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PD-L1 Multiplex and Quantitative Image Analysis for Molecular Diagnostics.

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  • 1Precision Medicine Centre of Excellence, The Patrick G Johnston Centre for Cancer Research, Queen's University, Belfast BT9 7AE, UK.

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|December 30, 2020
PubMed
Summary
This summary is machine-generated.

Multiplex immunofluorescence (mIF) with digital image analysis (DIA) accurately quantifies PD-L1 in non-small cell lung cancer (NSCLC). This validated workflow offers a rapid, comprehensive approach for biomarker analysis in clinical settings.

Keywords:
PD-L1high-throughput workflowimage analysismultiplexing

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Area of Science:

  • Oncology
  • Immunohistochemistry
  • Digital Pathology

Background:

  • Multiplex immunofluorescence (mIF) and digital image analysis (DIA) enable simultaneous detection and quantification of multiple biomarkers.
  • Accurate quantification of programmed death-ligand 1 (PD-L1) is crucial for predicting response to immunotherapy in non-small cell lung cancer (NSCLC).

Purpose of the Study:

  • To validate a clinical workflow for quantifying PD-L1 expression in NSCLC using mIF and DIA.
  • To assess the concordance between mIF-based DIA and traditional immunohistochemistry (IHC) for PD-L1 scoring.

Main Methods:

  • NSCLC samples were stained using a validated mIF panel.
  • Slides were scanned and analyzed using DIA software (QuPath).
  • PD-L1 quantification by mIF-DIA was compared with results from 3,3'-Diaminobenzidine (DAB) IHC.

Main Results:

  • Single-channel immunofluorescence showed concordance with single-plex IHC.
  • DIA facilitated quantification of various cell types and their marker expression.
  • Strong concordance (R² = 0.7323) was observed between DAB IHC and mIF slides, with 95% overall concordance between DIA on DAB IHC and mIF slides.
  • DIA on mIF slides demonstrated high comparability to DIA on DAB IHC slides, with less frequent overestimation of PD-L1 scores.

Conclusions:

  • DIA of mIF slides provides a rapid and highly comparable method to DIA on DAB IHC slides for PD-L1 quantification in NSCLC.
  • mIF combined with DIA enables comprehensive extraction of phenotypic data and microenvironmental details.
  • Further exploration of mIF's clinical relevance in immunotherapy-treated NSCLC cases is warranted.