Measuring CTL Lytic Granule Secretion and Target Cell Membrane Repair by Fluorescent Lipophilic Dye Uptake at the Lytic Synapse

Sabina Müller, Liza Filali, Marie Pierre Puissegur, Salvatore Valitutti*

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

Abstract

CD8+ cytotoxic T lymphocytes (CTL) play a key role in anti-tumor immune response. They are therefore at the heart of current immunotherapy protocols against cancer. Despite current strategies to potentiate CTL responses, cancer cells can resist CTL attack, thus limiting the efficacy of immunotherapies. To optimize immunotherapy, it is urgent to develop rapid assays allowing to assess CTL-cancer cell confrontation at the lytic synapse. In this chapter, we describe a flow cytometry-based method to simultaneously assess the extent of CTL activation and of tumor cell reparative membrane turnover in CTL/target cell conjugates. Such a method can be performed using a limited number of cells. It can therefore be employed in clinical settings when only a few patient-derived cells might be available.

Original languageEnglish
Pages (from-to)463-476
Number of pages14
JournalMethods in Molecular Biology
Volume2654
DOIs
Publication statusPublished - 28 Apr 2023

Keywords

  • Cytotoxic T lymphocytes
  • Flow cytometry
  • Fluorescent lipophilic dyes
  • Lytic synapse
  • Plasma membrane turnover
  • Time lapse video microscopy

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