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CEA/CD3-bispecific T cell-engaging (BiTE) antibody-mediated T lymphocyte cytotoxicity maximized by inhibition of both PD1 and PD-L1

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Abstract

Bispecific T cell-engaging (BiTE) antibodies recruit polyclonal cytotoxic T cells (CTL) to tumors. One such antibody is carcinoembryonic antigen (CEA) BiTE that mediates T cell/tumor interaction by simultaneously binding CD3 expressed by T cells and CEA expressed by tumor cells. A widely operative mechanism for mitigating cytotoxic T cell-mediated killing is the interaction of tumor-expressed PD-L1 with T cell-expressed PD-1, which may be partly reversed by PD-1/PD-L1 blockade. We hypothesized that PD-1/PD-L1 blockade during BiTE-mediated T cell killing would enhance CTL function. Here, we determined the effects of PD-1 and PD-L1 blockade during initial T cell-mediated killing of CEA-expressing human tumor cell lines in vitro, as well as subsequent T cell-mediated killing by T lymphocytes that had participated in tumor cell killing. We observed a rapid upregulation of PD-1 expression and diminished cytolytic function of T cells after they had engaged in CEA BiTE-mediated killing of tumors. T cell cytolytic activity in vitro could be maximized by administration of anti-PD-1 or anti-PD-L1 antibodies alone or in combination if applied prior to a round of T cell killing, but T cell inhibition could not be fully reversed by this blockade once the T cells had killed tumor. In conclusion, our findings demonstrate that dual blockade of PD-1 and PD-L1 maximizes T cell killing of tumor directed by CEA BiTE in vitro, is more effective if applied early, and provides a rationale for clinical use.

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Abbreviations

ALL:

Acute lymphoblastic leukemia

APC:

Allophycocyanin

BiTE:

Bispecific T cell engaging

CEA:

Carcinoembryonic antigen

CTLA-4:

Cytotoxic T lymphocyte-associated protein 4

EDTA:

Ethylenediaminetetraacetic acid

FITC:

Fluorescein isocyanate

mAb:

Monoclonal antibody

MDSC:

Myeloid-derived suppressor cells

MFI:

Mean fluorescence intensity

NOD/SCID:

Non-obese diabetic/severe combined immunodeficiency

PBMC:

Peripheral blood mononuclear cell

PD-1:

Programmed cell death 1

PD-L1:

Programmed death-ligand 1

PE:

Phycoerythrin

PerCP:

Peridinin chlorophyll protein

Treg:

Regulatory T cell

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Acknowledgments

Supported by a Grant from MedImmune LLC to Takuya Osada.

Conflict of interest

Scott A. Hammond is an employee of MedImmune LCC, which supplied CEA BiTE for this study. The other authors disclose no potential conflicts of interest.

Author information

Author notes

  1. Sandip P. Patel

    Present address: University of California (UC) San Diego Moores Cancer Center, La Jolla, CA, USA

Authors and Affiliations

  1. Section of Applied Therapeutics, Department of Surgery, Duke University Medical Center, 403 MSRB, Research Drive, Durham, NC, 27710, USA

    Takuya Osada, Koya Osada & H. Kim Lyerly

  2. Department of Medicine, Duke University Medical Center, 403 MSRB, Research Drive, Durham, NC, 27710, USA

    Sandip P. Patel & Michael A. Morse

  3. Duke Comprehensive Cancer Center, Duke University Medical Center, 403 MSRB, Research Drive, Durham, NC, 27710, USA

    Takuya Osada, Michael A. Morse & H. Kim Lyerly

  4. MedImmune LLC, One MedImmune Way, Gaithersburg, MD, 20878, USA

    Scott A. Hammond

  5. 443A MSRB 1, Research DR, Durham, NC, 27710, USA

    Takuya Osada

Authors
  1. Takuya Osada
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Corresponding author

Correspondence to Takuya Osada.

Additional information

BiTE ® is a registered trademark of Amgen.

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Osada, T., Patel, S.P., Hammond, S.A. et al. CEA/CD3-bispecific T cell-engaging (BiTE) antibody-mediated T lymphocyte cytotoxicity maximized by inhibition of both PD1 and PD-L1. Cancer Immunol Immunother 64, 677–688 (2015). https://doi.org/10.1007/s00262-015-1671-y

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  • DOI: https://doi.org/10.1007/s00262-015-1671-y

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