E3 ubiquitin ligases and immune tolerance: Targeting the immune synapse from within?

  • Irene Puga
  • Fernando Macian
Part of the Progress in Inflammation Research book series (PIR)


The success of adaptive immunity relies on the ability to eliminate invading pathogens without eliciting responses against the host. Unique antigen receptors are randomly generated and recognize both self and non-self antigens. Therefore, mechanisms of tolerance must be in place to control the activity of self-reactive lymphocytes. Negative selection in the thymus eliminates most of the developing thymocytes that can recognize self antigens 1, whereas mechanisms of peripheral tolerance prevent the surviving self-reactive cells from engaging in responses against self tissues. Self-reactive T cells can be suppressed by regulatory T cells, and also eliminated by clonal deletion or inactivated by a mechanism known as anergy 2. In anergic T cells, T cell receptor (TCR) signaling is blocked, and cells become unresponsive to subsequent stimulation events 3, 4, 5.


Cell Tolerance Cell Anergy Immune Synapse Anergy Induction Supramolecular Activation Cluster 


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  1. 1.
    Palmer E (2003) Negative selection — clearing out the bad apples from the T-cell repertoire. Nat Rev Immunol 3: 383–391PubMedCrossRefGoogle Scholar
  2. 2.
    Abbas AK, Lohr J, Knoechel B, Nagabhushanam V (2004) T cell tolerance and autoimmunity. Autoimmun Rev 3: 471–475PubMedCrossRefGoogle Scholar
  3. 3.
    Schwartz RH (2003) T cell anergy. Annu Rev Immunol 21: 305–334PubMedCrossRefGoogle Scholar
  4. 4.
    Macian F, Im SH, Garcia-Cozar FJ, Rao A (2004) T-cell anergy. Curr Opin Immunol 16: 209–216PubMedCrossRefGoogle Scholar
  5. 5.
    Appleman LJ, Boussiotis VA (2003) T cell anergy and costimulation. Immunol Rev 192: 161–180PubMedCrossRefGoogle Scholar
  6. 6.
    Greenwald RJ, Freeman GJ, Sharpe AH (2005) The B7 family revisited. Annu Rev Immunol 23: 515–548PubMedCrossRefGoogle Scholar
  7. 7.
    Acuto O, Mise-Omata S, Mangino G, Michel F (2003) Molecular modifiers of T cell antigen receptor triggering threshold: The mechanism of CD28 costimulatory receptor. Immunol Rev 192: 21–31PubMedCrossRefGoogle Scholar
  8. 8.
    Krogsgaard M, Huppa JB, Purbhoo MA, Davis MM (2003) Linking molecular and cellular events in T-cell activation and synapse formation. Semin Immunol 15: 307–315PubMedCrossRefGoogle Scholar
  9. 9.
    Dustin ML (2005) A dynamic view of the immunological synapse. Semin Immunol 17: 400–410PubMedCrossRefGoogle Scholar
  10. 10.
    Wulfing C, Davis MM (1998) A receptor/cytoskeletal movement triggered by costimulation during T cell activation. Science 282: 2266–2269PubMedCrossRefGoogle Scholar
  11. 11.
    Jenkins MK, Schwartz RH (1987) Antigen presentation by chemically modified splenocytes induces antigen-specific T cell unresponsiveness in vitro and in vivo. J Exp Med 165: 302–319PubMedCrossRefGoogle Scholar
  12. 12.
    Quill H, Schwartz RH (1987) Stimulation of normal inducer T cell clones with antigen presented by purified Ia molecules in planar lipid membranes: Specific induction of a long-lived state of proliferative nonresponsiveness. J Immunol 138: 3704–3712PubMedGoogle Scholar
  13. 13.
    Jenkins MK, Chen CA, Jung G, Mueller DL, Schwartz RH (1990) Inhibition of antigenspecific proliferation of type 1 murine T cell clones after stimulation with immobilized anti-CD3 monoclonal antibody. J Immunol 144: 16–22PubMedGoogle Scholar
  14. 14.
    Harding FA, McArthur JG, Gross JA, Raulet DH, Allison JP (1992) CD28-mediated signalling co-stimulates murine T cells and prevents induction of anergy in T-cell clones. Nature 356: 607–609PubMedCrossRefGoogle Scholar
  15. 15.
    Egen JG, Kuhns MS, Allison JP (2002) CTLA-4: New insights into its biological function and use in tumor immunotherapy. Nat Immunol 3: 611–618PubMedCrossRefGoogle Scholar
  16. 16.
    Waterhouse P, Penninger JM, Timms E, Wakeham A, Shahinian A, Lee KP, Thompson CB, Griesser H, Mak TW (1995) Lymphoproliferative disorders with early lethality in mice deficient in Ctla-4. Science 270: 985–988PubMedCrossRefGoogle Scholar
  17. 17.
    Krummel MF, Allison JP (1995) CD28 and CTLA-4 have opposing effects on the response of T cells to stimulation. J Exp Med 182: 459–465PubMedCrossRefGoogle Scholar
  18. 18.
    Perez VL, Van Parijs L, Biuckians A, Zheng XX, Strom TB, Abbas AK (1997) Induction of peripheral T cell tolerance in vivo requires CTLA-4 engagement. Immunity 6: 411–417PubMedCrossRefGoogle Scholar
  19. 19.
    Greenwald RJ, Boussiotis VA, Lorsbach RB, Abbas AK, Sharpe AH (2001) CTLA-4 regulates induction of anergy in vivo. Immunity 14: 145–155PubMedCrossRefGoogle Scholar
  20. 20.
    Rao A, Luo C, Hogan PG (1997) Transcription factors of the NFAT family: Regulation and function. Annu Rev Immunol 15: 707–747PubMedCrossRefGoogle Scholar
  21. 21.
    Hogan PG, Chen L, Nardone J, Rao A (2003) Transcriptional regulation by calcium, calcineurin, and NFAT. Genes Dev 17: 2205–2232PubMedCrossRefGoogle Scholar
  22. 22.
    Macian F (2005) NFAT proteins: Key regulators of T-cell development and function. Nat Rev Immunol 5: 472–484PubMedCrossRefGoogle Scholar
  23. 23.
    Jain J, McCaffrey PG, Miner Z, Kerppola TK, Lambert JN, Verdine GL, Curran T, Rao A (1993) The T-cell transcription factor NFATp is a substrate for calcineurin and interacts with Fos and Jun. Nature 365: 352–355PubMedCrossRefGoogle Scholar
  24. 24.
    Macian F, Lopez-Rodriguez C, Rao A (2001) Partners in transcription: NFAT and AP-1. Oncogene 20: 2476–2489PubMedCrossRefGoogle Scholar
  25. 25.
    Macian F, Garcia-Cozar F, Im SH, Horton HF, Byrne MC, Rao A (2002) Transcriptional mechanisms underlying lymphocyte tolerance. Cell 109: 719–731PubMedCrossRefGoogle Scholar
  26. 26.
    Heissmeyer V, Macian F, Varma R, Im SH, Garcia-Cozar F, Horton HF, Byrne MC, Feske S, Venuprasad K, Gu H et al (2005) A molecular dissection of lymphocyte unresponsiveness induced by sustained calcium signalling. Novartis Found Symp 267: 165–174; discussion 174–179PubMedGoogle Scholar
  27. 27.
    Bandyopadhyay S, Soto-Nieves N, Macian F (2007) Transcriptional regulation of T cell tolerance. Semin Immunol 19: 180–187PubMedCrossRefGoogle Scholar
  28. 28.
    Telander DG, Malvey EN, Mueller DL (1999) Evidence for repression of IL-2 gene activation in anergic T cells. J Immunol 162: 1460–1465PubMedGoogle Scholar
  29. 29.
