The data of CD4+, CD8+, and FOXP3+ TILs and PD-L1+ ICs were adopted from our previous studies [25 (link), 26 (link)] for all of the cases of pre-invasive carcinoma and 307 cases of invasive carcinoma. Immunohistochemical staining had been carried out using the following antibodies: CD4 (clone SP35; ready to use; Dako), CD8 (clone C8/144B; ready to use; Dako), FOXP3 (clone 236A/E7; 1:100; Abcam) and PD-L1 (clone E1L3N; 1:100; Cell Signaling, Danvers, MA, USA). CD4+, CD8+, and FOXP3+ T cells had been counted in intratumoral and stromal areas as absolute numbers per high-power field. Detailed information on the counting method of TILs is described in the previous studies [25 (link), 26 (link)]. For this study, CD4+, CD8+, and FOXP3+ TILs were dichotomized into high- and low-infiltration groups using cutoff values obtained by ROC curve analyses. PD-L1+ ICs were considered to be present when at least 1% of the tumor stromal area was occupied by PD-L1+ ICs.
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