Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1995 Dec 1;182(6):1997–2006. doi: 10.1084/jem.182.6.1997

Physical dissociation of the TCR-CD3 complex accompanies receptor ligation

PMCID: PMC2192252  PMID: 7500045

Abstract

Recent studies indicate that there may be functional uncoupling of the TCR-CD3 complex and suggest that the TCR-CD3 complex is composed of two parallel signal-transducing units, one made of gamma delta epsilon chains and the other of zeta chains. To elucidate the molecular mechanisms that may explain the functional uncoupling of TCR and CD3, we have analyzed their expression by using flow cytometry as well as immunochemical means both before and after stimulation with anti-TCR- beta, anti-CD3 epsilon, anti-CD2, staphylococcal enterotoxin B, and ionomycin. We present evidence that TCR physically dissociates from CD3 after stimulation of the TCR-CD3 complex. Stimulation with anti-CD3 resulted in down-modulation of TCR within 45 min whereas CD3 epsilon was still expressed on the cell surface as detected by flow cytometry. However, the cell surface expression of TCR and CD3 was not affected when cells were stimulated with anti-TCR-beta under the same conditions. In the case of anti-CD3 treatment of T cells, the TCR down- modulation appeared to be due to the internalization of TCR, as determined by immunoelectron microscopy. Immunochemical analysis of cells after stimulation with either anti-TCR or anti-CD3 mAbs revealed that the overall protein levels of TCR and CD3 were similar. More interestingly, the dissociation of the TCR-CD3 complex was observed with both treatments and occurred in a manner that the TCR and the associated TCR-zeta chain dissociated as a unit from CD3. These results provide the first report of physical dissociation of TCR and CD3 after stimulation through the TCR-CD3 complex. The results also suggest that the signal transduction pathway triggered by TCR may differ from that induced by CD3.

