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Journal of Cancer Research and Clinical Oncology

, Volume 144, Issue 7, pp 1279–1288 | Cite as

hIL-15-gene modified human natural killer cells (NKL-IL15) exhibit anti-human leukemia functions

Original Article – Cancer Research

Abstract

Purpose

Natural killer (NK) cells can kill transformed cells and represent anti-tumor activities for improving the immunotherapy of cancer. In previous works, we established human interleukin-15 (hIL-15) gene-modified NKL cells (NKL-IL15) and demonstrated their efficiency against human hepatocarcinoma cells (HCCs) in vitro and in vivo. To further assess the applicability of NKL-IL15 cells in adoptive cellular immunotherapy for human leukemia, here we report their natural cytotoxicity against leukemia in vitro and in vivo.

Methods

Flow cytometry, ELISA and MTT methods were performed for molecular expression, cell proliferation and cytotoxicity assays. Leukemia xenograft NOD/SCID mice were established by subcutaneous injection with K562 cells, and then treated with irradiated NKL cells.

Results

We found NKL-IL15 cells displayed a significant high cytolysis activity against both human leukemia cell lines and primary leukemia cells from patients, accompanied with up-regulated expression of molecules related to NK cell cytotoxicity such as perforin, granzyme B and NKp80. Moreover, cytokines secreted by NKL-IL15 cells, including TNF-α and IFN-γ, could induce the expression of NKG2D ligands on target cells, which increased the susceptibility of leukemia cells to NK cell-mediated cytolysis. Encouragingly, NKL-IL15 cells significantly inhibited the growth of leukemia cells in xenografted NOD/SCID mice and prolonged the survival of tumor-bearing mice dramatically. Furthermore, NKL-IL15 cells displayed stimulatory effects on hPBMCs, indicating the immunesuppressive status of leukemia patients could be improved by NKL-IL15 cell treatment.

Conclusions

These results provided evidence that IL-15 gene-modification could augment NK cell-mediated anti-human leukemia function, which would improve primary NK cell-based immunotherapy for leukemia in future.

Keywords

NK cell Interleukin 15 Gene modification Leukemia 

Notes

Funding

This work was supported by Grants from National Basic Research Program of China (No. 2013CB531503), National Natural Science Foundation of China (Nos. 81172789, 30972692).

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Medical Ethics Committee of Shandong University and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. This study was approved by the Institutional Review Board of Shandong University.

Informed consent

Informed consent was obtained from all individual participants and the parents of minor participant included in the study.

