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A TNFR2-Agonist Facilitates High Purity Expansion of Human Low Purity Treg Cells

Regulatory T cells (Treg) are important for immune homeostasis and are considered of great interest for immunotherapy. The paucity of Treg numbers requires the need for ex vivo expansion. Although therapeutic Treg flow-sorting is feasible, most centers aiming at Treg-based therapy focus on magnetic...

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Autores principales: He, Xuehui, Landman, Sija, Bauland, Stijn C. G., van den Dolder, Juliette, Koenen, Hans J. P. M., Joosten, Irma
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4880213/
https://www.ncbi.nlm.nih.gov/pubmed/27224512
http://dx.doi.org/10.1371/journal.pone.0156311
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author He, Xuehui
Landman, Sija
Bauland, Stijn C. G.
van den Dolder, Juliette
Koenen, Hans J. P. M.
Joosten, Irma
author_facet He, Xuehui
Landman, Sija
Bauland, Stijn C. G.
van den Dolder, Juliette
Koenen, Hans J. P. M.
Joosten, Irma
author_sort He, Xuehui
collection PubMed
description Regulatory T cells (Treg) are important for immune homeostasis and are considered of great interest for immunotherapy. The paucity of Treg numbers requires the need for ex vivo expansion. Although therapeutic Treg flow-sorting is feasible, most centers aiming at Treg-based therapy focus on magnetic bead isolation of CD4+CD25+ Treg using a good manufacturing practice compliant closed system that achieves lower levels of cell purity. Polyclonal Treg expansion protocols commonly use anti-CD3 plus anti-CD28 monoclonal antibody (mAb) stimulation in the presence of rhIL-2, with or without rapamycin. However, the resultant Treg population is often heterogeneous and pro-inflammatory cytokines like IFNγ and IL-17A can be produced. Hence, it is crucial to search for expansion protocols that not only maximize ex vivo Treg proliferative rates, but also maintain Treg stability and preserve their suppressive function. Here, we show that ex vivo expansion of low purity magnetic bead isolated Treg in the presence of a TNFR2 agonist mAb (TNFR2-agonist) together with rapamycin, results in a homogenous stable suppressive Treg population that expresses FOXP3 and Helios, shows low expression of CD127 and hypo-methylation of the FOXP3 gene. These cells reveal a low IL-17A and IFNγ producing potential and hardly express the chemokine receptors CCR6, CCR7 and CXCR3. Restimulation of cells in a pro-inflammatory environment did not break the stability of this Treg population. In a preclinical humanized mouse model, the TNFR2-agonist plus rapamycin expanded Treg suppressed inflammation in vivo. Importantly, this Treg expansion protocol enables the use of less pure, but more easily obtainable cell fractions, as similar outcomes were observed using either FACS-sorted or MACS-isolated Treg. Therefore, this protocol is of great interest for the ex vivo expansion of Treg for clinical immunotherapy.
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spelling pubmed-48802132016-06-09 A TNFR2-Agonist Facilitates High Purity Expansion of Human Low Purity Treg Cells He, Xuehui Landman, Sija Bauland, Stijn C. G. van den Dolder, Juliette Koenen, Hans J. P. M. Joosten, Irma PLoS One Research Article Regulatory T cells (Treg) are important for immune homeostasis and are considered of great interest for immunotherapy. The paucity of Treg numbers requires the need for ex vivo expansion. Although therapeutic Treg flow-sorting is feasible, most centers aiming at Treg-based therapy focus on magnetic bead isolation of CD4+CD25+ Treg using a good manufacturing practice compliant closed system that achieves lower levels of cell purity. Polyclonal Treg expansion protocols commonly use anti-CD3 plus anti-CD28 monoclonal antibody (mAb) stimulation in the presence of rhIL-2, with or without rapamycin. However, the resultant Treg population is often heterogeneous and pro-inflammatory cytokines like IFNγ and IL-17A can be produced. Hence, it is crucial to search for expansion protocols that not only maximize ex vivo Treg proliferative rates, but also maintain Treg stability and preserve their suppressive function. Here, we show that ex vivo expansion of low purity magnetic bead isolated Treg in the presence of a TNFR2 agonist mAb (TNFR2-agonist) together with rapamycin, results in a homogenous stable suppressive Treg population that expresses FOXP3 and Helios, shows low expression of CD127 and hypo-methylation of the FOXP3 gene. These cells reveal a low IL-17A and IFNγ producing potential and hardly express the chemokine receptors CCR6, CCR7 and CXCR3. Restimulation of cells in a pro-inflammatory environment did not break the stability of this Treg population. In a preclinical humanized mouse model, the TNFR2-agonist plus rapamycin expanded Treg suppressed inflammation in vivo. Importantly, this Treg expansion protocol enables the use of less pure, but more easily obtainable cell fractions, as similar outcomes were observed using either FACS-sorted or MACS-isolated Treg. Therefore, this protocol is of great interest for the ex vivo expansion of Treg for clinical immunotherapy. Public Library of Science 2016-05-25 /pmc/articles/PMC4880213/ /pubmed/27224512 http://dx.doi.org/10.1371/journal.pone.0156311 Text en © 2016 He et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
He, Xuehui
Landman, Sija
Bauland, Stijn C. G.
van den Dolder, Juliette
Koenen, Hans J. P. M.
Joosten, Irma
A TNFR2-Agonist Facilitates High Purity Expansion of Human Low Purity Treg Cells
title A TNFR2-Agonist Facilitates High Purity Expansion of Human Low Purity Treg Cells
title_full A TNFR2-Agonist Facilitates High Purity Expansion of Human Low Purity Treg Cells
title_fullStr A TNFR2-Agonist Facilitates High Purity Expansion of Human Low Purity Treg Cells
title_full_unstemmed A TNFR2-Agonist Facilitates High Purity Expansion of Human Low Purity Treg Cells
title_short A TNFR2-Agonist Facilitates High Purity Expansion of Human Low Purity Treg Cells
title_sort tnfr2-agonist facilitates high purity expansion of human low purity treg cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4880213/
https://www.ncbi.nlm.nih.gov/pubmed/27224512
http://dx.doi.org/10.1371/journal.pone.0156311
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