Determinants of variability of five programmed death ligand-1 immunohistochemistry assays in non-small cell lung cancer samples

Ross A. Soo, Joey Sze Yun Lim, Bernadette Reyna Asuncion, Zul Fazreen, Maria Cynthia Herrera, Mohd Feroz Mohd Omar, Nguyen Hoang Diem Phuong, Ju Ee Seet, Benhur Amanuel, Barry Iacopetta, David Byrne, Shona Hendry, Stephen Fox, Richie Soong

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Abstract

Programmed death ligand-1 (PD-L1) expression as determined by immunohistochemistry (IHC) is potentially predictive of clinical outcome. The aim of this study was to assess the concordance of reported PD-L1 IHC assays and investigate factors influencing variability. Consecutive sections from 20 non-small cell lung cancers (NSCLCs) comprising resection, core biopsy, cytology and pleural fluid samples underwent IHC with 5 different antibody/autostainer combinations: 22C3/Link48, 28-8/BOND-MAX, E1L3N/BOND-MAX, SP142/BenchMark and SP263/BenchMark. PDL1 RNA levels were assessed using RNAscope. The frequency of positive cases using scoring thresholds from clinical trials was 72%, 33%, 61%, 56%, and 33% for the 5 IHC protocols respectively, and 33% for RNAscope. Pairwise agreement on the classification of cases as positive or negative for PD-L1 expression ranged from 61%- 94%. On a continuous scale, the lowest correlation was between 28-8/BOND-MAX and SP142/BenchMark (R2=0.25) and highest was between 22C3/Link48 and E1L3N/ BOND-MAX (R2=0.71). When cases were ordered according to tumor cell (TC)%, a similar ranking of cases across IHC protocols could be observed, albeit with different quanta and limits of detection. Single-slide OPAL 7-color fluorescence IHC analysis revealed a high degree of co-localization of staining from the 5 PD-L1 antibodies. Using SP142 antibody in a BOND-MAX protocol led to increased TC% quanta, while retaining a similar ranking of samples according to TC%. The results of this study highlight tumor PD-L1 status can vary significantly according to IHC protocol. Protocoldependent staining intensities and nominated thresholds for positivity contribute to this variability, while the antibody used appears to be less of a factor.

Original languageEnglish
Pages (from-to)6841-6851
Number of pages11
JournalOncotarget
Volume9
Issue number6
DOIs
Publication statusPublished - 1 Jan 2018

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