Toxicol Pathol-2012-Peterson-186-204 PDF

Toxicologic Pathology
http://tpx.sagepub.com/
Regulatory T-Cells: Diverse Phenotypes Integral to Immune Homeostasis and Suppression

Richard A. Peterson
Toxicol Pathol 2012 40: 186 originally published online 5 January 2012
DOI: 10.1177/0192623311430693
The online version of this article can be found at:
http://tpx.sagepub.com/content/40/2/186
Published by:
http://www.sagepublications.com
On behalf of:
Society of Toxicologic Pathology
Additional services and information for Toxicologic Pathology can be found at:
Email Alerts: http://tpx.sagepub.com/cgi/alerts
Subscriptions: http://tpx.sagepub.com/subscriptions
Reprints: http://www.sagepub.com/journalsReprints.nav
Permissions: http://www.sagepub.com/journalsPermissions.nav
>> Version of Record - Mar 19, 2012

OnlineFirst Version of Record - Jan 5, 2012
What is This?
Downloaded from tpx.sagepub.com by guest on November 20, 2014
Toxicologic Pathology, 40: 186-204, 2012

Copyright # 2012 by The Author(s)
ISSN: 0192-6233 print / 1533-1601 online
DOI: 10.1177/0192623311430693
Regulatory T-Cells: Diverse Phenotypes Integral to

Immune Homeostasis and Suppression
RICHARD A. PETERSON
GlaxoSmithKline, Research Triangle Park, North Carolina, USA
ABSTRACT
Regulatory T-cells (TREG) are diverse populations of lymphocytes that regulate the adaptive immune response in higher vertebrates. TREG delete
autoreactive T-cells, induce tolerance, and dampen inflammation. TREG cell deficiency in humans (i.e., IPEX [Immunodysregulation, Polyendocrinopathy and Enteropathy, X-linked syndrome]) and animal models (e.g., Scurfy mouse) is associated with multisystemic autoimmune disease.
TREG in humans and laboratory animal species are similar in type and regulatory function. A molecular marker of and the cell lineage specification
factor for TREG is FOXP3, a forkhead box transcription factor. CD4 TREG are either natural (nTREG), which are thymus-derived
CD4CD25FOXP3 T-cells, or inducible (i.e., Tr1 cells that secrete IL-10, Th3 cells that secrete TGF-b and IL-10, and Foxp3 Treg). The proinflammatory Th17 subset has been a major focus of research. TH17 CD4 effector T-cells secrete IL-17, IL-21, and IL-22 in autoimmune and inflammatory disease, and are dynamically balanced with TREG cell development. Other lymphocyte subsets with regulatory function include: inducible
CD8 TREG, CD3CD4CD8 TREG (double-negative), CD4Va14 (NKTREG), and gd T-cells. TREG have four regulatory modes of action: secretion of inhibitory cytokines (e.g., IL-10 and TGF-b), granzyme-perforin-induced apoptosis of effector lymphocytes, depriving effector T-cells of
cytokines leading to apoptosis, or inhibition of dendritic cell function. The role of TREG in mucosal sites, inflammation/infection, pregnancy, and
cancer as well as a review of TREG as a modulatory target in drug development will be covered.
Keywords:
immunopathology; regulatory T-cells; lymphocytes; flow cytometry; Scurfy mouse; FOXP3; IPEX.
INTRODUCTION
TO
REGULATORY T-CELLS
delete autoreactive T-cells, induce tolerance, and dampen

inflammation (Akdis 2009; Ozdemir et al. 2009; Dittel 2008;
Orihara et al. 2008; Sakaguchi et al. 2008; Simpson 2008;
Bluestone and Tang 2005; Coutinho et al. 2005).
Over the past 40 years, the field of immunology has slowly
evolved, becoming much more complex than was initially
hypothesized in the 1960s. Between 1966 and 1968, several
groups identified B-lymphocytes as the source of the antibody
response and T-lymphocytes as critical in the delayed-type
hypersensitivity (DTH) response (Claman et al. 1966; Mosier
1967; Mitchell et al. 1968). In 1971, Gershon and Kondo identified negative interference on positively acting lymphocytes
during inflammation and referred to it as infectious immunological tolerance (Gershon and Kondo 1971). This was the
first indication that a suppressive process was active during
inflammation. The suppressor T-cell hypothesis was put
forth by two groups in the later years of the 1970s (Vadas et
al. 1976; Okumura et al. 1977). A series of negative results
from experiments conducted by several groups led to a loss
of confidence in the suppressor T-cell hypothesis, rejection
of the hypothesis by the field of immunology, and more
recently reacceptance (Kapp and Bucy 2008; Germain 2008;
Lehner 2008). The rejection of the suppressor T-cell hypothesis did not explain away the presence of a dampening of
immune responses commonly seen in the inflammatory milieu.
In 1989, Mosmann and Coffman identified the T-helper (TH)-1
and (TH)-2 CD4 T-cell subsets based upon the profiles of
cytokines secreted after activation. TH1 cells primarily secrete
IFN-g, IL-15, and TNF-a, while TH2 cells secrete IL-4, IL-5,
IL-6, and IL-13. Powrie et al. showed in severe combined
immunodeficient (SCID) mice that subsets of CD4 T-cells
This article will review the scientific literature and provide an

overview of Regulatory T-cell (TREG) characterization, function,
role in the immune response, at mucosal sites, during pregnancy
and in the tumor microenvironment, and as a target for modulation
in drug development. There has been enhanced focus on the TREG
in the field of immunology, which translates into a prominent
increase in the number of citations over the past decade (Figure 1).
TREG are diverse populations of lymphocytes that regulate
the adaptive immune response in higher vertebrates. TREG
The author declared no potential conflicts of interests with respect to the

authorship and/or publication of this article. The author received no financial
support for the research and/or authorship of this article.
Address correspondence to: Richard A. Peterson II, DVM, PhD, DACVP,
Safety Assessment, 9.3009.2D, GlaxoSmithKline, 5 Moore Drive, Research
Triangle Park, NC 27709; phone: (919) 315-2450; fax: (919) 483-6858;
e-mail: richard.a.peterson@gsk.com.
Abbreviations: A2A, adenosine receptor; APC, antigen presenting cell; BREG,
regulatory B-cell; CD, cluster of differentiation; CMV, cytomegalovirus; CTLA4, cytotoxic T-lymphocyte antigen-4; DN, double negative; DTH, delayed-type
hypersensitivity; EAE, experimental autoimmune encephalomyelitis; FIV, feline
immunodeficiency virus; FLG-2, fibrinogen-like protein-2; FOXP3, forkhead box
protein-3; GVHD, graft-versus-host disease; HCV, hepatitis C virus; HIV, human
immunodeficiency virus; IBD, inflammatory bowel disease; IFNg, interferon
gamma; IL, interleukin; IPEX, immunodysregulation polyendocrinopathy enteropathy X-linked syndrome; LAG3, lymphocyte-activating gene-3; MHC, major
histocompatability complex; miR, microRNA; MoAb, monoclonal antibody;
mTOR, mammalian target of rapamycin; NK cell, natural killer cell; NKTREG,
natural killer regulatory T-cell; SCID, severe combined immunodeficient; TCR,
T-cell receptor; TGF-b, transforming growth factor beta; TH0, T-helper 0 cell;
TH1, T-helper 1 cell; TH2, T-helper 2 cell; TH3, T-helper 3 cell; TH17, T-helper
17 cell; TREG, regulatory T-cell; Tr1, type 1 regulatory T-cells.
186
Vol. 40, No. 2, 2012
REGULATORY T-CELLS
FIGURE 1.There has been a steady increase in the number of regulatory T-cell (TREG) citations in the scientific literature from the mid1990s to the present (from Scopus). The key milestones in TREG
research are highlighted on the horizontal axis.
induce and protect from intestinal inflammation (Powrie et al.

1994). This once again defined a population of lymphocytes
with immunosuppressive actions, which was identified as a
population expressing CD45RBlow while the inducing cells
were CD45RBhigh. In 1995, Sakaguchi et al. found that immunologic self-tolerance was maintained by activated CD4 Tcells expressing CD25 (IL-2R alpha chain). Depletion of this
CD4CD25 population led to development of autoimmune disease (Sakaguchi et al. 1995). Two groups, Hori et al. and Fontenot
et al., identified a CD4CD25FOXP3 cell population that was
found to be thymically derived, natural TREG cells (Hori et al.
2003; Fontenot et al. 2003, 2005). FOXP3 has been shown to
be critical for regulatory action of most subtypes of TREG.
TYPES
AND
REGULATORY T-CELLS (TREG, NATURAL,

ADAPTIVE/INDUCED) CD4CD25 TREG
OF
Natural TREG are thymus-derived CD4CD25FOXP3 Tcells (Figure 2). They arise in the thymus during early stages of
human fetal development (gestational week 14) (Cupedo et al.
2005) and are resistant to thymic deletion (Lim et al. 2006).
nTREG differentiate from thymocytes that express TCRs with
an increased affinity for self-peptide-MHC complexes (Lim
et al. 2006; Maggi et al. 2005; Schwartz 2005). nTREG can suppress the following cell types: CD4 T-cells, dendritic cells,
CD8 T-cells, NKT cells, NK cells, monocytes/macrophages,
B-cells, mast cells, basophils, eosinophils, and osteoblasts
(Shevach et al. 2009; Lim et al. 2006). FOXP3 expression
is necessary for thymocytes to commit to the TREG lineage
(Lim et al. 2006). Resting nTREG are CD45RAFOXP3low,
while activated nTREG are CD45ROFOXP3high (Beissert
et al. 2006; Lim et al. 2006). Natural TREG also express the
following markers: T cell activation/differentiation markers
(CD45RO [memory phenotype; activated], CD45RB [resting],
CD25 [both]), adhesion molecules (CD62L [both], CD44
[both] and Integrin a4b7 [resting]), cytotoxic T lymphocyte-
187
associated protein-4 (CTLA-4; both), co-stimulatory molecule

CD28 (both), chemokine receptors (CCR7 [both], CXCR4
[both], CCR9 [resting]), glucocorticoid-induced TNFRrelated protein (GITR, both), OX40 (CD134, both), and folate
receptor-4 (FR4 in rodents, both) (Lim et al. 2006). IL-2 is essential for nTREG development, function, and homeostasis (i.e.,
CD25 is the a-chain of the high affinity IL2R) (Malek et al.
2008). Severe autoimmunity is seen in IL-2-, IL2Ra-, and IL2Rb-deficient mice, which also lack a normal number of TREG
cells (Malek et al. 2008). The nTREG profile in mammals is highly
conserved. Nonhuman primates have the same nTREG protein
expression and functional profile as that of humans including two
FOXP3 isoforms (Roncarolo and Battaglia 2007; Bloom 2006),
while in rodents there are only subtle differences primarily in the
type of granzyme expressed (i.e., B versus A) and only a single
FOXP3 isoform (Roncarolo and Battaglia 2007).
ADAPTIVE/INDUCED TREG
The two most common types of adaptive or induced TREG
are the T regulatory type 1 cells (Tr1) and TH3 TREG cells (Figure 2). Tr1 cells secrete IL-10 and also have an IL-10dependent induction process (Fujio et al. 2010; Beissert et al.
2006; Roncarolo et al. 2006). They are capable of secreting
high levels of IL-10 and TGF-b in the human and mouse and
also secrete low levels of IL-2, IL-5, and IFN-g (Fujio et al.
2010; Roncarolo et al. 2006). An important growth factor for
Tr1 cells is IL-15, which can support Tr1 cell proliferation even
without TCR activation (Fujio et al. 2010; Roncarolo et al.
2006). They are CD4, anergic, and proliferate poorly upon
antigen-specific activation which is likely due to the autocrine
production of IL-10 leading to suppression of proliferation
(Fujio et al. 2010; Roncarolo et al. 2006). The mechanism of
suppressive effects with Tr1 cells is soluble factor-based (i.e.,
IL-10), and the suppressive effects of Tr1 cells are negated
by anti-IL-10 neutralizing antibody in vitro (Fujio et al.
2010; Roncarolo et al. 2006). There is no specific marker for
Tr1 cells, although repressor of GATA-3 (ROG) shows potential utility (Fujio et al. 2010; Roncarolo et al. 2006), but it is not
specific for this cell population.
TH3 cells secrete TGF-b and IL-10 and express FOXP3
(Beissert et al. 2006). They are induced from nave CD4
T-cells by TGF-b and have an important role in oral tolerance
to non-self antigens and negating autoimmune reactions (Wan
and Flavell 2007). TH3 cells secrete TGF-b, which has immunosuppressive effects acting through a soluble-factor mechanism. TH3 cells have a reciprocal relationship with TH17 cells
(Korn et al. 2009; Nistala and Wedderburn 2009). TH17 cells
are highly proinflammatory and are thought to be important in
autoimmune disease (Abraham and Cho 2009; Crome et al.
2009; Korn et al. 2009; Nistala and Wedderburn 2009;
Romagnani et al. 2009); this will be discussed in detail later
in the article. There has been no specific surface marker
identified for TH3 cells, although expression of FOXP3 is
induced in TH3 cells (i.e., cannot differentiate from nTREG)
(Korn et al. 2009).
188
PETERSON
TOXICOLOGIC PATHOLOGY
FIGURE 2.TREG are either formed naturally by thymic differentiation (nTREG) or are induced in the periphery (iTREG) from nave T-cells (TH0).
Types of iTREG include TH3, Tr1, which are CD4CD25FOXP3, and CD4CD25-FOXP3 iTREG. The main effector CD4 subsets are TH1,
TH2, and TH17. The cytokines that are important in inducing these cells from TH0 cells and the cytokines that these cells secrete are listed.
OTHER T-CELLS WITH REGULATORY FUNCTION

