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. 1991 Jan 1;173(1):7–17. doi: 10.1084/jem.173.1.7

Structure of the T cell antigen receptor (TCR): two CD3 epsilon subunits in a functional TCR/CD3 complex

PMCID: PMC2118768  PMID: 1824636

Abstract

Transgenic mice carrying and expressing the human CD3 epsilon gene incorporate the corresponding protein product into T cell receptor (TCR)/CD3 complexes on thymocyte and T cell surfaces. The chimeric antigen receptors allow normal T cell development and selection of repertoires in vivo and are able to transduce activation signals in vitro. We have exploited the ability to distinguish mouse (m) and human (h)CD3 epsilon chains to analyze the stoichiometry of CD3 epsilon in transgenic mouse TCRs. Immunoprecipitation and fluorescence resonance energy transfer experiments demonstrate that such TCRs can contain both h- and mCD3 epsilon chains, implying that more than one CD3 epsilon subunit occurs per TCR. Antigen comodulation studies are consistent with a stochastic use of h- or mCD3 epsilon during receptor assembly, and further suggest a structure for the TCR/CD3 complex with two CD3 epsilon chains. The determination of CD3 epsilon subunit stoichiometry, together with existing biochemical data, allows the generation of a minimal model for the structure of the TCR and illustrates the potential value of the transgenic approach to the analysis of complex receptors.