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. Aoe T., Okamoto Y., Saito T. Activated macrophages induce structural abnormalities of the T cell receptor-CD3 complex. J Exp Med. 1995 May 1;181(5):1881–1886. doi: 10.1084/jem.181.5.1881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Becker M. L., Near R., Mudgett-Hunter M., Margolies M. N., Kubo R. T., Kaye J., Hedrick S. M. Expression of a hybrid immunoglobulin-T cell receptor protein in transgenic mice. Cell. 1989 Sep 8;58(5):911–921. doi: 10.1016/0092-8674(89)90943-4. [DOI] [PubMed] [Google Scholar]
  3. Blumberg R. S., Ley S., Sancho J., Lonberg N., Lacy E., McDermott F., Schad V., Greenstein J. L., Terhorst C. Structure of the T-cell antigen receptor: evidence for two CD3 epsilon subunits in the T-cell receptor-CD3 complex. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7220–7224. doi: 10.1073/pnas.87.18.7220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Born W., Miles C., White J., O'Brien R., Freed J. H., Marrack P., Kappler J., Kubo R. T. Peptide sequences of T-cell receptor delta and gamma chains are identical to predicted X and gamma proteins. Nature. 1987 Dec 10;330(6148):572–574. doi: 10.1038/330572a0. [DOI] [PubMed] [Google Scholar]
  5. Finkel T. H., Cambier J. C., Kubo R. T., Born W. K., Marrack P., Kappler J. W. The thymus has two functionally distinct populations of immature alpha beta + T cells: one population is deleted by ligation of alpha beta TCR. Cell. 1989 Sep 22;58(6):1047–1054. doi: 10.1016/0092-8674(89)90503-5. [DOI] [PubMed] [Google Scholar]
  6. Finkel T. H., Marrack P., Kappler J. W., Kubo R. T., Cambier J. C. Alpha beta T cell receptor and CD3 transduce different signals in immature T cells. Implications for selection and tolerance. J Immunol. 1989 May 1;142(9):3006–3012. [PubMed] [Google Scholar]
  7. Haughn L., Gratton S., Caron L., Sékaly R. P., Veillette A., Julius M. Association of tyrosine kinase p56lck with CD4 inhibits the induction of growth through the alpha beta T-cell receptor. Nature. 1992 Jul 23;358(6384):328–331. doi: 10.1038/358328a0. [DOI] [PubMed] [Google Scholar]
  8. Havran W. L., Poenie M., Kimura J., Tsien R., Weiss A., Allison J. P. Expression and function of the CD3-antigen receptor on murine CD4+8+ thymocytes. Nature. 1987 Nov 12;330(6144):170–173. doi: 10.1038/330170a0. [DOI] [PubMed] [Google Scholar]
  9. Jin Y. J., Clayton L. K., Howard F. D., Koyasu S., Sieh M., Steinbrich R., Tarr G. E., Reinherz E. L. Molecular cloning of the CD3 eta subunit identifies a CD3 zeta-related product in thymus-derived cells. Proc Natl Acad Sci U S A. 1990 May;87(9):3319–3323. doi: 10.1073/pnas.87.9.3319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. June C. H., Fletcher M. C., Ledbetter J. A., Samelson L. E. Increases in tyrosine phosphorylation are detectable before phospholipase C activation after T cell receptor stimulation. J Immunol. 1990 Mar 1;144(5):1591–1599. [PubMed] [Google Scholar]
  11. Kearse K. P., Roberts J. L., Munitz T. I., Wiest D. L., Nakayama T., Singer A. Developmental regulation of alpha beta T cell antigen receptor expression results from differential stability of nascent TCR alpha proteins within the endoplasmic reticulum of immature and mature T cells. EMBO J. 1994 Oct 3;13(19):4504–4514. doi: 10.1002/j.1460-2075.1994.tb06772.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kisielow P., Blüthmann H., Staerz U. D., Steinmetz M., von Boehmer H. Tolerance in T-cell-receptor transgenic mice involves deletion of nonmature CD4+8+ thymocytes. Nature. 1988 Jun 23;333(6175):742–746. doi: 10.1038/333742a0. [DOI] [PubMed] [Google Scholar]
  13. Koning F., Maloy W. L., Coligan J. E. The implications of subunit interactions for the structure of the T cell receptor-CD3 complex. Eur J Immunol. 1990 Feb;20(2):299–305. doi: 10.1002/eji.1830200211. [DOI] [PubMed] [Google Scholar]
  14. Kubo R. T., Born W., Kappler J. W., Marrack P., Pigeon M. Characterization of a monoclonal antibody which detects all murine alpha beta T cell receptors. J Immunol. 1989 Apr 15;142(8):2736–2742. [PubMed] [Google Scholar]
  15. Küster H., Thompson H., Kinet J. P. Characterization and expression of the gene for the human Fc receptor gamma subunit. Definition of a new gene family. J Biol Chem. 1990 Apr 15;265(11):6448–6452. [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Letourneur F., Klausner R. D. Activation of T cells by a tyrosine kinase activation domain in the cytoplasmic tail of CD3 epsilon. Science. 1992 Jan 3;255(5040):79–82. doi: 10.1126/science.1532456. [DOI] [PubMed] [Google Scholar]
  18. Levelt C. N., Mombaerts P., Iglesias A., Tonegawa S., Eichmann K. Restoration of early thymocyte differentiation in T-cell receptor beta-chain-deficient mutant mice by transmembrane signaling through CD3 epsilon. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11401–11405. doi: 10.1073/pnas.90.23.11401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Madrenas J., Wange R. L., Wang J. L., Isakov N., Samelson L. E., Germain R. N. Zeta phosphorylation without ZAP-70 activation induced by TCR antagonists or partial agonists. Science. 1995 Jan 27;267(5197):515–518. doi: 10.1126/science.7824949. [DOI] [PubMed] [Google Scholar]
  20. Meuer S. C., Fitzgerald K. A., Hussey R. E., Hodgdon J. C., Schlossman S. F., Reinherz E. L. Clonotypic structures involved in antigen-specific human T cell function. Relationship to the T3 molecular complex. J Exp Med. 1983 Feb 1;157(2):705–719. doi: 10.1084/jem.157.2.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Minami Y., Weissman A. M., Samelson L. E., Klausner R. D. Building a multichain receptor: synthesis, degradation, and assembly of the T-cell antigen receptor. Proc Natl Acad Sci U S A. 1987 May;84(9):2688–2692. doi: 10.1073/pnas.84.9.2688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mizoguchi H., O'Shea J. J., Longo D. L., Loeffler C. M., McVicar D. W., Ochoa A. C. Alterations in signal transduction molecules in T lymphocytes from tumor-bearing mice. Science. 1992 Dec 11;258(5089):1795–1798. doi: 10.1126/science.1465616. [DOI] [PubMed] [Google Scholar]
