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Cancer Immunology, Immunotherapy

, Volume 67, Issue 4, pp 575–587 | Cite as

Ex vivo-expanded NK cells from blood and ascites of ovarian cancer patients are cytotoxic against autologous primary ovarian cancer cells

  • Tina Nham
  • Sophie M. Poznanski
  • Isabella Y. Fan
  • Mira M. Shenouda
  • Marianne V. Chew
  • Amanda J. Lee
  • Fatemeh Vahedi
  • Yalda Karimi
  • Martin Butcher
  • Dean A. Lee
  • Hal Hirte
  • Ali A. Ashkar
Original Article

Abstract

Ovarian cancer (OC) is the leading cause of gynecological cancer-related death in North America. Most ovarian cancer patients (OCPs) experience disease recurrence after first-line surgery and chemotherapy; thus, there is a need for novel second-line treatments to improve the prognosis of OC. Although peripheral blood-derived NK cells are known for their ability to spontaneously lyse tumour cells without prior sensitization, ascites-derived NK cells (ascites-NK cells) isolated from OCPs exhibit inhibitory phenotypes, impaired cytotoxicity and may play a pro-tumourigenic role in cancer progression. Therefore, it is of interest to improve the cytotoxic effector function of impaired OCP ascites-NK cells at the tumour environment. We investigated the efficacy of using an artificial APC-based ex vivo expansion technique to generate cytotoxic, expanded NK cells from previously impaired OCP ascites-NK cells, for use in an autologous model of NK cell immunotherapy. We are the first to obtain a log-scale expansion of OCP ascites-NK cells that upregulate the surface expression of activating receptors NKG2D, NKp30, NKp44, produce robust amounts of anti-tumour cytokines in the presence of OC cells and mediate direct tumour cytotoxicity against ascites-derived, primary OC cells obtained from autologous patients. Our findings demonstrate that it is possible to generate cytotoxic OCP ascites-NK cells from previously impaired OCP ascites-NK cells, which presents a promising immunotherapeutic target for the second-line treatment of OC. Future work should focus on evaluating the in vivo efficacy of autologous NK cell immunotherapy through the intraperitoneal delivery of NK cell expansion factors to a preclinical xenograft mouse model of human OC.

Keywords

Ovarian cancer Natural killer cell cancer immunotherapy Ex vivo NK cell expansion Ascites Autologous natural killer cells 

Abbreviations

aAPC

Artificial antigen-presenting cell

CFSE

Carboxyfluorescein succinimidyl ester

E:T

Effector: target

HD

Healthy donor

IFN-γ

Interferon-γ

IL

Interleukin

NK cells

Natural killer cells

OC

Ovarian cancer

OCP

Ovarian cancer patient

OCP ascites-NK cells

NK cells derived from ovarian cancer patient ascites

PB

Peripheral blood

PB-NK cells

NK cells derived from peripheral blood

TNF-α

Tumor necrosis factor-α

VEGF

Vascular endothelial growth factor

Notes

Acknowledgements

We thank Dr. Karen Mossman for providing us with the OVCAR-8 cell line. We also thank Jann Ang and Natasha Kazhdan for aiding us in sample collection.

Funding

Funding for this research was supported by grants from the Juravinski Hospital and Cancer Centre Foundation, the Canadian Breast Cancer Foundation and the Canadian Institutes of Health Research Tier 1 Canada Research Chair awarded to Dr. Ali A. Ashkar.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Cell line authentication

The OVCAR-8 cell line was verified using the American Type Culture Collection® (ATCC®) Cell Line Authentication Service for which we received a Short Tandem Repeat Profile Report on September 5, 2017. The submitted OVCAR-8 sample is an exact match to the short tandem repeat profile for the cell line OVCAR-8 listed on the Expasy website: http://web.expasy.org/cellosaurus/CVCL_1629.

Supplementary material

262_2017_2112_MOESM1_ESM.pdf (333 kb)
Supplementary material 1 (PDF 332 KB)

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Copyright information

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

Authors and Affiliations

  • Tina Nham
    • 1
  • Sophie M. Poznanski
    • 1
  • Isabella Y. Fan
    • 1
  • Mira M. Shenouda
    • 1
  • Marianne V. Chew
    • 1
  • Amanda J. Lee
    • 1
  • Fatemeh Vahedi
    • 1
  • Yalda Karimi
    • 1
  • Martin Butcher
    • 2
  • Dean A. Lee
    • 3
  • Hal Hirte
    • 4
  • Ali A. Ashkar
    • 1
  1. 1.Department of Pathology and Molecular Medicine, McMaster Immunology Research CentreMcMaster UniversityHamiltonCanada
  2. 2.Juravinski Cancer Centre McMaster UniversityHamiltonCanada
  3. 3.The Research Institute at Nationwide Children’s HospitalOhio State UniversityColumbusUSA
  4. 4.Division of Medical Oncology, Department of OncologyJuravinski Cancer CentreHamiltonCanada

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