Immunol Res (2012) 53:25–40

DOI 10.1007/s12026-012-8293-7

SINGAPORE IMMUNOLOGY NETWORK

Activation and regulation of interferon-b

in immune responses
Wei-Xiang Sin • Peng Li • Joe Poh-Sheng Yeong •

Keh-Chuang Chin

Keh-Chuang Chin

Published online: 13 March 2012

Ó Springer Science+Business Media, LLC 2012

Abstract Interferons (IFNs) were discovered more than half a century ago, and extensive research has since identified
multifarious roles for type I IFN in human immune responses. Here, we review the functions of IFN-b in innate and
adaptive immunity. We also discuss the activation and influence of IFN-b on myeloid cell types, including monocytes and
dendritic cells, as well as address the effects of IFN-b on T cells and B cells. Findings from our own laboratory, which
explores the molecular mechanisms of IFN-b activation by LPS and viruses, as well as from other groups investigating the
regulation of IFN-b by viral proteins and endogenous factors are described. The effects of post-translational modifications
of the interferon regulatory factor (IRF)-3 on IFN-b induction are also highlighted. Many unanswered questions remain
concerning the regulation of the type I IFN response in inflammation, especially the role of transcription factors in the
modulation of inflammatory gene expression, and these questions will form the basis for exciting avenues of future
research.

Keywords Interferon-b  Interferon regulatory factors  Monocytes  Transcriptional regulation 

Post-translational modification

Introduction
More than 50 years ago, Isaac and Lindenmann identified
an inhibitory glycoprotein that ‘‘interfered’’ with influenza
virus replication and dissemination [1]. This novel glyco-
protein was named interferon (IFN), and since then, there
has been significant progress in our understanding of IFN
biology. To date, 10 major mammalian IFNs have been
described: IFN-a, IFN-b, IFN-c, IFN-d, IFN-e, IFN-f, IFN-
j, IFN-m, IFN-s, and IFN-x [2]. Among the seven IFNs
that have been identified in humans, this review focuses on
the regulation and functions of IFN-b, which was first
W.-X. Sin  P. Li  J. P.-S. Yeong  K.-C. Chin (&)
Laboratory of Gene Regulation and Inflammation, Singapore
Immunology Network (SIgN), Agency for Science, Technology
and Research (A*STAR), 8A Biomedical Grove, #04 Immunos,
Biopolis, Singapore 138648, Singapore
e-mail: kehchuang_chin@immunol.a-star.edu.sg
purified and characterized in the late 1970s and early 1980s
[3–5].
IFN-b has been implicated in many human diseases
related to inflammation, including autoimmune diseases
and various cancers. In 1993, IFN-b1a and IFN-b1b were
approved by the United States Food and Drug Adminis-
tration for the treatment of relapsing–remitting multiple
sclerosis (MS) [6], although the mechanisms of action and
efficacy of this treatment are still hotly debated and
intensely controversial [7, 8]. Recently, a 21-year follow-
up study provided important new evidence that IFN-b1b
reduced all-cause mortality in MS patients [9, 10].
Recent clinical and laboratory findings have shed new
light on the roles of IFN-b in human health and disease.
IFN-b exerts a wide range of biological activities in the
human immune system, including the initiation of anti-
viral protein synthesis, promotion of cytotoxic activity, and
driving the differentiation and maturation of certain leu-
kocytes [11–16]. The Laboratory of Gene Regulation and
Inflammation based within the Singapore Immunology

123
26
Network (SIgN)—a research institute under the auspices of
Singapore’s Agency for Science, Technology and Research
(A*STAR)—aims to improve our current knowledge of the
regulation and functions of IFN-b in inflammatory cells,
including human monocytes and dendritic cells (DCs),
which sit at the junction of innate and adaptive immunity.
The first part of this article reviews the roles of IFN-b in
the development and function of innate and adaptive
immune cells, focusing on key developments in the field
since 2007—the 50th anniversary of the ‘‘birth’’ of IFNs.
The second part describes our group’s research and related
investigations into the regulation of IFN-b production,
from transcription through to cellular signaling.
IFN-b in innate immune cells
Monocytes and macrophages
Although typically elicited by viruses, type I IFN can also
be induced by bacterial infections [17–19]. Regardless of
the type of pathogen encountered, macrophages are early
producers of IFN-b that primes other macrophages and
nearby immune cells to secrete a repertoire of pro-inflam-
matory cytokines [20–22]. IFN-b plays a pivotal role in
macrophage function by synergizing with signaling path-
ways downstream of pattern recognition receptors (PRRs),
including Toll-like receptors (TLRs), to induce optimal
production of pro-inflammatory cytokines. IFN-b synergy
with PRR signaling can be either beneficial or deleterious
to the host depending on the setting [23]. Previous reports
have identified starkly contrasting effects of IFN-b synergy
with PRR under different conditions, either increasing
resistance to infection and improving survival [24], or
alternatively, decreasing resistance to infection [25] and
impairing survival [26]. For example, in infection with
Listeria monocytogenes [25, 27, 28] or Tropheryma
whipplei [29], type I IFN production may be detrimental to
the host. Host defense critically depends on the anti-
microbial activities of macrophages [20, 30], which can
Table 1 Novel intracellular PRRs that induce IFN-b in monocytes and macrophages
PRR Ligand
NOD2 MDP, MTP
50 PPP RNA
DAI/DLM-1/ZBP1 Cytosolic DNA: poly(dA:dT)
AIM2-like receptors Cytosolic DNA: poly(dA:dT)
(Aim2, IFI16)
LRRFIP1 dsRNA, dsDNA
ND not done, HSV herpes simplex virus, VACV vaccinia virus, VSV vesicular stomatitis virus, ASC apoptosis-associated speck-like protein
containing a CARD, MAVS mitochondrial antiviral signaling protein, STING stimulator of interferon genes, TBK TANK-binding kinase
123
Singapore Immunology Network: SIgN (2012) 53:25–40
detect intracellular pathogen-associated molecular patterns
(PAMPs) via internalization into endosomal compartments
[31] followed by degradation in phago-lysosomes. Recent
infection studies using viruses and L. monocytogenes have
unveiled several novel cytosolic PRRs in macrophages,
namely NOD2 [30, 32], DAI [33], Aim2 [34, 35], and
LRRFIP1 [36], which can activate IFN-b with comparable
efficiency to more established TLRs, retinoic acid-induc-
ible gene-I (RIG-I), and melanoma differentiation-asso-
ciated gene-5 (Mda-5) (Table 1).
Nucleotide-binding oligomerization domain (NOD) pro-
teins comprise a family of NOD-like receptors (NLRs)—
intracellular PRRs that recognize peptidoglycan (PGN)
degradation products such as muramyl dipeptides (MDP)
and muramyl tripeptides (MTP). Herskovits and colleagues
showed that NOD2 is required for optimal induction of IFN-
b in macrophages after phagocytosis and degradation of
L. monocytogenes [30]. Recently, two other groups discov-
ered that Mycobacterium tuberculosis (Mtb) and respiratory
syncytial virus (RSV) stimulate the cytosolic NOD2 path-
way to initiate type I IFN expression, either by a TANK-
binding kinase 1 (TBK1)-IRF5-dependent mechanism in the
case of bacterial PGN [32] or via a mitochondrial anti-viral
signaling protein (MAVS)-IRF3-mediated pathway for sin-
gle-stranded RNA (ssRNA) [37].
DNA-dependent activator of IRFs (DAI, also known as
DLM-1 or ZBP1) is a cytosolic DNA sensor and activator
of the innate immune response [33]. Intracellular DAI
facilitates B-form DNA-mediated induction of IFN-b in a
TBK1-IRF3-dependent manner [33]. While Wang et al.
[38] reported that silencing of DAI mRNA expression
inhibits IFN-b activation, a separate group claimed that
DAI deficiency does not affect IFN-b production [39]. This
discrepancy may be due to the different cell types used in
these experiments.
Another new DNA sensor recently discovered in mac-
rophages and DCs is ‘‘absent in melanoma 2’’ (Aim2),
which was detected by screening for proteins that interact
with double-stranded DNA (dsDNA) and that can be
induced by IFN-b [40]. Aim2 localizes to the cell
Pathogen studied Signaling adaptor/ Reference
mediator
L. monocytogenes TBK1, IRF5 [30, 32, 37]
M. tuberculosis MAVS, IRF3
ND TBK1, IRF3 [33, 38]
HSV-1, VACV DNA ASC, Caspase-1, [34, 42]
STING, TBK1
VSV, L. monocytogenes b-catenin, IRF3 [36]
Singapore Immunology Network: SIgN (2012) 53:25–40
cytoplasm and binds dsDNA via its HIN200 domain
(hematopoietic IFN-inducible proteins with a 200 amino
acid repeat) [41], thus sensing DNA and inducing inflam-
masome-mediated production of pro-inflammatory cyto-
kines, especially IL-1b [34]. This study also revealed that
knocking down Aim2 resulted in a significant increase in
poly(dA:dT)-induced IFN-b, which may indicate that
Aim2 plays a redundant role or acts as a negative regulator
of the IFN-b response. Further investigations will be nec-
essary to address this question. IFI16 is another pyrin and
HIN domain (PYHIN)-containing protein that functions as
an innate sensor of intracellular DNA, and it has been
proposed that PYHIN proteins may represent a new family
of innate DNA sensors, named AIM2-like receptors
(ALRs) [42].
In addition, Yang et al. [36] identified a novel PRR that
can detect both dsDNA and dsRNA to trigger type I IFN
production in macrophages challenged with vesicular sto-
matitis virus (VSV) and L. monocytogenes. Leucine-rich
repeat flightless-interacting protein 1 (LRRFIP1) promoted
the activation of interaction partner b-catenin, which
enhanced IFN-b expression by binding to IRF3 and
recruiting histone acetyltransferase p300 to the IFNB
enhanceosome [36].
Interestingly, NOD2 [43], Aim2 [34, 40], and RIG-I [44]
are each induced by IFN-b itself, suggesting that they are
engaged in a positive feedback loop with this cytokine.
Upon pathogen stimulation, these PRRs activate IFN-b
secretion into the extracellular milieu where it binds to type
I IFN receptors (IFNARs) on the secreting cell, as well as
on neighboring cells, thus driving an autocrine/paracrine
loop that up-regulates a series of genes including NOD2
[43], Aim2 [34, 40], and RIG-I [44]. This sequence of
events presumably heightens immune surveillance in the
local micro-environment and increases cellular resistance
to infection.
A number of cytosolic PRRs have been implicated
in inflammasome complex formation and inflammatory
caspase-1 activation. Intriguingly, type I IFN signaling
downstream of the IFNAR was reported to inhibit NLRP1
and NLRP3 inflammasome activation, which reduced
mature IL-1b production in mouse and human monocytes/
macrophages pre-treated with IFN-b and then challenged
with lipopolysaccharide (LPS) and alum adjuvant [45].
This study also demonstrated that type I IFN signaling
induced IL-10 and reduced pro-IL-1b synthesis [45],
indicating that IFN-b treatment can suppress both pro-IL-
1b expression and processing to bioactive IL-1b [45].
Macrophage-derived cytokines including IFN-b play a
key role in the differentiation of immune effector cells,
thus helping to bridge the innate and adaptive immune
responses [46, 47]. Antigen-presenting cells (APCs) like
macrophages and DCs produce both type I and II IFNs, as
27
well as express high levels of MHC class I and II that
permit the activation of T-cell responses [48]. As described
below, IFN-b production by DCs helps to establish a crit-
ical link between innate and adaptive immunity, which is
required for the effective clearance of pathogens.
Dendritic cells (DCs)
DCs are archetypal sentinel cells that detect pathogens and
elicit immune responses [23]. DCs have long been known to
produce type I IFN in response to viruses, purified bacterial
DNA, and synthetic prokaryotic oligodeoxynucleotides
(ODN) [49], but until comparatively recently, only macro-
phages were thought to be capable of responding to whole
bacteria [50]. However, a novel role for DCs in innate
immunity has recently emerged with the report that con-
ventional DCs (cDCs), but not plasmacytoid DCs (pDCs) or
macrophages, were able to produce large amounts of IFN-b
following bacterial degradation in phago-lysosomes and
activation of TLR7, MyD88, and IRF1 [50].
The intriguing realization that APCs are the main
sources of IFN-b during infection [21, 51, 52] underscores
the pivotal role of IFN-b in adaptive immunity [53]. Using
a single-cell visualization method, Scheu et al. [51] char-
acterized the relative contributions of each cell type to in
vivo IFN-b production in response to different stimuli.
IFN-b induces DC up-regulation of MHC class I molecules
[54], enhances DC maturation and activation [55, 56], and
ultimately promotes Th1 responses [57, 58]. Type I IFN is
a major driver of DC maturation into immuno-stimulatory
cells and is required for the optimal generation of Th1
responses [14]. The high levels of type I IFNs secreted by
pDCs can activate CD8? T cells and support cross-pre-
sentation, while also enhancing CD4? Th1 development
and restricting differentiation toward Th2 and Th17 [59]
(Fig. 1).
The immuno-modulatory effects of IFN-b on DC-med-
iated T-cell responses provide possible targets for the
development of novel therapies. DC vaccination strategies
involve activating DCs with immunogenic antigens in vitro
to boost T-cell responses, followed by re-infusion of the
activated cells to generate protective immunity to cancers
or viral and bacterial infections. However, optimizing the
delivery and presentation of DCs remains a difficult chal-
lenge. Surprisingly, several recent studies have announced
unprecedented efficacy of DC-based vaccines in the pres-
ence of type I IFN, which may accentuate DC activation
[14]. TLR7 activation enhances the ability of migratory
DCs to acquire antigens provided that type I IFN is
available [60]. In a separate investigation, the TLR agonists
poly(I:C) and LPS together with type I and II IFNs gen-
erated mature DCs with high migratory capacity and IL-12
production [61]. As a result, type I IFN has now been
123
28 Singapore Immunology Network: SIgN (2012) 53:25–40