    Heissmeyer V, Macian F, Im SH, Varma R, Feske S, Venuprasad K, Gu H, Liu YC, Dustin ML, Rao A (2004) Calcineurin imposes T cell unresponsiveness through targeted proteolysis of signaling proteins. Nat Immunol 5: 255–265PubMedCrossRefGoogle Scholar
  30. 30.
    Zha Y, Marks R, Ho AW, Peterson AC, Janardhan S, Brown I, Praveen K, Stang S, Stone JC, Gajewski TF (2006) T cell anergy is reversed by active Ras and is regulated by diacylglycerol kinase-alpha. Nat Immunol 7: 1166–1173PubMedCrossRefGoogle Scholar
  31. 31.
    Olenchock BA, Guo R, Carpenter JH, Jordan M, Topham MK, Koretzky GA, Zhong XP (2006) Disruption of diacylglycerol metabolism impairs the induction of T cell anergy. Nat Immunol 7: 1174–1181PubMedCrossRefGoogle Scholar
  32. 32.
    Li L, Iwamoto Y, Berezovskaya A, Boussiotis VA (2006) A pathway regulated by cell cycle inhibitor p27(Kip1) and checkpoint inhibitor Smad3 is involved in the induction of T cell tolerance. Nat Immunol 7: 1157–1165PubMedCrossRefGoogle Scholar
  33. 33.
    Boussiotis VA, Freeman GJ, Taylor PA, Berezovskaya A, Grass I, Blazar BR, Nadler LM (2000) p27kip1 functions as an anergy factor inhibiting interleukin 2 transcription and clonal expansion of alloreactive human and mouse helper T lymphocytes. Nat Med 6: 290–297PubMedCrossRefGoogle Scholar
  34. 34.
    Powell JD, Lerner CG, Ewoldt GR, Schwartz RH (1999) The-180 site of the IL-2 promoter is the target of CREB/CREM binding in T cell anergy. J Immunol 163: 6631–6639PubMedGoogle Scholar
  35. 35.
    Bandyopadhyay S, Dure M, Paroder M, Soto-Nieves N, Puga I, Macian F (2007) Interleukin 2 gene transcription is regulated by Ikaros-induced changes in histone acetylation in anergic T cells. Blood 109: 2878–2886PubMedGoogle Scholar
  36. 36.
    Hundt M, Tabata H, Jeon MS, Hayashi K, Tanaka Y, Krishna R, De Giorgio L, Liu YC, Fukata M, Altman A (2006) Impaired activation and localization of LAT in anergic T cells as a consequence of a selective palmitoylation defect. Immunity 24: 513–522PubMedCrossRefGoogle Scholar
  37. 37.
    Hochstrasser M (2006) Lingering mysteries of ubiquitin-chain assembly. Cell 124: 27–34PubMedCrossRefGoogle Scholar
  38. 38.
    Fang S, Weissman AM (2004) A field guide to ubiquitylation. Cell Mol Life Sci 61: 1546–1561PubMedCrossRefGoogle Scholar
  39. 39.
    Ciechanover A (2005) Proteolysis:from the lysosome to ubiquitin and the proteasome. Nat Rev Mol Cell Biol 6: 79–87PubMedCrossRefGoogle Scholar
  40. 40.
    Schartner JM, Fathman CG, Seroogy CM (2007) Preservation of self: An overview of E3 ubiquitin ligases and T cell tolerance. Semin Immunol 19: 188–196PubMedCrossRefGoogle Scholar
  41. 41.
    Harper JW, Schulman BA (2006) Structural complexity in ubiquitin recognition. Cell 124: 1133–1136PubMedCrossRefGoogle Scholar
  42. 42.
    Nijman SM, Luna-Vargas MP, Velds A, Brummelkamp TR, Dirac AM, Sixma TK, Bernards R (2005) A genomic and functional inventory of deubiquitinating enzymes. Cell 123: 773–786PubMedCrossRefGoogle Scholar
  43. 43.
    Pickart CM, Fushman D (2004) Polyubiquitin chains: Polymeric protein signals. Curr Opin Chem Biol 8: 610–616PubMedCrossRefGoogle Scholar
  44. 44.
