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

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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.

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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

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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.

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Authors and Affiliations

  1. Department of Immunology, Imperial College London, Hammersmith Hospital, London, W12 0NN, UK

    David Coe, Shaima Begom, Caroline Addey, Matthew White, Julian Dyson & Jian-Guo Chai

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  1. David Coe
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  2. Shaima Begom
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  3. Caroline Addey
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  4. Matthew White
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  5. Julian Dyson
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  6. Jian-Guo Chai
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Corresponding author

Correspondence to Jian-Guo Chai.

Additional information

This work was supported by a Senior Cancer Research Fellowship to JGC from Cancer Research, UK.

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Coe, D., Begom, S., Addey, C. et al. Depletion of regulatory T cells by anti-GITR mAb as a novel mechanism for cancer immunotherapy. Cancer Immunol Immunother 59, 1367–1377 (2010). https://doi.org/10.1007/s00262-010-0866-5

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  • DOI: https://doi.org/10.1007/s00262-010-0866-5

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