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Augmented anti-tumor activity of NK-92 cells expressing chimeric receptors of TGF-βR II and NKG2D

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Abstract

The capacity of natural killer (NK) cells to kill tumor cells without specific antigen recognition provides an advantage over T cells and makes them potential effectors for tumor immunotherapy. However, the efficacy of NK cell adoptive therapy can be limited by the immunosuppressive tumor microenvironment. Transforming growth factor-β (TGF-β) is a potent immunosuppressive cytokine that can suppress NK cell function. To convert the suppressive signal induced by TGF-β to an activating signal, we genetically modified NK-92 cells to express a chimeric receptor with TGF-β type II receptor extracellular and transmembrane domains and the intracellular domain of NK cell-activating receptor NKG2D (TN chimeric receptor). NK-92 cells expressing TN receptors were resistant to TGF-β-induced suppressive signaling and did not down-regulate NKG2D. These modified NK-92 cells had higher killing capacity and interferon γ (IFN-γ) production against tumor cells compared with the control cells and their cytotoxicity could be further enhanced by TGF-β. More interestingly, the NK-92 cells expressing TN receptors were better chemo-attracted to the tumor cells expressing TGF-β. The presence of these modified NK-92 cells significantly inhibited the differentiation of human naïve CD4+ T cells to regulatory T cells. NK-92-TN cells could also inhibit tumor growth in vivo in a hepatocellular carcinoma xenograft tumor model. Therefore, TN chimeric receptors can be a novel strategy to augment anti-tumor efficacy in NK cell adoptive therapy.

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Abbreviations

α-MEM:

Minimum essential medium alpha

BFA:

Brefeldin A

DAP:

DNAX-activating protein

DMEM:

Dulbecco-modified eagle medium

FACS:

Fluorescence-activated cell sorter

FBS:

Fetal bovine serum

HLA:

Human leucocyte antigen

HRP:

Horseradish Peroxidase

IFN-γ:

Interferonγ

IgA:

Immunoglobulin A

IL-2:

Interleukin 2

KIR:

Killer cell Ig-like Receptors

LDH:

Lactic dehydrogenase

mAb:

Monoclonal antibody

MFI:

Mean fluorescent intensity

MHC:

Major histocompatibility complex

NK:

Natural killer

NK-92-TN:

NK-92 expressing extracellular and transmembrane domains of TGF-β receptor II with intracellular domain of NKG2D

NK-92-Vector:

NK-92 expressing vector

NKG2C:

NK Group 2 member C

NKG2D:

NK Group 2 member D

PCR:

Polymerase chain reaction

PE:

Phycoerythrin

PMA:

Phorbol myristate acetate

PVDF:

Polyvinylidene fluoride

RPMI:

Roswell Park Memorial Institute

RT-PCR:

Real-time PCR

SDS–PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

TGF-β:

Transforming growth factor-β

TGF-βR:

Transforming growth factor-β receptor

Treg:

Regulatory T

VSVG:

Vesicular stomatitis virus G

YFP:

Yellow fluorescent protein

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Acknowledgements

We thank Dr. Feili Gong for providing NK-92 cells and Dr. Yun Zhao for providing lenti-virus vectors. This work has been supported by grants from the National Natural Science Foundation of China (81273268, 81471586), the project funding from Suzhou city (SWG0904), Priority Academic Program Development of Jiangsu Higher Education Institutions, and the start-up grant from the National University of Singapore.

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Authors and Affiliations

  1. Institute of Blood and Marrow Transplantation, Department of Hematology, Collaborative Innovation Center of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, 215006, People’s Republic of China

    Zhongjuan Wang, Linghua Guo, Yuan Song, Yinsheng Zhang, Dandan Lin, Bo Hu & Yu Mei

  2. Immunology Programme, Life Sciences Institute and Department of Microbiology and Immunology, National University of Singapore, 28 Medical Dr., Singapore, 117456, Singapore

    Yu Mei, Dedy Sandikin & Haiyan Liu

Authors
  1. Zhongjuan Wang
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  2. Linghua Guo
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  3. Yuan Song
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  4. Yinsheng Zhang
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  9. Haiyan Liu
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Correspondence to Haiyan Liu.

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The authors declare no potential conflicts of interest.

Additional information

Z. Wang, L. Guo, and Y. Song contributed equally.

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Wang, Z., Guo, L., Song, Y. et al. Augmented anti-tumor activity of NK-92 cells expressing chimeric receptors of TGF-βR II and NKG2D. Cancer Immunol Immunother 66, 537–548 (2017). https://doi.org/10.1007/s00262-017-1959-1

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  • DOI: https://doi.org/10.1007/s00262-017-1959-1

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