    Ciechanover A, Heller H, Elias S, Haas AL, Hershko A (1980) ATP-dependent conjugation of reticulocyte proteins with the polypeptide required for protein degradation. Proc Natl Acad Sci USA 77: 1365–1368PubMedCrossRefGoogle Scholar
  45. 45.
    Hershko A, Ciechanover A, Heller H, Haas AL, Rose IA (1980) Proposed role of ATP in protein breakdown:conjugation of protein with multiple chains of the polypeptide of ATP-dependent proteolysis. Proc Natl Acad Sci USA 77: 1783–1786PubMedCrossRefGoogle Scholar
  46. 46.
    Ciechanover A, Elias S, Heller H, Ferber S, Hershko A (1980) Characterization of the heat-stable polypeptide of the ATP-dependent proteolytic system from reticulocytes. J Biol Chem 255: 7525–7528PubMedGoogle Scholar
  47. 47.
    Mukhopadhyay D, Riezman H (2007) Proteasome-independent functions of ubiquitin in endocytosis and signaling. Science 315: 201–205PubMedCrossRefGoogle Scholar
  48. 48.
    Chen ZJ (2005) Ubiquitin signalling in the NF-kappaB pathway. Nat Cell Biol 7: 758–765PubMedCrossRefGoogle Scholar
  49. 49.
    Wang HY, Altman Y, Fang D, Elly C, Dai Y, Shao Y, Liu YC (2001) Cbl promotes ubiquitination of the T cell receptor zeta through an adaptor function of Zap-70. J Biol Chem 276: 26004–26011PubMedCrossRefGoogle Scholar
  50. 50.
    Chiang YJ, Kole HK, Brown K, Naramura M, Fukuhara S, Hu RJ, Jang IK, Gutkind JS, Shevach E, Gu H (2000) Cbl-b regulates the CD28 dependence of T-cell activation. Nature 403: 216–220PubMedCrossRefGoogle Scholar
  51. 51.
    Krawczyk C, Bachmaier K, Sasaki T, Jones RG, Snapper SB, Bouchard D, Kozieradzki I, Ohashi PS, Alt FW, Penninger JM (2000) Cbl-b is a negative regulator of receptor clustering and raft aggregation in T cells. Immunity 13: 463–473PubMedCrossRefGoogle Scholar
  52. 52.
    Jeon M-S, Atfield A, Venuprasad K, Krawczyk C, Sarao R, Elly C, Yang C, Arya S, Bachmaier K, Su L (2004) Essential Role of the E3 Ubiquitin ligase Cbl-b in T cell anergy induction. Immunity 21: 167–177PubMedCrossRefGoogle Scholar
  53. 53.
    Wiedemann A, Muller S, Favier B, Penna D, Guiraud M, Delmas C, Champagne E, Valitutti S (2005) T-cell activation is accompanied by an ubiquitination process occurring at the immunological synapse. Immunol Lett 98: 57–61PubMedCrossRefGoogle Scholar
  54. 54.
    Fang D, Liu Y-C (2001) Proteolysis-independent regulation of PI3K by Cbl-b-mediated ubiquitination in T cells. Nat Immunol 2: 870–875PubMedCrossRefGoogle Scholar
  55. 55.
    Bustelo XR, Crespo P, Lopez-Barahona M, Gutkind JS, Barbacid M (1997) Cbl-b, a member of the Sli-1/c-Cbl protein family, inhibits Vav-mediated c-Jun N-terminal kinase activation. Oncogene 15: 2511–2520PubMedCrossRefGoogle Scholar
  56. 56.
    Safford M, Collins S, Lutz MA, Allen A, Huang C-T, Kowalski J, Blackford A, Horton MR, Drake C, Schwartz RH et al (2005) Egr-2 and Egr-3 are negative regulators of T cell activation. Nat Immunol 6: 472–480PubMedCrossRefGoogle Scholar
  57. 57.