Fig. 1 Roles of IFN-b in

Novel
different immune cells. IFN-b
intracellular
enhances DC maturation and
PRRs that
activation. Macrophage- and
induce IFN-β
DC-derived IFN-b promotes
Th1 responses and activates
CD8? T cells, as well as
enhances CD4? Th1
development while restricting
differentiation toward Th2 and
Th17. It also induces antibody IFNβ MΦ IFNβ
production and isotype class
switching by enhancing B-cell
development and differentiation
into plasma cells
Immune effector cells
IFNβ

DC T cell B cell

Maturation; ↑ Th1 development; ↑ Plasma cell

Activation; ↓ Th2 and Th17 differentiation;
DC-based differentiation; Activates plasma cells;
vaccine adjuvant Promotes Th1 Antibody production;
responses; Isotype class switching;
Activates CD8+ T cells; Auto-antibodies in
Autoimmune diseases autoimmune diseases

recognized as a vaccine adjuvant, and it may even be
possible to predict vaccine efficacy by assessing the level
of type I IFN induced [62].
IFN-b in adaptive immune cells
T cells
The ability of IFN-b to shape the adaptive immune
response by activating and priming T cells is well recog-
nized. Numerous studies have confirmed that the timing of
IFN-b exposure determines the positive or negative effect
on Th1 polarization [63], which is influenced by distinct
signal transducers and activators of transcription (STAT)
factors that are recruited upon type I IFN activation in DCs
[64]. Type I IFN can, therefore, influence T-cell activation
and differentiation directly, but can also modulate T-cell
response through effects on DCs (Fig. 1).
Although the mechanisms of action of IFN-b therapy
for relapsing–remitting MS remain unclear at this time,
effects on T-cell homeostasis seem likely to contribute to
the efficacy of this intervention. IFN-b ameliorates Th1
cell pathologies in MS patients by abrogating the pro-
inflammatory properties of IFN-c and IL-12 [65]. IFN-b
also restores the function of regulatory T cells in MS
patients by increasing the frequency and inhibitory
capacity of this population [66, 67]. In addition, IL-10/
IL-27-producing CD4? T cells are augmented after IFN-b
therapy, which may suppress effector T-cell responses and
constrain the expansion of Th17 cells [67, 68]. However,
a conflicting report instead indicated that IL-10 production
is unaltered and that IL-17 levels are decreased upon IFN-
b treatment in MS, leading the authors to conclude that
IFN-b is pro-inflammatory in Th17-induced experimental
autoimmune encephalomyelitis. These data indicate that
care must be exercised when administering IFN-b therapy
in the clinic before determining the disease subtype (Th1
vs. Th17) [69].
The therapeutic role of IFN-a in graft-versus-host dis-
ease (GVHD) and graft-versus-leukemia (GVL) has been
known for decades [70, 71], especially in the context of
relapsing chronic myeloid leukemia following bone mar-
row transplantation [72]. However, the mechanisms of
action remain obscure, and the application of type I IFN in
this setting remains controversial [73, 74]. A study by
Robb et al. [75] suggested that the efficacy of type I IFN in
protecting patients from GVHD and GVL can be attributed
to the suppression of donor CD4? T-cell proliferation and
differentiation. In contrast, CD8? T cell–dependent GVHD
and GVL responses were enhanced by the pleiotropic
effects of type I IFN. Further investigations will be nec-
essary to address this apparent dichotomy and to clarify the
complex role of type I IFN in this context.

123
Singapore Immunology Network: SIgN (2012) 53:25–40
B cells
Type I IFN is able to induce long-lived antibody produc-
tion and isotype class switching by inducing activation and
differentiation of plasma cells and memory B cells [53, 76].
Type I IFN enhances proliferation and prevents apoptosis
of primary B cells, even in the absence of mitogenic stimuli
[77], and generally enhances B cell development and
function [78] (Fig. 1).
A classical study published in 2002 demonstrated that
IFN-a/b increased survival and decreased apoptosis of B
cells, as well as amplified B-cell activation and antibody
responses upon B-cell receptor stimulation [79]. IFN-a/b
secretion by virus-activated pDCs was then shown to
induce the differentiation of CD40-activated B cells into
antibody-secreting plasma cells [80]. In line with these
findings, increasing evidence now links type I IFN with B
cell production of auto-antibodies and increased risk of
diseases such as systemic lupus erythematosus (SLE)
[81–83].
Activation of IFN-b
IFN-b can directly and indirectly influence the biological
functions of a plethora of innate and adaptive immune
cells, including monocytes, macrophages, DCs, T cells and
B cells. Type I IFN production can also be stimulated by
pathogen ligands from diverse microorganisms [84],
among which bacterial LPS and viruses are two of the most
well studied. Type I IFN has been shown to be critical for
the control of bacterial, viral, and even parasitic protozoan
infections [23]. The classical IFN-b signaling pathway
consists of membrane-associated or cytosolic receptors,
adaptor proteins in the cytosol, and transcription factors in
the nucleus.
Activation of IFN-b by LPS
PAMPs activate various membrane PRRs on the host cell,
among which TLRs are the most extensively studied. TLRs
can sense a variety of bacterial and viral ligands, which
include LPS (TLR4), PGN (TLR2), Pam3CSK4 (TLR1/2),
dsRNA and poly(I:C) (TLR3), ssRNA (TLR7/8), and CpG
ODN (TLR9) [85]. LPS is known to interact with TLR4 in
conjunction with CD14 and MD2 receptors on the host cell
membrane [86], which activates downstream MyD88-
dependent and MyD88-independent (TRIF-dependent)
signaling pathways. The MyD88-independent pathway
leads to type I IFN production, although there is also sig-
nificant cross-talk with the MyD88-dependent pathway
through NF-jB [86]. TRIF is a critical mediator in this
pathway, and TRAM and TBK1 are key adaptor molecules
29
in the cytoplasm. The kinase TBK1 phosphorylates tran-
scription factor IRF3 [87], which then dimerizes and
translocates into the nucleus to induce IFN-b expression
[88]. IFN-b is secreted into the extracellular milieu, where
it interacts with IFNARs on the host cell membrane to
activate downstream JAK-STAT signaling [89]. The
phosphorylation and activation of the kinases JAK1 and
Tyk2 lead to the phosphorylation and activation of the
transcription factors STAT1 and STAT2, which results in
the formation of transcriptional complexes that bind to
specific promoter regions of downstream cytokine genes
(known as IFN-stimulated response elements; ISREs).
STAT1/STAT2 binding to ISREs in the presence of path-
ogenic stimuli such as LPS and Sendai virus (SeV) cul-
minates in the induction of pro-inflammatory cytokines
including type I IFNs [90]. This signaling cascade, thus,
constitutes a positive feedback loop that amplifies the
production of type I IFNs.
TLR4 on the host cell membrane is a well-known
sensor of Gram-negative bacterial infections. However,
there are emerging reports that the Gram-negative bacteria
Francisella tularensis and Salmonella typhimurium acti-
vate cytosolic PRRs instead of TLR4 [91–94]. It was
reported that F. tularensis can activate the caspase-1 in-
flammasome via the intracellular sensor Aim2 and can
induce type I IFN and other pro-inflammatory cytokines
via an as yet unidentified sensor. Aim2 is now acknowl-
edged to function as a sensor of undigested viral DNA,
synthetic DNA, and DNA derived from killed bacteria
(presumably released from the phago-lysosome), ulti-
mately leading to inflammasome activation [34]. In vitro
and in vivo studies have now shown that this inflamma-
some activation is Aim2- and IRF3-dependent [95].
Although inflammasome activation depended on an intact
type I IFN response, F. tularensis-induced type I IFN
production required IRF3, but was independent of a
number of TLRs, RLRs (RIG-I like receptors), and NLRs
(including Aim2) [95]. More research will be needed to
unravel this complex relationship.
Activation of IFN-b by viruses
During viral infection, secreted IFN-b binds to IFNARs on
the surface of cells to trigger the expression of a large
number of anti-viral genes [96]. dsRNA is a PAMP derived
from viruses that can be detected by TLR3 to induce
activation of TRIF, which is also a key mediator of the LPS
response pathway. TRIF has been shown to bind to TLR3
by co-immunoprecipitation [97], and the same methodol-
ogy was employed to prove that the N-terminal domain of
TRIF interacts with TBK1 and TRAF6, while the C-ter-
minal domain of TRIF binds to the kinase RIP1 [98].
Furthermore, it was shown that TBK1 physically interacts
123
30 Singapore Immunology Network: SIgN (2012) 53:25–40

