The number of FoxP3-positive tumor-infiltrating lymphocytes in patients with synchronous bilateral breast cancer

  • Risa GotoEmail author
  • Yuko Hirota
  • Tomoyuki Aruga
  • Shinichiro Horiguchi
  • Sakiko Miura
  • Seigo Nakamura
  • Masafumi Takimoto
Original Article



In breast cancer, FoxP3-positive tumor-infiltrating lymphocytes (FoxP3+ TILs) vary depending on lymph node status, histological grade, and subtype. All these studies have compared the numbers of FoxP3+ TILs among different hosts, but recruitment of FoxP3+ TILs might depend on each individual’s immune environment and each tumor’s biological characteristics. In the present study, FoxP3+ TIL numbers were investigated in patients with synchronous bilateral breast cancer (SBBC) to determine the factors that affect FoxP3+ TIL recruitment in the same anti-tumor immune environment.


Patients diagnosed with SBBC who underwent curative surgery at two institutions were enrolled in this study. Patients who underwent primary systemic therapy or who were diagnosed with ductal carcinoma in situ or who had distant metastases at diagnosis were excluded. The average numbers of Foxp3+ TILs were determined from the scores of five high-power microscopic fields (HPF). The associations between Foxp3+ TIL numbers and the clinicopathological features of bilateral breasts in a single individual were examined.


Nuclear grade (NG) (p = 0.007) and subtype (p = 0.03), but not size (p = 0.18) and axillary lymph node (p = 0.23) were significantly associated with increase of FoxP3 + TIL numbers by univariate analysis. Further, only NG was a statistically significant clinicopathological factor for change in the number of FoxP3+ TILs by multivariate analysis (p = 0.046)


There was no relationship between FoxP3+ TIL numbers and cancer progression as reflected in tumor size and axillary lymph node in patients with SBBC. Aggressive biological factors, especially high NG, were significantly related to enhanced recruitment of FoxP3+ TILs.


Synchronous bilateral breast cancer Tumor-infiltrating lymphocytes FoxP3 Regulatory T cell Anti-tumor immunity environment 



We thank Ms. Tomoko Nagai, medical technician of department of pathology Showa University School of Medicine, for technical assistance with immunostaining method.


The present study was supported by Clinical Research Found of Tokyo Metropolitan Government (Grant no. H290303015).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

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. The study was approved by the ethics committee of Showa University Hospital (approval number #1969), and Tokyo Metropolitan Cancer and Infectious Diseases Center Komagome Hospital (approval number #1638).

Informed consent

Participants comprehensively provided their consent stating that the tissue samples from resected specimen may be used for various researches. The ethics committee of Showa University Hospital (approval number #1969), and Tokyo Metropolitan Cancer and Infectious Diseases Center Komagome Hospital (approval number #1638) approved the authors’ request for waiver of informed consent based on ethical consideration. All patients have the option to confirm ongoing studies on Showa University and Tokyo Metropolitan Cancer and Infectious Diseases Center Komagome Hospital web sites and may choose to opt out of consent at any time. The ethics committee approved this consent procedure.

Supplementary material

12282_2020_1049_MOESM1_ESM.tif (1.9 mb)
Supplementary Fig. 1 Flow diagram describe patient’s inclusion in present study file1 (TIF 1963 kb)
12282_2020_1049_MOESM2_ESM.tif (4.9 mb)
Supplementary Fig. 2 The median of FoxP3-positive tumor-infiltrating lymphocytes (FoxP3+ TILs) increased in order from luminal type with nuclear grade (NG) 1/2, luminal type with NG3, HER2 type, to basal-type (TIF 5000 kb)