Other T-cell populations that have been shown to have regulatory function include inducible CD8 TREG cells,
CD8CD28 TREG cells, and CD3CD4CD8 TREG
(double-negative [DN]). CD8 TREG (Kapp and Bucy 2008;
Beissert et al. 2006; Nakamura et al. 2003; Ke and Kapp
1996; Jiang et al. 1992) are CD8 T-cells that have regulatory
activity on CD4 TH1 cells. This TREG population inhibits
priming of CD8 T-cells, CD4 T-cells, and antibody
responses against oral antigens. They are induced by manipulation of co-stimulatory molecule interactions (i.e., CD137 and
CD40) primarily, although antigens introduced into immune
privileged sites (i.e., anterior chamber of eye leading to anterior
chamber-associated autoimmune deviation [ACAID]) elicit
CD8 TREG due to high levels of TGF-b and IL-10 at this site
(Biros 2008; Niederkorn 2008). They have also been shown to
arise spontaneously in Experimental Allergic Encephalitis
(EAE) and murine Ovalbumin-induced oral tolerance models
and in tumor-bearing hosts due to IL-10 secretion by APCs
in the neoplasm. They are well characterized in rodents, but
their importance in humans is uncertain. CD8CD28- TREG
(Cortesini et al. 2001; Chang et al. 2002) are FOXP3 and are
induced by inhibitory immunoglobulin-like transcript (ILT)-3
and ILT-4 receptor expression on dendritic cells and downregulation of CD80 and CD86. They interfere with the
CD28/B7 pathway with allogeneic antigens and CD154/
CD40 pathway with xenogeneic antigens. CD8CD28 TREG
regulate immune responses by direct cell-to-cell interactions
with dendritic cells; therefore they are directly tolerogenic.
This cell population has been described in rodents and humans.
CD3CD4CD8 (DN) TREG are TCRab, CD4, and CD8
(therefore DN). They promote the development of tolerance
and regulate autoimmunity (Strober et al. 1996). This cell population has been defined and characterized in heart xenotransplantation experiments (Chen et al. 2003, 2005; Zhang et al.
2006; Ma et al. 2008). DN TREG are poorly suppressive in nave
mice but are actively suppressive in recipients of heart xenografts co-transferred with donor DN Tregs. They have been
shown to prevent heart xenograft rejection (i.e., transplant tolerance). It has been shown that antigen presenting cells (APC)
from tolerant mice had down-regulation of MHCII, CD40, and
B7 molecules, and likely were in an immature state. The
Vol. 40, No. 2, 2012
REGULATORY T-CELLS
189
FIGURE 3.TREG can cause suppression of effector T-cells and dendritic cells through secretion of soluble factors including IL-10, TGF-b,
fibrinogen-like protein-2 (FLG-2), granzyme A or B and perforin, and adenosine. The figure shows the signaling pathways and the suppressive
effects on effector T-cells and dendritic cells.
significance of DN TREG in nontransplant scenarios is therefore

uncertain and more data is needed.
NKTREG cells are T-cells and APCs (Ikehara et al. 2000; Zeng
et al. 1999). The significance of NKTREG cells in immune regulation outside of xenotransplantation is uncertain.
CD4Va14 (NKTREG)
Natural killer T-cells coexpress NK receptors (i.e., NK1.1 or
CD161) as well as a conserved T-cell receptor (TCR) molecule
(Beissert et al. 2006), Va14. Va14 pairs with Vb8 in the mouse
and Vb11 in the human and are restricted by CD1d, a conserved
MHCI-type molecule that presents glycolipids. NKTREG cells
are generated in the thymus and can be CD4, CD8, DN, or
double positive (DP). In mice, DN NKTREG are the dominant
phenotype (Zeng et al. 1999); while in humans, CD4 NKTREG
are most efficient (Jiang et al. 2005). They have been studied primarily in rodent xenogeneic transplantation models (Ikehara et
al. 2000). They secrete IL-4, IL-10, TGF-b (paracrine), and
induce cytotoxicity by cell-to-cell contact, and the targets of
gd T-CELLS (MUCOSAL/INTRAEPITHELIAL)
Gamma delta (gd) T-cells, which express TCRgd and have a
mucosal distribution, are primarily suppressive and are associated with mucosal tolerance (Kapp and Ke 1997; Ke et al.
1997; Mowat 1994), but can also function to regulate autoimmunity (Weiner et al. 1994), elicit ACAID in the eye (Biros
2008), and function in tumor immunity (Seo et al. 1999). These
cells do not recognize peptides associated with MHC receptors
(Kronenberg 1994); alternatively they recognize cellular proteins including heat shock proteins (Munk et al. 1989) and nonclassic MHC molecules like CD1 (Brossay et al. 1998) and Qa1 (Vidovic et al. 1989). gd T-cells that infiltrate tumors have
190
PETERSON
FIGURE 4.TREG can cause suppression of effector T-cells and dendritic cells through cell contact-based mechanisms, including galectin-1 binding, CTLA-4 binding to CD80/86, lymphocyte activation gene-3 (LAG-3) binding to MHCII molecules, and neuropilin-1 binding. The figure
shows the ligands, receptors, and the suppressive effects on effector T-cells and dendritic cells.
cytokine profiles that are similar to Tr1 cells (i.e., IL-10 and
TGF-b) and inhibit the immune response against neoplasia
(Kapp et al. 2004; Seo and Tokura 1999; Seo et al. 1999; Kapp
and Ke 1997; Ke et al. 1997). Therefore, gd T-cells are a minor
population of mucosal and intratumoral T-cells with regulatory
activity and major roles in mucosal tolerance and tumor
immunity.
A NON-T LYMPHOCYTE SUBSET
WITH
REGULATORY FUNCTION
Another cell type exhibiting regulatory activity is the regulatory B-cell (BREG) which has been recently reviewed by Li et
al. (2011). BREG cells are beyond the scope of this review, but
are mentioned here briefly to showcase the diversity of lymphocytes with regulatory function in the immune system and
for completeness-sake. BREG cells have been shown in various
animal disease models, including collagen-induced arthritis
(Trentham et al. 1977), experimental autoimmune encephalitis
(EAE) (Wolf et al. 1996), nonobese diabetes mellitus (Tian et

al. 2001), inflammatory bowel disease (Mizoguchi et al. 2000),
and systemic lupus erythematosus (Lenert et al. 2005), and
function primarily to regulate T-cells, B-cells, and NKT cells.
BREG cells act by soluble factor-based means by secreting IL10 (Reigert and Bar-Or 2008), TGF-b (Takenoshita et al. 2002)
and producing anti-inflammatory antibodies (Diamond et al.
2003). Cell-cell interactions are an important mode of BREG
regulatory function and include CD1d binding (Yanaba et al.
2008) by CD1dhiCD5 cells and CD40/CD40 L interactions
(Quezada et al. 2004). More research into specific roles for
BREG is needed to more completely define this cell type.
TREG BASICS
TREG cells function at sites of inflammation in close spatial
proximity to effector T-cells, thus allowing direct interactions
between the two cell types (Askenasy et al. 2008). TREG cells
Vol. 40, No. 2, 2012
REGULATORY T-CELLS
191
FIGURE 5.TREG can cause suppression of effector T-cells by providing competition for growth factors (primarily IL-2), which is essential for
their survival in the periphery. TREG express multiple copies of IL-2R/CD25, which bind up local IL-2 competing with effector T-cells for
IL-2 stimulation. The figure shows the ligands, receptors, and the suppressive effects on effector T-cells.
after lymphopenia of numerous causes including those of a

physiologic and pathologic nature. Lymphopenia-induced lymphocyte proliferation does not increase the number of nave
lymphocytes, but instead the number of memory T-cells
increase, which might result in potential self-reactive clones
and subsequent autoimmunity. TREG have an important role
in regulating this process (Datta and Sarvetnick 2009).
EFFECTS
FIGURE 6.FOXP3 and CD3 dual immunofluorescence on tissue from

an area of inflammation. Note the green nuclear labeling (FOXP3) and
the red cytoplasmic labelling (CD3). The image shows a group of Tcells with several FOXP3 TREG cells in the field. 400 magnification.
are attracted by inflammatory signals, although they are less

chemotactic than effector T-cells (Siegmund et al. 2005).
When arriving at the site of inflammation, TREG downregulate chemotactic receptors and adhesive interactions to
arrest further migration (Siegmund et al. 2005). nTREG are
mitotically inactive under basal conditions (Kuniyasu et al.
2000). Activated TREG-mediated immune suppression is
antigen-nonspecific (i.e., not MHCII-restricted) (Bienvenu et
al. 2005), although induction of suppressor function in TREG
requires antigen-specific stimulation (i.e., via TCR) leading
to tissue-specific suppressive effects (bystander suppression) (Shevach 2009; Weiner 1997). TREG have also been associated with regulating lymphopenia-induced lymphocyte
proliferation, which is a normal homeostatic event that occurs
OF
ACUTE
AND
CHRONIC STRESS ON HUMAN TREG
beta 1-adrenergic and glucocorticoid a receptors are overexpressed in TREG versus effector T-cells (Freier et al. 2010; Atanackovic et al. 2006). These receptors likely mediate TREG
responses to stress (Freier et al. 2010; Atanackovic et al.
2006). TREG numbers have been shown to decrease with acute
physiologic stress (Freier et al. 2010; Atanackovic et al. 2006;
Atanackovic et al. 2002). Based on these findings, chronic
stress could exacerbate autoimmune disease and other inflammatory conditions (Freier et al. 2010).
MODES
OF
ACTION
OF
TREG REGULATION
TREG have primary effect on T-cells and/or dendritic cells

by three main regulatory modes of action: (1) soluble factors
(Figure 3), (2) cell-to-cell contact (Figure 4) and (3) competition for growth factors (local effect) (Figure 5). Soluble factors
include IL-10 and TGF-b with direct suppressive effects on
effector T-cells (Joetham et al. 2007; Annacker et al. 2003),
fibrinogen-like protein-2 (FLG-2) with apoptotic effects on
effector T-cells and prevention of maturation of dendritic cells
(Shalev et al. 2009), granzyme A/B and perforin apoptotic
effects on effector T-cells (Grossman et al. 2004), and production of adenosine by CD39/73 cleavage of ATP, which causes
cell cycle arrest in effector T-cells and prevention of maturation and decreased antigen presenting capability in dendritic
cells by binding to the A2A receptor on these cell types (Deaglio
192
PETERSON
FIGURE 7.Flow cytometry was performed on blood from male and female CD-IGS rats to evaluate the percentage of
CD4CD25CD62LFOXP3 (nTREG and iTREG) cells. The image highlights the gating strategy and steps used in this evaluation.
et al. 2007). Galectin-1 binding to effector T-cells and dendritic

