Tumour-infiltrating FOXP3(+) lymphocytes are associated with cytotoxic immune responses and good clinical outcome in oestrogen receptor-negative breast cancer - PubMed (original) (raw)

Tumour-infiltrating FOXP3(+) lymphocytes are associated with cytotoxic immune responses and good clinical outcome in oestrogen receptor-negative breast cancer

N R West et al. Br J Cancer. 2013.

Abstract

Background: Regulatory T cells (Tregs) are commonly identified by expression of the transcription factor FOXP3 and are conventionally thought to promote cancer progression by suppressing anti-tumour immune responses. We examined the relationship between FOXP3(+) tumour-infiltrating lymphocytes (TIL) and prognosis in oestrogen receptor (ER)-negative breast cancer, a tumour subtype with poor clinical outcome in which TIL are abundant.

Methods: FOXP3(+) and CD8(+) TIL were assessed by immunohistochemistry in a cohort of 175 ER- breast tumours. Results were confirmed in an independent data set of 78 ER- breast tumours with publically available gene expression data.

Results: High FOXP3(+) TIL levels were strongly associated with prolonged recurrence-free survival (HR=0.461, P=0.0002), particularly among basal-like tumours (HR=0.280, P=0.0001), for which FOXP3 status was independent of standard prognostic factors. Over 75% of FOXP3(+) TIL in triple negative breast tumours displayed a conventional CD4(+)CD25(+) Treg phenotype. Importantly, FOXP3(+) TIL were positively correlated with CD8(+) (cytotoxic) T cells (r(s)=0.76, P<0.0001), and were prognostically insignificant in tumours with low levels of CD8(+) TIL. These observations were confirmed in an independent cohort.

Conclusion: In contrast with current dogma, we show for the first time that FOXP3(+) TIL are associated with robust anti-tumour immunity and favourable prognosis in ER- breast cancer.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1

Figure 1

Association of FOXP3+ TIL with patient survival. (A) Representative immunohistochemical staining of FOXP3 (brown nuclei), with examples of intraepithelial (arrowhead) and stromal (arrows) FOXP3+ TIL. Original magnification, × 200. (B) Box plots representing FOXP3+ TIL levels among tumour epithelia and stroma. (C) Distribution of FOXP3+ TIL in cases with (_n_=93) or without (_n_=82) disease recurrence at final follow-up. (D) Association of FOXP3+ TIL with RFS in the training, validation, and combined cohorts (FOXP3 high, _n_=92; FOXP3 low, _n_=83).

Figure 2

Figure 2

Expression of CD4, CD25, and CD8 among FOXP3+ TIL in triple negative breast tumours. (A) Representative immunohistochemical (pseudocoloured) staining for FOXP3 (red), CD4 (green), and CD8 (blue). Examples of FOXP3+CD4+ (arrow), FOXP3–CD4+ (arrowhead), and FOXP3–CD8+ (asterisk) TIL are indicated. (B) Representative staining for FOXP3 (red), CD4 (green), and CD25 (blue). Examples of FOXP3+CD4+CD25+ (arrow) and FOXP3–CD4+CD25– (arrowhead) TIL are indicated. (C) Quantification of CD4 and CD8 expression and (D) CD4 and CD25 expression among FOXP3+ TIL for each tumour. Lines represent medians. *P<0.0001 vs all other groups, Mann–Whitney _U_-test.

Figure 3

Figure 3

Relationships between FOXP3+ TIL, cytotoxic T cells, and patient outcome. (A) Spearman’s correlation of FOXP3+ TIL vs total (left panel), intraepithelial (middle panel), and stromal (right panel) CD8+ TIL. The solid line represents the line of best fit, with 95% confidence intervals indicated as dashed lines. (B) RFS based on high (⩾upper quartile, _n_=37) or low (_n_=105) levels of intraepithelial CD8+ TIL (left panel), with box plots indicating FOXP3+ TIL levels in each group. (C, left panel) RFS among patients with high (⩾1, _n_=51) and low (<1, _n_=59) ratios of intraepithelial CD8+ to FOXP3+ TIL. Cases with no detectable FOXP3+ TIL were omitted. (C, right panel) Box plots representing FOXP3+ and intraepithelial CD8+ TIL levels in the indicated patient groups. (D) Association of FOXP3 (high _n_=30; low _n_=70) with RFS among patients with low levels of intraepithelial CD8+ TIL.

Figure 4

Figure 4

Association of Tregs with prognosis and immune parameters in an independent data set. (A) Five-year RFS of patients with high (_n_=38) and low (_n_=40) levels of CD25 expression. (B) Five-year RFS of patients with high (_n_=47) and low (_n_=47) levels of CD8A expression, with corresponding box plots indicating CD25 expression. (C) Association of CD25 (high _n_=11; low _n_=30) with RFS among patients with low CD8A expression. (D) Gene expression modules indicative of differentiated cytotoxic T cells, activated dendritic cells, inflammatory cytokines, and chemokines relative to CD25 status. In all panels, asterisks represent Mann–Whitney _U_-test significance levels as follows: ****P<0.0001, ***_P_=0.001–0.01, **_P_=0.01–0.05.

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References

    1. Allan SE, Crome SQ, Crellin NK, Passerini L, Steiner TS, Bacchetta R, Roncarolo MG, Levings MK (2007) Activation-induced FOXP3 in human T effector cells does not suppress proliferation or cytokine production. Int Immunol 19(4): 345–354 - PubMed
    1. Aruga T, Suzuki E, Saji S, Horiguchi S, Horiguchi K, Sekine S, Kitagawa D, Funata N, Toi M, Sugihara K, Kuroi K (2009) A low number of tumour-infiltrating FOXP3-positive cells during primary systemic chemotherapy correlates with favourable anti-tumour response in patients with breast cancer. Oncol Rep 22(2): 273–278 - PubMed
    1. Baker K, Lachapelle J, Zlobec I, Bismar TA, Terracciano L, Foulkes WD (2011) Prognostic significance of CD8+ T lymphocytes in breast cancer depends upon both oestrogen receptor status and histological grade. Histopathology 58(7): 1107–1116 - PubMed
    1. Bates GJ, Fox SB, Han C, Leek RD, Garcia JF, Harris AL, Banham AH (2006) Quantification of regulatory T cells enables the identification of high-risk breast cancer patients and those at risk of late relapse. J Clin Oncol 24(34): 5373–5380 - PubMed
    1. Blatner NR, Bonertz A, Beckhove P, Cheon EC, Krantz SB, Strouch M, Weitz J, Koch M, Halverson AL, Bentrem DJ, Khazaie K (2010) In colourectal cancer mast cells contribute to systemic regulatory T-cell dysfunction. Proc Natl Acad Sci USA 107(14): 6430–6435 - PMC - PubMed

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