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Cancer Immunology, Immunotherapy

, Volume 59, Issue 9, pp 1367–1377 | Cite as

Depletion of regulatory T cells by anti-GITR mAb as a novel mechanism for cancer immunotherapy

  • David Coe
  • Shaima Begom
  • Caroline Addey
  • Matthew White
  • Julian Dyson
  • Jian-Guo ChaiEmail author
Original Article

Abstract

In vitro, engagement of GITR on Treg cells by the agonistic anti-GITR mAb, DTA-1, appears to abrogate their suppressive function. The consequence of in vivo engagement of GITR by DTA-1 is, however, less clear. In this study, we show that Treg cells isolated from DTA-1-treated mice were as potent as those from untreated mice in suppressing conventional CD4 T cells in vitro, indicating that in vivo GITR ligation does not disable Treg cells. Treatment of Foxp3/GFP knock-in mice with DTA-1 led to a selective reduction of circulating Treg cells, suggesting that DTA-1 is a depleting mAb which preferentially targets Treg cells. In tumour-bearing mice, DTA-1-mediated depletion of Treg cells was most marked in tumours but not in tumour-draining lymph node. These features were confirmed in an adoptive transfer model using tumour antigen-specific Treg cells. Interestingly, Treg cells detected in tumour tissues expressed much higher levels of GITR than those in tumour-draining lymph nodes, indicating that the efficiency of depletion might be correlated with the level of GITR expression. Finally, in vivo labelling of GITR in naive or tumour-bearing mice demonstrated that Treg cells constitutively expressed higher levels of GITR than conventional T cells, independent of location and activation state, consistent with the preferential in vivo depletion of Tregs by DTA-1. Thus, depletion of Treg cells represents a previously unrecognised in vivo activity of DTA-1 which has important implications for the application of anti-GITR antibodies in cancer immunotherapy.

Keywords

Anti-GITR Regulatory T cells Depletion HY Tumour Therapy 

Abbreviations

dLN

Tumour-draining lymph node

pLN

Peripheral LN

nTreg

Natural CD4+CD25+Foxp3+ regulatory T cells

Mar

Marilyn mice

PBL

Peripheral blood lymphocytes

MFI

Mean florescence intensity

Foxp3/GFP

Foxp3/GFP knock-in mice

Notes

Acknowledgments

We thank Dr. Shimon Sakaguchi and Dr. Bernard Massile for providing DTA-1 hybridoma cells and Foxp3/GFP knockin mice, respectively. We also thank Dr. Elizabeth Simpson for critically reading the manuscript.

Supplementary material

262_2010_866_MOESM1_ESM.pdf (697 kb)
(PDF 696 kb)