cells resulting in cell cycle arrest and/or apoptosis (Garin et al.
2007), CTLA4 and CD80/86 binding to dendritic cells causing
decreased costimulation and decreased antigen presentation
(Read et al. 2000), lymphocyte activating gene-3 (LAG3 or
CD223, a CD4 homolog) binding to MHCII molecules on dendritic cells leading to prevention of maturation and a reduction in
antigen presentation capability (Workman and Vignali 2004),
and neuropilin-1 binding to dendritic cells resulting in decreased
antigen presentation (Sarris et al. 2008) are examples of cell
contact-based mediated effects of TREG. TREG can also deny
IL-2 to other inflammatory cells by binding the cytokine and acting as a sink leading to effector T-cell apoptosis (Pandiyan et
al. 2007). Therefore, TREG have a variety of mechanisms that
can be used to suppress an immune response and are able to use
all or a subset of these mechanisms simultaneously.
TREG MARKERS (IMMUNOHISTOCHEMISTRY, FLOW CYTOMETRY,
INTRAVITAL MICROSCOPY, AND MIRNA ANALYSIS)
Immunofluorescent and chromogenic IHC and flow cytometry (de Boer et al. 2007; Roncador et al. 2005) for the
following markers has been shown to be useful in characterizing TREG in tissue sections and isolated cells: FOXP3 (Figures
6, 7, 8, and 9), CD25 (alpha chain of IL2R) (Figures 7, 8, and
9), and CD62L (L-Selectin) (Figures 6 and 7). Intercellular
interactions of TREG cells and effector T-cells have been visualized in rodent models using intravital microscopy (Tang and
Krummel 2006). Differences in micro-RNA (miRNA) in
human TREG versus effector T-cells have been shown (Hezova
et al. 2010). There was a significant increase in miR-146a and
decreases in miR-20b, 31, 99a, 100, 125b, 151, 335, and 365 in
TREG when compared with effector T-cells. The pattern of
increased expression of miR-146a in combination with
decreases in the other miRNAs is a possible marker of TREG.
FOXP3: FUNCTION
AND
BIOLOGY
FOXP3, a forkhead box transcription factor, is a molecular

marker and cell lineage specification factor for TREG (Fontenot
et al. 2003; Hori et al. 2003). FOXP3 is critical for the thymic
differentiation of ab TCR thymocytes into nTREG (Sakaguchi
et al. 2008; Fontenot et al. 2003; Hori et al. 2003). The FOXP3
gene product is referred to as scurfin and functions by
Vol. 40, No. 2, 2012
REGULATORY T-CELLS
FIGURE 8.The graphs of flow cytometry data using markers useful in

characterizing TREG show the percentage of CD3CD4 cells,
CD62L cells in the CD3CD4population, CD25cells in the
CD3CD4 population, and FOXP3 cells in the CD3CD4 population cell subsets in male and female rats. There was no definitive difference in cell percentages between males and females in this
evaluation.
regulating a set of genes required for (1) the suppressor activity

of TREG, proliferative, and metabolic fitness of TREG and (2)
repressing alternative T cell differentiation pathways (Brunkow et al. 2001). It has been shown that a higher level of
expression of FOXP3 is sufficient for suppressive activity of
non-TREG T-cells in rodents (Sakaguchi et al. 2008; Lourenco
and La Cava 2011). Several pathways are important in the
induction of FOXP3 expression including TGF-b, IL-2, retinoic acid, 17-b-estradiol, and signaling through the
sphingosine-1-phosphate receptor 1 (S1PR1) (Figure 10).
TGF-b is essential for induction of FOXP3 expression in nave
T-cells (i.e., iTREG including TH3 cells), but in the human it
does not necessarily confer regulatory function (Lourenco and
La Cava 2011).
FOXP3 DEFICIENCY
The Scurfy Mouse
In mice (B.Cg-Foxp3sf), a frameshift mutation in the FOXP3
gene results in a truncated protein lacking the forkhead domain
and is responsible for the Scurfy phenotype (Clark et al.
1999; Ochs et al. 2002; Patel 2001). Scurfy is an X-linked
recessive mutation in the mouse that is lethal in hemizygous
males 16 to 25 days after birth (Ochs et al. 2002; Godfrey
et al. 1991). Scurfy mice have an overproliferation of CD4
T-lymphocytes and other cell types due to uncontrolled
cytokine secretion with extensive multiorgan infiltration due
to lack of functional TREG (Ochs et al. 2002; Godfrey et al.
1991). Lymphadenopathy, parenchymal organ infiltration, and
193
FIGURE 9.The graphs of flow cytometry data show the percentages of

FOXP3 cells in the CD4CD25 population and CD25 cells in the
CD4FOXP3 population. The majority (approximately 80%) of
CD4CD25 cells are also FOXP3, which would include nTREG and
TH3 cells. Only a minority (approximately 30%) of CD4FOXP3
cells are also CD25, which means approximately 70% of the
CD4FOXP3 population likely represents an iTREG cell population.
subcutaneous inflammatory cell infiltrates (e.g., ear pinnae) are

prominent (Figures 11 and 12) (Godfrey et al. 1991). The phenotype of Scurfy mice is similar to mouse models that lack
CTLA-4 or TGF-b, and in humans with immunodysregulation,
polyendocrinopathy and enteropathy, X-linked syndrome
(IPEX) (Clark et al. 1999; Ochs et al. 2002; Patel 2001).
IPEX in Humans
IPEX is a rare human multisystemic autoimmune disease
(X-linked: females are carriers, males have the disease) that
is linked to the dysfunction of the transcriptional activator
FOXP3 which results in dysfunction of regulatory T-cells with
subsequent autoimmunity (Ochs et al. 2002). Mutations in the
forkhead domain of FOXP3 disrupt FOXP3 protein: DNA
interactions (Ochs et al. 2002; Patel 2001). The disorder manifests with enteropathy, food allergy, massive lymphoproliferation, psoriasiform or eczematous dermatitis, nail dystrophy,
autoimmune endocrinopathies (primarily insulin-dependent
diabetes mellitus and autoimmune thyroid disease), Coombspositive anemia, autoimmune thrombocytopenia, autoimmune
neutropenia, tubular nephropathy, and autoimmune skin conditions such as alopecia universalis and bullous pemphigoid
(Ochs et al. 2002). Treatment for IPEX includes immunosuppressive agents (e.g., cyclosporin A, FK506) alone or in combination with steroids (Ochs et al. 2002). There has been
limited success in treating the syndrome with bone marrow
transplantation (Ochs et al. 2002).
194
PETERSON
FIGURE 10.The figure highlights several pathways of induction of FOXP3 expression including the ligands, receptors, and/or intermediary steps
in induction of gene expression. A schematic of the FOXP3 gene shows the promoters and important transcription factors that bind to the promoters. There are 11 coding exons in both the mouse and human FOXP3 genes.
TH17 CELLS AND TREG: A RECIPROCAL BALANCE

TH17 cells are a subpopulation of effector T cells that are
very pro-inflammatory and implicated in allergic/autoimmune
disease (Abraham and Cho 2009; Crome et al. 2009; Korn et al.
2009 and 2007; Nistala and Wedderburn 2009; Romagnani et
al. 2009). TH17 cells secrete IL-17, IL-22, IL-21 and IL-17F
(Korn et al. 2009, 2007; Veldhoen et al. 2006; Bettelli et al.
2006; Mangan et al. 2006). TGF-b is critical in induced/adaptive TREG cell (i.e., TH3 TREG) differentiation as well as development of TH17 effector cells (Wan and Flavell 2007) (Figure
13). Nave T-cells develop into a transitional cell-type that
expresses FOXP3 (TH3 TREG cell marker) and ROR-gt and
RORa (TH17 effector cell markers) in the presence of TGF-b
(Figure 14) (Nistala et al. 2009; Korn et al. 2007; Veldhoen
et al. 2006; Bettelli et al. 2006; Mangan et al. 2006). Further
exposure to TGF-b or IL-6/IL-21 in the inflammatory milieu
leads to TH3 cell differentiation or TH17 effector cell differentiation (Zhou et al. 2008; Korn et al. 2007; Veldhoen et al.
2006; Bettelli et al. 2006; Mangan et al. 2006). Therefore, a

reciprocal balance between TH17 cells and TH3 TREG cells
exists (Bettelli et al. 2006) where IL-6 and IL-21 inhibit TH3
cell differentiation to the benefit of TH17 cells (Korn et al.
2009, 2007) and TGF-b elicits TH3 cell differentiation to the
disadvantage of TH17 cells (Zhou et al. 2008).
ROLE OF TREG
IN
MUCOSAL SITES
Oral tolerance is an important mechanism in the mucosal

immune system whereby oral administration of a specific antigen induces unresponsiveness of systemic immune responses
with the same or different antigens (Chen et al. 2007; Weiner
et al. 1994). TGF-b is critical for the induction of oral tolerance
(Chen et al. 2007; Weiner et al. 1994). Low-dose of antigen
cells leads to dampening, a paracrine effect, of the immune
response due to TGF-b derived from nTREG, TH3, and Tr1 cells.
High-dose of an antigen is associated with mucosal T-cell
anergy and apoptosis (Chen et al. 2007; Weiner 1997; Weiner
Vol. 40, No. 2, 2012
REGULATORY T-CELLS
FIGURE 11.The Scurfy mouse has a frameshift mutation in the

forkhead box transcription factor, FOXP3. This results in early lethality due to unregulated inflammation (parenchymal organs, subcutis,
and lymphoid organs are the main sites of inflammatory cell accumulation) due to the lack of functional TREG cells. Note the gross pathology image of a Scurfy mouse with marked hepato- and splenomegaly
and evidence of subcutaneous lesions at the tail base.
et al. 1994). At mucosal sites, TREG are responsible for

bystander suppression, where TGF-b produced at mucosal
sites during development of oral tolerance regulates immune
responses on T-cells, B-cells, NK cells, macrophages, and dendritic cells in an antigen nonspecific manner (Weiner 1997).
Animal models of inflammatory bowel disease (IBD) have been
used to study TREG and mucosal tolerance (Blumberg 2009).
Researchers have adoptively transferred CD4CD45RBhigh
T-cells into SCID or Rag-1 mice (Powrie et al. 1994) or have
relied on the 2, 4,6-trinitrobenzene sulfonic acid (TNBS)-induced
colitis rodent model (Te Velde et al. 2006). A negative effect of
TREG in mucosal immunity is the secretion of TGF-b from TREG
in the face of IL-6 and IL-21 in the inflammatory milieu. TGF-b
can lead to differentiation of TH17 cells and subsequent inhibition
of TREG differentiation, resulting in more severe inflammation
(Korn et al. 2009, 2007). TREG have a critical role in tolerance and
bystander suppression of mucosal sites with resident gd
T-cells.
ROLE OF TREG IN INFLAMMATORY/INFECTIOUS DISEASE

