Regulatory T cells

Ethan Shevach and Todd Davidson

To avoid immune-mediated pathology and unrestricted organ transplantation, foetalmaternal tolerance and
clonal expansion of responder T cells, the immune even obesity. Defects in TReg cell function may be an
system has subsets of T cells, known as regulatory T important factor in the development of autoimmunity
(TReg) cells, that are dedicated to mediating immune or in the failure to control immunopathology, whereas
suppression. The most important subset of TReg cells overactive TReg cell function may contribute to the
expresses the transcription factor forkhead box P3 suppression of tumour immunity. Enhancement of TReg
(FOXP3). Both mice and humans with genetic cell function either pharmacologically or by cell-based
deficiencies of FOXP3 develop severe abnormalities of therapy may prove to be an adjunct to the treatment of

IMMUNOLOGY
immune homeostasis. TReg cells modulate the immune autoimmunity, whereas deletion or inactivation of TReg
response in numerous settings, including autoimmune cell function may facilitate the generation of tumour
disease, allergy, microbial infection, tumour immunity, immunity or enhance responses to weak vaccines.

Development and phenotype of regulatory T cells Function of FOXP3+ regulatory T cells

Many T cell types have immune regulatory function, but the two most important TReg cell subsets express the transcription FOXP3+ TReg cells have been shown to influence the outcome of immune responses in several tissues. For example, in the
factor FOXP3 and develop in the thymus or can be induced in peripheral sites including the mucosa-associated lymphoid intestine TReg cells have a key role in maintaining tissue homeostasis by inhibiting the overactivation of dendritic cells (DCs) and
tissue (MALT). Although expression of FOXP3 is considered a useful marker for these cell subpopulations in mice, FOXP3 effector T cells. In the case of autoimmunity, such as that depicted in the central nervous system (CNS), TReg cells can have a beneficial
expression may also be induced in human T cells that lack TReg cell function. However, functional activated human FOXP3+ effect by short circuiting the inflammatory loop of T cells and antigen-presenting cells (APCs). This same general mechanism can
TReg cells express a unique pattern of cell surface markers that can facilitate their isolation. A third important type of TReg cell have negative consequences in the setting of a tumour, in which TReg cells can inhibit the antitumour immune response, thereby
secretes the immunosuppressive cytokine interleukin-10 (IL-10) and may develop from conventional CD4+ T cells by activation in preventing tumour clearance. During infection TReg cells carry out a delicate balancing act; preventing immunity would lead to the
the presence of IL-10 or may develop from T helper 1 (TH1) or TH2 cell subsets. Other T cell subpopulations including natural killer T inability to clear the pathogen, whereas unrestrained immunity would lead to unwanted immune-mediated tissue destruction.
(NKT) cells, T cells and CD8+ T cells can also exert potent suppressor functions in certain settings. Although both human In each of the examples shown, we focus on the role of FOXP3+ TReg cells, although interactions between multiple immune
and mouse CD8+ T cells can be induced to express FOXP3, a suppressive function for these cells in vivo has yet to be clarified. cell types and indeed different types of regulatory T cell are also likely to be important in the regulation of immune responses.
6J[OWU 2TGXGPVKPICWVQKOOWPKV[
Inflammation FOXP3+IL-10 Regulation
MHC IL-2R IFN TReg cells in
(CD25) TH1 or
class II TH17 cell the periphery
Peptide CTLA4 IL-17
Uncommitted CD4 +
CNS
thymocyte thymocyte CD28 autoantigen
FOXP3 +
FOXP3 + TCR
Antigen
Self- presentation MHC Activation Damage TReg cell
reactive CD4+FOXP3+ class II to the CNS recruitment
TCR Thymic epithelial Unknown
IL-2 or natural TReg cell antigen
IL-15? cell or DC
DC
CD8+ thymocyte CD4+ thymocyte
Effector cell Damage IL-10 FOXP3+
2GTKRJGT[ recruitment to the CNS
FOXP3+IL-10+
/#.6 TReg cell
TH1 and TH17 cells Resolution of
IL-10 Natural in the periphery inflammation
APC CD4+ TR1 FOXP3+ T cell
Reg
IL-10
TGF, Naive T cell
CD8+ Naive Naive CD4+ IL-2, RA TGF
IL-10-secreting
CD4+ TReg cell
'UVCDNKUJKPIEJTQPKEKPHGEVKQP
T cell T cell
Hepatocyte TGF
CD4+ CD4+ Naive
Low dose antigen, T cell
TGF, IL-2 Naive T cell Hepatitis B TH17
IL-6, Induced
or C virus Naive TGF TReg cell
T cell NKT cell Type I IFNs T cell
FOXP3+/ FOXP3+ FOXP3+ TH3 MHC FOXP3+
CD4 + IL-4
TGF class II TH2
CD4 FOXP3
+ + FOXP3+
CD8+ TReg cell CD4+ FOXP3 + TGF-secreting Infected
induced TReg cell induced TReg cell TReg cell FOXP3+
TH3 cell TR1 cell tissue IL-12 Natural
TReg cell
DC MHC TH1
Plasticity in the periphery class I Naive
Under certain conditions, FOXP3+ TReg cells can TH1 cell Lethal infection, TReg cell B cell-derived TFH cell T cell
downregulate their expression of FOXP3, lose highly polarized signals in Pathogen
CD8+
suppressor functions and manifest some of the TH1 cell response Peyers patch
FOXP3 FOXP3+ FOXP3 persistence
functions of conventional effector TH1, TH2, TH17 Autoimmune CTL
microenvironment
and TFH cell subsets. The key causes of this loss
IL-6, Restricted
of FOXP3 expression include inflammatory ia tissue IL-10
IFN p en cy) IL-1, FOXP3+ IL-21 CTL
environments with high levels of cytokines that ho cien IL-23 IRF4 damage
m p fi
are normally involved in the induction of effector Ly 2 de in vitro TH2 cell
T cells, such as IL-6 and interferon- (IFN). In -
(IL
addition, TReg cell-specific deletion of certain /CKPVCKPKPIKPVGUVKPCNJQOGQUVCUKU
FOXP3 FOXP3 TH17 cell FOXP3
transcription factors that are shared between Antigen Gut lumen
Commensal bacteria TLR
TReg cells and effector cell subsets (for example, the ligand
TH2 cell-specific factor IRF4) results in impaired IEC
IFN, IL-17, IL-21 IL-17 IL-4, IL-5
suppression of TH2 cell responses by the TReg cells.

