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Upregulation of thioredoxin-1 in activated human NK cells confers increased tolerance to oxidative stress

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Abstract

Adoptive transfer of immune cells, such as T lymphocytes and NK cells, has potential to control cancer growth. However, this can be counteracted by immune escape mechanisms within the tumor microenvironment, including those mediated by reactive oxygen species (ROS). Here, we determined the levels of anti-oxidant molecules in NK cells and their capacity to overcome ROS-induced immune suppression. We investigated the effect of H2O2 on resting NK cells, IL-2-activated NK cells and NK cells expanded by coculture with the K562 leukemia cell line genetically modified to express membrane-bound IL-15 and 4-1BB ligand (K562-mb15-41BBL). Expression of anti-oxidant and anti-apoptotic genes was evaluated by expression array, and protein levels of anti-oxidant molecules by Western blot. Activated NK cells, IL-2-activated NK cells and NK cells expanded by K562-mb15-41BBL were significantly more resistant to H2O2-induced cell death than resting NK. Thioredoxin-1 (TXN1) and peroxiredoxin-1 (PRDX1) were also up-regulated in activated NK cells. Moreover, H2O2-induced cell death after IL-2 activation was significantly induced in the presence of an anti-TXN1-neutralising antibody. Collectively, these data document that activated NK cells can resist to H2O2-induced cell death by up-regulation of TXN1.

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Abbreviations

7-AAD:

7-Aminoactinomycin D

Akt:

Protein kinase B

ECL:

Enhanced chemiluminescence

Erk:

Extracellular signal–regulated kinase

Expanded NK:

NK cells expanded with K562 leukemia cell line genetically modified to express membrane-bound IL-15 and 4-1BB ligand for 7 days

IL-2 NK:

NK cells activated by several doses of IL-2 for 7 days

K562-mb15-41BBL:

K562 leukemia cell line genetically modified to express membrane-bound IL-15 and 4-1BB ligand

PRDX1:

Peroxiredoxin-1

Resting NK:

Resting NK cells

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

TXN1:

Thioredoxin-1

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Acknowledgements

This work was supported by a Clinician Scientist Award (CSA) and Clinician Scientist-Individual Research Grant (CS-IGR) from the National Medical Research Council of Singapore. We would like to thank Dr. Dario Campana (National University of Singapore, Singapore) for providing us with the K562-mb15-41BBL cell line.

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Authors and Affiliations

  1. Department of Surgery, National University of Singapore, 1E Kent Ridge Road, Singapore, Singapore

    Kousaku Mimura, Kensuke Shiraishi, Asim Shabbir, Jimmy So & Koji Kono

  2. Department of Hematology-Oncology, National University of Singapore, 1E Kent Ridge Road, Singapore, Singapore

    Ley-Fang Kua & Wei-Peng Yong

  3. Department of Pediatrics, National University of Singapore, 1E Kent Ridge Road, Singapore, Singapore

    Noriko Shimasaki

  4. Cancer Science Institute of Singapore, National University of Singapore, 14 Medical Drive, #12-01, Singapore, Singapore

    Shotaro Nakajima, Lim Kee Siang & Koji Kono

  5. Department of Surgery, Fujikawa Hospital, Kyonan Medical Center, 340-1 Kajikazawa, Fujikawa-cho, Minamikoma-gun, Yamanashi, Japan

    Kousaku Mimura & Kensuke Shiraishi

  6. Department of Organ Regulatory Surgery and Advanced Cancer Immunotherapy, Fukushima Medical University, 1 Hikarigaoka, Fukushima City, Fukushima, 960-1295, Japan

    Koji Kono

Authors
  1. Kousaku Mimura
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  2. Ley-Fang Kua
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  8. Jimmy So
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  9. Wei-Peng Yong
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Correspondence to Koji Kono.

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The authors declare that they have no conflict of interest.

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K. Mimura and L.-F. Kua equally contributed to this study.

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Mimura, K., Kua, LF., Shimasaki, N. et al. Upregulation of thioredoxin-1 in activated human NK cells confers increased tolerance to oxidative stress. Cancer Immunol Immunother 66, 605–613 (2017). https://doi.org/10.1007/s00262-017-1969-z

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  • DOI: https://doi.org/10.1007/s00262-017-1969-z

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