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A functional role for CD28 costimulation in tumor recognition by single-chain receptor-modified T cells

Cancer Gene Therapy volume 11, pages 371–379 (2004)Cite this article

Abstract

T cells engineered to express single-chain antibody receptors that incorporate TCR-ζ and cluster designation (CD)28 signaling domains (scFv-α-erbB2-CD28-ζ) can be redirected in vivo to cancer cells that lack triggering costimulatory molecules. To assess the contribution of CD28 signaling to the function of the scFv-CD28-ζ receptor, we expressed a series of mutated scFv-CD28-ζ receptors directed against erbB2. Residues known to be critical for CD28 signaling were mutated from tyrosine to phenylalanine at position 170 or proline to alanine at positions 187 and 190. Primary mouse T cells expressing either of the mutant receptors demonstrated impaired cytokine (IFN-γ and GM-CSF) production and decreased proliferation after antigen ligation in vitro and decreased antitumor efficacy in vivo compared with T cells expressing the wild-type scFv-CD28-ζ receptor, suggesting a key signaling role for the CD28 component of the scFv-CD28-ζ receptor. Importantly, cell surface expression, binding capacity and cytolytic activity mediated by the scFv-CD28-ζ receptor were not diminished by either mutation. Overall, this study has definitively demonstrated a functional role for the CD28 component of the scFv-CD28-ζ receptor and has shown that incorporation of costimulatory activity in chimeric scFv receptors is a powerful approach for improving adoptive cancer immunotherapy.

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Abbreviations

Bcl-XL:

B cell lymphoma/leukemia-x gene

CD:

cluster designation

LAK:

lymphokine activated killer

PI3 kinase:

phosphatidylinositol 3-kinase

scFv:

single chain variable domain

TAA:

tumor associated antigen

TIL:

tumor-infiltrating lymphocyte

VH:

variable heavy chain

VL:

variable light chain

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Acknowledgements

We thank Rachel Cameron and the staff of the Peter MacCallum Cancer Institute animal facilities for the caring and maintenance of mice used in this study. We also thank Dr Sarah Russell for intellectual input concerning biochemical experiments.

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Authors and Affiliations

  1. Cancer Immunology Program, Sir Donald and Lady Trescowthick Laboratories, Peter MacCallum Cancer Institute, St Andrews Place, East Melbourne, Victoria, Australia

    Maria Moeller, Nicole M Haynes, Joseph A Trapani, Michele WL Teng, Jacob T Jackson, Jane E Tanner, Michael H Kershaw, Mark J Smyth & Phillip K Darcy

  2. Rotary Bone Marrow Research Laboratory, Royal Melbourne Hospital, Parkville, Victoria, Australia

    Loretta Cerutti & Stephen M Jane

Authors
  1. Maria Moeller
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  2. Nicole M Haynes
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  3. Joseph A Trapani
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  4. Michele WL Teng
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  6. Jane E Tanner
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Corresponding author

Correspondence to Phillip K Darcy.

Additional information

MJS and JAT are currently supported by National Health and Medical Research Council of Australia (NH&MRC) Principal Research Fellowships. This study was supported by a Program Grant from the Susan G Komen Breast Cancer Foundation and a project grant from The Cancer Council Victoria.

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Moeller, M., Haynes, N., Trapani, J. et al. A functional role for CD28 costimulation in tumor recognition by single-chain receptor-modified T cells. Cancer Gene Ther 11, 371–379 (2004). https://doi.org/10.1038/sj.cgt.7700710

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