Hematoxylin & Eosin and immunohistochemical staining were performed on 10% formalin-fixed paraffin-embedded sections (5µm). The antibody panel included CD20, CD3, CD5 and PD-L1. Immunohistochemical staining was performed with an automated immunostainer ULTRA, Roche Diagnostics Corporation, Indianapolis, IN for CD20 (L26 clone; predilute; Roche), CD5 (SP19 clone; predilute; Roche) and CD3 (2GV6 clone; predilute; Ventana) or Bond-Max, Leica Biosystems, Buffalo Grove, IL for PD-L1(SP142; dilution 3; Spring Bioscience) using procedure per manufacturer’s instructions. Tumors were considered PD-L1 positive based on membrane staining in >5% of tumor cells as previously described by Herbst et al. 11 (link).
Protocol detail
Immunohistochemical Profiling of Tumors
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Antibody
Buffalo
Cells
Clone
Diagnostics
Dilution
Embedded paraffin
Eosin
Formalin
Hematoxylin
Membrane
Pd l1
Tumor
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Variable analysis
independent variables
- Formalin fixation
- Paraffin embedding
- Antibody panel (CD20, CD3, CD5, PD-L1)
- Membrane staining of PD-L1 in tumor cells (>5% positivity)
- 10% formalin fixation
- Paraffin embedding
- Tissue section thickness (5µm)
- Automated immunostaining procedure per manufacturer's instructions
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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