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Antibody–peptide–MHC fusion conjugates target non-cognate T cells to kill tumour cells

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Abstract

Attempts to generate robust anti-tumour cytotoxic T lymphocyte (CTL) responses using immunotherapy are frequently thwarted by exhaustion and anergy of CTL recruited to tumour. One strategy to overcome this is to retarget a population of virus-specific CTL to kill tumour cells. Here, we describe a proof-of-principle study using a bispecific conjugate designed to retarget ovalbumin (OVA)-specific CTL to kill tumour cells via CD20. A single-chain trimer (SCT) consisting of MHCI H-2Kb/SIINFEKL peptide/beta 2 microglobulin/BirA was expressed in bacteria, refolded and chemically conjugated to one (1:1; F2) or two (2:1; F3) anti-hCD20 Fab′ fragments. In vitro, the [SCT × Fab′] (F2 and F3) redirected SIINFEKL-specific OT-I CTL to kill CD20+ target cells, and in the presence of CD20+ target cells to provide crosslinking, they were also able to induce proliferation of OT-I cells. In vivo, activated OT-I CTL could be retargeted to kill [SCT × Fab′]-coated B cells from hCD20 transgenic (hCD20 Tg) mice and also EL4 and B16 mouse tumour cells expressing human CD20 (hCD20). Importantly, in a hCD20 Tg mouse model, [SCT × Fab′] administered systemically were able to retarget activated OT-I cells to deplete normal B cells, and their performance matched that of a bispecific antibody (BsAb) comprising anti-CD3 and anti-CD20. [SCT × Fab′] were also active therapeutically in an EL4 tumour model. Furthermore, measurement of serum cytokine levels suggests that [SCT × Fab′] are associated with a lower level of inflammatory cytokine release than the BsAb and so may be advantageous clinically in terms of reduced toxicity.

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Acknowledgments

We would like to thank all members of the Laboratory for their help and assistance with this project. Funding was provided by Tenovus Cardiff and Cancer Research UK.

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The authors confirm that there is no conflict of interest.

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Authors and Affiliations

  1. Antibody and Vaccine Group MP88, Cancer Sciences Unit, Faculty of Medicine, General Hospital, University of Southampton, Tremona Road, Southampton, SO16 6YD, UK

    Ben C. King, Angela D. Hamblin, Ruth R. French & Martin J. Glennie

  2. Southampton Cancer Research UK Centre, Cancer Sciences Unit, Faculty of Medicine, General Hospital, University of Southampton, Southampton, SO16 6YD, UK

    Leon R. Douglas & Peter W. M. Johnson

  3. Department of Medical Oncology, Charing Cross Hospital, Imperial College AHSC, London, W2 1NY, UK

    Philip M. Savage

  4. Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, 63110, USA

    Ted H. Hansen

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  1. Ben C. King
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  2. Angela D. Hamblin
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  3. Philip M. Savage
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  4. Leon R. Douglas
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  7. Peter W. M. Johnson
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Correspondence to Ruth R. French.

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Ben C. King and Angela D. Hamblin are joint first authors and contributed equally to this work.

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King, B.C., Hamblin, A.D., Savage, P.M. et al. Antibody–peptide–MHC fusion conjugates target non-cognate T cells to kill tumour cells. Cancer Immunol Immunother 62, 1093–1105 (2013). https://doi.org/10.1007/s00262-013-1408-8

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  • DOI: https://doi.org/10.1007/s00262-013-1408-8

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