with IRF3 and that macrophages from TBK1 knockout
mice exhibit defective IRF3 activation and IFN-b tran-
scription [87]. In this virus infection model, TBK1 and
IKKi phosphorylated IRF3, which either homodimerizes or
heterodimerizes with IRF7, translocates into the nucleus,
and binds to the IFN-b promoter region to up-regulate IFN-
b expression [87]. IRF7 is another member of the IRF
family of transcription factors that can be activated upon
virus infection [99]. It was previously reported that the
response to myxoma virus infection in mouse pDCs is
dependent on IRF7 but not on IRF3 [100]. Upon virus
infection, latent IRF7 in the cytoplasm is phosphorylated
by TBK1/IKKi and forms a homodimer, followed by
translocation into the nucleus to establish the IFNB en-
hanceosome together with IRF3 and other transcription
factors (Fig. 2).
Extensive studies on the chromatin structure and pro-
moter architecture of the IFNB gene have revealed that
IFN-b transcriptional activation requires the enhancer
region located immediately upstream of the core promoter
[101]. Virus infection results in the coordinate activation of
transcription factors that assemble on this IFN-b enhancer
region. NF-jB, ATF-2/c-Jun, PU.1, and IRFs have been
identified as possessing cognate binding sites on the IFN-b
promoter region, which regulate IFN-b mRNA transcrip-
tion: the binding of these regulators is concentrated on
*(-100)bp of the IFN-b genomic locus [102, 103]
(Fig. 2).
Type I IFN production differs in magnitude and kinetics
between different cell types. One of the research interests
of our laboratory has been to build a better understanding
of how differential IFN-b induction is achieved at the
molecular level. Other groups have previously reported the
initiation of IFN-b transcription at 6 h post-virus infection
in HeLa cells [104], while our group has observed early
expression of IFN-b in human blood monocytes within
only 1–2 h of exposure to SeV [105]. This disparity raised
the possibility that distinct myeloid-specific transcription
factor(s) may be involved in the rapid induction of IFN-b
in monocytes compared with non-myeloid cell types [105].
We determined that this monocyte-restricted phenomenon
can be attributed to the constitutive binding of IRF8 and
PU.1 to the ETS/IRF composite element (EICE) motif of
the IFN-b promoter, thus creating a preformed activation
complex on the IFN-b promoter, which facilitates rapid
recruitment of IRF3 via direct interactions with IRF8
(Fig. 2) [105]. In contrast to other ubiquitously expressed
IRF family members, IRF8 is expressed only in myeloid
cells. IRF8 (also known as interferon consensus sequence
binding protein; ICSBP) is a transcription factor protein
originally isolated based on recognition of the promoter
region of the H-2LD MHC class I gene [106]. siRNA-
mediated knockdown of IRF8 in monocytes dramatically
blocked SeV- and LPS-induced IFN-b transcription, while
ectopic expression of IRF8 in mouse 32D (IRF8-/-) cells
successfully rescued SeV-induced IFN-b transcription
[105]. Through these and other experiments, we have been
able to demonstrate that IRF8 directly synergizes with
IRF3 in monocytes to facilitate faster IFN-b transcription
after pathogenic stimulation.
Different patterns of type I IFN production can be
induced by different PRRs in response to viral infections.

a Classical model of the IFN -β enhanceosome in non-myeloid cells

C-Jun
ATF-2

IRF3 IRF7 IRF3 IRF7 p50 p65

-100 -50
5’..AATGACATAGGAAAACTGAAAGGGAGAAGTGAAAGTGGGAAATTCCTCTGA..3’
3’..TTACTGTATCCTTTTGACTTTCCCTCTTCACTTTCACCCTTTAAGGAGACT..5’
PRDIV PRDIII PRDI PRDII

b Hypothetical model of the IFN -β enhanceosome in myeloid cells

? ? IRF3 IRF8 PU.1

-100 -50
5’..AATGACATAGGAAAACTGAAAGGGAGAAGTGAAAGTGGGAAATTCCTCTGA..3’
3’..TTACTGTATCCTTTTGACTTTCCCTCTTCACTTTCACCCTTTAAGGAGACT..5’
EIRE EICE

Fig. 2 Models of the IFN-b enhanceosome in non-myeloid and
myeloid cells. a Assembly of transcription factors on the IFN-b
enhanceosome as determined from crystal structures. ATF-2/c-Jun,
IRF3/IRF7, and NF-jB bind cooperatively to the IFN-b enhancer:
ATF-2/c-Jun in complex with two IRF molecules on positive
regulatory domain (PRD) IV-PRDIII, and NF-jB in complex with
two IRF molecules on the PRDI and PRDII regions of the enhancer. b
Our laboratory determined that rapid induction of IFN-b in monocytes
can be attributed to the constitutive binding of IRF8 and PU.1 to the
ETS/IRF composite element (EICE) motif of the IFN-b promoter.
Putative IRF3 and IRF8 binding sites are underlined. The potential
involvement of other transcription factors at the ETS/IRF response
element (EIRE) motif within the PRDIII region as well as the PRDIV
region in monocytes remains to be confirmed

123
Singapore Immunology Network: SIgN (2012) 53:25–40 31

For example, West Nile virus activates TLR3 [107],
whereas VSV interacts with TLR7, and herpes simplex
virus (HSV)-1 and HSV-2 use TLR9 to activate type I IFN
signaling [108–110]. Besides membrane-associated TLRs,
members of the cytosolic RLR family can also be stimu-
lated by viruses. For instance, RIG-I is employed by
Newcastle disease virus (NDV) [111], while Mda-5 is
activated by Dengue virus [112]. This may be due to the
different infection strategies used: while some viruses bind
to cell membranes, other viruses gain entry into the cytosol
through fusion of the virus envelope with the plasma
membrane [113].
Activation of IFN-b by non-viral pathogens
Mtb is the causative agent of human tuberculosis [114], and
like many other pathogens, Mtb can induce production of
type I IFN. Previous studies have found that Mtb-induced
type I IFN is IRF3-independent, since IFN-b induction was
not defective in IRF3-deficient bone marrow-derived
macrophages (BMDMs) stimulated with mycobacterial
ligands [32]. The signal transduction pathway leading to
type I IFN induction is therefore likely to be different in
Mtb compared with other Gram-negative bacteria. Indeed,
while LPS interacts with TLR4 on the cell membrane, Mtb
instead binds to intracellular NOD2 [115]. NOD2 is located
in the cytoplasm and recognizes bacterial cell wall ligands
such as MDP [116]. The mycobacterial ESX1 system
causes phago-lysosomal escape and entry into the cytosol,
which is required for effective receptor-interacting protein-
2 (Rip2) activation downstream of NOD2 [117]. Conse-
quently, TBK1 becomes activated and phosphorylates
IRF5, which in turn translocates into the nucleus to induce
type I IFN. Experimental data have shown that type I IFN
production upon MDP stimulation is impaired in TBK1-
and IRF5-deficient macrophages, but not in IRF3-deficient
macrophages [32]. There are also reports that IRF1 is
involved in Mtb infection, although only demonstrated in
THP-1 cell lines [118]. However, for other strains of bac-
teria like L. monocytogenes, TBK1 has been shown to
activate type I IFN through IRF3 [119] (Table 2).
The protozoan parasite Plasmodium falciparum is the
etiological agent of malaria [120]. P. falciparum elicits
significant amounts of type I IFN from pDCs [121]. Plas-
modial DNA has been shown to act as a PAMP that
stimulates type I IFN production, since DNase treatment
ablates any stimulatory effect for this molecule. TLR9 is
believed to be the receptor activated by parasite DNA [122,
123], which contains a specific AT-rich DNA motif
responsible for that stimulation [124]. Stimulator of IFN
genes (STING), an ER-associated protein, forms a complex
with TBK1 to activate IRF3 and IRF7, leading to type I
IFN production. This STING-TBK1-IRF3/IRF7 pathway
contributes to DNA sensing in plasmodial infection [124]
(Table 2).
Regulation of IFN-b
The production of type I IFN by host cells in response to
pathogen exposure is critical in innate and adaptive
immunity (reviewed in [19]). However, dysregulated
expression of type I IFN can be detrimental to the host, and
systemic overproduction of type I IFN can lead to septic
shock syndrome in Gram-negative sepsis [23]. Further-
more, type I IFN and members of the IRF family have also
been implicated in the induction of autoimmune responses
and in the pathogenesis of diseases such as SLE and

Table 2 Pathogen-induced signaling pathways leading to IFN-b production

PAMP Type Transcription Knockout Phenotype Signaling Ref.
factor studied of knockout pathway
involved involved

dsRNA/Sendai Virus IRF3 IRF3-/- Inhibited downstream TLR3 [180]

virus dendritic cells genes
Encephalomyocarditis Virus IRF3, IRF7 IRF3-/-, Increased lethality TLR7 [181]
virus IRF7-/-mice
Mycobacterium Gram-negative IRF5 IRF5-/-BMDMs Impaired interferon NOD2 [32]
tuberculosis bacteria production
Plasmodium Malaria IRF3, IRF7 IRF3-/-, Resistant to malaria STING [124]
falciparum IRF7-/-BMDMs disease
Group B Gram-positive IRF1 IRF1-/- mice Susceptible to TLR7 [50]
Streptococcus bacteria infection
LPS Gram-negative IRF3 IRF3-/- mice Decreased downstream TLR4 [182]
bacteria cytokines
Listeria Gram-positive IRF3 IRF3-/- mice Increased lethality TLR-independent; [25]
monocytogenes bacteria TBK1-dependent

