Brightfield Multiplex Immunohistochemistry Assay for PD-L1 Evaluation in Challenging Melanoma Samples

Appl Immunohistochem Mol Morphol. 2024 Sep 1;32(8):389-394. doi: 10.1097/PAI.0000000000001219. Epub 2024 Aug 21.

Abstract

Targeting the PD1/PD-L1 immune checkpoint pathway has rapidly become a therapeutic strategy for melanoma patients. Indeed, the quantification of PD-L1 expression by immunohistochemistry (IHC) in melanoma samples is already required, in some contexts, to allow access to anti-PD-1/PD-L1 immunotherapy. Despite a rising demand for PD-L1 testing, paralleling increasing cumulative experience in its assessment and quantification, it is fair to recognize that PD-L1 evaluation in melanoma samples still presents some critical issues. The aim of this technical report is to develop and validate a multiplex double staining protocol for PD-L1/SOX10 in Ventana Benchmark Ultra for routine practice. Our results show that double labeling provides the necessary tools to identify PD-L1 + melanoma cells clearly. The simultaneous visualization of 2 different proteins targets allows the topographical relationship between the 2 labeling to be evaluated within the context of the tissue morphology. Future studies are needed to test this technique's real-world applicability and effectiveness in implementing interpathologist agreement.

MeSH terms

  • B7-H1 Antigen* / metabolism
  • Biomarkers, Tumor / metabolism
  • Humans
  • Immunohistochemistry* / methods
  • Melanoma* / diagnosis
  • Melanoma* / metabolism
  • Melanoma* / pathology
  • SOXE Transcription Factors / metabolism
  • Skin Neoplasms / diagnosis
  • Skin Neoplasms / metabolism
  • Skin Neoplasms / pathology

Substances

  • B7-H1 Antigen
  • CD274 protein, human
  • SOX10 protein, human
  • SOXE Transcription Factors
  • Biomarkers, Tumor