References

  1. Bray SM, Vujanovic L, Butterfield LH (2011) Dendritic cell-based vaccines positively impact natural killer and regulatory T cells in hepatocellular carcinoma patients. Clin Dev Immunol 2011:249–281.  https://doi.org/10.1155/2011/249281 CrossRefGoogle Scholar
  2. Cheng M, Chen Y, Xiao W et al (2013) NK cell-based immunotherapy for malignant diseases. Cell Mol Immunol 10:230–252  https://doi.org/10.1038/cmi.2013.10.CrossRefPubMedPubMedCentralGoogle Scholar
  3. Crucitti L, Crocchiolo R, Toffalori C et al (2015) Incidence, risk factors and clinical outcome of leukemia relapses with loss of the mismatched HLA after partially incompatible hematopoietic stem cell transplantation. Leukemia 29:1143–1152.  https://doi.org/10.1038/leu.2014.314 CrossRefPubMedGoogle Scholar
  4. de Thé H, Pandolfi PP, Chen Z (2017) Acute promyelocytic leukemia: a paradigm for oncoprotein-targeted cure. Cancer Cell 32(5):552–560.  https://doi.org/10.1016/j.ccell.2017.10.002 CrossRefPubMedGoogle Scholar
  5. Dubois S, Patel HJ, Zhang M et al (2008) Preassociation of IL-15 with IL-15R alpha-IgG1-Fc enhances its activity on proliferation of NK and CD8+/CD44high T cells and its antitumor action. J Immunol 180(4):2099–2106.  https://doi.org/10.4049/jimmunol.180.4.2099 CrossRefPubMedGoogle Scholar
  6. He YG, Mayhew E, Mellon J et al (2004) Expression and possible function of IL-2 and IL-15 receptors on human uveal melanoma cells. Investig Ophthalmol Vis Sci 45:4240–4246.  https://doi.org/10.1167/iovs.04-0599 CrossRefGoogle Scholar
  7. Jiang W, Zhang J, Tian Z (2008) Functional characterization of interleukin-15 gene transduction into the human natural killer cell line NKL. Cytotherapy 10:265–274.  https://doi.org/10.1080/14653240801965156 CrossRefPubMedGoogle Scholar
  8. Jiang W, Zhang C, Tian Z et al (2014) hIL-15 gene-modified human natural killer cells (NKL-IL15) augments the anti-human hepatocellular carcinoma effect in vivo. Immunobiology 219:547–553  https://doi.org/10.1016/j.imbio.2014.03.007 CrossRefPubMedGoogle Scholar
  9. Joyce S (2001) CD1d and natural T cells: how their properties jump-start the immune system. Cell Mol Life Sci 58:442–469.  https://doi.org/10.1007/PL00000869 CrossRefPubMedGoogle Scholar
  10. Kennedy MK, Glaccum M, Brown SN et al (2000) Reversible defects in natural killer and memory CD8 T cell lineages in IL-15 deficient mice. J Exp Med 191:771–780.  https://doi.org/10.1084/jem.191.5.771 CrossRefPubMedPubMedCentralGoogle Scholar
  11. Lion E, Willemen Y, Berneman ZN et al (2012) Natural killer cell immune escape in acute myeloid leukemia. Leukemia 26:2019–2026.  https://doi.org/10.1038/leu.2012.87 CrossRefPubMedGoogle Scholar
  12. Ljunggren HG, Malmberg KJ (2007) Prospects for the use of NK cells in immunotherapy of human cancer. Nat Rev Immunol 7:329–339.  https://doi.org/10.1038/nri2073 CrossRefPubMedGoogle Scholar
  13. Miller JS, Soignier Y, Panoskaltsis-Mortari A (2005) Successful adoptive transfer and in vivo expansion of human haploidentical NK cells in cancer patients. Blood 105:3051–3057.  https://doi.org/10.1182/blood-2004-07-2974 CrossRefPubMedGoogle Scholar
  14. Nagashima S, Mailliard R, Kashii Y et al (1998) Stable transduction of the interleukin-2 gene into human natural killer cell lines and their phenotypic and functional characterization in vitro and in vivo. Blood 91:3850–3861PubMedGoogle Scholar
  15. Ohteki T (2002) Critical Role for IL-15 in innate immunity. Curr Mol Med 2:371–380.  https://doi.org/10.2174/1566524023362519 CrossRefPubMedGoogle Scholar
  16. Pende D, Spaggiari GM, Marcenaro S et al (2005) Analysis of the receptor-ligand interactions in the natural killer-mediated lysis of freshly isolated myeloid or lymphoblastic leukemias: evidence for the involvement of the Poliovirus receptor (CD155) and Nectin-2 (CD112). Blood 105:2066–2073.  https://doi.org/10.1182/blood-2004-09-3548 CrossRefPubMedGoogle Scholar
  17. Pierson BA, Miller JS (1996) CD56+ bright and CD56+ dim natural killer cells in patients with chronic myelogenous leukemia progressively decrease in number, respond less to stimuli that recruit clonogenic natural killer cells, and exhibit decreased proliferation on a per cell basis. Blood 88:2279–2287PubMedGoogle Scholar
  18. Robertson MJ, Cochran KJ, Cameron C et al (1996) Characterization of a cell line, NKL, derived from an aggressive human natural killer cell leukemia. Exp Hematol 24:406–415PubMedGoogle Scholar
  19. Romanski A, Bug G, Becker S et al (2005) Mechanisms of resistance to natural killer cell-mediated cytotoxicity in acute lymphoblastic leukemia. Exp Hematol 33:344–352.  https://doi.org/10.1016/j.exphem.2004.11.006 CrossRefPubMedGoogle Scholar
  20. Romee R, Leong JW, Fehniger TA (2014) Utilizing cytokines to function-enable human NK cells for the immunotherapy of cancer. Scientifica (Cairo) 2014:205796.  https://doi.org/10.1155/2014/205796 PubMedCentralGoogle Scholar
  21. Ruggeri L, Capanni M, Urbani E et al (2002) Effectiveness of donor natural killer cell alloreactivity in mismatched hematopoietic transplants. Science 295:2097–2100.  https://doi.org/10.1126/science.1068440 CrossRefPubMedGoogle Scholar
  22. Stanietsky N, Mandelboim O et al (2010) Paired NK cell receptors controlling NK cytotoxicity. FEBS Lett 584:4895–4900  https://doi.org/10.1016/j.febslet.2010.08.047 CrossRefPubMedGoogle Scholar
  23. Sutlu T, Alici E (2009) Natural killer cell-based immunotherapy in cancer: current insights and future prospects. J Intern Med 26:154–181  https://doi.org/10.1111/j.1365-2796.2009.02121.x CrossRefGoogle Scholar
  24. Terme M, Ullrich E, Delahaye NF et al (2008) Natural killer cell-directed therapies: moving from unexpected results to successful strategies. Nat Immunol 9:486–494.  https://doi.org/10.1038/ni1580 CrossRefPubMedGoogle Scholar
  25. Willard VW, Klosky JL, Li C et al (2017) The impact of childhood cancer: perceptions of adult survivors. Cancer 123(9):1625–1634.  https://doi.org/10.1002/cncr.30514 CrossRefPubMedPubMedCentralGoogle Scholar
  26. Wodnar-Filipowicz A, Kalberer CP (2006) Function of natural killer cells in immune defence against human leukaemia. Swiss Med Wkly 136:359–364PubMedGoogle Scholar
  27. Xu D, Han Q, Hou Z et al (2016) miR-146a negatively regulates NK cell functions via STAT1 signaling. Cell Mol Immunol 14(8):712–720.  https://doi.org/10.1038/cmi.2015.113 CrossRefPubMedGoogle Scholar
  28. Yang Y, Han Q, Hou Z et al (2017) Exosomes mediate hepatitis B virus (HBV) transmission and NK-cell dysfunction. Cell Mol Immunol 14(5):465–475.  https://doi.org/10.1038/cmi.2016.24 CrossRefPubMedGoogle Scholar
  29. Zhang J, Sun R, Wei H et al (2004a) Characterization of interleukin-15 gene-modified human natural killer cells: implications for adoptive cellular immunotherapy. Haematologica 89:338–347PubMedGoogle Scholar
  30. Zhang J, Sun R, Wei H et al (2004b) Characterization of stem cell factor gene-modified human natural killer cell line, NK-92 cells: implication in NK cell-based adoptive cellular immunotherapy. Oncol Rep 11:1097–1106.  https://doi.org/10.3892/or.11.5.1097 PubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Institute of Immunopharmaceutical Sciences, School of Pharmaceutical SciencesShandong UniversityJinanChina

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