123
32
Sjogren’s syndrome [84, 125, 126]. Levels of type I IFN,
thus, need to be carefully regulated during the course of
infection. One of the research interests of our laboratory
has been to better understand how inflammatory processes
are regulated at the molecular level. These studies on
mechanisms that limit the expression of type I IFN may
lead to the design of better therapeutic interventions in
inflammatory and autoimmune diseases [127].
Regulation of IFN-b induction by viral proteins
Viruses have evolved a plethora of countermeasures to
subvert the innate anti-viral host response. For example,
Kaposi’s sarcoma-associated herpesvirus (KSHV) can
potently attenuate the production of type I IFN (reviewed
in [128, 129]). Several KSHV viral proteins have been
identified that suppress type I IFN induction at various
levels. For instance, at the post-translational level, ORF45
blocks IRF7 phosphorylation and nuclear translocation
[130, 131], and the ubiquitin E3 ligase replication tran-
scription activator (RTA) targets IRF7 for proteasomal
degradation [132]. At the transcriptional level, the viral
IRF homologues vIRF1 and vIRF3 interfere with the DNA
binding of IRF3 and IRF7, respectively [133, 134], and K-
bZIP, ORF36, and latency-associated nuclear antigen
(LANA-1) interfere with IFN-b promoter activation [135–
137] (Fig. 3).
Regulation of IRF7-mediated IFN-b induction
IRF7 is subject to multiple layers of regulation at the
transcriptional, translational, and post-translational levels
(Fig. 3). At the transcriptional level, the Epstein–Barr virus
(EBV) BRLF1 protein was shown to repress the tran-
scription of IRF7 and IRF3, leading to decreased IRF7 and
IRF3 mRNA and protein levels and reduced IFN-b
production [138]. ATF4 is a component of the cellular
integrated stress response that can also inhibit IRF7 tran-
scription to reduce type I IFN induction [139]. Interest-
ingly, IRF7 can in turn regulate ATF4 levels and activity,
suggesting a cross-talk between the IFN response and
cellular integrated stress response [139].
IRF7 translation was recently found to be inhibited by
the translational repressors 4E-BP1 and 4E-BP2 [140].
Accordingly, 4E-BP1 and 4E-BP2 knockout mice show
enhanced type I IFN production in pDCs and increased
resistance to VSV [140]. Moreover, 4E-BP1- and 4E-BP2-
deficient mouse embryonic fibroblasts exhibited enhanced
IRF7 translation and type I IFN responses, resulting in
increased resistance to encephalomyocarditis virus and
VSV [140].
Numerous post-translational modifications can regulate
IRF7 activity: modification of IRF7 phosphorylation status
123
Singapore Immunology Network: SIgN (2012) 53:25–40
and nuclear translocation by the EBV oncoprotein latent
membrane protein-1 (LMP1) [141]; polyubiquitination of
IRF7 and subsequent proteasomal degradation by TRIM21
[142]; SUMO modification of IRF7 transcriptional activity
mediated by PIAS1 [143, 144]; and lysine acetylation
effects on IRF7 DNA binding as mediated by p300/CREB-
binding protein-associated factor (PCAF) and GCN5 [145].
Recently, TRIM28 was found to act as a IRF7-specific
SUMO E3 ligase (analogous to PIAS1) that attenuates
transcriptional activity and abrogates IFNA1, IFNB, and
ISRE promoter activation and gene expression [146]. MafB
is another molecule that was found to inhibit IRF7-medi-
ated transcription by disrupting the binding of IRF7 to
target gene promoters [147]. Interested readers are directed
to the comprehensive review of Ning et al. [99]. We shall
discuss in greater detail the modulation of IRF3 activity in
the following sections.
Regulation of IRF3-mediated IFN-b induction
by endogenous factors
Many groups have studied IRF3-mediated IFN-b activation
following TLR3 ligation. IRF3 can be regulated by myriad
post-translational modifications, including phosphor-
ylation, ubiquitination, SUMOylation, ISGylation, and
S-glutathionylation (Fig. 3). The growing list of proteins
that modulate IRF3 activation, though by no means
exhaustive, currently includes A20, Atg5-Atg12, CYLD,
DAK, DUBA, gC1qR, ISG15, ISG56, LGP2, NLRX1,
optineurin, OTUB1/2, Pin1, RBCK1, RNF5, RNF125,
SARM, SHIP-1, SIKE, SINTBAD, TAG, TRIAD3A, and
TRIM21 [148–152].
Phosphorylation of IRF3
The cytoplasmic ubiquitin-editing protein A20 (also known
as TNFAIP3) is induced by TLR ligands and can down-
regulate IRF3-dependent gene expression to reduce
inflammation (reviewed in [153]). A20 is induced upon
SeV infection [154] and functions as a negative regulator
of TLR3-induced IRF3 activation and IFN-b transcription.
A20 was initially shown to interact with TRIF and TBK1-
IKKi to inhibit activation of ISRE and the IFNB promoter
[154, 155]. Recently, the adaptor protein Tax1-binding
protein 1 (TAX1BP1) was found to be required for A20
inhibition of TBK1-IKKi by limiting Lys[63]-linked
polyubiquitination of TBK1-IKKi [149]. Finally, A20-
binding inhibitor of NF-jB 1 (ABIN1) has been identified
as a novel player that cooperates with TAX1BP1 and
TBK1-IKKi in recruiting A20 proteins to form a complex
with TBK1-IKKi [156]. While additional novel mediators
may yet be identified, the prevailing hypothesis is that the
trimolecular complex comprising ABIN1-TAX1BP1-A20
Singapore Immunology Network: SIgN (2012) 53:25–40 33

Cell membrane
dsRNA dsRNA
Endosome

TLR3
RIG-I /
MDA-5

TRIF MAVS

A20, TAX1BP1, ABIN1

(TBK1-IKKi K63-linked EBV
polyubiquitination) TBK1 IKKi
LMP1 (IRF7 K63-linked
polyubiquitination)
IRF3 post-translational modifications

Pin1, TRIM21/Ro52, RAUL, IRF3 EBV

Caspase-8, RBCK1, Cullin-1 BRLF1 (IRF7 transcription)
(IRF3 polyubiquitination) IRF7
KSHV
SUMO1, SUMO2, SUMO3 RTA (IRF7 polyubiquitination)
(IRF3 SUMOylation);
SENP2 (IRF3 de-SUMOylation) ATF4 (IRF7 transcription);
4E-BP1, 4E-BP2 (IRF7 translation);
Herc5 (IRF3 ISGylation) TRIM21, RAUL (IRF7 polyubiquitination);
SUMO1, SUMO2, SUMO3,
GRX-1 (IRF3 de-glutathionylation) PIAS1, TRIM28 (IRF7 SUMOylation)

A20, TAX1BP1, ABIN1 P P P P P P EBV

(IRF3 phosphorylation) LMP1
IRF3
IRF3

IRF3

IRF7

IRF7
IRF7

(IRF7 phosphorylation)
KSHV
ORF45
(IRF7 phosphorylation)
IRF3-CL (IRF3 nuclear translocation)
Nuclear membrane

KSHV
vIRF1 (IRF3 DNA binding); KSHV
IFN-β
K-bZIP, ORF36, LANA-1 vIRF3
(promoter activation) (IRF7 DNA binding)

E2F1, PPAR-γ, MafB PCAF, GCN5, MafB

(promoter activation) (IRF7 DNA binding)

Fig. 3 Positive and negative regulators of IFN-b induction. Viral proteasomal degradation, while their phosphorylation status affects
proteins and endogenous factors regulate IFN-b induction through their nuclear translocation and DNA binding. IFN-b induction is also
different post-translational modifications of IRF3 (left; shaded boxes) regulated at the level of promoter activation by various transcription
and IRF7 (right). Polyubiquitination of the IRFs leads to their factors

is sufficient to disrupt TBK1-IKKi Lys[63]-linked poly-
ubiquitination, decrease IRF3 phosphorylation, and depress
virus or poly(I:C)-induced IFN-b promoter activation.
Lin et al. probed the signaling pathway upstream of IRF3
and concluded that the virus-inducible, NF-jB-dependent
E3 ligase activity of A20 completely blocked RIG-I-medi-
ated IFNB, ISRE, and IFNA4 promoter activation, but only
partially inhibited TLR3-TRIF- and TBK1-IKKi-mediated
ISRE promoter activation. A20 blocks RIG-I-induced IRF3
phosphorylation, homodimerization, and DNA binding via
TRIF protein degradation and can block MAVS/VISA/IPS-
1/Cardif-mediated ISRE promoter activation in reporter
gene assays [157]. The authors, therefore, contend that A20
inhibition acts upstream of TBK1-IKKi and impacts more
greatly on RIG-I signaling than on TLR3-TRIF signaling
[157].
It was recently discovered that A20 is also a negative
regulator of IRF7. The EBV oncoprotein LMP1 induces

