Protocol to assess the suppression of T-cell proliferation by human MDSC

Annika M. Bruger, Christophe Vanhaver, Kirsten Bruderek, Giada Amodio, Ece Tavukçuoğlu, Günes Esendagli, Silvia Gregori, Sven Brandau, Pierre van der Bruggen

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Inhibition of T-cell proliferation is the most common approach to assess human myeloid-derived suppressor cell (MDSC) functions. However, diverse methodologies hinder the comparison of results obtained in different laboratories. In this chapter, we present a T-cell proliferation assay procedure based on allogeneic MDSC and T-cells that is potentially suitable to multi-center studies. The T-cells are isolated from non-cancerous donors and frozen for later use in different research groups. We observed that pure thawed T-cells showed poor proliferative capacities. To retain proliferation, T-cell-autologous mature dendritic cells are supplemented after thawing. MDSC are isolated from clinical samples and represent the sole variant between assays. Flow cytometry is used to assess T-cell proliferation by the dilution of a tracking dye.

Original languageEnglish
Title of host publicationMethods in Enzymology
EditorsLorenzo Galluzzi, Nils-Petter Rudqvist
PublisherAcademic Press Inc.
Pages155-192
Number of pages38
ISBN (Print)9780128186756
DOIs
Publication statusPublished - 2020

Publication series

NameMethods in Enzymology
Volume632
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • MDSC
  • Mye-EUNITER
  • Suppression assay
  • T lymphocytes
  • T-cell proliferation
  • T-cells
  • Tumor

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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