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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: Chapter in Book/Report/Conference proceedingChapterpeer-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
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages463-476
Number of pages14
DOIs
Publication statusPublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2654
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

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

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