123
34
Lys[63]-linked polyubiquitination of IRF7 [158], but can
simultaneously induce A20, which decreases the poly-
ubiquitination of IRF7 to reduce transcriptional activation
of the IFN-a promoter [158]. In this way, A20 seems to
function as a negative regulator that balances the positive
signaling of LMP1-IRF7 activation.
Ubiquitination of IRF3
Ye et al. [127] showed that the degradation of IRF3 is
responsible for switching off IFN-b production after acute
viral infection, with no significant contribution from tran-
scriptional repressors. The peptidyl-prolyl isomerase Pin1
interacts with phosphorylated IRF3 to promote polyubiq-
uitination and proteasomal degradation of IRF3, thereby
suppressing TLR3- and RIG-I-mediated IFN-b production
following poly(I:C) stimulation or NDV infection [159].
At this point, it is interesting to highlight two conflicting
reports that describe the role of the E3 ubiquitin ligase
TRIM21 (also known as Ro52) in IRF3-mediated IFN-b
induction. Higgs et al. [160] demonstrated a negative reg-
ulatory role for TRIM21, which promoted polyubiquitina-
tion and proteasomal degradation of IRF3 to inhibit
poly(I:C)-induced activation of the IFN-b promoter
downstream of TLR3 stimulation. However, Yang et al.
[148] instead demonstrated a positive regulatory role for
TRIM21, which interfered with Pin1-IRF3 interactions to
prevent IRF3 polyubiquitination and proteasomal degra-
dation, thereby enhancing IFN-b production in response to
SeV infection. The authors attributed this discrepancy to
differing experimental conditions yielding different out-
comes [148]. In Higgs’ study, TRIM21 inhibited IFN-b
promoter activation after 48-h SeV infection of 293T cells
[160], while in Yang’s study, TRIM21 increased IFN-b
promoter activation after 9-h SeV infection of HEK293
cells [148]. Small hairpin (sh)RNA knockdown of TRIM21
led to an increase in IFN-b transcript levels after 48-h
poly(I:C) stimulation of 293T cells in Higgs’ article [160],
while siRNA knockdown of TRIM21 resulted in reduced
IFN-b transcript levels after 9-h SeV infection of HEK293
cells in Yang’s report [148]. Although cell type–specific
responses to different stimuli may account for some of
these observations, it is interesting to speculate that
unknown time critical factors may also contribute to the
disparate actions of TRIM21 after infection.
Another recent report from Higgs et al. [142] revealed
that TRIM21 negatively regulates the stability and activity
of IRF7 protein in a similar fashion to IRF3. TRIM21 also
promotes the polyubiquitination and subsequent proteaso-
mal degradation of IRF7, resulting in decreased IRF7
protein expression and decreased transactivation of the
IFNA4 promoter downstream of TLR7/9 stimulation [142].
TRIM21 has been also found to enhance IRF8-dependent
123
Singapore Immunology Network: SIgN (2012) 53:25–40
expression of pro-inflammatory cytokine genes such as IL-
12p40 following IFN-c/CpG stimulation of macrophages
[161]. TRIM21 mediates Lys[63]-ubiquitination of IRF8 to
enhance the expression of IL-12p40 in myeloid cells [161].
In contrast, Cbl-mediated ubiquitination of IRF8 can lead
to the destabilization and degradation of IRF8, in a manner
analogous to Pin1-mediated ubiquitination of IRF3 [162].
RAUL is another HECT-type ubiquitin E3 ligase that has
been implicated in the proteasomal degradation of IRF3.
RAUL inhibits type I IFN production by catalyzing the
polyubuquitination and proteasomal degradation of IRF3
and IRF7 [163]. RAUL is positively regulated by a deubiq-
uitinating enzyme ubiquitin-specific processing protease 7
(Usp7) [163]. Interestingly, the KSHV protein RTA medi-
ates RAUL deubiquitination via Usp7, thereby enhancing
RAUL degradation of IRF3 and IRF7 [163]. RIG-I-induced
caspase-8 cleaves IRF3 into an intermediary that is amena-
ble to polyubiquitination and proteasomal degradation
[164]. Other ubiquitin ligases implicated in the proteasomal
degradation of IRF3 include RBCK1 and Cullin-1.
SUMOylation of IRF3
Ubiquitin is structurally and functionally related to another
post-translational modification tag, small ubiquitin-related
modifier (SUMO). SUMOylation has been shown to serve
dual functions by antagonizing polyubiquitination and
proteasomal degradation on the one hand, but also syner-
gizing with E3 ubiquitin ligase recruitment to enhance this
pathway of degradation in other contexts (reviewed in [165,
166]). Kubota et al. [143] found that TLR and RIG-I/MDA5
activation upon VSV infection leads to TRIF- and MAVS/
VISA/IPS-1/Cardif-mediated SUMO1, SUMO2, and
SUMO3 modification of mouse IRF3 and IRF7 and that this
SUMOylation negatively regulates type I IFN gene
expression. On the other hand, Ran et al. [167] showed that
the deSUMOylating enzyme Sentrin/SUMO-specific pro-
tease 2 (SENP2) caused the deSUMOylation, polyubiqui-
tination, and proteasomal degradation of IRF3 to impair
virus-induced IFN-b transcription. These variable findings
could perhaps be attributed to species-specific differences,
but they may also indicate cross-talk between the SU-
MOylation and ubiquitination pathways acting on IRF3.
ISGylation of IRF3
In 2006, our laboratory and a separate group made the
almost simultaneous discovery that interferon-induced
HECT-type E3 protein ligase Herc5 is involved in the
conjugation of ubiquitin-like protein ISG15 to protein tar-
gets in human cells [168, 169]. One such protein target has
now been identified as IRF3 [170]. Lu et al. [170] illus-
trated that ISGylation of IRF3 in response to IFN treatment
Singapore Immunology Network: SIgN (2012) 53:25–40
and NDV infection prevents proteasomal degradation and
augments nuclear translocation of phosphorylated IRF3 to
support IFN-b promoter activation. Shi et al. [171] further
demonstrated that Herc5-mediated IRF3 ISGylation dis-
rupts the Pin1-IRF3 interaction to inhibit IRF3 poly-
ubiquitination and degradation by proteasomes. Herc5 is,
therefore, a positive regulator of IRF3-mediated IFN-b
promoter activation and IFN-b mRNA expression.
S-glutathionylation of IRF3
S-glutathionylation is another crucial post-translational
modification of IRF3 that can regulate IFN-b activation.
IRF3 is S-glutathionylated in non-infected cells, and
deglutathionylation of IRF3 catalyzed by glutaredoxin-1
(GRX-1) following viral infection is required for the
recruitment of transcription co-activator CBP-p300 [172].
However, phosphorylation, dimerization, and nuclear
translocation of IRF3 do not appear to depend on GRX-1-
mediated deglutathionylation of IRF3 [172].
Other regulators of IRF3: E2F1 and IRF3-CL
Xu et al. [173] characterized the IRF3 promoter and found
that it contains E2F transcription factor binding sites
(TFBS). Mutation of the E2F TFBS increased IRF3 pro-
moter activation, while over-expression of E2F1 decreased
IRF3 promoter activation, thereby implicating E2F1 in the
negative regulation of IRF3 gene expression [174]. In
human monocytic cells stimulated with LPS, our laboratory
previously found that TLR4-induced NF-jB recruits E2F1,
both of which then cooperatively bind to RelA and E2F1
binding sites in NF-jB target genes, such as TNF and IL1B
[175]. It is therefore interesting to speculate that E2F1 may
positively regulate NF-jB target gene expression, while
also negatively regulating IRF3-mediated gene expression.
Very recently, an alternative splicing isoform of IRF3
was cloned that shares a common N-terminus with IRF3,
but exhibits a unique C-terminus of 125 amino acids [176].
The mRNA transcript and protein of this isoform, termed
IRF3-CL, was detected in many cell lines [176]. IRF3-CL
was also found to inhibit SeV-induced IRF3 nuclear
translocation and IFN-b induction [176]. Together with
other alternative splicing isoforms such as IRF-3a [177]
and IRF3-nirs3 [178], IRF3-CL may represent a general
mechanism of reducing IRF3 activity [176].
Novel transcriptional regulators at the IFN-b promoter:
PPAR-c and MafB
Zhao et al. [156] recently reported that the peroxisome
proliferator-activated receptor (PPAR)-c agonist troglit-
azone negatively regulates LPS- and poly(I:C)-induced
35
IFN-b mRNA transcription and protein secretion in primary
peritoneal macrophages. PPAR-c acts as a transcriptional
repressor by inhibiting TLR4- and TLR3-induced IRF3
transcriptional activation—preventing IRF3 binding to the
IFN-b promoter and impairing IFN-b signaling [156].
Transcription factor MafB is involved in the establish-
ment and maintenance of the myeloid lineage and has been
identified as a negative regulator of IFN-b induction at the
IFNB gene promoter [147]. MafB blocks the type I IFN
amplification loop by interacting with IRF3 and interfering
with the recruitment of co-activators to reduce IRF3 tran-
scriptional activity [147].
Finally, a fascinating function of the IFN-b enhancer as
a ‘‘signal amplifier’’ has been described. It was demon-
strated that virus infection leads to IFN-b enhancer asso-
ciation with NF-jB at a different chromosomal genomic
region to trigger mono-allelic IFN-b gene expression [104].
Thereafter, an autocrine/paracrine loop of IFN-b signaling
through the IFNAR induced IRF7 expression, promoted
IFNB enhanceosome assembly on the remaining alleles,
and resulted in multi-allelic IFN-b expression [104]. In a
separate study, LPS stimulation of TLR4 drove activated
NF-jB RelA recruitment from the cytosol to the IFN-b
locus at a cluster of novel NF-jB sites downstream of the
human IFN-b gene, remote from the IFNB enhanceosome
region, which by itself was insufficient for maximal IFN-b
induction upon LPS stimulation [179].
Concluding remarks
More than half a century of research has spawned a huge
body of data describing the multifarious roles of type I IFN
in human health and disease. As much as we now know
about the beneficial and detrimental effects of IFN-b,
an in-depth understanding of the underlying molecular
mechanisms of IFN-b responses is still lacking. Scientific
efforts in this area are complicated by the complex inter-
play between multiple signaling pathways and numerous
regulatory factors that modulate the actions of IFN-b. From
a survey of the current literature, it is clear that many
determinants influence the magnitude and kinetics of the
