Background:
Older age is associated with decreased immune function but younger patients present with more aggressive, higher-grade breast cancer (BC)1, and poorer response to immune checkpoint blockade2. We therefore investigated age-related differences in systemic immunity of early-stage breast cancer patients.
Methods:
We collected peripheral blood mononuclear cells (PBMCs) from 132 early-stage BC patients, and subdivided them into young (≤39 years), middle-aged (40-59 years), and older (≥60 years) groups. Age-matched healthy donors were also analysed (n=15 per group). We investigated peripheral blood immune phenotypes of patients by age with multiparameter flow cytometric analysis, and correlations between peripheral blood and tumour through T cell receptor sequencing (TCR-seq) using genomic DNA.
Results:
Median age of our patient cohort was 48 (range 24-83), with 32 young (24%), 75 middle aged (57%), and 25 older (19%) patients. Subtype distribution was predominantly ER+/HER2- (51%), with samples mostly node positive (52%) and grade 3 (70%), and a median tumor size of 27.5 mm. Flow cytometry data showed significantly more CD8+ memory T cell subsets with older age (P<0.0001). Moreover, an increased inflammatory phenotype with significantly higher IFN-γ+ and TNF-α+ CD8+ T cells in older BC patients compared with healthy donors (TNF-α P=0.01) and younger BC patients (P<0.0001; both TNF-α and IFN-γ) was observed. There was also significantly decreased PD-1 expression in peripheral blood CD8+ T cells in older BC patients compared to their healthy counterparts (P=0.03). Through TCR-seq analysis, we identified a 10.5 fold increase in peripheral hyperexpanded TCRs (P<0.0001) and a 57% increase in overlap of TCRs from tumour-enriched clones and PBMCs (P=0.03) in older patients (n=6) compared with young (n=5). Clonal expansion of virus-specific T cells (e.g. EBV and CMV) in breast tumour tissue was also observed in both young and older patients. Due to low power, we did not segregate by subtype.
Conclusions:
This study revealed significant differences in peripheral T cell immune responses to early-stage breast cancer with age which may account for the differential responses observed in young and older patients to immune checkpoint blockade. Further phenotypic and functional analysis of T cells from patients’ tumours is clearly warranted.