Both natural and induced TREG are important in suppressing
inflammation in infectious disease (Figure 15; Abraham and
Medhitzov 2011; Liu et al. 2010; Jager and Kuchroo 2010;
Scholzen et al. 2009; de Boer et al. 2007; Belkaid and Rouse
2005). nTREG are activated by microbial infections and products of the resulting inflammatory milieu. TREG express tolllike receptors 4, 5, 7, and 8, allowing the cells to respond to
bacterial products such as lipopolysaccharide leading to
increased TREG survival and proliferation (Caramahlo et al.
2003). Probiotics have been shown to elicit activation of TREG,
which then suppress TH1-mediated colitis in the TNBS mouse
colitis model (Di Giacinto et al. 2005). Microbes are also able
195
FIGURE 12.Note the prominent mononuclear inflammatory cell accumulation in the dermis and upper subcutis from a Scurfy mouse.
Hematoxylin and eosin, 200 magnification.
to manipulate antigen-presenting cells leading to priming/activation of TREG and elicit secretion of chemokines and cytokines that favor induction, priming, recruitment, and survival
of TREG, thus using TREG for their own benefit (Cabrera et al.
2004; Hesse et al. 2004; Caramahlo et al. 2003; Belkaid et al.
2002). These microbial organisms include virus (HIV [Aandahl
et al. 2004], HCV [Cabrera et al. 2004], CMV [Aandahl et al.
2004], FIV [Vahlenkamp et al. 2004], Herpes simplex virus
[Suvas et al. 2004]), bacteria (Helicobacter spp. [Lundgren et
al. 2003; Kullberg et al. 2002] and Listeria monocytogenes
[Kursar et al. 2002]), fungi (Candida albicans [Montagnoli et
al. 2002] and Pneumocystis carinii [Hori et al. 2002]), and protozoal (Leishmania major [Belkaid et al. 2002], Schistosoma
mansoni [Hesse et al. 2004] and Plasmodium spp. [Scholzen
et al. 2009]). TREG can become infected with viral pathogens
(e.g., HIV and FIV), and it is uncertain if they remain suppressive when infected (Joshi et al. 2004; Oswald-Richter et al.
2004). Animal infection models and infected humans show
accumulation of TREG at the site of infection with leishmaniasis, herpes simplex virus, and schistosomiasis (Hesse et al.
2004; Suvas et al. 2004; Belkaid et al. 2002), in the peripheral
blood with HIV and HCV (Oswald-Richter et al. 2004) and in
lymphoid organs of HIV patients (Andersson et al. 2005). A
good example of the role of TREG in infection would be Plasmodium spp. that causes malaria in human patients and rodent
models and is associated with increased numbers of TREG (i.e.,
natural and induced), which have a direct association with the
parasitic load in the host. TREG accumulate in areas of inflammation induced by the organisms and inhibit TH1 responses
(i.e., both protective and pathologic) primarily by the suppressive effects of IL-10 and TGF-b, and ultimately result in either
clearance of the organism and resolution or clinical cases of
malaria depending on the level of TREG cell induction and the
organisms manipulation of this lymphocyte population
(Scholzen et al. 2009). TREG has also been shown to be a factor
196
PETERSON
FIGURE 13.There is a reciprocal relationship between TH17 (proinflammatory effector T-cell subset that are associated with autoimmune disease) and TH3 cells (iTREG that are CD4CD25FOXP3), which is elicited during differentiation of nave TH0 cells, but final differentiation
depends on the cytokine milieu. TGF-b is critical in differentiation of both cell types but depends on the cytokine milieu. The schematic shows the
cytokine pathways in the differentiation of TH17 and TH3 cells from TH0 effector cells.
in the development of silica-induced pulmonary fibrosis in a

mouse model. Depletion of TREG leads to the maintenance of
a TH1 response and subsequent attenuation of fibrosis versus
the shift to a TH2 response that normally predisposes to development of fibrosis in this model (Liu et al. 2010). The balance
between effector T-cells and TREG is critical in determining the
immune response to pathogens (Figure 16).
ROLE OF TREG
IN
ORGAN TRANSPLANTATION
TREG have been shown to have a pivotal role in transplant

tolerance leading to graft acceptance and prevention of rejection in xenotransplantation (Muller et al. 2009; Zhang et al.
2009; Le and Chao 2007; Yong et al. 2007). Adoptive transfer
of TREG into mice has been shown to inhibit, delay, and prevent
development of graft versus host disease (GVHD) (Taylor et al.
2002; Cohen et al. 2002) as well as to prevent allograft (i.e.,
pancreatic islet cell transplant) rejection (Battaglia et al.

2006; Gregori et al. 2001). Post-transplant immunosuppressive
drugs have been shown to have variable effects on TREG.
Mycophenolate mofetil increases TREG numbers and induces
transplant tolerance when given with vitamin D (Han et al.
2005; Huang et al. 2003; Gregori et al. 2001), calcineurin inhibitors (e.g., cyclosporine, tacrolimus, sirolimus) decrease TREG
and abrogate transplant tolerance (Shoji et al. 2005; Larsen et
al. 1996), and Rapamycin and methylprednisolone do not seem
to have effects on TREG or transplant tolerance (Blaha et al.
2003).
ROLE OF TREG IN PREGNANCY
Natural TREG, CD8 TREG, TH3 cells, and Tr1 cells have
been shown to be important in immune tolerance during pregnancy (Guerin et al. 2009; Trowsdale and Betz 2006; Aluvihare
Vol. 40, No. 2, 2012
REGULATORY T-CELLS
197
FIGURE 14.This schematic focuses on the differentiation pathways of TH0 to TH17 and TH3 cells showing a transitional state (FOXP3, ROR-gt
and RORa cell) where addition of certain cytokines/factors drives differentiation to TH3 cells (FOXP3) leading to suppression or TH17 cells
(ROR-gt and RORa) resulting in inflammation/autoimmune disease.
et al. 2004). IL-2/STAT5 signaling and 17-b-estradiol (E2)

during pregnancy cause induction of FOXP3 and increased
TREG numbers (i.e., humans and animal models) by induction
of FOXP3 iTREG and TREG cell expansion (Fainboim and
Arruvito 2011). Fetal alloantigen (Zhao et al. 2007) and placental trophoblast antigens such as heat shock protein-60, carcinoembryonic antigen, CD274 (i.e., ligand for programmed
cell death-1 receptor), and human leukocyte antigen-G
(Taglauer et al. 2008; Yang et al. 2006; Shao et al. 2005;
LeMaoult et al. 2004) are also important in TREG cell induction/activation and expansion during early pregnancy. Paternal
alloantigens and TGF-b in seminal fluid can also induce/activate TREG (Robertson et al. 2009; Robertson and Sharkey
2001). During the course of pregnancy, there are increased
numbers of CD4CD25 cells in the circulation in early pregnancy, reaching an apex during the second trimester and declining until the postpartum period where they reach a number that
is nearly at the prepregnancy level (Heikkinen et al. 2004).
TREG are significantly decreased in deciduas from spontaneous
vaginal deliveries when compared with Caesarean section,

indicating a potential role of declining TREG in fetal delivery
(Xhao et al. 2007; Sindram-Trujillo et al. 2004). The ovary normally contains a population of thymic-derived nTREG (Samy et
al. 2005), and the testis has been shown to have a locally
induced iTREG population that are specifically targeted to testicular antigens and contributes to testicular immune privilege
(Nasr et al. 2005). TREG have been shown to be necessary for
immune tolerance of the conceptus, oocytes, and spermatozoa
(Trowsdale and Betz 2006). There are increased numbers of
TREG in the blood, decidual tissue, and lymph nodes draining
the uterus (Tilburgs et al. 2008; Thuere et al. 2007). TREG cell
depletion and/or decreased function have been shown to lead to
loss of pregnancy in rodent models (Aluvihare et al. 2004) and
in patients who have miscarried (Yang et al. 2008; Sasaki et al.
2004). Infertility, miscarriage, and pre-eclampsia have been
linked with decreased TREG number and/or function (Guerin
et al. 2009; Tilburgs et al. 2008; Thuere et al. 2007; Trowsdale
and Betz 2006; Aluvihare et al. 2004).
198
PETERSON
FIGURE 15.During an infection with a pathogenic organism complex networks develop where nTREG undergo antigen-specific expansion and Tr1
and iTREG cells undergo pathogen-specific differentiation in the presence of tolerogenic cytokines (IL-10 and TGF-b). All three types of TREG
cells then work to suppress effector T-cells in the inflammatory milieu by soluble factors, cell-to-cell contact, or competing for IL-2. nTREG can
also induce infectious tolerance in dendritic cells. Together these result in suppression of immune responses to pathogens. Pathogens can also use
TREG for their own benefit to evade the immune response.
ROLE OF TREG IN CANCER

TREG have been shown to dampen the cytotoxic immune
response (CD8/NK cell) as well as suppressing overall level
of host immune response to the neoplasm in cancer. Researchers have looked at numbers and location of TREG and the effects
on cancer patient outcome (Menetrier-Caux et al. 2009). They
showed that TREG are present in a large panel of solid tumors in
humans. TREG in pulmonary, pancreatic, gastric, hepatic, and
ovarian carcinomas have been correlated with a negative outcome; while in B-cell lymphoma, head and neck cancer and
colonic carcinoma TREG have been associated with a positive
patient outcome (Menetrier-Caux et al. 2009). TREG had no
effect on survival with prostatic, renal, and squamous cell carcinomas (Menetrier-Caux et al. 2009). In breast cancer, TREG in
peri-tumoral lymphoid aggregates were correlated with a
negative outcome, while intra-tumoral TREG were associated

with a positive outcome. (Menetrier-Caux et al. 2009) The
results of vaccine-induced cancer T-cell mediated immunotherapy (i.e., cancer vaccine therapy) have proven to be disappointing (Welters et al. 2008) due to the vaccines inability to
induce effector T-cells, TREG already present in the tumor, and/
or induction of TREG by the vaccine itself. Based on these findings, the success of cancer vaccine therapy will depend on the
balance between TREG and effector cells at the start of therapy
(Welters et al. 2008). Neoplastic TREG cells in Sezary Syndrome (leukemic variant of cutaneous T-cell lymphoma in the
human) express variant splices of FOXP3. The neoplastic cells
with variant FOXP3 are able to function as TREG and work to
dampen the host immune response against the neoplastic
T-lymphocytes (Krejsgaard et al. 2008).
Vol. 40, No. 2, 2012
REGULATORY T-CELLS
199
FIGURE 16.The balance between effector cells and TREG cells in the inflammatory milieu dictate the course of and ultimate resolution or persistence of the inflammatory response to an infectious agent. There are advantages and disadvantages for both the host and microbe if there is an
imbalance in the numbers/function of effector cells and TREG.
TREG AS
MODULATORY TARGET
IN
DRUG DEVELOPMENT
Pharmacological agents have been developed that control

TREG number and/or function (Ohkura et al. 2011; Nizar et
al. 2010; Thorburn and Hansbro 2010; Nandakumar et al.
2009; Tao et al. 2007; Yang et al. 2006). Molecules that
increase TREG number include rTGF-b, FTY720 (Fingolimod,
a S1P1 modulator), retinoic acids, histone deacetylase inhibitors, and probiotics. TREG suppressive function is augmented
with the following agents: recombinant IL-2, Rapamycin
(mTOR pathway inhibitor) in type-1-diabetes mellitus and
renal transplantation, and histone deacetylase inhibitors. Histone deacetylase inhibitors that are also known as chromatin
modifying agents (e.g., HDAC9) lead to increased TREG number and function by a combination of FOXP3 protein acetylation, TREG chromatin remodeling, and promotion of TREG
development (Tao et al. 2007). While agents that block the
TGF-b and/or CTLA-4 signaling pathways decrease TREG
number and/or function. Aromatase inhibitors (e.g., Letrozole)
used for estrogen modulation in cancer chemotherapy regimens
and recombinant IL-2 lead to decreased TREG numbers.
Biopharmaceutical agents have been developed to modulate
TREG (Khan et al. 2011; Ohkura et al. 2011; De Serres et al.
2011; Weber et al. 2009) including Tremelimumab (CTLA-4
modulation) in advanced melanoma which leads to increased

effector T-cell numbers with no change in TREG number,
CD25 modulation by Daclizumab in multiple sclerosis (MS)
and advanced breast cancer, Denileukin diftitox as a cancer
vaccine, LMB-2 and RFT5-SMPY-dgA in advanced melanoma
decrease TREG number, anti-GITR MoAb (DTA1) which leads
to decreased TREG number, agonist anti-OX40 MoAb (OX86)
and anti-FR4 MoAb which decrease TREG number, and Alemtuzumab (anti-CD52 MoAb) which increases TREG number.
Adoptive transfer of TREG has been evaluated in clinical
trials of patients that have received human stem cell transplants
to induce transplant tolerance, patients with GVHD, and type 1
diabetes mellitus patients to ameliorate autoimmune disease
with a good patient safety profile and showed that the procedure is feasible, yet efficacy still needs to be shown convincingly (Riley et al. 2009; Trzonkowski et al. 2009).
CONCLUSIONS
TREG have both positive and negative functions in the
immune system. Positive aspects of TREG include suppression
of immune responses including autoimmunity, maintaining
immune homeostasis, as well as tolerance in xenotransplantation, at mucosal sites, and during pregnancy. Negative aspects
200
PETERSON
of TREG can include manipulation by some infectious agents to

