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Fig. 3 | Journal for ImmunoTherapy of Cancer

Fig. 3

From: Resistance to anti-PD-1-based immunotherapy in basal cell carcinoma: a case report and review of the literature

Fig. 3

Representative staining patterns of the formalin-fixed, paraffin-embedded primary BCC lesion with granzyme-B (a), FOXP3 (b), CD8 (c), CD3 (d) and CD11b (e) specific mAbs. Number of positive cells were enumerated in an entire lesion and reported as an absolute number. f Representative staining patterns of the formalin-fixed, paraffin-embedded primary BCC lesion with HLA class I antigen-specific mAbs. Tumor tissue sections were immunohistochemically (IHC) stained with a pool of mouse HLA-A–specific mAb HCA2 and HLA-B/C-specific mAb HC10 (ratio, 1:1). mAb HCA2 recognizes β2m-free HLA-A (excluding -A24), -B7301, and -G heavy chains; mAb HC10 recognizes β2m-free HLA-A3, −A10, −A28, −A29, −A30, −A31, −A32, −A33, and all β2m-free -HLA-B (excluding -B5702, -B5804, and -B73) and -HLA-C heavy chains [53,54,55]. IHC staining was performed as described previously [56]. Staining of infiltrating immune cells was used as an internal positive control. The staining with HLA class I antigen–specific mAbs on tumor cells was scored as negative because HLA class I antigen score in an entire lesion was 0. g Representative staining patterns of the formalin-fixed, paraffin-embedded primary BCC lesion with β2m-specific mAb NAMB-1 [57]. IHC staining was performed as described previously [56]. Staining of infiltrating immune cells or fibroblasts was used as an internal positive control. The staining with β2m-specific mAb on tumor cells was scored as negative because β2m score in an entire lesion was 0. h Representative staining patterns of the formalin-fixed, paraffin-embedded primary BCC lesion with PD-L1-specific mAb (left panel). PD-L1 IHC staining was performed utilizing the automated PD-L1 IHC assay (PD-L1 IHC 28–8 pharmDx SK005) with Dako’s Autostainer Link 48 [58]. This test is a complementary diagnostic for nivolumab. MCF7 and NCI-H226 cell lines were used as a negative (upper and right panel) and positive control (bottom and right panel), respectively, accordingly to manufacturer kit instructions. PD-L1 expression was scored as negative because PD-L1 score in an entire lesion was < 1%, both on tumor cells and immune cells. Slides were reviewed and enumerated by an experienced pathologist (GB). i Representative staining patterns of the formalin-fixed, paraffin-embedded primary BCC lesion with hematoxylin and eosin (H&E). Magnification is indicated

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