2JGPQV[RKEOCTMGTUQH(1:2 TGIWNCVQT[6EGNNU M cell

Markers shared by FOXP3+ TReg cells Markers preferentially expressed by Markers specifically expressed by
and conventional activated CD4+ activated mouse FOXP3+ TReg cells activated human FOXP3+ TReg cells
T cells (mice and humans) FOXP3 Latent TGF FOXP3hi TGF RA Inflammatory DC
CD103+ DC
CD25 CD127low CD103 Latent TGF
GITR CTLA4 GARP
CD45RBlow (mice only) Subpopulations of human FOXP3+ TReg cells CD121a (IL-1R1) TGF, RA, IL-2 IL-12, IL-6, IL-23
CD45RO (humans only) CD45RA+FOXP3low (naive) CD121b (IL-1R2)
Folate receptor 4 (mice only) CD45RAFOXP3hi (activated)
CD45RAFOXP3low (cytokine-secreting)
FOXP3+ CD4+ TGF, CD4+
/GEJCPKUOUQHCEVKQPQH(1:2 TGIWNCVQT[6EGNNU IL-10?
TReg cell Naive Naive
Suppressive cytokines and secreted molecules Cytolysis T cell T cell
Perforin
Granzyme A pore
Membrane- Contact-dependent
or granzyme B FOXP3+ FOXP3
tethered TGF inhibition IL-10, TGF
Cell cycle arrest Lamina propria
TGF TReg cell TH1 or TH17 cell
TReg cell FOXP3+
IL-35 IL-10 CD4+ or CD8+
Granzymes effector T cell Apoptosis
2TQOQVKPIVWOQWTRTQITGUUKQP
CD4+ or CD8+ FOXP3+ and perforin
Galectin 1 IL-10 effector T cell Tumour-associated
macrophage or DC IDO
CD45 CD43 TReg cell TReg cell FOXP3+ Activation CD4+ Effector T cells
TGF
Apoptosis Co-stimulation blockade CCR4 CD8+ CD8+
CCL22 FOXP3+
DC
Recruitment
Control of tumour
Metabolic disruption Targeting DCs by other mechanisms
Tryptophan Proliferation FOXP3+
catabolites Co-stimulation
IDO Apoptosis
blockade FOXP3+
Tryptophan
CD80 or CD86 Increased TGF,
depletion Trogocytosis? FOXP3+
CTLA4 interaction TGF IL-10
MHC CD80 or Transendocytosis
Inhibition of with DCs CD86 CD4+
CD39 A2AR class II of CD80/CD86
TReg cell FOXP3+ DC maturation T cell TGF
TCR CTLA4
Adenosine and decreased
CD73 T cell activation DC
FOXP3+ Inhibition of Conversion
IL-2R DC maturation
Apoptosis owing to LAG3 Tumour cell
(CD25) TReg cell
cytokine deprivation MHC class II FOXP3+
IL-2 Neuropilin 1
molecule

