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

doi:10.1016/bs.mie.2019.05.046

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

Division of Basic Oncology (Interdisciplinary)

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

26 Citations (Scopus)

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 language	English
Title of host publication	Tumor Immunology and Immunotherapy - Cellular Methods Part B
Editors	Lorenzo Galluzzi, Nils-Petter Rudqvist
Publisher	Academic Press Inc.
Pages	155-192
Number of pages	38
ISBN (Print)	9780128186756
DOIs	https://doi.org/10.1016/bs.mie.2019.05.046
Publication status	Published - 2020

Publication series

Name	Methods in Enzymology
Volume	632
ISSN (Print)	0076-6879
ISSN (Electronic)	1557-7988

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

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

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10.1016/bs.mie.2019.05.046

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Bruger, A. M., Vanhaver, C., Bruderek, K., Amodio, G., Tavukçuoğlu, E., Esendagli, G., Gregori, S., Brandau, S., & van der Bruggen, P. (2020). Protocol to assess the suppression of T-cell proliferation by human MDSC. In L. Galluzzi, & N.-P. Rudqvist (Eds.), Tumor Immunology and Immunotherapy - Cellular Methods Part B (pp. 155-192). (Methods in Enzymology; Vol. 632). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2019.05.046

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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.",

keywords = "MDSC, Mye-EUNITER, Suppression assay, T lymphocytes, T-cell proliferation, T-cells, Tumor",

author = "Bruger, \{Annika M.\} and Christophe Vanhaver and Kirsten Bruderek and Giada Amodio and Ece Tavuk{\c c}uoğlu and G{\"u}nes Esendagli and Silvia Gregori and Sven Brandau and \{van der Bruggen\}, Pierre",

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year = "2020",

doi = "10.1016/bs.mie.2019.05.046",

language = "English",

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series = "Methods in Enzymology",

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Bruger, AM, Vanhaver, C, Bruderek, K, Amodio, G, Tavukçuoğlu, E, Esendagli, G, Gregori, S, Brandau, S & van der Bruggen, P 2020, Protocol to assess the suppression of T-cell proliferation by human MDSC. in L Galluzzi & N-P Rudqvist (eds), Tumor Immunology and Immunotherapy - Cellular Methods Part B. Methods in Enzymology, vol. 632, Academic Press Inc., pp. 155-192. https://doi.org/10.1016/bs.mie.2019.05.046

Protocol to assess the suppression of T-cell proliferation by human MDSC. / Bruger, Annika M.; Vanhaver, Christophe; Bruderek, Kirsten et al.
Tumor Immunology and Immunotherapy - Cellular Methods Part B. ed. / Lorenzo Galluzzi; Nils-Petter Rudqvist. Academic Press Inc., 2020. p. 155-192 (Methods in Enzymology; Vol. 632).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

TY - CHAP

T1 - Protocol to assess the suppression of T-cell proliferation by human MDSC

AU - Bruger, Annika M.

AU - Vanhaver, Christophe

AU - Bruderek, Kirsten

AU - Amodio, Giada

AU - Tavukçuoğlu, Ece

AU - Esendagli, Günes

AU - Gregori, Silvia

AU - Brandau, Sven

AU - van der Bruggen, Pierre

PY - 2020

Y1 - 2020

N2 - 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.

AB - 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.

KW - MDSC

KW - Mye-EUNITER

KW - Suppression assay

KW - T lymphocytes

KW - T-cell proliferation

KW - T-cells

KW - Tumor

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T3 - Methods in Enzymology

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BT - Tumor Immunology and Immunotherapy - Cellular Methods Part B

A2 - Galluzzi, Lorenzo

A2 - Rudqvist, Nils-Petter

PB - Academic Press Inc.

ER -

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

Abstract

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UN SDGs

Keywords

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