IFN-b response in different physiological and pathological
settings. Such context-specific factors include: the cell type
in which IFN-b is induced; the species of pathogen
invading the host cell; the duration of infection; and the
local microenvironment where the infection occurs. It is
fascinating that the strength and longevity of IFN-b syn-
thesis are so finely attuned to every nuance of pathogenic
or inflammatory stimuli received by the host cell. Out-
standing questions include: the timing and cell types in
which IFN-b is induced; how signaling pathways are
activated and repressed to fine-tune IFN-b expression;
123
36
which molecules are involved in the spatio-temporal reg-
ulation of IFN-b; and why indeed are such tight controls
required for the regulation of IFN-b production. Many
unanswered questions remain regarding the role of tran-
scription factors in regulating the type I IFN response in
inflammation, and these quandaries will provide the plat-
form for exciting avenues of future research.
Acknowledgments The authors thank Dr Neil McCarthy for criti-
cally reviewing the manuscript. W.-X.S. and J.P.-S.Y. are supported
by the Agency for Science, Technology and Research (A*STAR)
graduate scholarships. Work in the Laboratory of Gene Regulation
and Inflammation is supported by the Singapore Immunology
Network (SIgN), Agency for Science, Technology and Research
(A*STAR) (Grant 06-001; K.-C.C.).
References
1. Isaacs A, Lindenmann J. Virus interference. 1. The interferon. P R
Soc Lond B Biol. 1957;147:258–267.
2. Pestka S. The interferons: 50 years after their discovery, there is
much more to learn. J Biol Chem. 2007;282:20047–51.
3. Friesen HJ, et al. Purification and molecular characterization of
human fibroblast interferon. Arch Biochem Biophys. 1981;206:
432–50.
4. Rubinstein M, Rubinstein S, Familletti PC, Miller RS, Waldman
AA, Pestka S. Human leukocyte interferon: production, purifi-
cation to homogeneity, and initial characterization. Proc Natl
Acad Sci U S A. 1979;76:640–4.
5. Stein S, Kenny C, Friesen HJ, Shively J, Del Valle U, Pestka S.
NH2-terminal amino acid sequence of human fibroblast inter-
feron. Proc Natl Acad Sci U S A. 1980;77:5716–9.
6. Borden EC, et al. Interferons at age 50: past, current and future
impact on biomedicine. Nat Rev Drug Discov. 2007;6:975–90.
7. de Andres C, et al. Interferon beta-1a therapy enhances CD4?
regulatory T-cell function: an ex vivo and in vitro longitudinal
study in relapsing-remitting multiple sclerosis. J Neuroimmunol.
2007;182:204–11.
8. Ebers GC, et al. Long-term follow-up of the original interferon-
beta1b trial in multiple sclerosis: design and lessons from a 16-
year observational study. Clin Ther. 2009;31:1724–36.
9. Goodin DS, et al. Predictive value of baseline and short-term
outcomes on mortality in multiple sclerosis: data from the
interferon beta-1b 21-year long-term follow-up study. 21st
Meeting of the European Neurological Society (ENS)
2011;Abstract P585.
10. Ebers GC, et al. Cause of death in patients with multiple scle-
rosis: the 21-year long-term follow-up study. 21st Meeting of
the European Neurological Society (ENS) 2011;Abstracts P429.
11. Chiang EY, et al. Targeted depletion of lymphotoxin-alpha-
expressing TH1 and TH17 cells inhibits autoimmune disease.
Nat Med. 2009;15:766–73.
12. Ito T, Amakawa R, Inaba M, Ikehara S, Inaba K, Fukuhara S.
Differential regulation of human blood dendritic cell subsets by
IFNs. J Immunol. 2001;166:2961–9.
13. Kito T, Kuroda E, Yokota A, Yamashita U. Enhancement of
macrophage cytotoxicity against murine gliomas by interferon
beta: increase in nitric oxide production in response to glioma-
derived soluble factors. J Neurosurg. 2002;97:619–26.
14. Longhi MP, et al. Dendritic cells require a systemic type I
interferon response to mature and induce CD4? Th1 immunity
with poly IC as adjuvant. J Exp Med. 2009;206:1589–602.
123
Singapore Immunology Network: SIgN (2012) 53:25–40
15. Nguyen KB, et al. Coordinated and distinct roles for IFN-alpha
beta, IL-12, and IL-15 regulation of NK cell responses to viral
infection. J Immunol. 2002;169:4279–87.
16. Su L, David M. Inhibition of B cell receptor-mediated apoptosis
by IFN. J Immunol. 1999;162:6317–21.
17. Sing A, et al. Bacterial induction of beta interferon in mice is a
function of the lipopolysaccharide component. Infect Immun.
2000;68:1600–7.
18. Parker D, Prince A. Type I interferon response to extracellular
bacteria in the airway epithelium. Trends Immunol. 2011;
32:582–8.
19. Perry AK, Chen G, Zheng D, Tang H, Cheng G. The host type I
interferon response to viral and bacterial infections. Cell Res.
2005;15:407–22.
20. Mancuso G, et al. Type I IFN signaling is crucial for host
resistance against different species of pathogenic bacteria. J
Immunol. 2007;178:3126–33.
21. Utaisincharoen P, Anuntagool N, Limposuwan K, Chaisuriya P,
Sirisinha S. Involvement of beta interferon in enhancing
inducible nitric oxide synthase production and antimicrobial
activity of Burkholderia pseudomallei-infected macrophages.
Infect Immun. 2003;71:3053–7.
22. Gautier G, et al. A type I interferon autocrine-paracrine loop is
involved in Toll-like receptor-induced interleukin-12p70 secre-
tion by dendritic cells. J Exp Med. 2005;201:1435–46.
23. Decker T, Muller M, Stockinger S. The yin and yang of type I
interferon activity in bacterial infection. Nat Rev Immunol.
2005;5:675–87.
24. Mahieu T, Libert C. Should we inhibit type I interferons in
sepsis? Infect Immun. 2007;75:22–9.
25. O’Connell RM, et al. Type I interferon production enhances
susceptibility to Listeria monocytogenes infection. J Exp Med.
2004;200:437–45.
26. Thomas KE, Galligan CL, Newman RD, Fish EN, Vogel SN.
Contribution of interferon-beta to the murine macrophage
response to the toll-like receptor 4 agonist, lipopolysaccharide. J
Biol Chem. 2006;281:31119–30.
27. Zwaferink H, Stockinger S, Hazemi P, Lemmens-Gruber R,
Decker T. IFN-b increases listeriolysin O-induced membrane
permeabilization and death of macrophages. J Immunol. 2008;
180:4116–23.
28. Carrero JA, Calderon B, Unanue ER. Type I interferon sensi-
tizes lymphocytes to apoptosis and reduces resistance to listeria
infection. J Exp Med. 2004;200:535–40.
29. Al Moussawi K, Ghigo E, Kalinke U, Alexopoulou L, Mege J-L,
Desnues B. Type I interferon induction is detrimental during
infection with the Whipple’s disease bacterium, Tropheryma
whipplei. Plos Pathog. 2010;6:e1000722.
30. Herskovits AA, Auerbuch V, Portnoy DA. Bacterial ligands
generated in a phagosome are targets of the cytosolic innate
immune system. Plos Pathog. 2007;3:e51.
31. Gruenberg J, van der Goot FG. Mechanisms of pathogen entry
through the endosomal compartments. Nat Rev Mol Cell Biol.
2006;7:495–504.
32. Pandey AK, et al. NOD2, RIP2 and IRF5 play a critical role in
the type I interferon response to Mycobacterium tuberculosis.
PLoS Pathog. 2009;5:e1000500.
33. Takaoka A, et al. DAI (DLM-1/ZBP1) is a cytosolic DNA
sensor and an activator of innate immune response. Nature.
2007;448:501–5.
34. Hornung V, et al. AIM2 recognizes cytosolic dsDNA and forms
a caspase-1-activating inflammasome with ASC. Nature. 2009;
458:514–8.
35. Roberts TL, et al. HIN-200 proteins regulate caspase activation
in response to foreign cytoplasmic DNA. Science. 2009;323:
1057–60.
Singapore Immunology Network: SIgN (2012) 53:25–40
36. Yang P, et al. The cytosolic nucleic acid sensor LRRFIP1
mediates the production of type I interferon via a beta-catenin-
dependent pathway. Nat Immunol. 2010;11:487–94.
37. Sabbah A, et al. Activation of innate immune antiviral responses
by Nod2. Nat Immunol. 2009;10:1073–80.
38. Wang ZC, et al. Regulation of innate immune responses by DAI
(DLM-1/ZBP1) and other DNA-sensing molecules. Proc Natl
Acad Sci U S A. 2008;105:5477–82.
39. Lippmann J, et al. IFNb responses induced by intracellular
bacteria or cytosolic DNA in different human cells do not
require ZBP1 (DLM-1/DAI). Cell Microbiol. 2008;10:2579–88.
40. Burckstummer T, et al. An orthogonal proteomic-genomic
screen identifies AIM2 as a cytoplasmic DNA sensor for the
inflammasome. Nat Immunol. 2009;10:266–72.
41. Ludlow LEA, Johnstone RW, Clarke CJP. The HIN-200 family:
more than interferon-inducible genes? Exp Cell Res. 2005;308:
1–17.
42. Unterholzner L, et al. IFI16 is an innate immune sensor for
intracellular DNA. Nat Immunol. 2010;11:997–1004.
43. Kim YG, et al. Viral infection augments Nod1/2 signaling to
potentiate lethality associated with secondary bacterial infec-
tions. Cell Host Microbe. 2011;9:496–507.
44. Hu JZ, et al. Role of cell-to-cell variability in activating a
positive feedback antiviral response in human dendritic cells.
Plos One 2011;6:e16614.
45. Guarda G, et al. Type I interferon inhibits interleukin-1 production
and inflammasome activation. Immunity. 2011;34:213–23.
46. O’Riordan M, Yi CH, Gonzales R, Lee KD, Portnoy DA. Innate
recognition of bacteria by a macrophage cytosolic surveillance
pathway. Proc Natl Acad Sci U S A. 2002;99:13861–6.
47. Mocci S, Dalrymple SA, Nishinakamura R, Murray R. The
cytokine stew and innate resistance to L-monocytogenes.
Immunol Rev. 1997;158:107–14.
48. Sen GC. Viruses and interferons. Annu Rev Microbiol. 2001;
55:255–81.
49. Tailor P, Tamura T, Ozato K. IRF family proteins and type I
interferon induction in dendritic cells. Cell Res. 2006;16:134–
40.
50. Mancuso G, et al. Bacterial recognition by TLR7 in the lyso-
somes of conventional dendritic cells. Nat Immunol. 2009;10:
587–94.
51. Scheu S, Dresing P, Locksley RM. Visualization of IFNbeta
production by plasmacytoid versus conventional dendritic cells
under specific stimulation conditions in vivo. Proc Natl Acad Sci
U S A. 2008;105:20416–21.
52. Cervantes-Barragan L, et al. Control of coronavirus infection
through plasmacytoid dendritic-cell-derived type I interferon.
Blood. 2007;109:1131–7.
53. Le Bon A, Schiavoni G, D’Agostino G, Gresser I, Belardelli F,
Tough DF. Type I interferons potently enhance humoral
immunity and can promote isotype switching by stimulating
dendritic cells in vivo. Immunity. 2001;14:461–70.
54. Lindahl P, Gresser I, Leary P, Tovey M. Interferon treatment of
mice: enhanced expression of histocompatibility antigens on