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Selected References

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  1. Alarcon B., Berkhout B., Breitmeyer J., Terhorst C. Assembly of the human T cell receptor-CD3 complex takes place in the endoplasmic reticulum and involves intermediary complexes between the CD3-gamma.delta.epsilon core and single T cell receptor alpha or beta chains. J Biol Chem. 1988 Feb 25;263(6):2953–2961. [PubMed] [Google Scholar]
  2. Alexander D. R., Cantrell D. A. Kinases and phosphatases in T-cell activation. Immunol Today. 1989 Jun;10(6):200–205. doi: 10.1016/0167-5699(89)90325-3. [DOI] [PubMed] [Google Scholar]
  3. Baniyash M., Garcia-Morales P., Bonifacino J. S., Samelson L. E., Klausner R. D. Disulfide linkage of the zeta and eta chains of the T cell receptor. Possible identification of two structural classes of receptors. J Biol Chem. 1988 Jul 15;263(20):9874–9878. [PubMed] [Google Scholar]
  4. Berkhout B., Alarcon B., Terhorst C. Transfection of genes encoding the T cell receptor-associated CD3 complex into COS cells results in assembly of the macromolecular structure. J Biol Chem. 1988 Jun 15;263(17):8528–8536. [PubMed] [Google Scholar]
  5. Beverley P. C., Callard R. E. Distinctive functional characteristics of human "T" lymphocytes defined by E rosetting or a monoclonal anti-T cell antibody. Eur J Immunol. 1981 Apr;11(4):329–334. doi: 10.1002/eji.1830110412. [DOI] [PubMed] [Google Scholar]
  6. Bill J., Kanagawa O., Woodland D. L., Palmer E. The MHC molecule I-E is necessary but not sufficient for the clonal deletion of V beta 11-bearing T cells. J Exp Med. 1989 Apr 1;169(4):1405–1419. doi: 10.1084/jem.169.4.1405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bonifacino J. S., Chen C., Lippincott-Schwartz J., Ashwell J. D., Klausner R. D. Subunit interactions within the T-cell antigen receptor: clues from the study of partial complexes. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6929–6933. doi: 10.1073/pnas.85.18.6929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brenner M. B., Trowbridge I. S., Strominger J. L. Cross-linking of human T cell receptor proteins: association between the T cell idiotype beta subunit and the T3 glycoprotein heavy subunit. Cell. 1985 Jan;40(1):183–190. doi: 10.1016/0092-8674(85)90321-6. [DOI] [PubMed] [Google Scholar]
  9. Cantrell D. A., Davies A. A., Crumpton M. J. Activators of protein kinase C down-regulate and phosphorylate the T3/T-cell antigen receptor complex of human T lymphocytes. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8158–8162. doi: 10.1073/pnas.82.23.8158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Clevers H., Alarcon B., Wileman T., Terhorst C. The T cell receptor/CD3 complex: a dynamic protein ensemble. Annu Rev Immunol. 1988;6:629–662. doi: 10.1146/annurev.iy.06.040188.003213. [DOI] [PubMed] [Google Scholar]
  11. Evans G. A., Lewis K. A., Lawless G. M. Molecular organization of the human CD3 gene family on chromosome 11q23. Immunogenetics. 1988;28(5):365–373. doi: 10.1007/BF00364236. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Forni L., de Petris S. Use of fluorescent antibodies in the study of lymphoid cell membrane molecules. Methods Enzymol. 1984;108:413–425. doi: 10.1016/s0076-6879(84)08108-8. [DOI] [PubMed] [Google Scholar]
  14. Geisler C., Kuhlmann J., Rubin B. Assembly, intracellular processing, and expression at the cell surface of the human alpha beta T cell receptor/CD3 complex. Function of the CD3-zeta chain. J Immunol. 1989 Dec 15;143(12):4069–4077. [PubMed] [Google Scholar]
  15. Havran W. L., Allison J. P. Developmentally ordered appearance of thymocytes expressing different T-cell antigen receptors. Nature. 1988 Sep 29;335(6189):443–445. doi: 10.1038/335443a0. [DOI] [PubMed] [Google Scholar]
  16. Hein W. R., Tunnacliffe A. Characterization of the CD3 gamma and delta invariant subunits of the sheep T cell antigen receptor. Eur J Immunol. 1990 Jul;20(7):1505–1511. doi: 10.1002/eji.1830200715. [DOI] [PubMed] [Google Scholar]
  17. Hochstenbach F., Brenner M. B. T-cell receptor delta-chain can substitute for alpha to form a beta delta heterodimer. Nature. 1989 Aug 17;340(6234):562–565. doi: 10.1038/340562a0. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Kanagawa O., Palmer E., Bill J. The T cell receptor V beta 6 domain imparts reactivity to the Mls-1a antigen. Cell Immunol. 1989 Apr 1;119(2):412–426. doi: 10.1016/0008-8749(89)90255-4. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Krissansen G. W., Verbi W., Totty N. F., Crumpton M. J. Purification and identification by amino acid sequence analysis of the subunits of the CD3 antigen of human tonsil T-lymphocytes. Mol Immunol. 1987 Oct;24(10):1069–1079. doi: 10.1016/0161-5890(87)90075-7. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Leo O., Foo M., Sachs D. H., Samelson L. E., Bluestone J. A. Identification of a monoclonal antibody specific for a murine T3 polypeptide. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1374–1378. doi: 10.1073/pnas.84.5.1374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Letourneur F., Mattei M. G., Malissen B. The mouse CD3-gamma, -delta, and -epsilon genes reside within 50 kilobases on chromosome 9, whereas CD3-zeta maps to chromosome 1, band H. Immunogenetics. 1989;29(4):265–268. doi: 10.1007/BF00717911. [DOI] [PubMed] [Google Scholar]