avoid the hosts immune response and in cancer TREG can suppress the bodys cytotoxic immune response to neoplastic cells.
The balance between TREG and effector cells determines the
fate of an immune response. The fact that the immune system
has so many sources of regulatory cells which overlap functionally but have slightly different induction/activation pathways
underscores the importance of TREG in host survival.
ACKNOWLEDGMENTS
I would like to acknowledge the following researchers:
Virginia Godfrey, DVM, PhD, DACVP, from the University
of North Carolina at Chapel Hill for her kind donation
of the Scurfy mouse images; Caroline Genell from the
GlaxoSmithKline, Safety Assessment, Immunotoxicology
Group for the rat TREG flow cytometry data; and John
Spaull of the GlaxoSmithKline MCT Cell Biology Group
for the FOXP3 immunofluorescence images.
REFERENCES
Aandahl, E. M., Michaelsson, J., Moretto, W. J., Hecht, F. M., and Nixon, D. F.
(2004). Human CD4CD25 regulatory T cells control T-cell responses to
Human Immunodeficiency Virus and Cytomegalovirus antigens. J Virol
78, 24549.
Abraham, C., and Cho, J. (2009). Interleukin-23/TH 17 pathways and inflammatory bowel disease. Inflamm Bowel Dis 15(7), 1090100.
Abraham, C., and Medzhitov, R. (2011). Interactions between the host innate
immune system and microbes in inflammatory bowel disease. Gastroenterol 140, 172937.
Akdis, M. (2009). Immune tolerance in allergy. Curr Opin Immunol 21, 7007.
Aluvihare, V. R., Kallikourdis, M., and Betz, A. G. (2004). Regulatory T cells
mediate maternal tolerance to the fetus. Nat Immunol 5, 26671.
Andersson, J., Boasso, A., Nilsson, J., Zhang, R., Shire, N. J., Lindback, S.,
Shearer, G. M., and Chougnet, C. A. (2005). Cutting edge: The prevalence
of regulatory T cells in lymphoid tissue is correlated with viral load in HIVinfected patients. J Immunol 174(6), 31437.
Annacker, O., Asseman, C., Read, S., and Powrie, F. (2003). Interleukin-10 in
the regulation of T cell-induced colitis. J Autoimmun 20, 2779.
Askenasy, N., Kaminitz, A., and Yarkoni, S. (2008). Mechanisms of T regulatory cell function. Autoimmunity Rev 7, 3705.
Atanackovic, D., Brunner-Weinzieri, M. C., Kroger, H., Serke, S., and Deter,
H. C. (2002). Acute physiological stress simultaneously alters hormone
levels, recruitment of lymphocyte subsets, and production of reactive oxygen species. Immunol Invest 31(2), 7391.
Atanackovic, D., Schnee, B., Schuch, G., Faltz, C., Schulze, J., Weber, C. S.,
Schafhausen, P., Bartels, K., Bokermeyer, C., Brunner-Weinzieri, M. C.,
and Deter, H. C. (2006). Acute physiological stress alerts the adaptive
immune response: stress-induced mobilization of effector T cells. J Neuroimmunol 176, 14152.
Battaglia, M., Stabilini, A., Migliavacca, B., Horejs-Hoeck, J., Kaupper, T.,
and Roncarolo, M. G. (2006). Rapamycin promotes expansion of functional
CD4CD25FOXP regulatory T cells of both healthy subjects and type 1
diabetic patients. J Immunol 177(12), 833847.
Beissert, S., Schwartz, A., and Schwartz, T. (2006). Regulatory T cells. J Invest
Dermatol 126, 1524.
Belkaid, Y., Piccirillo, A. C., Mendez, S., Shevack, E., and Sacks, D. L. (2002).
CD4CD25 regulatory T cells control Leishmania major persistence and
immunity. Nature 420, 5027.
Belkaid, Y., and Rouse, B. T. (2005). Natural regulatory T cells in infectious
disease. Nature Immunol 6(4), 35360.
Bettelli, E., Carrier, Y., Gao, W., Korn, T., and Strom, T. B. (2006). Reciprocal
developmental pathways for the generation of pathogenic effector TH 17
and regulatory T cells. Nature 441, 2358.
Bienvenu, B., Martin, B., Auffray, C., Cordier, C., Becourt, C., and Lucas, B.
(2005). Peripheral CD8CD25 T lymphocytes from MHC class IIdeficient mice exhibit regulatory activity. J Immunol 175, 24653.
Biros, D. (2008). Anterior chamber-associated immune deviation. Vet Clin
Small Anim 38, 30921.
Blaha, P., Bigenzahn, S., Koporc, Z., Scmid, M., Langer, F., and Selzer, E.
(2003). The influence of immunosuppressive drugs on tolerance induction
through bone marrow transplantation with costimulation blockade. Blood
101(7), 288693.
Bloom, D. (2006). CD4CD25FOXP3 regulatory T cells in nonhuman primates. Transplant Rev 20, 1215.
Bluestone, J. A., and Tang, Q. (2005). How do CD4CD25 regulatory T cells
control autoimmunity? Curr Opin Immunol 17, 63842.
Blumberg, R. S. (2009). Inflammation in the intestinal tract: Pathogenesis and
treatment. Dig Dis 27, 45564.
Brossay, L., Tangri, S., Bix, M., Cardell, S., Locksley, R. M., and Kronenberg,
M. (1998). Mouse CD1-autoreactive T cells have diverse patterns of reactivity to CD1 targets. J Immunol 160, 36818.
Brunkow, M. E., Jeffery, E. W., Hjerrild, K. A., Paeper, B., Clark, L. B.,
Yasayko, S.-A., Wilkinson, J. E., Galas, D., Ziegler, S. F., and Ramsdell,
F. (2001). Disruption of a new forkhead/winged helix protein, scurfin,
results in the fatal lymphoproliferative disorder of the Scurfy mouse. Nat
Genetics 27, 6873.
Cabrera, R., Tu, Z., Xu, Y., Firpi, R. J., Rosen, H. R., Liu, C., and Nelson,
D. R. (2004). An immunomodulatory role for CD4CD25 regulatory
T lymphocytes in Hepatitis C Virus infection. Hepatology 40,
106271.
Caramahlo, I., Lopes-Carvahlo, T., Ostler, D., Zelenay, S., Haury, M., and
Demengeot, J. (2003). Regulatory T cells selectively express Toll-Like
Receptors and are activated by lipolysaccharide. J Exp Med 197,
40311.
Chang, C. C., Ciubotariu, R., and Manavalan, J. S. (2002). Tolerization of dendritic cells by T(s) cells: The crucial role of inhibitory receptors ILT3 and
ILT4. Nature Immunol 3, 23743.
Chen, W., Ford, M. S., and Young, K. J. (2003). Role of double-negative regulatory T cells in long-term cardiax xenograft survival. J Immunol 170,
184653.
Chen, W., Perruche, S., and Li, J. (2007). CD4CD25 T regulatory cells and
TGF-b in mucosal immune system: The good and the bad. Curr Medicinal
Chem 14, 22459.
Chen, W., Zhou, D., and Torrealba, J. R. (2005). Donor lymphocyte infusion
induces long-term donor-specific cardiac xenograft survival through activation of recipient double-negative regulatory T cells. J Immunol 175,
340916.
Claman, H. N., Chaperon, E. A., and Triplett, R. F. (1966). Thymus-marrow
cell combinations. Synergism in antibody production. Proc Soc Exp Biol
Med 122, 116771.
Clark, L. B., Appleby, M. W., Brunkow, M. E., Wilkinson, J. E., Ziegler, S. F.,
and Ramsdell, F. (1999). Cellular and molecular characterization of the
Scurfy mouse mutant. J Immunol 162, 254654.
Cohen, J. L., Trenado, A., Vasey, D., Klatzmann, D., and Salomon, B. L.
(2002). CD4CD25 immunoregulatory T cells: New therapeutics for
graft-versus-host disease. J Exp Med 196, 4016.
Cortesini, R., LeMaoult, J., Ciubotariu, R., and Suciu Foca Cortesini, N.
(2001). CD8CD28 T suppressor cells and the induction of antigenspecific, antigen-presenting cell-mediated suppression of TH reactivity.
Immunol Rev 182, 2016.
Coutinho, A., Caramalho, I., Seixas, E., and Demengeot, J. (2005). Thymic
commitment of regulatory T cells is a pathway of TCR-dependent selection
that isolates repertoires undergoing positive or negative selection. Curr Top
Microbiol Immunol 293, 4371.
Crome, S. Q., Wang, A. Y., and Levings, M. K. (2009). Translational minireview series on T H 17 cells: Function and regulation of human T helper
17 cells in health and disease. Clin Exp Immunol 159, 10919.
Vol. 40, No. 2, 2012
REGULATORY T-CELLS
Cupedo, T., Nagasawa, M., Weijer, K., Blom, B., and Spits, H. (2005).
Development and activation of regulatory T cells in the human fetus. Eur
J Immunol 35, 38390.
Datta, S., and Sarvetnick, N. (2009). Lymphocyte proliferation in immunemediated diseases. Trends Immunol 30(9), 4308.
Deaglio, S., et al. (2007). Adenosine generation catalyzed by CD39 and CD73
expressed on regulatory T cells mediates immune suppression. J Exp Med
204, 125765.
De Boer, O. J., van der Loos, C. M., Teeling, P., van der Wal, A. C., and
Teunissen, M. B. M. (2007). Immunohistocehmical analysis of regulatory
T cell markers FOXP3 and GITR on CD4CD25 T cells in normal skin
and inflammatory dermatoses. J Histochem and Cytochem 55(9), 8918.
De Serres, S. A., Yeung, M. Y., Mfarrej, B. G., and Najafian, N. (2011).
Effect of biologic agents on regulatory T cells. Transplantation Rev
25(3), 1106.
Diamond, M. S., Shrestha, B., Marri, A., Mahan, D., and Engle, M. (2003). B
cells and antibody play critical roles in the immediate defense of disseminated infection by West Nile Encephalitis Virus. J Virol 77(4), 257886.
Di Giacinto, C., Marinaro, M., Sanchez, M., Strober, W., and Boirivant, M.
(2005). Probiotics ameliorate recurrent TH 1-mediated murine colitis by
inducing IL-10 and IL-10-dependent TGF-b-bearing regulatory cells. J
Immunol 174, 323746.
Dittel, B. N. (2008). CD4 T cells: Balancing the coming and going of
autoimmune-mediated inflammation in the CNS. Brain, Behavior, and
Immunity 22, 42130.
Fainboim, L., and Arruvito, L. (2011). Mechanisms involved in the expansion
of TREG during pregnancy: role of IL-2/STAT5 signalling. Journal of
Repro Immunol 88, 938.
Fontenot, J. D., Gavin, M. A., and Rudensky, A. Y. (2003). FOXP3 programs
the development and function of CD4CD25 regulatory T-cells. Nat
Immunol 4, 3306.
Fontenot, J. D., and Rudensky, A. Y. (2005). A well adapted regulatory contrivance: Regulatory T cell development and the forkhead family transcription factor FOXP3. Nature Immunol 6(4), 3317.
Freir, E., Weber, C. S., Nowottne, U., Horn, C., Bartels, K., Meyer, S., Hildebrandt, Y., Luetkens, T., Cao, Y., Pabst, C., Muzzulini, J., Schnee, B., Brunner-Weinzieri, M. C., Marangalo, M., Bokemeyer, C., Deter, H.-C., and
Atanackovic, D. (2010). Decrease of CD4FOXP3 T regulatory cells in
the peripheral blood of human subjects undergoing a mental stressor.
Psychoneuroendocrinol 35, 66373.
Fujio, K., Okamura, T., and Yamamoto, K. (2010). The family of IL-10secreting CD4 T cells. Advances in Immunology 105, 99129.
Garin, M. I., et al. (2007). Galectin-1: A key effector of regulation mediated by
CD4CD25 T cells. Blood 109, 205865.
Germain, R. N. (2008). Special regulatory T cell review: A rose by any other
name: from suppressor T cells to TREG, approbation to unbridled enthusiasm. Immunol 123, 207.
Godfrey, V. L., Wilkinson, J. E., and Russell, L. B. (1991). X-linked lymphoreticular disease in the Scurfy (sf) mutant mouse. Am J Pathol 138(6),
137987.
Gregori, S., Casorati, M., Amuchastegui, S., Smiroldo, S., Davalli, A. M., and
Adorini, L. (2001). Regulatory T cells induced by 1-Alpha-25Dihydroxyvitamin D3 and Mycophenolate Mofetil treatment mediate
transplantation tolerance. J Immunol 167(4), 194553.
Greshon, R. K., and Kondo, K. (1971). Infectious immunological tolerance.
Immunology 21, 90314.
Grossman, W. J., et al. (2004). Differential expression of Granzymes A and B
in human cytotoxic lymphocyte subsets and T regulatory cells. Blood 104,
28408.
Guerin, L. R., Prins, J. R., and Robertson, S. A. (2009). Regulatory T-cells and
immune tolerance in pregnancy: A new target for infertility treatment?
Human Repro Update 15(5), 51735.
Han, C. H., Li, H. F., Wang, Y. X., Zhang, M., Wang, Y., and Yin, M. (2005).
The influence of Mycophenolate Mofetil upon the maturation and allostimulatory activity of cultured dendritic cell progenitors and the effects of
tolerance induction in allograft recipients. Zhonghua YiXue ZaZhi 85(19),
132732.
201
Heikkinen, J., Mottonen, M., Alanen, A., and Lassila, O. (2004). Phenotypic
characterization of regulatory T cells in the human decidua. Clin Exp
Immunol 136, 3738.
Hesse, M., Piccirillo, C. A., Belkaid, Y., Prufer, J., Mentink-Kane, M., Leusink,
M., Cheever, A. W., and Wynn, T. A. (2004). The pathogenesis of schistosomiasis is controlled by cooperating IL-10-producing innate effector and
regulatory T cells. J Immunol 172, 315766.
Hezova, R., Slaby, O., Faltejskova, P., Mikulkova, Z., Buresova, I., Muthu
Raja, K. R., Hodek, J., Ovesna, J., and Michalek, J. (2010). MicroRNA342, microRNA-191, and microRNA-510 are differentially expressed in
T regulatory cells of type 1 diabetic patients. Cellular Immunol 260, 704.
Hori, S., Carvahlo, T. L., and Demengeot, J. (2002). CD25CD4 regulatory T
cells suppress CD4 T cell-mediated pulmonary hyperinflammation driven
by Pneumocystis carinii in immunodeficient mice. Eur J Immunol 32,
128291.
Hori, S., Nomura, T., and Sakaguchi, S. (2003). Control of regulatory T cell
development by the transcription factor FOXP3. Science 299, 105761.
Huang, W. H., Yan, Y., Li, J., De Boer, B., House, A. K., and Bishop, G. A.
(2003). A short course of Mycophenolate immunosuppression inhibits
rejection, but not tolerance, of rat liver allografts in association with inhibition of Interleukin-4 and alloantibody responses. Transplantation 76(8),
115965.
Ikehara, Y., Yasunami, Y., and Kodama, S. (2000). CD4 Valpha14 natural
killer T cells are essential for acceptance of rat islet xenografts in mice.
J Clin Invest 105, 17617.
Jager, A., and Kuchroo, V. K. (2010). Effector and regulatory T-cell subsets in
autoimmunity and tissue inflammation. Scand J Immunol 72, 17384.
Jiang, S., Game, D. S., and Davies, D. (2005). Activated CD1d-restricted
natural killer T cells secrete IL-2: Innate help for CD4CD25 regulatory
T cells? Eur J Immunol 35, 1193200.
Jiang, H., Zhang, S. I., and Pernis, B. (1992). Role of CD8 T cells in murine
experimental allergic encephalomyelitis. Science 256, 12135.
Joetham, A., et al. (2007). Naturally occurring lung CD4CD25 T cell regulation of airway allergic responses depends on IL-10 induction of TGF-b. J
Immunol 178, 143342.
Joshi, A., Vahlenkamp, T. W., Garg, H., Tompkins, W. A. F., and Tompkins,
M. B. (2004). Preferential replication of FIV in activated CD4CD25 T
cells independent of cellular proliferation. Virol 321, 30722.
Kapp, J. A., and Bucy, R. P. (2008). CD8 suppressor T cells resurrected.
Human Immunol 69, 71520.
Kapp, J. A., Kapp, L. M., and McKenna, K. C. (2004 ). gd T cells play an essential role in several forms of tolerance. Immunol Res 29(1), 93102.
Kapp, J. A., and Ke, Y. (1997). The role of gd TCR-bearing T cells in oral tolerance. In Proceedings of the 72nd Forum in Immunology: Tolerance and
Immunity in the Mucosal Immune System, pp. 5617.
Ke, Y., and Kapp, J. A. (1996). Oral antigen inhibits priming of CD8 CTL,
CD4 T cells, and antibody responses while activating CD8 suppressor
T cells. J Immunol 156, 91621.
Ke, Y., Pearce, K., Lake, J. P., Ziegler, H. K., and Kapp, J. A. (1997). gd T lymphocytes regulate the induction and maintenance of oral tolerance. J Immunol 158, 36108.
Khan, S., Burt, D. J., Ralph, C., Thistlethwaite, F. C., Hawkins, R. E., and Elkord, E. (2011). Tremelimumab (anti-CTLA-4) mediates immune responses
mainly by direct activation of T effector cells rather than by affecting T regulatory cells. Clinical Immunol 138, 8596.
Korn, T., Bettelli, E., Gao, W., Awasthi, A., and Jager, A. (2007). IL-21 initiates an alternative pathway to induce proinflammatory TH 17 cells. Nature
448, 4847.
Korn, T., Bettelli, E., Oukka, M., and Kuchroo, V. K. (2009). IL-17 and TH 17
cells. Annu Rev Immunol 27, 485517.
Krejsgaard, T., Gjerdrum, L. M., Ralfkiaer, E., Lauenborg, B., Eriksen, K. W.,
Mathiesen, A.-M., Bovin, L. F., Gniadecki, R., Geisler, C., Ryder, L. P.,
Zhang, Q., Wasik, M. A., Odum, N., and Woetmann, A. (2008). Malignant
TREG express low molecular splice forms of FOXP3 in Sezary syndrome.
Leukemia 22, 22309.
Kronenberg, M. (1994). Antigens recognized by gd T cells. Curr Opin Immunol 6, 6471.
202
PETERSON
Kullberg, M. C., Jankovic, D., Gorelick, P. L., Caspar, P., Letterio, J. J.,
Cheever, A. W., and Sher, A. (2002). Bacteria-triggered CD4 T regulatory
cells suppress Helicobacter hepaticus-induced colitis. J Exp Med 196,
50515.
Kuniyasu, Y., Takahashi, T., Itoh, M., Shimizu, J., Toda, G., and Sakaguchi, S.
(2000). Naturally anergic and suppressive CD25CD4 T cells as a functionally and phenotypically distinct immunoregulatory T cell population.
Int Immunol 12, 114555.
Kursar, M., Bonhagen, K., Fensterle, J., Kohler, A., Hurwitz, R., Kamradt,
T., Kaufman, S. H. E., and Mittrucker, H.-W. (2002). Regulatory
CD4CD25 T cells restrict memory CD8 T cell responses. J Exp
Med 196, 158592.
Larsen, C. P., Elwood, E. T., Alexander, D. Z., Ritchie, S. C., Hendrix, R., and
Tucker-Burden, C. (1996). Long-term acceptance of skin and cardiac allografts after blocking CD40 and CD28 pathways. Nature 381(6581), 4348.
Le, N. T., and Chao, N. (2007). Regulating regulatory T cells. Bone Marrow
Transplant 39, 19.
Lehner, T. (2008). Special regulatory T cell review: The resurgence of the concept of contrasuppression in immunoregulation. Immunol 123, 404.
LeMaoult, J., Krawice-Radanne, I., Dausset, J., and Carosella, E. D. (2004).
HLA-GI-expressing antigen presenting cells induce immunosuppressive
CD4 T cells. Proc Natl Acad Sci USA 101, 70649.
Lenert, P., Brummel, R., Field, E. H., and Ashman, R. F. (2005). TLR-9 activation of marginal zone B cells in lupus mice regulates immunity through
increased IL-10 production. J Clin Immunol 25(1), 29.
Li, B., and Greene, M. I. (2008). Special regulatory T cell review: FOXP3 biochemistry in regulatory T cellshow diverse signals regulate suppression.
Immunol 123, 179.
Li, X., Braun, J., and Wei, B. (2011). Regulatory B cells in autoimmune diseases and mucosal immune homeostasis. Autoimmunity 44(1), 5868.
Lim, H. W., Broxmeyer, H. E., and Kim, C. H. (2006). Regulation of trafficking receptor expression in human forkhead box P3 regulatory T cells.
J Immunol 177, 84051.
Liston, A., Nutsch, K. M., Farr, A. G., Lund, J. M., Rasmussen, J. P., Koni, P.
A., and Rudensky, A. Y. (2008). Differentiation of regulatory FOXP3 T
cells in the thymic cortex. Proc Nat Acad Sciences 105(33), 119038.
Liu, F., Liu, J., Chen, Y., Song, L., He, Q., and Chen, J. (2010).
CD4CD25FOXP3 regulatory T cell depletion may attenuate the development of silica-induced lung fibrosis in mice. PLoS ONE 5(11), 111.
Lourenco, E. V., and La Cava, A. (2011). Natural regulatory T cells in autoimmunity. Autoimmunity 44(1), 3342.
Lundgren, A., Suri-Payer, E., Enarsson, K., Svennerholm, A. M., and Lundin,
B. S. (2003). Helicobacter pylori-specific CD4CD25 high regulatory cells
suppress memory T-cell responses to H. pylori in infected individuals.
Infect Immun 71, 175562.
Ma, C. S., Chew, G. Y. J., and Simpson, N. (2008). Deficiency of Th17 cells in
hyper IgE syndrome due to mutations in STAT3. J Exp Med 205, 1551.
Maggi, E., Cosmi, L., Liotta, F., Romagnani, P., Romagnani, S., and Annunziato, F. (2005). Thymic regulatory T cells. Autoimmunity Rev 4, 57986.
Malek, T. R., Yu, A., Zhu, L., Matsutani, T., Adeegbe, D., and Bayer, A. L.
(2008). IL-2 family of cytokines in T regulatory cell development and
homeostasis. J Clin Immunol 28, 6359.
Mangan, P. R., Harrington, L. E., OQuinn, D. B., Helms, W. S., and Bullard,
D. C. (2006). Transforming growth factor-b induces development of the
TH 17 lineage. Nature 441, 2314.
Menetrier-Caux, C., Gobert, M., and Caux, C. (2009). Differences in tumor
regulatory T-cell localization and activation status impact patient outcome.
Cancer Res 69, 78958.
Mitchell, G. F., and Miller, J. F. (1968). Cell to cell interaction in the immune
response. II. The source of hemolysin-forming cells in irradiated mice
given bone marrow and thymus or thoracic duct lymphocytes. J Exp Med
128, 82137.
Mizoguchi, E., Mizoguchi, A., Preffer, F. I., and Bhan, A. K. (2000). Regulatory role of mature B cells in a murine model of inflammatory bowel disease. Int Immunol 12(5), 597605.
Montagnoli, C., Bacci, A., Bozza, S., Gaziano, R., Mosci, P., Sharpe, A. H.,
and Romani, L. (2002). B7/CD28-dependent CD4CD25 regulatory
T-cells are essential components of the memory-protective immunity to

