Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

The breast tumor microenvironment alters the phenotype and function of natural killer cells

Cellular & Molecular Immunology volume 13, pages 628–639 (2016)Cite this article

Abstract

Natural killer (NK) cells are innate immune cells with the ability to identify and eliminate transformed cells. However, within tumors, many studies have described NK cells as non-functional. The developmental stage of tumor-associated NK cells and how this may relate to functionality has not been explored. We examined the developmental state of NK cells from polyoma middle T antigen (pyMT) transgenic mouse (MMTV-pMT) breast tumors. In pyMT tumors, NK cells were immature as evidenced by their decreased expression of DX5 and their CD27lowCD11blow phenotype. These immature NK cells also had increased expression of NKG2A and expressed low levels of NKp46, perforin, and granzyme B. In contrast, splenic NK cells isolated from the same mice maintained their maturity and their expression of activation markers. To delineate whether the tumor microenvironment directly alters NK cells, we adoptively transferred labeled NK cells and followed their activation status in both the spleen and the tumor. NK cells that arrived at the tumor had half the expression of NKp46 within three days of transfer in comparison to those which arrived at the spleen. In an effort to modify the tumor microenvironment and assess the plasticity of intratumoral NK cells, we treated pyMT tumors with IL-12 and anti-TGF-β. After one week of treatment, the maturity of tumor-associated NK cells was increased; thus, indicating that these cells possess the ability to mature and become activated. A better understanding of how NK cells are modified by the tumor microenvironment will help to develop strategies aimed at bolstering immune responses against tumors.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. Srivastava S, Lundqvist A, Childs RW . Natural killer cell immunotherapy for cancer: a new hope. Cytotherapy 2008; 10: 775–783.

    Article  CAS  Google Scholar 

  2. Kiessling R, Klein E, Wigzell H . “Natural” killer cells in the mouse. I. Cytotoxic cells with specificity for mouse Moloney leukemia cells. Specificity and distribution according to genotype. Eur J Immunol 1975; 5: 112–117.

    Article  CAS  Google Scholar 

  3. Gillgrass A, Ashkar A . Stimulating natural killer cells to protect against cancer: recent developments. Expert Rev Clin Immunol 2011; 7: 367–382.

    Article  CAS  Google Scholar 

  4. Vivier E, Colonna M . (Eds.). Immunobiology of Natural Killer Cell Receptors. Springer-Verlag Berlin Heidelberg, 2006, Vol. VIII, p. 284.

    Google Scholar 

  5. Di Santo JP . Natural killer cell developmental pathways: a question of balance. Annu Rev Immunol 2006; 24: 257–286.

    Article  CAS  Google Scholar 

  6. Puzanov IJ, Bennett M, Kumar V . IL-15 can substitute for the marrow microenvironment in the differentiation of natural killer cells. J Immunol 1996; 157: 4282–4285.

    CAS  PubMed  Google Scholar 

  7. Huntington ND, Vosshenrich CA, Di Santo JP . Developmental pathways that generate natural-killer-cell diversity in mice and humans. Nat Rev Immunol 2007; 7: 703–714.

    Article  CAS  Google Scholar 

  8. Chiossone L et al. Maturation of mouse NK cells is a 4-stage developmental program. Blood 2009; 113: 5488–5496.

    Article  CAS  Google Scholar 

  9. Fu B et al. CD11b and CD27 reflect distinct population and functional specialization in human natural killer cells. Immunology 2011; 133: 350–359.

    Article  CAS  Google Scholar 

  10. Kim S et al. In vivo natural killer cell activities revealed by natural killer cell-deficient mice. Proc Natl Acad Sci U S A 2000; 97: 2731–2736.

    Article  CAS  Google Scholar 

  11. Street SE et al. Host perforin reduces tumor number but does not increase survival in oncogene-driven mammary adenocarcinoma. Cancer Res 2007; 67: 5454–5460.

    Article  CAS  Google Scholar 

  12. Mamessier E et al. Human breast cancer cells enhance self tolerance by promoting evasion from NK cell antitumor immunity. J Clin Invest 2011; 121: 3609–3622.

    Article  CAS  Google Scholar 

  13. Richards J et al. Tumor growth decreases NK and B cells as well as common lymphoid progenitor. PLoS One 2008; 3: e3180.

    Article  Google Scholar 

  14. Richards JO et al. Tumor growth impedes natural-killer-cell maturation in the bone marrow. Blood 2006; 108: 246–252.

    Article  CAS  Google Scholar 

  15. Xia J et al. Prevention of spontaneous breast carcinoma by prophylactic vaccination with dendritic/tumor fusion cells. J Immunol 2003; 170: 1980–1986.

    Article  CAS  Google Scholar 

  16. De Colvenaer V et al. CD27-deficient mice show normal NK-cell differentiation but impaired function upon stimulation. Immunol Cell Biol 2011; 89: 803–811.

    Article  CAS  Google Scholar 

  17. Rosmaraki EE et al. Identification of committed NK cell progenitors in adult murine bone marrow. Eur J Immunol 2001; 31: 1900–1909.

    Article  CAS  Google Scholar 

  18. Waldhauer I, Steinle A . NK cells and cancer immunosurveillance. Oncogene 2008; 27: 5932–5943.

    Article  CAS  Google Scholar 

  19. Vivier E et al. Functions of natural killer cells. Nat Immunol 2008; 9: 503–510.

    Article  CAS  Google Scholar 

  20. Nausch N, Cerwenka A . NKG2D ligands in tumor immunity. Oncogene 2008; 27: 5944–5958.

    Article  CAS  Google Scholar 

  21. Gasser S et al. The DNA damage pathway regulates innate immune system ligands of the NKG2D receptor. Nature 2005; 436: 1186–1190.

    Article  CAS  Google Scholar 

  22. Halfteck GG et al. Enhanced in vivo growth of lymphoma tumors in the absence of the NK-activating receptor NKp46/NCR1. J Immunol 2009; 182: 2221–2230.

    Article  CAS  Google Scholar 

  23. Arteaga CL et al. Anti-transforming growth factor (TGF)-beta antibodies inhibit breast cancer cell tumorigenicity and increase mouse spleen natural killer cell activity. Implications for a possible role of tumor cell/host TGF-beta interactions in human breast cancer progression. J Clin Invest 1993; 92: 2569–2576.

    Article  CAS  Google Scholar 

  24. Marcenaro E et al. IL-12 or IL-4 prime human NK cells to mediate functionally divergent interactions with dendritic cells or tumors. J Immunol 2005; 174: 3992–3998.

    Article  CAS  Google Scholar 

  25. Yuzhalin AE, Kutikhin AG . Interleukin-12: clinical usage and molecular markers of cancer susceptibility. Growth Factors 2012; 30: 176–191.

    Article  CAS  Google Scholar 

  26. Carrega P et al. Natural killer cells infiltrating human nonsmall-cell lung cancer are enriched in CD56 bright CD16(-) cells and display an impaired capability to kill tumor cells. Cancer 2008; 112: 863–875.

    Article  Google Scholar 

  27. Konjevic G et al. Investigation of NK cell function and their modulation in different malignancies. Immunol Res 2012; 52: 139–156.

    Article  CAS  Google Scholar 

  28. Gogali F et al. CD3(-)CD16(-)CD56(bright) immunoregulatory NK cells are increased in the tumor microenvironment and inversely correlate with advanced stages in patients with papillary thyroid cancer. Thyroid 2013; 23: 1561–1568.

    Article  CAS  Google Scholar 

  29. Garcia-Iglesias T et al. Low NKp30, NKp46 and NKG2D expression and reduced cytotoxic activity on NK cells in cervical cancer and precursor lesions. BMC Cancer 2009; 9: 186.

    Article  Google Scholar 

  30. Platonova S et al. Profound coordinated alterations of intratumoral NK cell phenotype and function in lung carcinoma. Cancer Res 2011; 71: 5412–5422.

    Article  CAS  Google Scholar 

  31. Narni-Mancinelli E et al. Fate mapping analysis of lymphoid cells expressing the NKp46 cell surface receptor. Proc Natl Acad Sci U S A 2011; 108: 18324–18329.

    Article  CAS  Google Scholar 

  32. Gill S et al. Rapid development of exhaustion and downregulation of eomesodermin limits the anti-tumor activity of adoptively transferred murine natural killer cells. Blood 2012; 119: 5758–5768.

    Article  CAS  Google Scholar 

  33. Wilson EB et al. Human tumour immune evasion via TGF-beta blocks NK cell activation but not survival allowing therapeutic restoration of anti-tumour activity. PLoS One 2011; 6: e22842.

    Article  CAS  Google Scholar 

  34. Allan DS et al. TGF-beta affects development and differentiation of human natural killer cell subsets. Eur J Immunol 2010; 40: 2289–2295.

    Article  CAS  Google Scholar 

  35. Li J, Hunter CA, Farrell JP . Anti-TGF-beta treatment promotes rapid healing of Leishmania major infection in mice by enhancing in vivo nitric oxide production. J Immunol 1999; 162: 974–979.

    CAS  PubMed  Google Scholar 

  36. Loza MJ et al. Expression of type 1 (interferon gamma) and type 2 (interleukin-13, interleukin-5) cytokines at distinct stages of natural killer cell differentiation from progenitor cells. Blood 2002; 99: 1273–1281.

    Article  CAS  Google Scholar 

  37. Bruno A et al. The proangiogenic phenotype of natural killer cells in patients with non-small cell lung cancer. Neoplasia 2013; 15: 133–142.

    Article  CAS  Google Scholar 

  38. Lima PD et al. Heterogeneity in composition of mouse uterine natural killer cell granules. J Leukoc Biol 2012; 92: 195–204.

    Article  CAS  Google Scholar 

  39. Jin J et al. CD11b(-)CD27(-) NK cells are associated with the progression of lung carcinoma. PLoS One 2013; 8: e61024.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by a grant from the Canadian Breast Cancer Foundation (CBCF). Amy Gillgrass was supported by a PhD. Fellowship from CBCF.

Author information

Author notes

  1. Tamara Krneta and Amy Gillgrass: Both Tamara Krneta and Amy Gillgrass contributed equally to this work.

Authors and Affiliations

  1. Department of Pathology and Molecular Medicine, McMaster Immunology Research Center and Institute for Infectious Disease, McMaster University, Hamilton, ON, Canada

    Tamara Krneta, Amy Gillgrass, Marianne Chew & Ali A. Ashkar

Authors
  1. Tamara Krneta
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Amy Gillgrass
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Marianne Chew
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ali A. Ashkar
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ali A. Ashkar.

Ethics declarations

Competing interests

All authors declare no competing financial interests.

Additional information

Supplementary information accompanies the paper on Cellular & Molecular Immunology website (http://www.nature.com/cmi).

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Krneta, T., Gillgrass, A., Chew, M. et al. The breast tumor microenvironment alters the phenotype and function of natural killer cells. Cell Mol Immunol 13, 628–639 (2016). https://doi.org/10.1038/cmi.2015.42

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/cmi.2015.42

Keywords

This article is cited by

Search

Advanced search

Quick links