  25. Merćep M., Bonifacino J. S., Garcia-Morales P., Samelson L. E., Klausner R. D., Ashwell J. D. T cell CD3-zeta eta heterodimer expression and coupling to phosphoinositide hydrolysis. Science. 1988 Oct 28;242(4878):571–574. doi: 10.1126/science.2845582. [DOI] [PubMed] [Google Scholar]
  26. Merćep M., Weissman A. M., Frank S. J., Klausner R. D., Ashwell J. D. Activation-driven programmed cell death and T cell receptor zeta eta expression. Science. 1989 Dec 1;246(4934):1162–1165. doi: 10.1126/science.2531464. [DOI] [PubMed] [Google Scholar]
  27. Meuer S. C., Acuto O., Hussey R. E., Hodgdon J. C., Fitzgerald K. A., Schlossman S. F., Reinherz E. L. Evidence for the T3-associated 90K heterodimer as the T-cell antigen receptor. Nature. 1983 Jun 30;303(5920):808–810. doi: 10.1038/303808a0. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. 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]
  30. Mittler R. S., Goldman S. J., Spitalny G. L., Burakoff S. J. T-cell receptor-CD4 physical association in a murine T-cell hybridoma: induction by antigen receptor ligation. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8531–8535. doi: 10.1073/pnas.86.21.8531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Roosnek E., Lanzavecchia A. Triggering T cells by otherwise inert hybrid anti-CD3/antitumor antibodies requires encounter with the specific target cell. J Exp Med. 1989 Jul 1;170(1):297–302. doi: 10.1084/jem.170.1.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sancho J., Chatila T., Wong R. C., Hall C., Blumberg R., Alarcon B., Geha R. S., Terhorst C. T-cell antigen receptor (TCR)-alpha/beta heterodimer formation is a prerequisite for association of CD3-zeta 2 into functionally competent TCR.CD3 complexes. J Biol Chem. 1989 Dec 5;264(34):20760–20769. [PubMed] [Google Scholar]
  33. 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]
  34. Sussman J. J., Bonifacino J. S., Lippincott-Schwartz J., Weissman A. M., Saito T., Klausner R. D., Ashwell J. D. Failure to synthesize the T cell CD3-zeta chain: structure and function of a partial T cell receptor complex. Cell. 1988 Jan 15;52(1):85–95. doi: 10.1016/0092-8674(88)90533-8. [DOI] [PubMed] [Google Scholar]
  35. Szöllösi J., Damjanovich S., Goldman C. K., Fulwyler M. J., Aszalos A. A., Goldstein G., Rao P., Talle M. A., Waldmann T. A. Flow cytometric resonance energy transfer measurements support the association of a 95-kDa peptide termed T27 with the 55-kDa Tac peptide. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7246–7250. doi: 10.1073/pnas.84.20.7246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Szöllösi J., Mátyus L., Trón L., Balázs M., Ember I., Fulwyler M. J., Damjanovich S. Flow cytometric measurements of fluorescence energy transfer using single laser excitation. Cytometry. 1987 Mar;8(2):120–128. doi: 10.1002/cyto.990080204. [DOI] [PubMed] [Google Scholar]
  37. Transy C., Moingeon P. E., Marshall B., Stebbins C., Reinherz E. L. Most anti-human CD3 monoclonal antibodies are directed to the CD3 epsilon subunit. Eur J Immunol. 1989 May;19(5):947–950. doi: 10.1002/eji.1830190525. [DOI] [PubMed] [Google Scholar]
  38. Tunnacliffe A., Olsson C., Buluwela L., Rabbitts T. H. Organization of the human CD3 locus on chromosome 11. Eur J Immunol. 1988 Oct;18(10):1639–1642. doi: 10.1002/eji.1830181027. [DOI] [PubMed] [Google Scholar]
  39. Tunnacliffe A., Olsson C., de la Hera A. The majority of human CD3 epitopes are conferred by the epsilon chain. Int Immunol. 1989;1(5):546–550. doi: 10.1093/intimm/1.5.546. [DOI] [PubMed] [Google Scholar]
  40. Van Wauwe J. P., De Mey J. R., Goossens J. G. OKT3: a monoclonal anti-human T lymphocyte antibody with potent mitogenic properties. J Immunol. 1980 Jun;124(6):2708–2713. [PubMed] [Google Scholar]
  41. Weissman A. M., Frank S. J., Orloff D. G., Merćep M., Ashwell J. D., Klausner R. D. Role of the zeta chain in the expression of the T cell antigen receptor: genetic reconstitution studies. EMBO J. 1989 Dec 1;8(12):3651–3656. doi: 10.1002/j.1460-2075.1989.tb08539.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Weissman A. M., Hou D., Orloff D. G., Modi W. S., Seuanez H., O'Brien S. J., Klausner R. D. Molecular cloning and chromosomal localization of the human T-cell receptor zeta chain: distinction from the molecular CD3 complex. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9709–9713. doi: 10.1073/pnas.85.24.9709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Williams A. F., Barclay A. N. The immunoglobulin superfamily--domains for cell surface recognition. Annu Rev Immunol. 1988;6:381–405. doi: 10.1146/annurev.iy.06.040188.002121. [DOI] [PubMed] [Google Scholar]
  44. de la Hera A., Toribio M. L., Martinez C. Delineation of human thymocytes with or without functional potential by CD1-specific antibodies. Int Immunol. 1989;1(5):496–502. doi: 10.1093/intimm/1.5.496. [DOI] [PubMed] [Google Scholar]
  45. von Boehmer H., Teh H. S., Kisielow P. The thymus selects the useful, neglects the useless and destroys the harmful. Immunol Today. 1989 Feb;10(2):57–61. doi: 10.1016/0167-5699(89)90307-1. [DOI] [PubMed] [Google Scholar]

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