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

作者： Clémence Granier1, Emeline Vinatier2,3,4, Elia Colin2,3, Marion Mandavit2,3, Charles Dariane2,3,5, Virginie Verkarre6, Lucie Biard7, Rami El Zein2, Corinne Lesaffre2, Isabelle Galy-Fauroux2, Hélène Roussel2,3,6, Eléonore De Guillebon8, Charlotte Blanc2,3, Antonin Saldmann4, Cécile Badoual2,6, Alain Gey*4, Éric Tartour*2,4 1PARCC-INSERM U970,Université Paris Descartes, 2INSERM U970,Université Paris Descartes, 3Equipe Labellisée Ligue Contre le Cancer, 4Department of Immunology,Hopital Européen Georges Pompidou, 5Department of Medical Urology,Hopital Européen Georges Pompidou, 6Department of Pathology,Hopital Européen Georges Pompidou, 7Université Paris Diderot Paris 7, 8Department of medical oncology,Hopital Européen Georges Pompidou

简介：

Overview

This study presents an innovative multiplexed immunofluorescence analysis method for automatic counting of tumor immune cells based on their phenotype. The technique is designed to analyze the tumor microenvironment and identify prognostic biomarkers for immunotherapy response.

Key Study Components

Area of Science

• Immunology
• Oncology
• Cell Biology

Background

• The tumor microenvironment plays a crucial role in cancer progression and treatment response.
• CD8 + T cells are important for immune response in tumors.
• Existing methods for cell counting can be time-consuming and require extensive data processing.
• This study aims to streamline the analysis of immune cell phenotypes.

Purpose of Study

• To develop a reliable method for automatic counting of immune cells in tumor samples.
• To facilitate large cohort analyses in cancer research.
• To improve the understanding of immune interactions within the tumor microenvironment.

Methods Used

• Multiplexed immunofluorescence for cell phenotype analysis.
• In situ fluorescence multispectral imaging for automatic counting.
• Preparation of tissue sections with hydrophobic barriers for imaging.
• Data processing techniques to analyze cell populations.

Main Results

• The method allows for efficient counting of CD8 + T cell subpopulations.
• Results indicate potential prognostic markers derived from the tumor microenvironment.
• The technique is reproducible and suitable for large-scale studies.
• Improved understanding of immune cell dynamics in tumors.

Conclusions

• This innovative method enhances the analysis of tumor immune cells.
• It provides a framework for identifying biomarkers for immunotherapy.
• The approach can be adapted for various research applications in oncology.

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