Candida albicans. J Immunol 169, 6298308.
Mosier, D. E. (1967). A requirement for two cell types for antibody formation
in vitro. Science 158, 15735.
Mowat, A. M. (1994). Oral tolerance and regulation of immunity to dietary
antigens. In Handbook of Mucosal Immunology (P. L. Ogra, J. Mestecky,
M. E. Lamm, W. Strober, J. R. McGhee, J. Bienenstock, eds.), pp. 185201.
Academic Press, San Diego, CA.
Muller, Y. D., Golshayan, D., Ehirchiou, D., Wekerle, T., Seebach, J. D., and
Buhler, L. H. (2009). T regulatory cells in xenotransplantation. Xenotransplantation 16, 1218.
Munk, M. E., Schoel, B., Modrow, S., Karr, R. W., Young, R. A., and Kaufmann, S. H. E. (1989). T lymphocytes from healthy individuals with specificity to self-epitopes shared by the mycobacterial and human
65-Kilodalton heat shock protein. J Immunol 143, 28449.
Nakamura, T., Sonoda, K. H., Faunce, D. E., Gumperz, J., Yamamura, T.,
Miyake, S., and Stein-Streilein, J. (2003). CD4 NKT cells, but not conventional CD4 T cells, are required to generate efferent CD8 T regulatory cells following antigen inoculation in an immune-privileged site. J
Immunol 171, 126671.
Nandakumar, S., Miller, C. W. T., and Kumarguru, U. (2009). T regulatory
cells: An overview and intervention techniques to modulate allergy outcome. Clin Mol Allergy 7, 511.
Nasr, I. W., Wang, Y., Gao, G., Deng, S., Diggs, L., Rothstein, D. M., Tellides,
G., Lakkis, F. G., and Dai, Z. (2005). Testicular immune privilege promotes
transplantation tolerance by altering the balance between memory and regulatory T cells. J Immunol 174, 61618.
Niederkorn, J. Y. (2008). Emerging concepts in CD8 T regulatory cells. Curr
Opin Immunol 20, 32731.
Nistala, K., and Wedderburn, L. R. (2009). TH 17 and regulatory T cells: Rebalancing pro- and anti-inflammatory forces in autoimmune arthritis. Rheumatol 48, 6026.
Nizar, S., Meyer, B., Galustian, C., Kumar, D., and Dalgleish, A. (2010). T regulatory cells, the evolution of targeted immunotherapy. Biochim Biophys
Acta 1806, 717.
Ochs, H. D., Khattri, R., Bennett, C. L., and Brunkow, M. E. (2002). Immune
dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome and
the Scurfy mutant mouse. Immunol Allergy Clin of North America 22,
35768.
OConnor, R. A., and Anderton, S. M. (2008). FOXP3 regulatory T cells in
the control of experimental CNS autoimmune disease. J Neuroimmunol
193, 111.
Ohkura, N., Hamaguchi, M., and Sakaguchi, S. (2011). FOXP3 regulatory T
cells: Control of FOXP3 expression by pharmacological agents. Trends
Pharmacol Sci 32(3), 15866.
Okumura, K., Takemori, T., Tokuhisa, T., and Tada, T. (1977). Specific enrichment of the suppressor T cell bearing I-J determinants: Parallel functional
and serological characterizations. J Exp Med 146, 123445.
Orihara, K., Nakae, S., Pawankar, R., and Saito, H. (2008). Role of regulatory
and proinflammatory T cell populations in allergic diseases. WAO J 1,
914.
Oswald-Richter, K., Grill, S. M., Shariat, N., Leelawong, M., Sundrud, M. S.,
Haas, D. W., and Unutmaz, D. (2004). HIV infection of naturally occurring
and genetically reprogrammed human regulatory T-cells. PLOS Biology
2(7), 95566.
Ozdemir, C., Akdis, M., and Akdis, C. A. (2009). T regulatory cells and their
counterparts: Masters of immune regulation. Clin Exp Allergy 39, 62639.
Pandiyan, P., Zheng, L., Ishihara, S., Reed, J., and Lenardo, M. J. (2007).
CD4CD25FOXP3 regulatory T cells induce cytokine deprivationmediated apoptosis of effector CD4 T cells. Nature Immunol 8, 135362.
Patel, D. D. (2001). Escape from tolerance in the human X-linked
autoimmunity-allergic dysregulation syndrome and the Scurfy mouse. J
Clin Invest 107(2), 1557.
Powrie, F., Leach, M. W., Mauze, S., Menon, S., Caddle, L. B., and Coffman,
R. L. (1994). Inhibition of TH1 responses prevents inflammatory bowel disease in SCID mice reconstituted with CD45RB high CD4 T cells. Immunity
1(7), 55362.
Vol. 40, No. 2, 2012
REGULATORY T-CELLS
Quezada, S. A., Jarvinen, L. Z., Lind, E. F., and Noelle, R. J. (2004). CD40/
CD154 interactions at the interface of tolerance and immunity. Ann Rev
Immunol 22(1), 30728.
Read, S., Malstrom, V., and Powrie, F. (2000). Cytotoxic T lymphocyteassociated antigen-4 plays an essential role in the function of CD25CD4
regulatory cells that control intestinal inflammation. J Exp Med 192,
295302.
Reigert, A., and Bar-Or, A. (2008). B-cell-derived Interleukin-10 in autoimmune disease: Regulating the regulators. Nat Rev Immunol 8(6), 486.
Riley, J. L., June, C. H., and Blazar, B. R. (2009). Human T regulatory cell
therapy: Take a billion or so and call me in the morning. Immunity 30,
65665.
Robertson, S. A., Guerin, L. R., Bromfield, J. J., Branson, K. M., Ahistrom, A.
C., and Care, A. S. (2009). Seminal fluid drives expansion of the
CD4CD25 T regulatory cell pool and induces tolerance to paternal
alloantigens in mice. Biol Reprod 80(5), 103645.
Robertson, S. A., and Sharkey, D. J. (2001). The role of semen in induction of
maternal immune tolerance to pregnancy. Semin Immunol 13, 24354.
Romagnani, S., Maggi, E., Liotta, F., Cosmi, L., and Annunziato, F. (2009).
Properties and origin of human TH 17 cells. Mol Immunol 47, 37.
Roncador, G., Brown, P. J., Maestre, L., Hue, S., Martinez-Torrecuadrada, J. L.,
Ling, K.-L, Pratap, S., Toms, C., Fox, B. C., Cerundolo, V., Powrie, F., and
Banham, A. H. (2005). Analysis of FOXP3 protein expression in human
CD4CD25 regulatory T cells at the single-cell level. Eur J Immunol 35,
168191.
Roncarolo, M.-G., and Battaglia, M. (2007). Regulatory T-cell immunotherapy
for tolerance to self antigens and alloantigens in humans. Nature Rev:
Immunol 7, 58598.
Roncarolo, M.-G., Gregori, S., Battaglia, M., Bacchetta, R., Fleischhauer, K.,
and Levings, M. K. (2006). Interleukin-10-secreting type 1 regulatory T
cells in rodents and humans. Immunol Rev 212, 2850.
Sakaguchi, S. (2005). Naturally arising FOXP3-expressing CD25CD4 regulatory T cells in immunological tolerance to self and non-self. Nature
Immunol 6(4), 34552.
Sakaguchi, S., Sakaguchi, N., Asano, M., Itoh, M., and Toda, M. (1995).
Immunologic self-tolerance maintained by activated T cells expressing
IL-2 recpetor alpha-chains (CD25). Breakdown of a single mechanism of
self-tolerance causes various autoimmune diseases. J Immunol 155,
115164.
Sakaguchi, S., Yamaguchi, T., Nomura, T., and Ono, M. (2008). Regulatory T
cells and immune tolerance. Cell 133, 77587.
Samy, E. T., Parker, L. A., Sharp, C. P., and Tung, K. S. K. (2005). Continuous
control of autoimmune disease by antigen-dependent polyclonal
CD4CD25 regulatory T cells in the regional lymph node. J Exp Med
202, 77181.
Sarris, M., Andersen, K. G., Randow, F., Mayr, L., and Betz, A. G. (2008).
Neuropilin-1 expression on regulatory T cells enhances their interactions
with dendritic cells during antigen recognition. Immunity 28, 40213.
Sasaki, Y., Sakai, M., Miyazaki, S., Higuma, S., Shiozaki, A., and Saito, S.
(2004). Decidual and peripheral blood CD4CD25 regulatory T cells in
early pregnancy subjects and spontaneous abortion cases. Mol Hum Reprod
10, 34753.
Scholzen, A., Minigo, G., and Plebanski, M. (2009). Heroes or villains? T regulatory cells in malaria infection. Trends in Parasitol 26 (1), 1625.
Schwartz, R. H. (2005). Natural regulatory T cells and self-tolerance. Nature
Immunol 6(4), 32730.
Seo, N., and Tokura, Y. (1999). Downregulation of innate and acquired antitumor immunity by bystander gd and ab T lymphocytes with TH2 or Tr1
cytokine profiles. J Interferon and Cytokine Res 19, 55561.
Seo, N., Tokura, Y., Takigawa, M., and Egawa, K. (1999). Depletion of IL-10
and TGF-b producing regulatory gd T cells by administering a
Daunomycin-conjugated specific monoclonal antibody in early tumor
lesions augments the activity of CTLs and NK cells. J Immunol 163, 2429.
Shalev, I., Wong, K. M., Foerster, K., Zhu, Y., Chan, C., Maknojia, A., Zhang,
J., and Levy, G. (2009). The novel CD4CD25 regulatory T-cell effector
molecule fibrinogem-like protein-2 contributes to the outcome of murine
fulminant viral hepatitis. Hepatology 49(2), 38797.
203
Shao, L., Jacobs, A. R., Johnson, V. V., and Mayer, L. (2005). Activation of
CD8 regulatory T-cells by human placental trophoblasts. J Immunol
174, 753947.
Shevach, E. M. (2009). Mechanisms of FOXP3 T regulatory cell-mediated
suppression. Immunity 30, 63645.
Shoji, T., Muniappan, A., Guenther, D. A., Wain, J. C., Houser, S. L., and
Hoerbelt, R. (2005). Long-term acceptance of porcine pulmonary allografts
without chronic rejection. Transplant Proc 37(1), 724.
Siegmund, K., Feuerer, M., Siewert, C., et al. (2005). Migration matters: Regulatory T-cell compartmentalization determines suppressive activity in
vivo. Blood 106, 3097104.
Simpson, E. (2008). Special regulatory T-cell review: Regulation of immune
responses-examining the role of T cells. Immunol 123, 136.
Sindram-Trujillo, A. P., and Scherjon, S. A., van Hulst-van Miert, P. P., Kanhai, H. H., Roelen, D. L., and Claas, F. H. (2004). Comparison of decidual
leukocytes following spontaneous vaginal delivery and elective caesarean
section in uncomplicated human term pregnancy. J Reprod Immunol 62,
12537.
Strober, S., Cheng, L., and Zeng, D. (1996). Double-negative (CD4CD8
alpha beta) T cells which promote tolerance induction and regulate autoimmunity. Immunol Rev 149, 21730.
Suvas, S., Azkur, A. K., Kim, B. S., Kumaraguru, U., and Rouse, B. T. (2004).
CD4CD25 regulatory T cells control the severity of viral immunoinflammatory lesions. J Immunol 172, 412332.
Taglauer, S., Trikhacheva, A. S., Slusser, J. G., and Petroff, M. G. (2008).
Expression and function of PDCDI at the human maternal-fetal interface.
Biol Reprod 79, 5629.
Takenoshita, S., Fukushima, T., Kumamoto, K., and Iwadate, M. (2002). The
role of TGF-b in digestive organ disease. J Gastroenterol 37(12), 991.
Tang, Q., and Krummel, M. F. (2006). Imaging the function of regulatory
T-cells in vivo. Curr Opin Immunol 18, 496502.
Tao, R., de Zoeten, E. F., Ozkaynak, E., Wang, L., Li, B., Greene, M. I., Wells,
A. D., and Hancock, W. W. (2007). Histone deacetylase inhibitors and
transplantation. Curr Opin Immunol 19, 58995.
Taylor, P. A., Lees, C. J., and Blazar, B. R. (2002). The infusion of ex vivo activated and expanded CD4CD25 immune regulatory cells inhibits graftversus-host disease lethality. Blood 99, 34939.
Te Velde, A. A., Verstege, M. I., and Hommes, D. W. (2006). Critical appraisal
of the current practice in murine TNBS-induced colitis. Inflammatory
Bowel Diseases 12(10), 9959.
Thorburn, A. N., and Hansbro, P. M. (2010). Harnessing regulatory T cells to
suppress asthma: From potential to therapy. Am J Respir Cell Mol Biol 43,
5119.
Thuere, C., Zenclussen, M. L., Shumacher, A., Langwisch, S., Schulte-Wrede,
U., Teles, A., Paeschke, S., Volk, H. D., and Zenclussen, A. C. (2007).
Kinetics of regulatory T cells during murine pregnancy. Am J Reprod
Immunol 58, 51423.
Tian, J., Zekzer, D., Hanssen, L., Lu, Y., Olcott, A., and Kaufman, D. L.
(2001). Lipopolysaccharide-activated B cells down-regulate TH1 immunity
and prevent autoimmune diabetes in non-obese diabetic mice. J Immunol
167(2), 10819.
Tilburgs, T., Roelen, D. L., van der Mast, B. J., de Groot-Swings, G. M., Kleijburg,
C., Scherjon, S. A., and Claas, F. H. (2008). Evidence for a selective migration
of fetus-specific CD4CD25 bright regulatory T cells from the peripheral
blood to the decidua in human pregnancy. J Immunol 180, 573745.
Trentham, D. E., Townes, A. S., and Kang, A. H. (1977). Autoimmunity to type
II collagen in an experimental model of arthritis. J Exp Med 146(3),
85768.
Trowsdale, J., and Betz, A. G. (2006). Mothers little helpers: Mechanisms of
maternal-fetal tolerance. Nat Immunol 7, 2416.
Trzonkowski, P., Bieniaszewska, M., Juscinska, J., Dobyszuk, A., Krzystyniak,
A., Marek, N., Mysliwska, J., and Hellmann, A. (2009). First-in-man clinical results of the treatment of patients with graft versus host disease with
human ex vivo expanded CD4CD25CG127 T regulatory cells. Clinical
Immunol 133, 226.
Vadas, M. A., Miller, J. F., McKenzie, I. F, Chism, S. E., Shen, F. W., Boyse, E.
A., Gamble, J. R., and Whitelaw, A. M. (1976). Ly and Ia antigen
204
PETERSON
phenotypes of T cells involved in delayed-type hypersensitivity and in

