmembrane calcium flux and intracellular calcium mobilization after differentiation antigen cluster 2 (E rosette receptor) or 3 (T3) stimulation of human lymphocytes. J Clin Invest. 1986 Apr;77(4):1224–1232. doi: 10.1172/JCI112425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Katada T., Ui M. Direct modification of the membrane adenylate cyclase system by islet-activating protein due to ADP-ribosylation of a membrane protein. Proc Natl Acad Sci U S A. 1982 May;79(10):3129–3133. doi: 10.1073/pnas.79.10.3129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kroczek R. A., Gunter K. C., Seligmann B., Shevach E. M. Induction of T cell activation by monoclonal anti-Thy-1 antibodies. J Immunol. 1986 Jun 15;136(12):4379–4384. [PubMed] [Google Scholar]
  22. Lassing I., Lindberg U. Specific interaction between phosphatidylinositol 4,5-bisphosphate and profilactin. Nature. 1985 Apr 4;314(6010):472–474. doi: 10.1038/314472a0. [DOI] [PubMed] [Google Scholar]
  23. Laub F., Kaplan M., Gitler C. Actin polymerization accompanies Thy-1-capping on mouse thymocytes. FEBS Lett. 1981 Feb 9;124(1):35–38. doi: 10.1016/0014-5793(81)80048-8. [DOI] [PubMed] [Google Scholar]
  24. Ledbetter J. A., June C. H., Martin P. J., Spooner C. E., Hansen J. A., Meier K. E. Valency of CD3 binding and internalization of the CD3 cell-surface complex control T cell responses to second signals: distinction between effects on protein kinase C, cytoplasmic free calcium, and proliferation. J Immunol. 1986 Jun 1;136(11):3945–3952. [PubMed] [Google Scholar]
  25. MacDonald H. R., Bron C., Rousseaux M., Horvath C., Cerottini J. C. Production and characterization of monoclonal anti-Thy-1 antibodies that stimulate lymphokine production by cytolytic T cell clones. Eur J Immunol. 1985 May;15(5):495–501. doi: 10.1002/eji.1830150514. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Meuer S. C., Hussey R. E., Fabbi M., Fox D., Acuto O., Fitzgerald K. A., Hodgdon J. C., Protentis J. P., Schlossman S. F., Reinherz E. L. An alternative pathway of T-cell activation: a functional role for the 50 kd T11 sheep erythrocyte receptor protein. Cell. 1984 Apr;36(4):897–906. doi: 10.1016/0092-8674(84)90039-4. [DOI] [PubMed] [Google Scholar]
  28. O'Flynn K., Zanders E. D., Lamb J. R., Beverley P. C., Wallace D. L., Tatham P. E., Tax W. J., Linch D. C. Investigation of early T cell activation: analysis of the effect of specific antigen, interleukin 2 and monoclonal antibodies on intracellular free calcium concentration. Eur J Immunol. 1985 Jan;15(1):7–11. doi: 10.1002/eji.1830150103. [DOI] [PubMed] [Google Scholar]
  29. Oettgen H. C., Terhorst C., Cantley L. C., Rosoff P. M. Stimulation of the T3-T cell receptor complex induces a membrane-potential-sensitive calcium influx. Cell. 1985 Mar;40(3):583–590. doi: 10.1016/0092-8674(85)90206-5. [DOI] [PubMed] [Google Scholar]
  30. Okajima F., Ui M. ADP-ribosylation of the specific membrane protein by islet-activating protein, pertussis toxin, associated with inhibition of a chemotactic peptide-induced arachidonate release in neutrophils. A possible role of the toxin substrate in Ca2+-mobilizing biosignaling. J Biol Chem. 1984 Nov 25;259(22):13863–13871. [PubMed] [Google Scholar]
  31. Putney J. W., Jr A model for receptor-regulated calcium entry. Cell Calcium. 1986 Feb;7(1):1–12. doi: 10.1016/0143-4160(86)90026-6. [DOI] [PubMed] [Google Scholar]
  32. Rabinovitch P. S., June C. H., Grossmann A., Ledbetter J. A. 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. 1986 Aug 1;137(3):952–961. [PubMed] [Google Scholar]
  33. Smith C. D., Cox C. C., Snyderman R. Receptor-coupled activation of phosphoinositide-specific phospholipase C by an N protein. Science. 1986 Apr 4;232(4746):97–100. doi: 10.1126/science.3006254. [DOI] [PubMed] [Google Scholar]
  34. Tse A. G., Barclay A. N., Watts A., Williams A. F. A glycophospholipid tail at the carboxyl terminus of the Thy-1 glycoprotein of neurons and thymocytes. Science. 1985 Nov 29;230(4729):1003–1008. doi: 10.1126/science.2865810. [DOI] [PubMed] [Google Scholar]
  35. Ui M., Okajima F., Itoh H. ADP-ribosylation of the inhibitory guanine nucleotide regulatory protein (Ni) as a possible mechanism underlying development of beta-adrenergic responses during primary culture of rat hepatocytes. Adv Cyclic Nucleotide Protein Phosphorylation Res. 1985;19:195–205. [PubMed] [Google Scholar]
  36. Van Dop C., Yamanaka G., Steinberg F., Sekura R. D., Manclark C. R., Stryer L., Bourne H. R. ADP-ribosylation of transducin by pertussis toxin blocks the light-stimulated hydrolysis of GTP and cGMP in retinal photoreceptors. J Biol Chem. 1984 Jan 10;259(1):23–26. [PubMed] [Google Scholar]
  37. Weiss A., Imboden J., Shoback D., Stobo J. Role of T3 surface molecules in human T-cell activation: T3-dependent activation results in an increase in cytoplasmic free calcium. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4169–4173. doi: 10.1073/pnas.81.13.4169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weiss M. J., Daley J. F., Hodgdon J. C., Reinherz E. L. Calcium dependency of antigen-specific (T3-Ti) and alternative (T11) pathways of human T-cell activation. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6836–6840. doi: 10.1073/pnas.81.21.6836. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Yelton D. E., Desaymard C., Scharff M. D. Use of monoclonal anti-mouse immunoglobulin to detect mouse antibodies. Hybridoma. 1981;1(1):5–11. doi: 10.1089/hyb.1.1981.1.5. [DOI] [PubMed] [Google Scholar]

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