Affiliations STEMCELL Technologies Starting Phenotype of cells STEMCELL Cell Isolation Kit STEMCELL Catalog
sample number
Ethan Shevach and Todd Davidson are at the Regulatory T cells (Tregs) comprise only a small fraction of total or CD4+CD127lowCD49d- T cells are pre-enriched by RosetteSep,
Human
Laboratory of Immunology, National Institute of CD4+ T cells in human peripheral blood and mouse spleen, and which combines a normal FicollTM density centrifugation step with an
Allergy and Infectious Diseases, National Institutes Whole blood CD4+CD25+ T cells Complete Kit for Human CD4+CD25+ T Cells 15862
therefore must be highly enriched to evaluate their suppressive antibody-mediated specific cell enrichment procedure. CD25bright
of Health, Bethesda, Maryland 20892, USA. function and therapeutic potential. Since Tregs lack a unique cell T cells are then positively selected from the pre-enriched cells using CD4 CD127 CD25 T cells
+ low +
Complete Kit for Human CD4 CD127 CD25 Regulatory T Cells
+ low +
15861
e-mail: eshevach@niaid.nih.gov surface marker and often share phenotypic similarities with our column-free immunomagnetic cell separation system, EasySep, CD4+CD127lowCD49d-CD25+ Complete Kit for Human CD4+ CD127lowCD49d-CD25+ 15864
T cells Regulatory T Cells
The authors declare no competing financial interests. activated T cells, isolation of highly purified Tregs is typically or our fully automated cell separator, RoboSep. For researchers
PBMC CD4+CD25+ T cells EasySep/RoboSep Human CD4+CD25+ T Cell Isolation Kit 18062
difficult and time consuming, often requiring multiple steps. working with human PBMCs or mouse spleen samples, our EasySep
To meet the needs of Treg researchers, STEMCELL Technologies kits provide a fast, easy and gentle method to isolate highly purified CD4 CD127 T cells
+ low
EasySep/RoboSep Human CD4 CD127 T Cell Enrichment Kit
+ low
19231
Edited by Lucy Bird and Kirsty Minton;
copyedited by Gemma Ryan; has developed a full range of optimized Treg isolation kits that Tregs. Positively selected cells express high levels of FOXP3 and are CD4+CD127lowCD49d- T cells EasySep/RoboSep Human CD4+CD127lowCD49d-Regulatory 19232
T Cell Enrichment Kit
designed by Simon Bradbrook. addresses these specific challenges. suitable for immediate downstream experiments, including flow
Our Complete Kits for the isolation of Tregs provide the fastest cytometry, in vitro expansion or suppression assays. Mouse
(2010) Macmillan Publishers Ltd. All rights reserved. and easiest method to isolate highly purified human Tregs directly For more information about our complete range of cell isolation Spleen or CD4+CD25+ T cells EasySep Mouse CD4+CD25+ Regulatory T Cell Isolation Kit 19782
http://www.nature.com/nri/posters/tregcells other tissues
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