Abstract
Regulatory T cells (Treg) are a subset of T lymphocytes that in humans represent less than the 10 % of circulating CD4+ T cells. Treg are specialized in the inhibition of the immune responses and play a crucial role in the maintenance of immunological tolerance. Several lines of evidence clearly documented the role of Treg in restraining antitumor immune responses. For this reason, antitumor immunotherapy approaches have been recently associated with drug treatments aimed at depleting Treg or blocking their functions. A summary of the currently used in vivo approaches to limit Treg expansion in cancer patients is here provided. A comprehensive phenotypic and functional monitoring of Treg is crucial for the precise assessment of the effects that these different drug treatments exert on Treg. In this chapter, we will provide guidelines for an accurate ex vivo identification of human Treg. Due to the phenotypic and functional heterogeneity, intrinsic plasticity, and the lack of a unique marker exclusively expressed by human Treg, the clear-cut identification of this T cell subset requires the expert usage of multiparametric flow cytometry analysis (FACS). In this view, a combination of phenotypic and functional assessment of Treg is mandatory. In this chapter, we will describe the most reliable methods to identify and monitor the modulation of human Treg in patients undergoing immunological or drug-based treatments. Protocols to measure ex vivo the suppressive functions of Treg are also provided.
| Original language | English |
|---|---|
| Pages (from-to) | 201-221 |
| Number of pages | 21 |
| Journal | Methods in molecular biology (Clifton, N.J.) |
| Volume | 1139 |
| DOIs | |
| Publication status | Published - 2014 |
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Keywords
- CFSE
- Immunomagnetic isolation
- Intracellular staining
- Multiparametric FACS analysis
- Regulatory T cells
- Suppression assay
- Treg depletion
ASJC Scopus subject areas
- Molecular Biology
- Genetics
- Medicine(all)
Cite this
Monitoring the frequency and function of regulatory T cells and summary of the approaches currently used to inhibit regulatory T cells in cancer patients. / Camisaschi, Chiara; Tazzari, Marcella; Rivoltini, Licia; Castelli, Chiara.
In: Methods in molecular biology (Clifton, N.J.), Vol. 1139, 2014, p. 201-221.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Monitoring the frequency and function of regulatory T cells and summary of the approaches currently used to inhibit regulatory T cells in cancer patients
AU - Camisaschi, Chiara
AU - Tazzari, Marcella
AU - Rivoltini, Licia
AU - Castelli, Chiara
PY - 2014
Y1 - 2014
N2 - Regulatory T cells (Treg) are a subset of T lymphocytes that in humans represent less than the 10 % of circulating CD4+ T cells. Treg are specialized in the inhibition of the immune responses and play a crucial role in the maintenance of immunological tolerance. Several lines of evidence clearly documented the role of Treg in restraining antitumor immune responses. For this reason, antitumor immunotherapy approaches have been recently associated with drug treatments aimed at depleting Treg or blocking their functions. A summary of the currently used in vivo approaches to limit Treg expansion in cancer patients is here provided. A comprehensive phenotypic and functional monitoring of Treg is crucial for the precise assessment of the effects that these different drug treatments exert on Treg. In this chapter, we will provide guidelines for an accurate ex vivo identification of human Treg. Due to the phenotypic and functional heterogeneity, intrinsic plasticity, and the lack of a unique marker exclusively expressed by human Treg, the clear-cut identification of this T cell subset requires the expert usage of multiparametric flow cytometry analysis (FACS). In this view, a combination of phenotypic and functional assessment of Treg is mandatory. In this chapter, we will describe the most reliable methods to identify and monitor the modulation of human Treg in patients undergoing immunological or drug-based treatments. Protocols to measure ex vivo the suppressive functions of Treg are also provided.
AB - Regulatory T cells (Treg) are a subset of T lymphocytes that in humans represent less than the 10 % of circulating CD4+ T cells. Treg are specialized in the inhibition of the immune responses and play a crucial role in the maintenance of immunological tolerance. Several lines of evidence clearly documented the role of Treg in restraining antitumor immune responses. For this reason, antitumor immunotherapy approaches have been recently associated with drug treatments aimed at depleting Treg or blocking their functions. A summary of the currently used in vivo approaches to limit Treg expansion in cancer patients is here provided. A comprehensive phenotypic and functional monitoring of Treg is crucial for the precise assessment of the effects that these different drug treatments exert on Treg. In this chapter, we will provide guidelines for an accurate ex vivo identification of human Treg. Due to the phenotypic and functional heterogeneity, intrinsic plasticity, and the lack of a unique marker exclusively expressed by human Treg, the clear-cut identification of this T cell subset requires the expert usage of multiparametric flow cytometry analysis (FACS). In this view, a combination of phenotypic and functional assessment of Treg is mandatory. In this chapter, we will describe the most reliable methods to identify and monitor the modulation of human Treg in patients undergoing immunological or drug-based treatments. Protocols to measure ex vivo the suppressive functions of Treg are also provided.
KW - CFSE
KW - Immunomagnetic isolation
KW - Intracellular staining
KW - Multiparametric FACS analysis
KW - Regulatory T cells
KW - Suppression assay
KW - Treg depletion
UR - http://www.scopus.com/inward/record.url?scp=84909633833&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84909633833&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-0345-0_18
DO - 10.1007/978-1-4939-0345-0_18
M3 - Article
C2 - 24619682
AN - SCOPUS:84909633833
VL - 1139
SP - 201
EP - 221
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -