Crucial Contributions by T Lymphocytes (Effector, Regulatory, and Checkpoint Inhibitor) and Cytokines (TH1, TH2, and TH17) to a Pathological Complete Response Induced by Neoadjuvant Chemotherapy in Women with Breast Cancer

Figure 7

IL-17 (a, b), TGF-β (c, d), and PD-L1 (e, f) expression in the sections of LLABCs, using IHC staining, at 400x magnification. Briefly, heat-mediated antigen retrieval was performed using citrate buffer, pH 6 (20 mins). The sections were then incubated with polyclonal Abs to IL-17 (Abcam, ab9565) at a 1 : 100 dilution for 30 mins at RT, MAbs to TGF-β (Abcam, ab64715) at a concentration of 12 μg/mL overnight at 4°C, and polyclonal Abs to PDL1 (Abcam, ab58810) at a concentration of 2.5 μg/mL for 15 mins at RT, respectively. Polymeric HRP-linker antibody conjugate was used as secondary antibody. DAB chromogen was used to visualize the staining. The sections were counterstained with haematoxylin. (a, c, e) Low level of expression; (b, d, f) high level of expression. The score (% of positive cells (brown membrane/cytoplasmic-stained tumour and immune cells) × intensity of staining (1 to 3)) was used to assess the level of expression; low was ≤100 and high was >100. Scoring performed on whole tissue section (>10 HPFs); Tu: tumour and Ly: lymphocyte.

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