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Multiplexed Immunofluorescence Analysis and Quantification of Intratumoral PD-1+ Tim-3+ CD8+ T Cells
Published on: February 8, 2018
Multicentric Analytical and Inter-observer Comparability of Four Clinically Developed Programmed Death-ligand 1
Ulrich Sommer1, Markus Eckstein2, Johannes Ammann3
1Institute of Pathology, Universitätsklinikum Carl Gustav Carus Dresden, Dresden, Germany.
Background:
Previous studies have suggested increased clinical benefit with inhibition of programmed death-ligand 1 (PD-L1)/programmed death-1 in patients with PD-L1-positive locally advanced/metastatic renal cell carcinoma (RCC). We examined the analytical and inter-observer comparability of PD-L1-positivity across 4 clinically developed immunohistochemistry assays in clear-cell RCC (CCRCC).
Materials And Methods:
Randomly selected archived, formalin-fixed, paraffin-embedded nephrectomy specimens from 201 patients with locally advanced CCRCC were screened using VENTANA SP142. From these, 30 cases were selected based on their tumor-infiltrating immune cell (IC) PD-L1 status (PD-L1-IC-positivity of < 1%, 1%-5%, or > 5%; 10 cases each). These cases were stained for PD-L1 using VENTANA SP142 and SP263, and DAKO 22C3 and 28-8, and scored for PD-L1 expression on IC and tumor cells (TC) by trained readers at 5 sites.
Results:
Adjusted mean percentages of PD-L1-IC-positivity and PD-L1-TC-positivity varied from 4.0% to 4.9% and from 1.3% to 10.7%, respectively, between assays. Inter-assay differences in PD-L1-IC-positivity were small and non-significant (P = .1938 to .9963); for PD-L1-TC-positivity, significant differences were observed between VENTANA SP142 and the other assays (P ≤ .0001) and between VENTANA SP263 and DAKO 28-8 (P = .0248). Intra-class correlation values showed moderate-to-high inter-reader agreement for each assay for PD-L1-IC-positivity and for 3 assays for PD-L1-TC-positivity.
Conclusions:
In this first multicenter analytical comparison study of PD-L1 assays in CCRCC, PD-L1-positivity could be assessed reproducibly using all 4 assays for IC and for 3 of the 4 assays for TC.
Insights
This study compared four programmed death-ligand 1 (PD-L1) immunohistochemistry assays in clear-cell renal cell carcinoma (CCRCC). All assays showed reproducible assessment of PD-L1 on immune cells, with three also reproducible on tumor cells.
Area of Science:
- Oncology
- Immunotherapy
- Diagnostic Pathology
Background:
- Previous studies indicate clinical benefit from programmed death-ligand 1 (PD-L1) and programmed death-1 inhibition in advanced renal cell carcinoma (RCC).
- PD-L1 expression is a key biomarker for patient selection in targeted therapies.
- The comparability of different PD-L1 assays in clear-cell RCC (CCRCC) requires thorough investigation.
Purpose of the Study:
- To evaluate the analytical and inter-observer comparability of four clinically utilized immunohistochemistry assays for PD-L1 detection in CCRCC.
- To assess the reliability of PD-L1 scoring on both immune cells (IC) and tumor cells (TC) across different assays.
Main Methods:
- A multicenter study involving 30 CCRCC specimens selected based on PD-L1 immune cell status.
- Staining for PD-L1 was performed using four assays: VENTANA SP142, SP263, and DAKO 22C3, 28-8.
- Trained readers at five sites scored PD-L1 expression on IC and TC.
Main Results:
- Inter-assay differences in PD-L1 immune cell (IC) positivity were minimal and not statistically significant.
- Significant differences were observed in PD-L1 tumor cell (TC) positivity between VENTANA SP142 and other assays, and between VENTANA SP263 and DAKO 28-8.
- Moderate-to-high inter-reader agreement was achieved for PD-L1-IC and for PD-L1-TC with three of the four assays.
Conclusions:
- This is the first multicenter study comparing PD-L1 assays in CCRCC.
- All four assays demonstrated reproducible assessment of PD-L1 positivity on immune cells.
- Three of the four assays provided reproducible assessment of PD-L1 positivity on tumor cells.
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