Analysis of miR-146a and miR-142-3p as Potential Markers of Freshly Isolated or In Vitro-Expanded Human Treg cells

K Holmstrøm; A E Pedersen; M Gad

doi:10.1111/sji.12517

Analysis of miR-146a and miR-142-3p as Potential Markers of Freshly Isolated or In Vitro-Expanded Human Treg cells

K Holmstrøm, A E Pedersen, M Gad

Section 01 - Prosthetics

3 Citations (Scopus)

Abstract

Regulatory CD4(+) T cells (Tregs) are pivotal for prevention of autoimmunity. The use of Tregs is therefore of increasing interest in in vitro drug screening assays as well as for a cytotherapy per se against autoimmune disorders. For both purposes, in vitro expansion of peripheral blood Tregs is necessary and there is an increasing need to identify novel markers that can discriminate natural thymic-derived Tregs (tTregs) from other T cell subsets, and ideally, such markers should be stably expressed during in vitro expansion procedures. We screened for novel miRNAs differentially expressed in tTregs and identified miR-146a and 142-3p as possible candidates. We analysed freshly isolated naïve and activated tTregs and non-Treg subsets after or prior to in vitro expansion. We observed a tTreg-specific profile of these miRNAs together with FOXP3 and Helios in freshly isolated tTregs, but observed a decline in the same markers in activated tTregs as opposed to naïve tTregs. In vitro-expanded Tregs could be identified based on FOXP3 expression, but with loss of a discriminate profile for miRNA candidates and a decline in FOXP3 when activated tTregs were expanded. Our data demonstrate miR-146a and 142-3p as potential miRNA markers for discrimination between non-Treg cells and tTregs, but these miRNAs are not stable markers for in vitro-expanded Treg cells. In addition, the loss of FOXP3 in expansion of activated tTregs has implication for in vitro use of this cell subset in immunopharmacological assays and cytotherapy as FOXP3 is pivotal for suppressive function.

Original language	English
Journal	Scandinavian Journal of Immunology
Volume	85
Issue number	2
Pages (from-to)	113-121
Number of pages	9
ISSN	0300-9475
DOIs	https://doi.org/10.1111/sji.12517
Publication status	Published - 1 Feb 2017

Keywords

Biomarkers
Cell Proliferation
Cells, Cultured
Flow Cytometry
Forkhead Transcription Factors
Gene Expression Profiling
Humans
Ikaros Transcription Factor
Lymphocyte Activation
MicroRNAs
Oligonucleotide Array Sequence Analysis
Reverse Transcriptase Polymerase Chain Reaction
T-Lymphocytes, Regulatory
Journal Article

Access to Document

10.1111/sji.12517

sji.12517Final published version, 454 KB

Cite this

@article{7896321794734181a9788bdded4b6f63,

title = "Analysis of miR-146a and miR-142-3p as Potential Markers of Freshly Isolated or In Vitro-Expanded Human Treg cells",

abstract = "Regulatory CD4(+) T cells (Tregs) are pivotal for prevention of autoimmunity. The use of Tregs is therefore of increasing interest in in vitro drug screening assays as well as for a cytotherapy per se against autoimmune disorders. For both purposes, in vitro expansion of peripheral blood Tregs is necessary and there is an increasing need to identify novel markers that can discriminate natural thymic-derived Tregs (tTregs) from other T cell subsets, and ideally, such markers should be stably expressed during in vitro expansion procedures. We screened for novel miRNAs differentially expressed in tTregs and identified miR-146a and 142-3p as possible candidates. We analysed freshly isolated na{\"i}ve and activated tTregs and non-Treg subsets after or prior to in vitro expansion. We observed a tTreg-specific profile of these miRNAs together with FOXP3 and Helios in freshly isolated tTregs, but observed a decline in the same markers in activated tTregs as opposed to na{\"i}ve tTregs. In vitro-expanded Tregs could be identified based on FOXP3 expression, but with loss of a discriminate profile for miRNA candidates and a decline in FOXP3 when activated tTregs were expanded. Our data demonstrate miR-146a and 142-3p as potential miRNA markers for discrimination between non-Treg cells and tTregs, but these miRNAs are not stable markers for in vitro-expanded Treg cells. In addition, the loss of FOXP3 in expansion of activated tTregs has implication for in vitro use of this cell subset in immunopharmacological assays and cytotherapy as FOXP3 is pivotal for suppressive function.",

keywords = "Biomarkers, Cell Proliferation, Cells, Cultured, Flow Cytometry, Forkhead Transcription Factors, Gene Expression Profiling, Humans, Ikaros Transcription Factor, Lymphocyte Activation, MicroRNAs, Oligonucleotide Array Sequence Analysis, Reverse Transcriptase Polymerase Chain Reaction, T-Lymphocytes, Regulatory, Journal Article",