suppression. J Exp Med 144, 109.
Vahlenkamp, T. W., Tompkins, M. B., and Tompkins, W. A. (2004). Feline
immunodeficiency virus infection phenotypically and functionally activates immunosuppressive CD4CD25 T regulatory cells. I Immunol
172, 475261.
Veldhoen, M., Hocking, R. J., Atkins, C. J., Locksley, R. M., and Stockinger, B.
(2006). TGF-b in the context of an inflammatory cytokine milieu supports de
novo differentiation of IL-17-producing T cells. Immunity 24, 17989.
Vidovic, D., Roglic, M., McKune, K., Guerder, S., MacKay, C., and Dembic,
Z. (1989). Qa-1 restricted recognition of foreign antigen by a gd T cell
hybridoma. Nature 340, 64650.
Vignali, D. A. A., Collison, L. W., and Workman, C. J. (2008). How regulatory
T cells work. Nature Rev Immunol 8, 52332.
Von Boehmer, H. (2005). Mechanisms of suppression by suppressor T cells.
Nature Immunol 6(4), 33844.
Wan, Y. Y., and Flavell, R. A. (2007). Yin-yang functions of transforming
growth factor-b and T regulatory cells in immune regulation. Immunol Rev
220, 199213.
Weaver, C. T., Harrington, L. E., Mangan, P. R., Gavriell, M., and Murphy, K.
M. (2006). TH 17 an effector CD4 T cell lineage with regulatory T cell ties.
Immunity 24, 67788.
Weber, C. A., Mehta, P. J., Ardito, M., Moise, L., Martin, B., and De Groot, A.
S. (2009). T cell epitope: Friend or foe? Immunogenicity of biologics in
context. Adv Drug Delivery Rev 61, 96576.
Weiner, H. L. (1997). Oral tolerance: Immune mechanisms and treatment of
autoimmune diseases. Immunology Today 18(7), 33543.
Weiner, H. L., Friedman, A., Miller, A., Khoury, S. J., al-Sabbagh, A., and Santos, L. (1994). Oral tolerance: Immunologic mechanisms and treatment of
animal and human organ-specific autoimmune disease by oral administration of autoantigens. Ann Rev Immunol 12, 80937.
Welters, M. J. P., Piersma, S. J., and van der Burg, S. H. (2008). T regulatory
cells in tumour-specific vaccination strategies. Expert Opin Biol Ther 8(9),
136579.
Wirnsberger, G., Hinterberger, M., and Klein, L. (2011). Regulatory T-cell differentiation versus clonal deletion of autoreactive thymocytes. Immunol
and Cell Biol 89, 4553.
Wolf, S. D., Dittel, B. N., Hardardottir, F., and Janeway, C. A. (1996).

