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Multiplexed Immunofluorescence Analysis and Quantification of Intratumoral PD-1+ Tim-3+ CD8+ T Cells

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Published on: February 8, 2018

In Situ Detection of Programmed Cell Death Protein 1 and Programmed Death Ligand 1 Interactions as a Functional

Amanda Lindberg¹, Lars Muhl², Hui Yu¹

¹Department of Immunology, Genetics, and Pathology, Uppsala University, Uppsala, Sweden.

Journal of Thoracic Oncology : Official Publication of the International Association for the Study of Lung Cancer

|January 1, 2025

Summary

Detecting programmed cell death protein 1 (PD1)-programmed death ligand 1 (PD-L1) interactions offers more precise lung cancer treatment insights than standard assays. This method can predict immune checkpoint inhibitor (ICI) response and identify resistance mechanisms.

Keywords:

Immune checkpoint inhibitor Programmed cell death protein 1 Programmed death ligand 1 Proximity ligation assay non–small cell lung cancer

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Area of Science:

Oncology
Immunology
Biomarker Discovery

Background:

Immune checkpoint inhibitors (ICIs) have revolutionized lung cancer therapy but show variable efficacy.
Current biomarkers like programmed death ligand 1 (PD-L1) expression have limited predictive power.
Assessing direct programmed cell death protein 1 (PD1)-PD-L1 interactions may offer superior prediction of treatment response.

Purpose of the Study:

To evaluate the utility of detecting PD1-PD-L1 interactions as a biomarker for ICI treatment in various cancers.
To compare the predictive value of PD1-PD-L1 interactions with standard PD-L1 expression assays.
To identify mechanisms of ICI resistance in non-responding patients.

Main Methods:

A proximity ligation assay was used to detect PD1-PD-L1 interactions in diagnostic samples from 16 cancer types.
Analysis included early-stage and advanced non-small cell lung cancer (NSCLC) patient cohorts, with and without ICI treatment.
RNA sequencing was employed to investigate resistance mechanisms.

Main Results:

PD1-PD-L1 interactions were detected in a subset of NSCLC cases with PD-L1 and PD1 expression, with lower levels in EGFR-mutated tumors.
Higher PD1-PD-L1 interaction levels correlated with complete response and improved survival in ICI-treated NSCLC patients.
Non-responders with high interactions showed additional expression of immune mediators like EOMES and HAVCR1.

Conclusions:

Assessing active immune pathways via PD1-PD-L1 interactions provides more precise ICI treatment guidance than static biomarker quantification.
This functional diagnostic approach is applicable to small biopsies and can be extended to other immune checkpoints.
Findings suggest combined diagnostic and therapeutic strategies are needed to overcome ICI resistance.