Effect of neoadjuvant chemotherapy on tumor-infiltrating lymphocytes and PD-L1 expression in breast cancer and its clinical significance

Abstract Background The effects of neoadjuvant chemotherapy on immune markers remain largely unknown. The specific aim of this study was to assess stromal tumor-infiltrating lymphocytes (TILs) and programmed death ligand 1 (PD-L1) protein expression in a cohort of breast cancer patients treated with neoadjuvant chemotherapy. Methods Using quantitative immunofluorescence, we investigated stromal TILs and PD-L1 protein expression in pre-treatment and residual breast cancer tissue from a Yale Cancer Center patient cohort of 58 patients diagnosed with breast cancer from 2003 to 2009 and treated with neoadjuvant chemotherapy. We compared the TIL count and PD-L1 status in paired pre-treatment and residual cancer tissues and correlated changes and baseline levels with survival. Results Of the 58 patients, 46 (79.3%) had hormone-positive and 34 (58.6%) had node-positive breast cancer. Eighty-six percent of residual cancer tissues had TIL infiltration and 17% had PD-L1 expression. There was a trend for higher TIL counts in postchemotherapy compared to prechemotherapy samples (p = 0.09). Increase in TIL count was associated with longer 5-year recurrence-free survival (p = 0.02, HR = 3.9, 95% CI = 1.179–15.39). PD-L1 expression (both stromal and tumor cells) was significantly lower in post-treatment samples (p = 0.001). Change in PD-L1 expression after therapy or TILs and PD-L1 expression in the posttreatment samples did not correlate with survival. Conclusions Increase in stromal TILs in residual cancer compared to pretreatment tissue is associated with improved recurrence-free survival. Despite a trend for increasing TIL counts, PD-L1 expression decreased in residual disease compared to pretreatment samples

Additional file 4: Figure S2. of Effect of neoadjuvant chemotherapy on tumor-infiltrating lymphocytes and PD-L1 expression in breast cancer and its clinical significance

Showing PD-L1 expression in post-neoadjuvant breast cancer specimens. A Distribution of maximal scores of PD-L1 (SP142 antibody) in the tumor (red) and stromal (blue) compartments. The cutoff was set at 500 AQUA units (QIF). B Linear regression of stromal versus tumor PD-L1 AQUA (QIF) scores. (TIF 148 kb

Additional file 2: Figure S1. of Effect of neoadjuvant chemotherapy on tumor-infiltrating lymphocytes and PD-L1 expression in breast cancer and its clinical significance

Showing HES of TILs at baseline and post-treatment. A Baseline HES of a case with moderate TIL infiltration at baseline and increased TIL counts following treatment. B Matched post-neoadjuvant HES of the baseline biopsy shown in (A). C Baseline HES of a case that displayed decrease TIL counts following treatment. D. Matched post-neoadjuvant HES of the baseline biopsy shown in (C). 20× Magnification, bar = 200 μm. (TIF 948 kb

Additional file 5: Figure S3. of Effect of neoadjuvant chemotherapy on tumor-infiltrating lymphocytes and PD-L1 expression in breast cancer and its clinical significance

Showing SP142 antibody validation and reproducibility. A Regressions in QIF scores (average) between staining performed in different days in serial sections of a lung cancer TMA (245). B Representative immunostaining for PD-L1 SP142 antibody in control tissues (placenta in upper panel and lung in lower panel) using QIF (left panel; SP142 in Cy5, red and cytokeratin mask in Cy3, green) and conventional IHC staining with DAB (right panel). (TIF 583 kb