Experimental autoimmune encephalomyelitis induction in genetically
B-cell deficient mice. J Exp Med 184(6), 22718.
Workman, C. J., and Vignali, D. A. A. (2004). Negative regulation of T cell
homeostasis by LAG-3 (CD223). J Immunol 174, 68895.
Xhao, J. X., Zeng, Y. Y., and Liu, Y. (2007). Fetal alloantigen is responsible for
the expansion of the CD4CD25 regulatory T cell pool during pregnancy.
J Reprod Immunol 75, 7181.
Yanaba, K., Bouaziz, J.-D., Haas, K. M., Poe, J. C., Fujimoto, M., and Tedder,
T. F. (2008). A regulatory B cell subset with a unique CD1d hi CD5 phenotype controls T cell-dependent inflammatory responses. Immunity 28(5),
639.
Yang, H., Qiu, L., Chen, G., Ye, Z., Lu, C., and Lin, Q. (2008). Proportional
change of CD4 CD25 regulatory T cells in decidua and peripheral
blood in unexplained recurrent spontaneous abortion patients. Fertil Steril
89, 65661.
Yang, K., Li, D., Luo, M., and Hu, Y. (2006). Generation of HSP60-specific
regulatory T cells and effect on atherosclerosis. Cell Immunol 243, 905.
Yong, Z., Chang, L., Mei, Y. X., and Yi, L. (2007). Role and mechanisms of
CD4 CD25 regulatory T cells in the induction and maintenance of
transplantation tolerance. Transplant Immunol 17, 1209.
Zeng, D., Lewis, D., and Dejbakhsh-Jones, S. (1999). Bone marrow NK1.1
and NK1.1 T cells reciprocally regulate acute graft versus host disease.
J Exp Med 189, 107381.
Zhang, G. Y., Hu, M., Wang, Y. M., and Alexander, S. I. (2009). FOXP3 as a
marker of tolerance induction versus rejection. Curr Opin Organ Transplant 14, 405.
Zhang, Z. X., Ma, Y., and Wang, H. (2006). Double-negative T cells, activated by xenoantigen, lyse autologous B and T cells using a perforin/
granzyme-dependent, Fas-Fas ligand-independent pathway. J Immunol
177, 69209.
Zhao, J. X., Zeng, Y. Y., and Liu, Y. (2007). Fetal alloantigen is responsible for
the expansion of the CD4CD25 regulatory T cell pool during pregnancy.
J Reprod Immunol 75, 7181.
Zhou, L., Lopes, J. E., Chong, M. M., Ivanov, I. I., and Min, R. (2008).
TGF-b-induced FOXP3 inhibits TH 17 cell differentiation by antagonizing
RORgt function. Nature 453, 23640.
For reprints and permissions queries, please visit SAGEs Web site at http://www.sagepub.com/journalsPermissions.nav.

Toxicol Pathol-2012-Peterson-186-204 PDF

Загружено:

Сведения о документе

Исходное описание:

Оригинальное название

Авторское право

Доступные форматы

Поделиться этим документом

Поделиться или встроить документ

Параметры публикации

Этот документ был вам полезен?

Это неприемлемый материал?

Авторское право:

Доступные форматы

Toxicol Pathol-2012-Peterson-186-204 PDF

Загружено:

Авторское право:

Доступные форматы

Toxicologic Pathology

Regulatory T-Cells: Diverse Phenotypes Integral to Immune Homeostasis and Suppression

Society of Toxicologic Pathology

>> Version of Record - Mar 19, 2012

Downloaded from tpx.sagepub.com by guest on November 20, 2014

Toxicologic Pathology, 40: 186-204, 2012

Regulatory T-Cells: Diverse Phenotypes Integral to

delete autoreactive T-cells, induce tolerance, and dampen

This article will review the scientific literature and provide an

The author declared no potential conflicts of interests with respect to the

Downloaded from tpx.sagepub.com by guest on November 20, 2014

Vol. 40, No. 2, 2012

induce and protect from intestinal inflammation (Powrie et al.

REGULATORY T-CELLS (TREG, NATURAL,

associated protein-4 (CTLA-4; both), co-stimulatory molecule

Downloaded from tpx.sagepub.com by guest on November 20, 2014

OTHER T-CELLS WITH REGULATORY FUNCTION

Downloaded from tpx.sagepub.com by guest on November 20, 2014

Vol. 40, No. 2, 2012

significance of DN TREG in nontransplant scenarios is therefore

Downloaded from tpx.sagepub.com by guest on November 20, 2014

(EAE) (Wolf et al. 1996), nonobese diabetes mellitus (Tian et

Downloaded from tpx.sagepub.com by guest on November 20, 2014

Vol. 40, No. 2, 2012

after lymphopenia of numerous causes including those of a

FIGURE 6.FOXP3 and CD3 dual immunofluorescence on tissue from

are attracted by inflammatory signals, although they are less

CHRONIC STRESS ON HUMAN TREG

TREG have primary effect on T-cells and/or dendritic cells

Downloaded from tpx.sagepub.com by guest on November 20, 2014

et al. 2007). Galectin-1 binding to effector T-cells and dendritic

FOXP3, a forkhead box transcription factor, is a molecular

Downloaded from tpx.sagepub.com by guest on November 20, 2014

Vol. 40, No. 2, 2012

FIGURE 8.The graphs of flow cytometry data using markers useful in

regulating a set of genes required for (1) the suppressor activity

FIGURE 9.The graphs of flow cytometry data show the percentages of

subcutaneous inflammatory cell infiltrates (e.g., ear pinnae) are

Downloaded from tpx.sagepub.com by guest on November 20, 2014

TH17 CELLS AND TREG: A RECIPROCAL BALANCE

2006; Bettelli et al. 2006; Mangan et al. 2006). Therefore, a

Oral tolerance is an important mechanism in the mucosal

Downloaded from tpx.sagepub.com by guest on November 20, 2014

Vol. 40, No. 2, 2012

FIGURE 11.The Scurfy mouse has a frameshift mutation in the

et al. 1994). At mucosal sites, TREG are responsible for

ROLE OF TREG IN INFLAMMATORY/INFECTIOUS DISEASE

Downloaded from tpx.sagepub.com by guest on November 20, 2014

in the development of silica-induced pulmonary fibrosis in a

TREG have been shown to have a pivotal role in transplant

pancreatic islet cell transplant) rejection (Battaglia et al.

Downloaded from tpx.sagepub.com by guest on November 20, 2014

Vol. 40, No. 2, 2012

et al. 2004). IL-2/STAT5 signaling and 17-b-estradiol (E2)

vaginal deliveries when compared with Caesarean section,

Downloaded from tpx.sagepub.com by guest on November 20, 2014

ROLE OF TREG IN CANCER

negative outcome, while intra-tumoral TREG were associated

Downloaded from tpx.sagepub.com by guest on November 20, 2014

Vol. 40, No. 2, 2012

Pharmacological agents have been developed that control

modulation) in advanced melanoma which leads to increased