  1. 1.
    Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144(5):646–74.CrossRefGoogle Scholar
  2. 2.
    Pardoll DM. The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer. 2012;12:252–64.CrossRefGoogle Scholar
  3. 3.
    Fridman WH, Zitvogel L, Sautès-Fridman C, Kroemer G. The immune contexture in cancer prognosis and treatment. Nat Rev Clin Oncol. 2017;14(12):717–34.CrossRefGoogle Scholar
  4. 4.
    Wing K, Sakaguchi S. Regulatory T cells exert checks and balances on self tolerance and autoimmunity. Nat Immunol. 2010; 11(1):7–13CrossRefGoogle Scholar
  5. 5.
    Levings MK, Sangregorio R, Human RMG. cd25(+)cd4(+) t regulatory cells suppress naive and memory T cell proliferation and can be expanded invitro without loss of function. J Exp Med. 2001;193(11):1295–302.CrossRefGoogle Scholar
  6. 6.
    Ng WF, Duggan PJ, Ponchel F, Matarese G, Lombardi G, Edwards AD, et al. Human CD4(+)CD25(+) cells: a naturally occurring population of regulatory T cells. Blood. 2001;98(9):2736–44.CrossRefGoogle Scholar
  7. 7.
    Shevach EM. CD4+ CD25+ suppressor T cells: more questions than answers. Nat Rev Immunol. 2002;2(6):389–400.CrossRefGoogle Scholar
  8. 8.
    Bach JF. Regulatory T cells under scrutiny. Nat Rev Immunol. 2003;3(3):189–98.CrossRefGoogle Scholar
  9. 9.
    Coffer PJ, Burgering BM. Forkhead-box transcription factors and their role in the immune system. Nat Rev Immunol. 2004;4(11):889–99.CrossRefGoogle Scholar
  10. 10.
    Hori S, Nomura T, Sakaguchi S. Control of regulatory T cell development by the transcription factor Foxp3. Sci (N Y). 2003;299(5609):1057–61.CrossRefGoogle Scholar
  11. 11.
    Sakaguchi S, Ono M, Setoguchi R, Yagi H, Hori S, Fehervari Z, et al. Foxp3+ CD25+ CD4+ natural regulatory T cells in dominant self-tolerance and autoimmune disease. Immunol Rev. 2006;212:8–27.CrossRefGoogle Scholar
  12. 12.
    deLeeuw RJ, Kost SE, Kakal JA, Nelson BH. The prognostic value of FoxP3+ tumor-infiltrating lymphocytes in cancer: a critical review of the literature. Clin Cancer Res. 2012;18(11):3022–9.CrossRefGoogle Scholar
  13. 13.
    Sakaguchi S. Naturally arising Foxp3-expressing CD25 + CD4+ regulatory T cells in immunological tolerance to self and non-self. Nat Immunol. 2005;6(4):345–52.CrossRefGoogle Scholar
  14. 14.
    Jiang D, Gao Z, Cai Z, Wang M, He J. Clinicopathological and prognostic significance of FOXP3+ tumor infiltrating lymphocytes in patients with breast cancer: a meta-analysis. BMC Cancer. 2015;15:727.CrossRefGoogle Scholar
  15. 15.
    Nichol AM, Yerushalmi R, Tyldesley S, Lesperance M, Bajdik CD, Speers C, et al. A case-match study comparing unilateral with synchronous bilateral breast cancer outcomes. J Clin Oncol. 2011;29(36):4763–8.CrossRefGoogle Scholar
  16. 16.
    Verkooijen HM, Chatelain V, Fioretta G, Vlastos G, Rapiti E, Sappino AP, et al. Survival after bilateral breast cancer: results from a population-based study. Breast Cancer Res Treat. 2007;105(3):347–57.CrossRefGoogle Scholar
  17. 17.
    Kurebayashi J, Miyosi Y, Ishikawa T, Saji S, Sugie T, Suzuki T, et al. Clinicopathological characteristics of breast cancer and trends in the management of breast cancer patients in Japan: Based on the Breast Cancer Registry of the Japanese Breast Cancer Society between 2004 and 2011. Breast Cancer. 2015;22(3):235–44.CrossRefGoogle Scholar
  18. 18.
    Japanese Breast Cancer Society. General rules for clinical and pathological recording of breast cancer, the 17 Kanehara & Co. Ltd, Tokyo; 2012.Google Scholar
  19. 19.
    Wolff AC, Hammond ME, Hicks DG, Dowsett M, McShane LM, Allison KH, et al. Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline update. J Clin Oncol. 2013;31(31):3997–4013.CrossRefGoogle Scholar
  20. 20.
    Aruga T, Suzuki E, Saji S, Horiguchi S, Horiguchi K, Kitagawa D, et al. A low number of tumor-infiltrating FOXP3-positive cells during primary systemic chemotherapy correlates with favorable anti-tumor response in patients with breast cancer. Oncol Rep. 2009;22:273–8.PubMedGoogle Scholar
  21. 21.
    Kanda Y. Investigation of the freely available easy-to-use software ‘EZR’ for medical statistics. Bone Marrow Transpl. 2013;48(3):452–8.CrossRefGoogle Scholar
  22. 22.
    Padmanabhan N, Subramanyan A, Radhakrishna S. Synchronous bilateral breast cancers. J Clin Diagn Res. 2015;9(9):XC05–8.Google Scholar
  23. 23.
    Renz DM, Böttcher J, Baltzer PA, Dietzel M, Vag T, Gajda M, et al. The contralateral synchronous breast carcinoma: a comparison of histology, localization, and magnetic resonance imaging characteristics with the primary index cancer. Breast Cancer Res Treat. 2010;120(2):449–59.CrossRefGoogle Scholar
  24. 24.
    Baker B, Morcos B, Daoud F, Sughayer M, Shabani H, Salameh H, et al. Histo-biological comparative analysis of bilateral breast cancer. Med Oncol. 2013;30(4):711.CrossRefGoogle Scholar
  25. 25.
    Liu F, Lang R, Zhao J, Zhang X, Fan Y, et al. CD8+ cytotoxic T cell and FOXP3+ regulatory T cell infiltration in relation to breast cancer survival and molecular subtypes. Breast Cancer Res Treat. 2011;130(2):645–55.CrossRefGoogle Scholar
  26. 26.
    Martin F, Ladoire S, Mignot G, Apetoh L, Ghiringhelli G. Human FOXP3 and cancer. Oncogene. 2010;29:4121–9.CrossRefGoogle Scholar

Copyright information

© The Japanese Breast Cancer Society 2020

Authors and Affiliations

  1. 1.Department of Breast Surgical OncologyTokyo Metropolitan Cancer and Infectious Diseases Center Komagome HospitalTokyoJapan
  2. 2.Department of PathologyShowa UniversityTokyoJapan
  3. 3.Department of PathologyShowa University Koto Toyosu HospitalTokyoJapan
  4. 4.Department of PathologyTokyo Metropolitan Cancer and Infectious Diseases Center Komagome HospitalTokyoJapan
  5. 5.Department of Surgery, Division of Breast Surgical OncologyShowa UniversityTokyoJapan

Personalised recommendations