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Mechanistic analysis of the antitumor efficacy of human natural killer cells against breast cancer cells

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Abstract

We investigated the role of human natural killer (NK) cells in the peripheral blood (PB) and liver in controlling breast cancer. The proportion of NK cells among liver mononuclear cells was significantly higher than among PB mononuclear cells. Liver NK cells inductively expressed higher levels of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) than PB NK cells in response to interleukin-2 (IL-2). Liver NK cells displayed higher cytotoxicity against various breast cancer cell lines (MDA-MB231, MDA-MB453, MDA-MB468, and MCF-7) after IL-2 stimulation than did PB NK cells. Anti-HER2 monoclonal antibody (mAb) promoted the cytotoxicity of both the types of NK cells toward HER2-expressing cell lines. All breast cancer cell lines highly expressed death-inducing TRAIL receptors, death receptor 4, but did not express death-inhibitory receptors (DcR1 and DcR2). Both PB and liver NK cell-induced cytotoxicity was inhibited partially by anti-TRAIL mAb and more profoundly by the combination of anti-TRAIL mAb and concanamycin A, indicating that TRAIL and perforin are involved. IL-2-stimulated liver and PB NK cells exhibited upregulated expression of CXCR3, which bind to the chemokines CXCL9, CXCL10, and CXCL11 secreted by breast cancer cells. We also found that IFN-γ promoted the production of CXCL10 from breast cancer cells. The results of this study show that IFN-γ secreted from NK cells likely promotes the production of CXCL10 from breast cancer cells, which in turn accelerates the migration of CXCR3-expressing NK cells into the tumor site. These findings suggest the possibility of a therapeutic approach by either activation of endogenous PB and liver NK cells or adoptive transfer of in vitro-activated autologous NK cells.

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Abbreviations

NK:

Natural killer

TRAIL:

TNF-related apoptosis-inducing ligand

PBMC:

Peripheral blood mononuclear cell

LMNC:

Liver mononuclear cell

mAbs:

Monoclonal antibodies

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Acknowledgments

We thank Drs. Kohei Ishiyama, and Masahiro Ohira for their advice and encouragement and Drs. Doskali Marlen, Yuka Igarashi and Nabin Basnet, and Ms. Yuko Ishida and Ms. Midori Kiyokawa for their expert technical assistance. This work was supported by a Grant-in-Aid for Scientific Research (A) from the Japan Society for the Promotion of Science and a Grant-in-Aid for the Research on Hepatitis and BSE from the Japanese Ministry of Health, Labour and Welfare.

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Authors and Affiliations

  1. Division of Frontier Medical Science, Department of Surgery, Programs for Biomedical Research, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan

    Keiko Kajitani, Yuka Tanaka & Hideki Ohdan

  2. Department of Anatomical Pathology, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan

    Koji Arihiro

  3. Division of Frontier Nursing Science, Department of Health Care for Adult, Graduate School of Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan

    Tsuyoshi Kataoka

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  1. Keiko Kajitani
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  2. Yuka Tanaka
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  4. Tsuyoshi Kataoka
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  5. Hideki Ohdan
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Correspondence to Hideki Ohdan.

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Fig. 1

The results of 4 other similar experiments using liver NK cells to that shown in Fig. 7. Isolated liver NK cell populations were used as effector cells (E) in assays of cytotoxicity against the target (T) MDA-MB231 human breast cancer cell line. Cytotoxicity assays were performed at an E/T ratio of 10:1, 5:1, or 2.5:1 in the presence or absence of anti-TRAIL (N2B2) mAb (10 μg/mL) and/or concanamycin A (CMA) (50 nmol/L). Data are the average ± SEM values from triplicate sample. Error bars not shown appear within the data point. (TIFF 103 kb)

Fig. 2

The results of 4 other similar experiments using PB NK cells to that shown in Fig. 7. Isolated PB NK cell populations were used as effector cells (E) in assays of cytotoxicity against the target (T) MDA-MB231 human breast cancer cell line. Cytotoxicity assays were performed at an E/T ratio of 10:1, 5:1, or 2.5:1 in the presence or absence of anti-TRAIL (N2B2) mAb (10 μg/mL) and/or concanamycin A (CMA) (50 nmol/L). Data are the average ± SEM values from triplicate sample. Error bars not shown appear within the data point. (TIFF 102 kb)

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Kajitani, K., Tanaka, Y., Arihiro, K. et al. Mechanistic analysis of the antitumor efficacy of human natural killer cells against breast cancer cells. Breast Cancer Res Treat 134, 139–155 (2012). https://doi.org/10.1007/s10549-011-1944-x

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  • DOI: https://doi.org/10.1007/s10549-011-1944-x

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