In the present study, we use high-dimensional flow cytometry, functional assays, T-cell receptor (TCR) repertoire analysis, and RNA sequencing to characterize the immune phenotype, function and dynamics of Tregs present in paired I and NI TDLNs, and tumor; and identified CD80-expressing Tregs as a subset of Tregs associated with bad prognosis in breast cancer patients. The gene discussed is CD80; the disease is breast cancer.