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Immune checkpoint inhibitors enhance cytotoxicity of cytokine-induced killer cells against human myeloid leukaemic blasts

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Abstract

We studied whether blockade of inhibitory receptors on cytokine-induced killer (CIK) cells by immune checkpoint inhibitors could increase its anti-tumour potency against haematological malignancies. CIK cultures were generated from seven normal donors and nine patients with acute myeloid leukaemia (AML), acute lymphoblastic leukaemia (ALL) or multiple myeloma (MM). The inhibitory receptors B and T lymphocyte attenuator, CD200 receptor, lymphocyte activation gene-3 (LAG-3) and T cell immunoglobulin and mucin-domain-containing-3 (TIM-3) were present at variable percentages in most CIK cultures, while cytotoxic T lymphocyte-associated protein 4 (CTLA-4), programmed death-1 (PD-1) and killer cell immunoglobulin-like receptors (KIR2DL1/2/3) were expressed at low level in most cultures. Without blockade, myeloid leukaemia cells were susceptible to autologous and allogeneic CIK-mediated cytotoxicity. Blockade of KIR, LAG-3, PD-1 and TIM-3 but not CTLA-4 resulted in remarkable increase in killing against these targets, even in those with poor baseline cytotoxicity. ALL and MM targets were resistant to CIK-mediated cytotoxicity, and blockade of receptors did not increase cytotoxicity to a meaningful extent. Combination of inhibitors against two receptors did not further increase cytotoxicity. Interestingly, potentiation of CIK killing by blocking antibodies was not predicted by expression of receptors on CIK and their respective ligands on the targets. Compared to un-activated T and NK cells, blockade potentiated the cytotoxicity of CIK cells to a greater degree and at a lower E:T ratio, but without significant increase in cytotoxicity against normal white cell. Our findings provide the basis for clinical trial combining autologous CIK cells with checkpoint inhibitors for patients with AML.

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Abbreviations

ALL:

Acute lymphoblastic leukaemia

AML:

Acute myeloid leukaemia

BTLA:

B and T lymphocyte attenuator

CD200R:

CD200 receptor

CIK:

Cytokine-induced killer

CLL:

Chronic lymphocytic leukaemia

CML:

Chronic myeloid leukaemia

Cr:

Chromium

CTLA-4:

Cytotoxic T lymphocyte-associated protein-4

FSC:

Forward scatter

HVEM:

Herpes virus entry mediator

KIR:

Killer cell immunoglobulin-like receptor

LAG-3:

Lymphocyte activation gene-3

MM:

Multiple myeloma

PD-1:

Programmed death-1

SSC:

Side scatter

TIM-3:

T-cell immunoglobulin and mucin-domain-containing-3

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Acknowledgments

This project is funded by the Singapore General Hospital (SGH) Research Grant (2014). We thank the SGH Department of Haematology Tissue Repository for provision of patients’ samples; Bristol-Myers Squibb for their kind gifts of the mAbs ipilimumab, nivolumab, lirilumab and BMS-986016; and the SingHealth Legal Department for their assistance in the agreement for transfer of these mAbs.

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  1. Department of Haematology, Singapore General Hospital, Level 3, Academia, 20, College Road, Singapore, 169856, Singapore

    Su Li Poh & Yeh Ching Linn

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Correspondence to Yeh Ching Linn.

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Poh, S.L., Linn, Y.C. Immune checkpoint inhibitors enhance cytotoxicity of cytokine-induced killer cells against human myeloid leukaemic blasts. Cancer Immunol Immunother 65, 525–536 (2016). https://doi.org/10.1007/s00262-016-1815-8

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