    Zhang J, Bardos T, Li D, Gal I, Vermes C, Xu J, Mikecz K, Finnegan A, Lipkowitz S, Glant TT (2002) Cutting edge: Regulation of T cell activation threshold by CD28 costimulation through targeting Cbl-b for ubiquitination. J Immunol 169: 2236–2240PubMedGoogle Scholar
  58. 58.
    Li D, Gal I, Vermes C, Alegre ML, Chong AS, Chen L, Shao Q, Adarichev V, Xu X, Koreny T et al (2004) Cutting edge: Cbl-b:one of the key molecules tuning CD28-and CTLA-4-mediated T cell costimulation. J Immunol 173: 7135–7139PubMedGoogle Scholar
  59. 59.
    Elly C, Witte S, Zhang Z, Rosnet O, Lipkowitz S, Altman A, Liu YC (1999) Tyrosine phosphorylation and complex formation of Cbl-b upon T cell receptor stimulation. Oncogene 18: 1147–1156PubMedCrossRefGoogle Scholar
  60. 60.
    Bachmaier K, Krawczyk C, Kozieradzki I, Kong YY, Sasaki T, Oliveira-dos-Santos A, Mariathasan S, Bouchard D, Wakeham A, Itie A et al (2000) Negative regulation of lymphocyte activation and autoimmunity by the molecular adaptor Cbl-b. Nature 403: 211–216PubMedCrossRefGoogle Scholar
  61. 61.
    Naramura M, Jang IK, Kole H, Huang F, Haines D, Gu H (2002) c-Cbl and Cbl-b regulate T cell responsiveness by promoting ligand-induced TCR down-modulation. Nat Immunol 3: 1192–1199PubMedCrossRefGoogle Scholar
  62. 62.
    Kitaura Y, Jang IK, Wang Y, Han YC, Inazu T, Cadera EJ, Schlissel M, Hardy RR, Gu H (2007) Control of the B cell-intrinsic tolerance programs by ubiquitin ligases Cbl and Cbl-b. Immunity 26: 567–578PubMedCrossRefGoogle Scholar
  63. 63.
    Wohlfert EA, Callahan MK, Clark RB (2004) Resistance to CD4+CD25+ regulatory T cells and TGF-β in Cbl-b−/− mice. J Immunol 173: 1059–1065PubMedGoogle Scholar
  64. 64.
    Perry WL, Hustad CM, Swing DA, O’Sullivan TN, Jenkins NA, Copeland NG (1998) The itchy locus encodes a novel ubiquitin protein ligase that is disrupted in a18H mice. Nat Genet 18: 143–146PubMedCrossRefGoogle Scholar
  65. 65.
    Gao M, Labuda T, Xia Y, Gallagher E, Fang D, Liu YC, Karin M (2004) Jun turnover is controlled through JNK-dependent phosphorylation of the E3 ligase Itch. Science 306: 271–275PubMedCrossRefGoogle Scholar
  66. 66.
    Rossi M, Aqeilan RI, Neale M, Candi E, Salomoni P, Knight RA, Croce CM, Melino G (2006) The E3 ubiquitin ligase Itch controls the protein stability of p63. Proc Natl Acad Sci USA 103: 12753–12758PubMedCrossRefGoogle Scholar
  67. 67.
    Rossi M, De Laurenzi V, Munarriz E, Green DR, Liu YC, Vousden KH, Cesareni G, Melino G (2005) The ubiquitin-protein ligase Itch regulates p73 stability. EMBO J 24: 836–848PubMedCrossRefGoogle Scholar
  68. 68.
    Gallagher E, Gao M, Liu YC, Karin M (2006) Activation of the E3 ubiquitin ligase Itch through a phosphorylation-induced conformational change. Proc Natl Acad Sci USA 103: 1717–1722PubMedCrossRefGoogle Scholar
  69. 69.
    Yang C, Zhou W, Jeon MS, Demydenko D, Harada Y, Zhou H, Liu YC (2006) Negative regulation of the E3 ubiquitin ligase itch via Fyn-mediated tyrosine phosphorylation. Mol Cell 21: 135–141PubMedCrossRefGoogle Scholar
  70. 70.