author = "K Holmstr{\o}m and Pedersen, {A E} and M Gad",

note = "{\textcopyright} 2016 The Foundation for the Scandinavian Journal of Immunology.",

year = "2017",

month = feb,

day = "1",

doi = "10.1111/sji.12517",

language = "English",

volume = "85",

pages = "113--121",

journal = "Scandinavian Journal of Immunology",

issn = "0300-9475",

publisher = "Wiley-Blackwell",

number = "2",

}

TY - JOUR

T1 - Analysis of miR-146a and miR-142-3p as Potential Markers of Freshly Isolated or In Vitro-Expanded Human Treg cells

AU - Holmstrøm, K

AU - Pedersen, A E

AU - Gad, M

PY - 2017/2/1

Y1 - 2017/2/1

N2 - Regulatory CD4(+) T cells (Tregs) are pivotal for prevention of autoimmunity. The use of Tregs is therefore of increasing interest in in vitro drug screening assays as well as for a cytotherapy per se against autoimmune disorders. For both purposes, in vitro expansion of peripheral blood Tregs is necessary and there is an increasing need to identify novel markers that can discriminate natural thymic-derived Tregs (tTregs) from other T cell subsets, and ideally, such markers should be stably expressed during in vitro expansion procedures. We screened for novel miRNAs differentially expressed in tTregs and identified miR-146a and 142-3p as possible candidates. We analysed freshly isolated naïve and activated tTregs and non-Treg subsets after or prior to in vitro expansion. We observed a tTreg-specific profile of these miRNAs together with FOXP3 and Helios in freshly isolated tTregs, but observed a decline in the same markers in activated tTregs as opposed to naïve tTregs. In vitro-expanded Tregs could be identified based on FOXP3 expression, but with loss of a discriminate profile for miRNA candidates and a decline in FOXP3 when activated tTregs were expanded. Our data demonstrate miR-146a and 142-3p as potential miRNA markers for discrimination between non-Treg cells and tTregs, but these miRNAs are not stable markers for in vitro-expanded Treg cells. In addition, the loss of FOXP3 in expansion of activated tTregs has implication for in vitro use of this cell subset in immunopharmacological assays and cytotherapy as FOXP3 is pivotal for suppressive function.

AB - Regulatory CD4(+) T cells (Tregs) are pivotal for prevention of autoimmunity. The use of Tregs is therefore of increasing interest in in vitro drug screening assays as well as for a cytotherapy per se against autoimmune disorders. For both purposes, in vitro expansion of peripheral blood Tregs is necessary and there is an increasing need to identify novel markers that can discriminate natural thymic-derived Tregs (tTregs) from other T cell subsets, and ideally, such markers should be stably expressed during in vitro expansion procedures. We screened for novel miRNAs differentially expressed in tTregs and identified miR-146a and 142-3p as possible candidates. We analysed freshly isolated naïve and activated tTregs and non-Treg subsets after or prior to in vitro expansion. We observed a tTreg-specific profile of these miRNAs together with FOXP3 and Helios in freshly isolated tTregs, but observed a decline in the same markers in activated tTregs as opposed to naïve tTregs. In vitro-expanded Tregs could be identified based on FOXP3 expression, but with loss of a discriminate profile for miRNA candidates and a decline in FOXP3 when activated tTregs were expanded. Our data demonstrate miR-146a and 142-3p as potential miRNA markers for discrimination between non-Treg cells and tTregs, but these miRNAs are not stable markers for in vitro-expanded Treg cells. In addition, the loss of FOXP3 in expansion of activated tTregs has implication for in vitro use of this cell subset in immunopharmacological assays and cytotherapy as FOXP3 is pivotal for suppressive function.

KW - Biomarkers

KW - Cell Proliferation

KW - Cells, Cultured

KW - Flow Cytometry

KW - Forkhead Transcription Factors

KW - Gene Expression Profiling

KW - Humans

KW - Ikaros Transcription Factor

KW - Lymphocyte Activation

KW - MicroRNAs

KW - Oligonucleotide Array Sequence Analysis

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - T-Lymphocytes, Regulatory

KW - Journal Article

U2 - 10.1111/sji.12517

DO - 10.1111/sji.12517

M3 - Journal article

C2 - 27943367

SN - 0300-9475

VL - 85

SP - 113

EP - 121

JO - Scandinavian Journal of Immunology

JF - Scandinavian Journal of Immunology

IS - 2

ER -

Analysis of miR-146a and miR-142-3p as Potential Markers of Freshly Isolated or In Vitro-Expanded Human Treg cells

Abstract

Keywords

Access to Document

Fingerprint

Cite this