References

  1. 1.
    Nocentini G, Giunchi L, Ronchetti S et al (1997) A new member of the tumor necrosis factor/nerve growth factor receptor family inhibits T cell receptor-induced apoptosis. Proc Natl Acad Sci USA 94:6216–6221CrossRefPubMedGoogle Scholar
  2. 2.
    Gurney AL, Marsters SA, Huang RM et al (1999) Identification of a new member of the tumor necrosis factor family and its receptor, a human ortholog of mouse GITR. Curr Biol 9:215–218CrossRefPubMedGoogle Scholar
  3. 3.
    Kwon B, Yu KY, Ni J et al (1999) Identification of a novel activation-inducible protein of the tumor necrosis factor receptor superfamily and its ligand. J Biol Chem 274:6056–6061CrossRefPubMedGoogle Scholar
  4. 4.
    Shimizu J, Yamazaki S, Takahashi T et al (2002) Stimulation of CD25(+)CD4(+) regulatory T cells through GITR breaks immunological self-tolerance. Nat Immunol 3:135–142CrossRefPubMedGoogle Scholar
  5. 5.
    McHugh RS, Whitters MJ, Piccirillo CA et al (2002) CD4+ CD25+ immunoregulatory T cells: gene expression analysis reveals a functional role for the glucocorticoid-induced TNF receptor. Immunity 16:311–323CrossRefPubMedGoogle Scholar
  6. 6.
    Ji HB, Liao G, Faubion WA et al (2004) Cutting edge: the natural ligand for glucocorticoid-induced TNF receptor-related protein abrogates regulatory T cell suppression. J Immunol 172:5823–5827PubMedGoogle Scholar
  7. 7.
    Tone M, Tone Y, Adams E et al (2003) Mouse glucocorticoid-induced tumor necrosis factor receptor ligand is costimulatory for T cells. Proc Natl Acad Sci USA 100:15059–15064CrossRefPubMedGoogle Scholar
  8. 8.
    Ronchetti S, Zollo O, Bruscoli S et al (2004) GITR, a member of the TNF receptor superfamily, is costimulatory to mouse T lymphocyte subpopulations. Eur J Immunol 34:613–622CrossRefPubMedGoogle Scholar
  9. 9.
    Kohm AP, Williams JS, Miller SD (2004) Cutting edge: ligation of the glucocorticoid-induced TNF receptor enhances autoreactive CD4+ T cell activation and experimental autoimmune encephalomyelitis. J Immunol 172:4686–4690PubMedGoogle Scholar
  10. 10.
    Kanamaru F, Youngnak P, Hashiguchi M et al (2004) Costimulation via glucocorticoid-induced TNF receptor in both conventional and CD25+ regulatory CD4+ T cells. J Immunol 172:7306–7314PubMedGoogle Scholar
  11. 11.
    Stephens GL, McHugh RS, Whitters MJ et al (2004) Engagement of glucocorticoid-induced TNFR family-related receptor on effector T Cells by its ligand mediates resistance to suppression by CD4+ CD25+ T cells. J Immunol 173:5008–5020PubMedGoogle Scholar
  12. 12.
    Shevach EM, Stephens GL (2006) The GITR-GITRL interaction: co-stimulation or contrasuppression of regulatory activity? Nat Rev Immunol 6:613–618CrossRefPubMedGoogle Scholar
  13. 13.
    Nocentini G, Riccardi C (2005) GITR: a multifaceted regulator of immunity belonging to the tumor necrosis factor receptor superfamily. Eur J Immunol 35:1016–1022CrossRefPubMedGoogle Scholar
  14. 14.
    Turk MJ, Guevara-Patiño JA, Rizzuto GA et al (2004) Concomitant tumor immunity to a poorly immunogenic melanoma is prevented by regulatory T cells. J Exp Med 200:771–782CrossRefPubMedGoogle Scholar
  15. 15.
    Ko K, Yamazaki S, Nakamura K et al (2005) Treatment of advanced tumors with agonistic anti-GITR mAb and its effects on tumor-infiltrating Foxp3 + CD25+ CD4+ regulatory T cells. J Exp Med 202:885–891CrossRefPubMedGoogle Scholar
  16. 16.
    Ramirez-Montagut T, Chow A, Hirschhorn-Cymerman D et al (2006) Glucocorticoid-induced TNF receptor family related gene activation overcomes tolerance/ignorance to melanoma differentiation antigens and enhances antitumor immunity. J Immunol 176:6434–6442PubMedGoogle Scholar
  17. 17.
    Cohen AD, Diab A, Perales MA et al (2006) Agonist anti-GITR antibody enhances vaccine-induced CD8(+) T-cell responses and tumor immunity. Cancer Res 66:4904–4912CrossRefPubMedGoogle Scholar
  18. 18.
    Zhou P, L’italien L, Hodges D et al (2007) Pivotal roles of CD4+ effector T cells in mediating agonistic anti-GITR mAb-induced-immune activation and tumor immunity in CT26 tumors. J Immunol 179:7365–7375PubMedGoogle Scholar
  19. 19.
    Valzasina B, Guiducci C, Dislich H et al (2005) Triggering of OX40 (CD134) on CD4(+)CD25+ T cells blocks their inhibitory activity: a novel regulatory role for OX40 and its comparison with GITR. Blood 105:2845–2851CrossRefPubMedGoogle Scholar
  20. 20.
    Sharma S, Dominguez AL, Manrique SZ et al (2008) Systemic targeting of CpG-ODN to the tumor microenvironment with anti-neu-CpG hybrid molecule and T regulatory cell depletion induces memory responses in BALB-neuT tolerant mice. Cancer Res 68:7530–7540CrossRefPubMedGoogle Scholar
  21. 21.
    Suvas S, Kim B, Sarangi PP et al (2005) In vivo kinetics of GITR and GITR ligand expression and their functional significance in regulating viral immunopathology. J Virol 79:11935–11942CrossRefPubMedGoogle Scholar
  22. 22.