lymphoid cells. Proc Natl Acad Sci U S A. 1976;73:1284–7.
55. Santini SM, et al. Type I interferon as a powerful adjuvant for
monocyte-derived dendritic cell development and activity in vitro
and in Hu-PBL-SCID mice. J Exp Med. 2000;191:1777–88.
56. Luft T, et al. Type I IFNs enhance the terminal differentiation of
dendritic cells. J Immunol. 1998;161:1947–53.
57. Wenner CA, Guler ML, Macatonia SE, O’Garra A, Murphy
KM. Roles of IFN-gamma and IFN-alpha in IL-12-induced T
helper cell-1 development. J Immunol. 1996;156:1442–7.
58. Ito T, et al. Plasmacytoid dendritic cells regulate Th cell
responses through OX40 ligand and type I IFNs. J Immunol.
2004;172:4253–9.
37
59. Huber JP, Farrar JD. Regulation of effector and memory T-cell
functions by type I interferon. Immunology. 2011;132:466–74.
60. Kastenmuller K, et al. Protective T cell immunity in mice fol-
lowing protein-TLR7/8 agonist-conjugate immunization
requires aggregation, type I IFN, and multiple DC subsets. J Clin
Invest. 2011;121:1782–96.
61. Nguyen-Pham TN, et al. Type I and II interferons enhance
dendritic cell maturation and migration capacity by regulating
CD38 and CD74 that have synergistic effects with TLR ago-
nists. Cell Mol Immunol. 2011;8:341–7.
62. Rajagopal D, Paturel C, Morel Y, Uematsu S, Akira S, Diebold
SS. Plasmacytoid dendritic cell-derived type I interferon is
crucial for the adjuvant activity of Toll-like receptor 7 agonists.
Blood. 2010;115:1949–57.
63. Nagai T, Devergne O, Mueller TF, Perkins DL, van Seventer
JM, van Seventer GA. Timing of IFN-beta exposure during
human dendritic cell maturation and naive Th cell stimulation
has contrasting effects on Th1 subset generation: a role for IFN-
beta-mediated regulation of IL-12 family cytokines and IL-18 in
naive Th cell differentiation. J Immunol. 2003;171:5233–43.
64. Longman RS, Braun D, Pellegrini S, Rice CM, Darnell RB,
Albert ML. Dendritic-cell maturation alters intracellular sig-
naling networks, enabling differential effects of IFN-alpha/beta
on antigen cross-presentation. Blood. 2007;109:1113–22.
65. Nagai T, Devergne O, van Seventer GA, van Seventer JM.
Interferon-beta mediates opposing effects on interferon-gamma-
dependent Interleukin-12 p70 secretion by human monocyte-
derived dendritic cells. Scand J Immunol. 2007;65:107–17.
66. Korporal M, et al. Interferon beta-induced restoration of regu-
latory T-cell function in multiple sclerosis is prompted by an
increase in newly generated naive regulatory T cells. Arch
Neurol-Chicago 2008;65:1434–1439.
67. Martin-Saavedra FM, Gonzalez-Garcia C, Bravo B, Ballester S.
Beta interferon restricts the inflammatory potential of CD4?
cells through the boost of the Th2 phenotype, the inhibition of
Th17 response and the prevalence of naturally occurring T
regulatory cells. Mol Immunol. 2008;45:4008–19.
68. Zhang X, Markovic-Plese S. Interferon beta inhibits the Th17
cell-mediated autoimmune response in patients with relapsing-
remitting multiple sclerosis. Clin Neurol Neurosurg. 2010;112:
641–5.
69. Axtell RC, et al. T helper type 1 and 17 cells determine efficacy
of interferon-beta in multiple sclerosis and experimental
encephalomyelitis. Nat Med. 2010;16:406–12.
70. Steegmann JL, et al. Interferon alpha for chronic myeloid leu-
kemia relapsing after allogeneic bone marrow transplantation.
Bone Marrow Transplant. 1999;23:483–8.
71. Streetly M, Kazmi M, Radia D, Hoyle C, Schey SA. Second
autologous transplant with cyclosporin/interferon alpha-induced
graft versus host disease for patients who have failed first-line
consolidation. Bone Marrow Transplant. 2004;33:1131–5.
72. Porter DL, Roth MS, Mcgarigle C, Ferrara JLM, Antin JH.
Induction of graft-versus-host disease as immunotherapy for
relapsed chronic myeloid-leukemia. N Engl J Med. 1994;
330:100–6.
73. Thornley TB, et al. Type 1 IFN mediates cross-talk between
innate and adaptive immunity that abrogates transplantation
tolerance. J Immunol. 2007;179:6620–9.
74. Serrano J, Prieto E, Mazarbeitia F, Roman A, Llamas P, Tomas
JF. Atypical chronic graft-versus-host disease following inter-
feron therapy for chronic myeloid leukaemia relapsing after
allogeneic BMT. Bone Marrow Transplant. 2001;27:85–7.
75. Robb RJ, et al. Type I-IFNs control GVHD and GVL responses
after transplantation. Blood. 2011;118:3399–409.
76. Slifka MK, Antia R, Whitmire JK, Ahmed R. Humoral immu-
nity due to long-lived plasma cells. Immunity. 1998;8:363–72.
123
38
77. Ruuth K, Carlsson L, Hallberg B, Lundgren E. Interferon-alpha
promotes survival of human primary B-lymphocytes via phos-
phatidylinositol 3-kinase. Biochem Biophys Res Commun.
2001;284:583–6.
78. Oka H. Regulation of human B-cell responsiveness by inter-
feron-alpha—interferon-alpha-mediated suppression of B-cell
function is reversed through direct interactions between mono-
cytes and B-cells. Cell Immunol. 1993;146:238–48.
79. Braun D, Caramalho I, Demengeot J. IFN-a/b enhances BCR-
dependent B cell responses. Int Immunol. 2002;14:411–9.
80. Jego G, Palucka AK, Blanck J-P, Chalouni C, Pascual V, Ban-
chereau J. Plasmacytoid dendritic cells induce plasma cell dif-
ferentiation through Type I interferon and interleukin 6.
Immunity. 2003;19:225–34.
81. Niewold TB, Clark DN, Salloum R, Poole BD. Interferon alpha
in systemic lupus erythematosus. J Biomed Biotechnol. 2010;
2010:948364.
82. Thibault DL, Graham KL, Lee LY, Balboni I, Hertzog PJ, Utz
PJ. Type I interferon receptor controls B-cell expression of
nucleic acid-sensing Toll-like receptors and autoantibody pro-
duction in a murine model of lupus. Arthritis Res Ther. 2009;
11:R112.
83. de Goer de Herve MG, et al. Interferon-alpha triggers B cell
effector 1 (Be1) commitment. Plos One 2011;6:e19366.
84. Theofilopoulos AN, Baccala R, Beutler B, Kono DH. Type I
interferons (alpha/beta) in immunity and autoimmunity. Annu
Rev Immunol. 2005;23:307–36.
85. Kang JY, Lee JO. Structural biology of the Toll-like receptor
family. Annu Rev Biochem. 2011;80:917–41.
86. Akira S, Takeda K. Toll-like receptor signalling. Nat Rev
Immunol. 2004;4:499–511.
87. Fitzgerald KA, et al. IKKepsilon and TBK1 are essential com-
ponents of the IRF3 signaling pathway. Nat Immunol. 2003;4:
491–6.
88. Sharma S, tenOever BR, Grandvaux N, Zhou GP, Lin R, Hiscott
J. Triggering the interferon antiviral response through an IKK-
related pathway. Science. 2003;300:1148–51.
89. Uze G, Schreiber G, Piehler J, Pellegrini S. The receptor of the
type I interferon family. Curr Top Microbiol Immunol. 2007;
316:71–95.
90. Schindler C, Levy DE, Decker T. JAK-STAT signaling: from
interferons to cytokines. J Biol Chem. 2007;282:20059–63.
91. Mariathasan S, Weiss DS, Dixit VM, Monack DM. Innate
immunity against Francisella tularensis is dependent on the
ASC/caspase-1 axis. J Exp Med. 2005;202:1043–9.
92. Henry T, Brotcke A, Weiss DS, Thompson LJ, Monack DM.
Type I interferon signaling is required for activation of the in-
flammasome during Francisella infection. J Exp Med. 2007;
204:987–94.
93. Mariathasan S, et al. Cryopyrin activates the inflammasome in
response to toxins and ATP. Nature. 2006;440:228–32.
94. Lara-Tejero M, et al. Role of the caspase-1 inflammasome in
Salmonella typhimurium pathogenesis. J Exp Med. 2006;203:
1407–12.
95. Fernandes-Alnemri T, et al. The AIM2 inflammasome is critical
for innate immunity to Francisella tularensis. Nat Immunol.
2010;11:385–93.
96. Stetson DB, Medzhitov R. Type I interferons in host defense.
Immunity. 2006;25:373–81.
97. Jiang Z, Mak TW, Sen G, Li X. Toll-like receptor 3-mediated
activation of NF-kappaB and IRF3 diverges at Toll-IL-1
receptor domain-containing adapter inducing IFN-beta. Proc
Natl Acad Sci U S A. 2004;101:3533–8.
98. Han KJ, Su X, Xu LG, Bin LH, Zhang J, Shu HB. Mechanisms
of the TRIF-induced interferon-stimulated response element and
123
Singapore Immunology Network: SIgN (2012) 53:25–40
NF-kappaB activation and apoptosis pathways. J Biol Chem.
2004;279:15652–61.
99. Ning S, Pagano JS, Barber GN. IRF7: activation, regulation,
modification and function. Genes Immun. 2011;12:399–414.
100. Dai P, et al. Myxoma virus induces type I interferon production in
murine plasmacytoid dendritic cells via a TLR9/MyD88-, IRF5/
IRF7-, and IFNAR-dependent pathway. J Virol. 2011;85:10814–25.
101. Smale ST, Fisher AG. Chromatin structure and gene regulation
in the immune system. Annu Rev Immunol. 2002;20:427–62.
102. Panne D, Maniatis T, Harrison SC. An atomic model of the
interferon-beta enhanceosome. Cell. 2007;129:1111–23.
103. Ford E, Thanos D. The transcriptional code of human IFN-b
gene expression. Biochimica et Biophysica Acta (BBA)—Gene
Regul Mech. 2010;1799:328–336.
104. Apostolou E, Thanos D. Virus infection induces NF-kappaB-
dependent interchromosomal associations mediating monoall-
elic IFN-beta gene expression. Cell. 2008;134:85–96.
105. Li P, et al. IRF8 and IRF3 cooperatively regulate rapid inter-
feron-beta induction in human blood monocytes. Blood.
2011;117:2847–54.
106. Driggers PH, et al. An interferon gamma-regulated protein that
binds the interferon-inducible enhancer element of major his-
tocompatibility complex class I genes. Proc Natl Acad Sci U S
A. 1990;87:3743–7.
107. Wang T, Town T, Alexopoulou L, Anderson JF, Fikrig E, Flavell
RA. Toll-like receptor 3 mediates West Nile virus entry into the
brain causing lethal encephalitis. Nat Med. 2004;10:1366–73.
108. Hemmi H, et al. The roles of two IkappaB kinase-related kinases
in lipopolysaccharide and double stranded RNA signaling and
viral infection. J Exp Med. 2004;199:1641–50.
109. Krug A, Luker GD, Barchet W, Leib DA, Akira S, Colonna M.
Herpes simplex virus type 1 activates murine natural interferon-
producing cells through toll-like receptor 9. Blood. 2004;103:
1433–7.
110. Lund J, Sato A, Akira S, Medzhitov R, Iwasaki A. Toll-like
receptor 9-mediated recognition of Herpes simplex virus-2 by
plasmacytoid dendritic cells. J Exp Med. 2003;198:513–20.
111. Yoneyama M, et al. The RNA helicase RIG-I has an essential
function in double-stranded RNA-induced innate antiviral
responses. Nat Immunol. 2004;5:730–7.
112. Nasirudeen AM, Wong HH, Thien P, Xu S, Lam KP, Liu DX.
RIG-I, MDA5 and TLR3 synergistically play an important role