    Mouchantaf R, Azakir BA, McPherson PS, Millard SM, Wood SA, Angers A (2006) The ubiquitin ligase Itch is auto-ubiquitylated in vivo and in vitro but is protected from degradation by interacting with the deubiquitylating enzyme FAM/USP9X. J Biol Chem 281: 38738–38747PubMedCrossRefGoogle Scholar
  71. 71.
    Fang D, Elly C, Gao B, Fang N, Altman Y, Joazeiro C, Hunter T, Copeland N, Jenkins N, Liu Y-C (2002) Dysregulation of T lymphocyte function in itchy mice: A role for Itch in TH2 differentiation. Nat Immunol 3: 281–287PubMedCrossRefGoogle Scholar
  72. 72.
    Marchese A, Raiborg C, Santini F, Keen JH, Stenmark H, Benovic JL (2003) The E3 ubiquitin ligase AIP4 mediates ubiquitination and sorting of the G protein-coupled receptor CXCR4. Dev Cell 5: 709–722PubMedCrossRefGoogle Scholar
  73. 73.
    Qiu L, Joazeiro C, Fang N, Wang HY, Elly C, Altman Y, Fang D, Hunter T, Liu YC (2000) Recognition and ubiquitination of Notch by Itch, a hect-type E3 ubiquitin ligase. J Biol Chem 275: 35734–35737PubMedCrossRefGoogle Scholar
  74. 74.
    Matesic LE, Haines DC, Copeland NG, Jenkins NA (2006) Itch genetically interacts with Notch1 in a mouse autoimmune disease model. Hum Mol Genet 15: 3485–3497PubMedCrossRefGoogle Scholar
  75. 75.
    Anandasabapathy N, Ford GS, Bloom D, Holness C, Paragas V, Seroogy C, Skrenta H, Hollenhorst M, Fathman CG, Soares L (2003) GRAIL: An E3 ubiquitin ligase that inhibits cytokine gene transcription is expressed in anergic CD4+ T cells. Immunity 18: 535–547PubMedCrossRefGoogle Scholar
  76. 76.
    Soares L, Seroogy C, Skrenta H, Anandasabapathy N, Lovelace P, Chung CD, Engleman E, Fathman CG (2004) Two isoforms of otubain 1 regulate T cell anergy via GRAIL. Nat Immunol 5: 45–54PubMedCrossRefGoogle Scholar
  77. 77.
    Seroogy CM, Soares L, Ranheim EA, Su L, Holness C, Bloom D, Fathman CG (2004) The gene related to anergy in lymphocytes, an E3 ubiquitin ligase, is necessary for anergy induction in CD4 T cells. J Immunol 173: 79–85PubMedGoogle Scholar
  78. 78.
    Su L, Lineberry N, Huh Y, Soares L, Fathman CG (2006) A novel E3 ubiquitin ligase substrate screen identifies Rho guanine dissociation inhibitor as a substrate of gene related to anergy in lymphocytes. J Immunol 177: 7559–7566PubMedGoogle Scholar
  79. 79.
    MacKenzie DA, Schartner J, Lin J, Timmel A, Jennens-Clough M, Fathman CG, Seroogy CM (2007) GRAIL is up-regulated in CD4+ CD25+ T regulatory cells and is sufficient for conversion of T Cells to a regulatory phenotype. J Biol Chem 282: 9696–9702PubMedCrossRefGoogle Scholar
  80. 80.
    Kostianovsky AM, Maier LM, Baecher-Allan C, Anderson AC, Anderson DE (2007) Up-regulation of gene related to anergy in lymphocytes is associated with notch-mediated human T cell suppression. J Immunol 178: 6158–6163PubMedGoogle Scholar

Copyright information

© Birkhäuser Verlag Basel/Switzerland 2008

Authors and Affiliations

  • Irene Puga
    • 1
  • Fernando Macian
    • 1
  1. 1.Department of PathologyAlbert Einstein College of MedicineBronxUSA

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