    Wang Y, Kissenpfennig A, Mingueneau M et al (2008) Th2 lymphoproliferative disorder of LatY136F mutant mice unfolds independently of TCR-MHC engagement and is insensitive to the action of Foxp3 + regulatory T cells. J Immunol 180:1565–1575PubMedGoogle Scholar
  23. 23.
    Grandjean I, Duban L, Bonney EA et al (2003) Are major histocompatibility complex molecules involved in the survival of naive CD4+ T Cells? J Exp Med 198:1089–1102CrossRefPubMedGoogle Scholar
  24. 24.
    Summerhayes IC, Franks LM (1979) Effects of donor age on neoplastic transformation of adult mouse bladder epithelium in vitro. J Natl Cancer Inst 62:1017–1023PubMedGoogle Scholar
  25. 25.
    Gulbenkian AR, Egan RW, Fernandez X et al (1992) Interleukin-5 modulates eosinophil accumulation in allergic guinea pig lung. Am Rev Respir Dis 146:263–266PubMedGoogle Scholar
  26. 26.
    Lowenthal JW, Corthesy P, Tougne C et al (1985) High and low affinity IL 2 receptors: analysis by IL 2 dissociation rate and reactivity with monoclonal anti-receptor antibody PC61. J Immunol 135:3988–3994PubMedGoogle Scholar
  27. 27.
    Chai JG, Tsang JY, Lechler R et al (2002) CD4+ CD25+ T cells as immunoregulatory T cells in vitro. Eur J Immunol 32:2365–2375CrossRefPubMedGoogle Scholar
  28. 28.
    Yang AS, Monken CE, Lattime EC (2003) Intratumoral vaccination with vaccinia-expressed tumor antigen and granulocyte macrophage colony-stimulating factor overcomes immunological ignorance to tumor antigen. Cancer Res 63:6956–6961PubMedGoogle Scholar
  29. 29.
    Fontenot JD, Rasmussen JP, Williams LM et al (2005) Regulatory T cell lineage specification by the forkhead transcription factor Foxp3. Immunity 22:329–341CrossRefPubMedGoogle Scholar
  30. 30.
    Piconese S, Valzasina B, Colombo MP (2008) OX40 triggering blocks suppression by regulatory T cells and facilitates tumor rejection. J Exp Med 205:825–839CrossRefPubMedGoogle Scholar
  31. 31.
    Lutsiak ME, Tagaya Y, Adams AJ et al (2008) Tumor-induced impairment of TCR signaling results in compromised functionality of tumor-infiltrating regulatory T cells. J Immunol 180:5871–5881PubMedGoogle Scholar
  32. 32.
    Zou W (2006) Regulatory T cells, tumour immunity and immunotherapy. Nat Rev Immunol 6:295–307CrossRefPubMedGoogle Scholar
  33. 33.
    Qin FX (2009) Dynamic behavior and function of Foxp3 + regulatory T cells in tumor bearing host. Cell Mol Immunol 6:3–13CrossRefPubMedGoogle Scholar
  34. 34.
    Sakaguchi S (2004) Naturally arising CD4+ regulatory T cells for immunological self-tolerance and negative control of immune responses. Annu Rev Immunol 22:531–562CrossRefPubMedGoogle Scholar
  35. 35.
    Ghiringhelli F, Puig PE, Roux S et al (2005) Tumor cells convert immature myeloid dendritic cells into TGF-beta-secreting cells inducing CD4+ CD25+ regulatory T cell proliferation. J Exp Med 202:919–929CrossRefPubMedGoogle Scholar
  36. 36.
    Sharma MD, Baban B, Chandler P et al (2007) Plasmacytoid dendritic cells from mouse tumor-draining lymph nodes directly activate mature Tregs via indoleamine 2, 3-dioxygenase. J Clin Invest 117:2570–2582CrossRefPubMedGoogle Scholar
  37. 37.
    Tang Q, Bluestone J (2008) The Foxp3 + regulatory T cell: a jack of all trades, master of regulation. Nat Rev Immunol 9:239–244CrossRefGoogle Scholar
  38. 38.
    Mempel TR, Pittet MJ, Khazaie K et al (2006) Regulatory T cells reversibly suppress cytotoxic T cell function independent of effector differentiation. Immunity 25:129–141CrossRefPubMedGoogle Scholar
  39. 39.
    Khazaie K, von Boehmer H (2006) The impact of CD4+ CD25+ Treg on tumor specific CD8+ T cell cytotoxicity and cancer. Semin Cancer Biol 16:124–136CrossRefPubMedGoogle Scholar
  40. 40.
    Cao X, Cai SF, Fehniger TA et al (2007) Granzyme B and perforin are important for regulatory T cell-mediated suppression of tumour clearance. Immunity 27:635–646CrossRefPubMedGoogle Scholar
  41. 41.
    You S, Poulton L, Cobbold S et al (2009) Key role of the GITR/GITRLigand pathway in the development of murine autoimmune diabetes: a potential therapeutic target. PLoS One 4:7848CrossRefGoogle Scholar
  42. 42.
    Yu P, Lee Y, Liu W et al (2005) Intratumor depletion of CD4+ cells unmasks tumor immunogenicity leading to the rejection of late-stage tumors. J Exp Med 201:779–791CrossRefPubMedGoogle Scholar
  43. 43.
    Larsen CP, Elwood ET, Alexander DZ et al (1996) Long-term acceptance of skin and cardiac allografts after blocking CD40 and CD28 pathways. Nature 381:434–438CrossRefPubMedGoogle Scholar
  44. 44.
    Monk NJ, Hargreaves RE, Marsh JE et al (2003) Fc-dependent depletion of activated T cells occurs through CD40L-specific antibody rather than costimulation blockade. Nat Med 10:1275–1280CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • David Coe
    • 1
  • Shaima Begom
    • 1
  • Caroline Addey
    • 1
  • Matthew White
    • 1
  • Julian Dyson
    • 1
  • Jian-Guo Chai
    • 1
    Email author
  1. 1.Department of ImmunologyImperial College London, Hammersmith HospitalLondonUK

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