  23. Mombaerts P., Clarke A. R., Rudnicki M. A., Iacomini J., Itohara S., Lafaille J. J., Wang L., Ichikawa Y., Jaenisch R., Hooper M. L. Mutations in T-cell antigen receptor genes alpha and beta block thymocyte development at different stages. Nature. 1992 Nov 19;360(6401):225–231. doi: 10.1038/360225a0. [DOI] [PubMed] [Google Scholar]
  24. Mombaerts P., Iacomini J., Johnson R. S., Herrup K., Tonegawa S., Papaioannou V. E. RAG-1-deficient mice have no mature B and T lymphocytes. Cell. 1992 Mar 6;68(5):869–877. doi: 10.1016/0092-8674(92)90030-g. [DOI] [PubMed] [Google Scholar]
  25. Nakayama T., Kubo R. T., Kishimoto H., Asano Y., Tada T. Biochemical identification of I-J as a novel dimeric surface molecule on mouse helper and suppressor T cell clones. Int Immunol. 1989;1(1):50–58. doi: 10.1093/intimm/1.1.50. [DOI] [PubMed] [Google Scholar]
  26. Newell M. K., Haughn L. J., Maroun C. R., Julius M. H. Death of mature T cells by separate ligation of CD4 and the T-cell receptor for antigen. Nature. 1990 Sep 20;347(6290):286–289. doi: 10.1038/347286a0. [DOI] [PubMed] [Google Scholar]
  27. Rozdzial M. M., Kubo R. T., Turner S. L., Finkel T. H. Developmental regulation of the TCR zeta-chain. Differential expression and tyrosine phosphorylation of the TCR zeta-chain in resting immature and mature T lymphocytes. J Immunol. 1994 Aug 15;153(4):1563–1580. [PubMed] [Google Scholar]
  28. Saito T., Weiss A., Miller J., Norcross M. A., Germain R. N. Specific antigen-Ia activation of transfected human T cells expressing murine Ti alpha beta-human T3 receptor complexes. Nature. 1987 Jan 8;325(7000):125–130. doi: 10.1038/325125a0. [DOI] [PubMed] [Google Scholar]
  29. Samelson L. E., Harford J. B., Klausner R. D. Identification of the components of the murine T cell antigen receptor complex. Cell. 1985 Nov;43(1):223–231. doi: 10.1016/0092-8674(85)90027-3. [DOI] [PubMed] [Google Scholar]
  30. Sano K., Fujisawa I., Abe R., Asano Y., Tada T. MHC-restricted minimal regulatory circuit initiated by a class II-autoreactive T cell clone. J Exp Med. 1987 May 1;165(5):1284–1295. doi: 10.1084/jem.165.5.1284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shinkai Y., Alt F. W. CD3 epsilon-mediated signals rescue the development of CD4+CD8+ thymocytes in RAG-2-/- mice in the absence of TCR beta chain expression. Int Immunol. 1994 Jul;6(7):995–1001. doi: 10.1093/intimm/6.7.995. [DOI] [PubMed] [Google Scholar]
  32. Shinkai Y., Koyasu S., Nakayama K., Murphy K. M., Loh D. Y., Reinherz E. L., Alt F. W. Restoration of T cell development in RAG-2-deficient mice by functional TCR transgenes. Science. 1993 Feb 5;259(5096):822–825. doi: 10.1126/science.8430336. [DOI] [PubMed] [Google Scholar]
  33. Shinkai Y., Rathbun G., Lam K. P., Oltz E. M., Stewart V., Mendelsohn M., Charron J., Datta M., Young F., Stall A. M. RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangement. Cell. 1992 Mar 6;68(5):855–867. doi: 10.1016/0092-8674(92)90029-c. [DOI] [PubMed] [Google Scholar]
  34. Sloan-Lancaster J., Shaw A. S., Rothbard J. B., Allen P. M. Partial T cell signaling: altered phospho-zeta and lack of zap70 recruitment in APL-induced T cell anergy. Cell. 1994 Dec 2;79(5):913–922. doi: 10.1016/0092-8674(94)90080-9. [DOI] [PubMed] [Google Scholar]
  35. Staerz U. D., Rammensee H. G., Benedetto J. D., Bevan M. J. Characterization of a murine monoclonal antibody specific for an allotypic determinant on T cell antigen receptor. J Immunol. 1985 Jun;134(6):3994–4000. [PubMed] [Google Scholar]
  36. Tada T., Hu F. Y., Kishimoto H., Furutani-Seiki M., Asano Y. Molecular events in the T cell-mediated suppression of the immune response. Ann N Y Acad Sci. 1991 Dec 30;636:20–27. doi: 10.1111/j.1749-6632.1991.tb33434.x. [DOI] [PubMed] [Google Scholar]
  37. Ullman K. S., Northrop J. P., Verweij C. L., Crabtree G. R. Transmission of signals from the T lymphocyte antigen receptor to the genes responsible for cell proliferation and immune function: the missing link. Annu Rev Immunol. 1990;8:421–452. doi: 10.1146/annurev.iy.08.040190.002225. [DOI] [PubMed] [Google Scholar]
  38. Wegener A. M., Letourneur F., Hoeveler A., Brocker T., Luton F., Malissen B. The T cell receptor/CD3 complex is composed of at least two autonomous transduction modules. Cell. 1992 Jan 10;68(1):83–95. doi: 10.1016/0092-8674(92)90208-t. [DOI] [PubMed] [Google Scholar]
  39. Weissman A. M., Baniyash M., Hou D., Samelson L. E., Burgess W. H., Klausner R. D. Molecular cloning of the zeta chain of the T cell antigen receptor. Science. 1988 Feb 26;239(4843):1018–1021. doi: 10.1126/science.3278377. [DOI] [PubMed] [Google Scholar]
  40. Yagita H., Nakamura T., Asakawa J., Matsuda H., Tansyo S., Iigo Y., Okumura K. CD2 expression in murine B cell lineage. Int Immunol. 1989;1(1):94–98. doi: 10.1093/intimm/1.1.94. [DOI] [PubMed] [Google Scholar]
  41. Yorifuji H., Hirokawa N. Cytoskeletal architecture of neuromuscular junction: localization of vinculin. J Electron Microsc Tech. 1989 Jun;12(2):160–171. doi: 10.1002/jemt.1060120210. [DOI] [PubMed] [Google Scholar]
  42. de la Hera A., Müller U., Olsson C., Isaaz S., Tunnacliffe A. Structure of the T cell antigen receptor (TCR): two CD3 epsilon subunits in a functional TCR/CD3 complex. J Exp Med. 1991 Jan 1;173(1):7–17. doi: 10.1084/jem.173.1.7. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES