Cancer Immunology, Immunotherapy

, Volume 68, Issue 4, pp 577–585 | Cite as

A potential role for peripheral natural killer cell activity induced by preoperative chemotherapy in breast cancer patients

  • Ryungsa KimEmail author
  • Ami Kawai
  • Megumi Wakisaka
  • Yuri Funaoka
  • Naomi Yasuda
  • Masayuki Hidaka
  • Yukitaka Morita
  • Shoichro Ohtani
  • Mitsuya Ito
  • Koji Arihiro
Original Article


Tumor-infiltrating lymphocytes are an important prognostic factor after neoadjuvant chemotherapy (NAC) in patients with breast cancer. Natural killer (NK) cells play critical roles in antitumor immune surveillance. Here, we assessed the relationship between peripheral natural killer (pNK) cell activity, tumor microenvironmental factors (TMEFs), and the therapeutic efficacy of preoperative chemotherapy in patients with breast cancer. In a cohort of 39 patients diagnosed with stage II–IV breast cancer who received NAC, we measured pNK cell activity by chromium release assay and assessed TMEF levels by next-generation sequencing. Following NAC, pNK cell activity was increased in 24/39 patients but decreased in 15/39 patients. Increased pNK cell activity following preoperative chemotherapy was associated significantly with the disappearance of axillary lymph node metastasis (Ax+; p = 0.0235). Increased pNK cell activity remained significantly associated with the disappearance of Ax+ in multivariate logistic regression analysis (OR 5.41, 95% CI 1.19–24.52, p = 0.0283). A Grade 2 or higher effect of NAC was associated with high pre-NAC cytotoxic T lymphocyte-associated protein 4 (CTLA-4) levels (p = 0.0281) and elevated post-NAC NK (p = 0.0005) cells and transforming growth factor-beta (TGF-β; p = 0.0350) levels. The disappearance of Ax+ was associated with high pre-NAC CTLA-4 levels (p = 0.0278) and elevated CD4 levels after NAC (p = 0.0250). The systemic activation of pNK cells after NAC may improve metastatic tumor elimination in patients with breast cancer owing to a release from local immunosuppression, and immune activation in the tumor microenvironment.


Breast cancer Preoperative chemotherapy Immune response Peripheral natural killer cell (pNK) activity Transforming growth factor β Tumor microenvironment 



Axillary lymph node metastasis


Confidence interval


Cytotoxic T lymphocyte-associated protein 4






Formalin-fixed paraffin embedded


Human epidermal growth factor receptor-2


Nanoparticle albumin-bound-paclitaxel


Neoadjuvant chemotherapy


Odds ratio


Peripheral natural killer


Tumor microenvironmental factor


Triple negative





The authors thank the patients and their families for their participation in the study. The authors also thank SRL. Inc. (Tokyo, Japan) for the measurement of pNK cell activity.

Author contributions

Conception and design: RK. Collection and assembly of data: RK, AK, MW, YF, NY, MH, YM, SO, MI, KA. Data analysis and interpretation: RK, AK, MH, YM. Manuscript writing: RK, YM. Final approval of manuscript: all authors.


No specific funding was received for this study.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

This study was approved by the research ethics committee of the Hiroshima Mark Clinic on July 1, 2012. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.


  1. 1.
    Denkert C, von Minckwitz G, Darb-Esfahani S, Lederer B, Heppner BI, Weber KE, Budczies J, Huober J, Klauschen F, Furlanetto J, Schmitt WD, Blohmer JU, Karn T, Pfitzner BM, Kümmel S, Engels K, Schneeweiss A, Hartmann A, Noske A, Fasching PA, Jackisch C, van Mackelenbergh M, Sinn P, Schem C, Hanusch C, Untch M, Loibl S (2018) Tumour-infiltrating lymphocytes and prognosis in different subtypes of breast cancer: a pooled analysis of 3771 patients treated with neoadjuvant therapy. Lancet Oncol 19:40–50. CrossRefGoogle Scholar
  2. 2.
    Shanker A, Marincola FM (2011) Cooperativity of adaptive and innate immunity: implications for cancer therapy. Cancer Immunol Immunother 60:1061–1074. CrossRefGoogle Scholar
  3. 3.
    Seo AN, Lee HJ, Kim EJ, Kim HJ, Jang MH, Lee HE, Kim YJ, Kim JH, Park SY (2013) Tumour-infiltrating CD8+ lymphocytes as an independent predictive factor for pathological complete response to primary systemic therapy in breast cancer. Br J Cancer 109:2705–2713. CrossRefGoogle Scholar
  4. 4.
    Verma C, Kaewkangsadan V, Eremin JM, Cowley GP, Ilyas M, El-Sheemy MA, Eremin O (2015) Natural killer (NK) cell profiles in blood and tumour in women with large and locally advanced breast cancer (LLABC) and their contribution to a pathological complete response (PCR) in the tumour following neoadjuvant chemotherapy (NAC): differential restoration of blood profiles by NAC and surgery. J Transl Med 13:180. CrossRefGoogle Scholar
  5. 5.
    Muraro E, Comaro E, Talamini R, Turchet E, Miolo G, Scalone S, Militello L, Lombardi D, Spazzapan S, Perin T, Massarut S, Crivellari D, Dolcetti R, Martorelli D (2015) Improved Natural Killer cell activity and retained anti-tumor CD8(+) T cell responses contribute to the induction of a pathological complete response in HER2-positive breast cancer patients undergoing neoadjuvant chemotherapy. J Transl Med 13:204. CrossRefGoogle Scholar
  6. 6.
    Brierley JD, Gospodarowicz MK, Wittekind C (eds) (2017) TNM classification of malignant tumours, 8th edn. Wiley-Blackwell, HobokenGoogle Scholar
  7. 7.
    Kurosumi M (2006) Significance and problems in evaluations of pathological responses to neoadjuvant therapy for breast cancer. Breast Cancer 13:254–259. CrossRefGoogle Scholar
  8. 8.
    Rouzier R, Extra JM, Klijanienko J, Falcou MC, Asselain B, Vincent-Salomon A, Vielh P, Bourstyn E (2002) Incidence and prognostic significance of complete axillary downstaging after primary chemotherapy in breast cancer patients with T1 to T3 tumors and cytologically proven axillary metastatic lymph nodes. J Clin Oncol 20:1304–1310CrossRefGoogle Scholar
  9. 9.
    Diaz-Botero S, Espinosa-Bravo M, Gonçalves VR, Esgueva-Colmenarejo A, Peg V, Perez J, Cortes J, Rubio IT (2016) Different prognostic implications of residual disease after neoadjuvant treatment: Impact of Ki 67 and site of response. Ann Surg Oncol 23:3831–3837CrossRefGoogle Scholar
  10. 10.
    Hayashi N, Takahashi Y, Matsuda N, Tsunoda H, Yoshida A, Suzuki K, Nakamura S, Yamauchi H (2018) The prognostic effect of changes in tumor stage and nodal status after neoadjuvant chemotherapy in each primary breast cancer subtype. Clin Breast Cancer 18:e219–e229. CrossRefGoogle Scholar
  11. 11.
    Slaney CY, Rautela J, Parker BS (2013) The emerging role of immunosurveillance in dictating metastatic spread in breast cancer. Cancer Res 73:5852–5857. CrossRefGoogle Scholar
  12. 12.
    Mamessier E, Sylvain A, Thibult ML, Houvenaeghel G, Jacquemier J, Castellano R, Gonçalves A, André P, Romagné F, Thibault G, Viens P, Birnbaum D, Bertucci F, Moretta A, Olive D (2011) Human breast cancer cells enhance self tolerance by promoting evasion from NK cell antitumor immunity. J Clin Invest 121:3609–3622. CrossRefGoogle Scholar
  13. 13.
    Pickup M, Novitskiy S, Moses HL (2013) The roles of TGFβ in the tumour microenvironment. Nat Rev Cancer 13:788–799. CrossRefGoogle Scholar
  14. 14.
    Kerdiles Y, Ugolini S, Vivier E (2013) T cell regulation of natural killer cells. J Exp Med 210:1065–1068. CrossRefGoogle Scholar
  15. 15.
    Ghiringhelli F, Menard C, Terme M, Flament C, Taieb J, Chaput N, Puig PE, Novault S, Escudier B, Vivier E, Lecesne A, Robert C, Blay JY, Bernard J, Caillat-Zucman S, Freitas A, Tursz T, Wagner-Ballon O, Capron C, Vainchencker W, Martin F, Zitvogel L (2005) CD4+ CD25+ regulatory T cells inhibit natural killer cell functions in a transforming growth factor-beta-dependent manner. J Exp Med 202:1075–1085CrossRefGoogle Scholar
  16. 16.
    Spitzer MH, Carmi Y, Reticker-Flynn NE, Kwek SS, Madhireddy D, Martins MM, Gherardini PF, Prestwood TR, Chabon J, Bendall SC, Fong L, Nolan GP, Engleman EG (2017) Systemic immunity is required for effective cancer immunotherapy. Cell 168:487–502. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Breast SurgeryHiroshima Mark ClinicHiroshimaJapan
  2. 2.Genetic Testing Gene ResearchHiroshimaJapan
  3. 3.Department of Breast SurgeryHiroshima City HospitalHiroshimaJapan
  4. 4.Department of Anatomical PathologyHiroshima University HospitalHiroshimaJapan

Personalised recommendations