in restriction of dengue virus infection. PLoS Negl Trop Dis.
2011;5:e926.
113. Hogle JM. Poliovirus cell entry: common structural themes in
viral cell entry pathways. Annu Rev Microbiol. 2002;56:677–
702.
114. Rohde K, Yates RM, Purdy GE, Russell DG. Mycobacterium
tuberculosis and the environment within the phagosome.
Immunol Rev. 2007;219:37–54.
115. Sirard JC, Vignal C, Dessein R, Chamaillard M. Nod-like
receptors: cytosolic watchdogs for immunity against pathogens.
PLoS Pathog. 2007;3:e152.
116. Girardin SE, et al. Peptidoglycan molecular requirements
allowing detection by Nod1 and Nod2. J Biol Chem. 2003;
278:41702–8.
117. van der Wel N, et al. M. tuberculosis and M. leprae translocate
from the phagolysosome to the cytosol in myeloid cells. Cell.
2007;129:1287–98.
118. Qiao Y, et al. Host defense responses to infection by Myco-
bacterium tuberculosis. Induction of IRF-1 and a serine protease
inhibitor. J Biol Chem. 2002;277:22377–85.
119. Stetson DB, Medzhitov R. Recognition of cytosolic DNA acti-
vates an IRF3-dependent innate immune response. Immunity.
2006;24:93–103.
Singapore Immunology Network: SIgN (2012) 53:25–40
120. Clark IA, Alleva LM, Mills AC, Cowden WB. Pathogenesis of
malaria and clinically similar conditions. Clin Microbiol Rev.
2004;17:509–39.
121. Voisine C, Mastelic B, Sponaas AM, Langhorne J. Classical
CD11c? dendritic cells, not plasmacytoid dendritic cells, induce
T cell responses to Plasmodium chabaudi malaria. Int J Parasi-
tol. 2010;40:711–9.
122. Khor CC, et al. A Mal functional variant is associated with
protection against invasive pneumococcal disease, bacteremia,
malaria and tuberculosis. Nat Genet. 2007;39:523–8.
123. Parroche P, et al. Malaria hemozoin is immunologically inert but
radically enhances innate responses by presenting malaria DNA
to Toll-like receptor 9. Proc Natl Acad Sci U S A. 2007;
104:1919–24.
124. Sharma S, et al. Innate immune recognition of an AT-rich stem-
loop DNA motif in the Plasmodium falciparum genome.
Immunity. 2011;35:194–207.
125. Sozzani S, Type I. Interferons in systemic autoimmunity.
Autoimmunity. 2010;43:196–203.
126. Salloum R, Niewold TB. Interferon regulatory factors in human
lupus pathogenesis. Transl Res. 2011;157:326–31.
127. Ye J, Maniatis T. Negative regulation of interferon-b gene
expression during acute and persistent virus infections. PLoS
ONE. 2011;6:e20681.
128. Sathish N, Yuan Y. Evasion and subversion of interferon-med-
iated antiviral immunity by Kaposi’s sarcoma-associated her-
pesvirus: an overview. J Virol. 2011;85:10934–44.
129. Rezaee SAR, Cunningham C, Davison AJ, Blackbourn DJ.
Kaposi’s sarcoma-associated herpesvirus immune modulation:
an overview. J Gen Virol. 2006;87:1781–804.
130. Sathish N, Zhu FX, Golub EE, Liang Q, Yuan Y. Mechanisms
of autoinhibition of IRF-7 and a probable model for inactivation
of IRF-7 by Kaposi’s sarcoma-associated herpesvirus protein
ORF45. J Biol Chem. 2011;286:746–56.
131. Zhu FX, King SM, Smith EJ, Levy DE, Yuan Y. A Kaposi’s sar-
coma-associated herpesviral protein inhibits virus-mediated induc-
tion of type I interferon by blocking IRF-7 phosphorylation and
nuclear accumulation. Proc Natl Acad Sci U S A. 2002;99:5573–8.
132. Yu Y, Wang SE, Hayward GS. The KSHV immediate-early
transcription factor RTA encodes ubiquitin E3 ligase activity
that targets IRF7 for proteosome-mediated degradation. Immu-
nity. 2005;22:59–70.
133. Lin R, et al. HHV-8 encoded vIRF-1 represses the interferon
antiviral response by blocking IRF-3 recruitment of the CBP/
p300 coactivators. Oncogene. 2001;20:800–11.
134. Joo CH, Shin YC, Gack M, Wu L, Levy D, Jung JU. Inhibition
of interferon regulatory factor 7 (IRF7)-mediated interferon
signal transduction by the Kaposi’s sarcoma-associated herpes-
virus viral IRF homolog vIRF3. J Virol. 2007;81:8282–92.
135. Lefort S, Soucy-Faulkner A, Grandvaux N, Flamand L. Binding
of Kaposi’s sarcoma-associated herpesvirus K-bZIP to inter-
feron-responsive factor 3 elements modulates antiviral gene
expression. J Virol. 2007;81:10950–60.
136. Hwang S, et al. Conserved herpesviral kinase promotes viral
persistence by inhibiting the IRF-3-mediated type I interferon
response. Cell Host Microbe. 2009;5:166–78.
137. Cloutier N, Flamand L. Kaposi sarcoma-associated herpesvirus
latency-associated nuclear antigen inhibits interferon (IFN) b
expression by competing with IFN regulatory factor-3 for
binding to IFNB promoter. J Biol Chem. 2010;285:7208–21.
138. Bentz GL, Liu R, Hahn AM, Shackelford J, Pagano JS. Epstein–
Barr virus BRLF1 inhibits transcription of IRF3 and IRF7 and
suppresses induction of interferon-b. Virology. 2010;402:121–8.
139. Liang Q, Deng H, Sun C-W, Townes TM, Zhu F. Negative
regulation of IRF7 activation by activating transcription factor 4
suggests a cross-regulation between the IFN responses and
39
the cellular integrated stress responses. J Immunol. 2011;186:
1001–10.
140. Colina R, et al. Translational control of the innate immune
response through IRF-7. Nature. 2008;452:323–8.
141. Zhang L, Pagano JS. Interferon regulatory factor 7 mediates
activation of Tap-2 by Epstein-Barr virus latent membrane
protein 1. J Virol. 2001;75:341–50.
142. Higgs R, et al. Self protection from anti-viral responses—Ro52
promotes degradation of the transcription factor IRF7 down-
stream of the viral toll-like receptors. PLoS ONE. 2010;5:e11776.
143. Kubota T, et al. Virus infection triggers SUMOylation of IRF3
and IRF7, leading to the negative regulation of type I interferon
gene expression. J Biol Chem. 2008;283:25660–70.
144. Chang T-H, et al. Ebola Zaire virus blocks type I interferon
production by exploiting the host SUMO modification machin-
ery. PLoS Pathog. 2009;5:e1000493.
145. Caillaud A, et al. Acetylation of interferon regulatory factor-7
by p300/CREB-binding protein (CBP)-associated factor (PCAF)
impairs its DNA binding. J Biol Chem. 2002;277:49417–21.
146. Liang Q, et al. Tripartite motif-containing protein 28 is a small
ubiquitin-related modifier E3 ligase and negative regulator of
IFN regulatory factor 7. J Immunol. 2011;187:4754–63.
147. Kim H, Seed B. The transcription factor MafB antagonizes
antiviral responses by blocking recruitment of coactivators to
the transcription factor IRF3. Nat Immunol. 2010;11:743–50.
148. Yang K, et al. TRIM21 is essential to sustain IFN regulatory
factor 3 activation during antiviral response. J Immunol. 2009;
182:3782–92.
149. Parvatiyar K, Barber GN, Harhaj EW. TAX1BP1 and A20
inhibit antiviral signaling by targeting TBK1-IKKi kinases. J
Biol Chem. 2010;285:14999–5009.
150. Yang Y-K, et al. ARF-like protein 16 (ARL16) inhibits RIG-I by
binding with its C-terminal domain in a GTP-dependent manner.
J Biol Chem. 2011;286:10568–80.
151. Gabhann JN, et al. Absence of SHIP-1 results in constitutive
phosphorylation of tank-binding kinase 1 and enhanced TLR3-
dependent IFN-b production. J Immunol. 2010;184:2314–20.
152. Siednienko J, Gajanayake T, Fitzgerald KA, Moynagh P, Mig-
gin SM. Absence of MyD88 results in enhanced TLR3-depen-
dent phosphorylation of IRF3 and increased IFN-b and
RANTES production. J Immunol. 2011;186:2514–22.
153. Verstrepen L, Verhelst K, van Loo G, Carpentier I, Ley SC, Beyaert
R. Expression, biological activities and mechanisms of action of
A20 (TNFAIP3). Biochem Pharmacol. 2010;80:2009–20.
154. Wang Y-Y, Li L, Han K-J, Zhai Z, Shu H-B. A20 is a potent
inhibitor of TLR3- and Sendai virus-induced activation of NF-
jB and ISRE and IFN-b promoter. FEBS Lett. 2004;576:86–90.
155. Saitoh T, et al. A20 is a negative regulator of IFN regulatory
factor 3 signaling. J Immunol. 2005;174:1507–12.
156. Zhao W, et al. Peroxisome proliferator-activated receptor c
negatively regulates IFN-b production in toll-like receptor
(TLR) 3- and TLR4-stimulated macrophages by preventing
interferon regulatory factor 3 binding to the IFN-b promoter. J
Biol Chem. 2011;286:5519–28.
157. Lin R, et al. Negative regulation of the retinoic acid-inducible
gene I-induced antiviral state by the ubiquitin-editing protein
A20. J Biol Chem. 2006;281:2095–103.
158. Ning S, Pagano JS. The A20 deubiquitinase activity negatively
regulates LMP1 activation of IRF7. J Virol. 2010;84:6130–8.
159. Saitoh T, et al. Negative regulation of interferon-regulatory
factor 3-dependent innate antiviral response by the prolyl
isomerase Pin1. Nat Immunol. 2006;7:598–605.
160. Higgs R, Gabhann JN, Larbi NB, Breen EP, Fitzgerald KA,
Jefferies CA. The E3 ubiquitin ligase Ro52 negatively regulates
IFN-b production post-pathogen recognition by polyubiquitin-
mediated degradation of IRF3. J Immunol. 2008;181:1780–6.
123
40
161. Kong HJ, et al. Cutting edge: autoantigen Ro52 is an interferon
inducible E3 ligase that ubiquitinates IRF-8 and enhances cyto-
kine expression in macrophages. J Immunol. 2007;179:26–30.
162. Xiong H, et al. Ubiquitin-dependent degradation of interferon
regulatory factor-8 mediated by Cbl down-regulates interleukin-
12 expression. J Biol Chem. 2005;280:23531–9.
163. Yu Y, Hayward GS. The ubiquitin E3 ligase RAUL negatively
regulates type I interferon through ubiquitination of the tran-
scription factors IRF7 and IRF3. Immunity. 2010;33:863–77.
164. Sears N, Sen GC, Stark GR, Chattopadhyay S. Caspase-8-
mediated cleavage inhibits IRF-3 protein by facilitating its
proteasome-mediated degradation. J Biol Chem. 2011;286:
33037–44.
165. Praefcke GJK, Hofmann K, Dohmen RJ. SUMO playing tag
with ubiquitin. Trends Biochem Sci. 2011;37:23–31.
166. Denuc A, Marfany G. SUMO and ubiquitin paths converge.
Biochem Soc Trans. 2010;38:34–9.
167. Ran Y, et al. SENP2 negatively regulates cellular antiviral
response by deSUMOylating IRF3 and conditioning it for
ubiquitination and degradation. J Mol Cell Biol. 2011;3:283–92.
168. Dastur A, Beaudenon S, Kelley M, Krug RM, Huibregtse JM.
Herc5, an interferon-induced HECT E3 enzyme, is required for
conjugation of ISG15 in human cells. J Biol Chem. 2006;
281:4334–8.
169. Wong JJY, Pung YF, Sze NS-K, Chin K-C. HERC5 is an IFN-
induced HECT-type E3 protein ligase that mediates type I IFN-
induced ISGylation of protein targets. Proc Natl Acad Sci.
2006;103:10735–40.
170. Lu G, et al. ISG15 enhances the innate antiviral response by
inhibition of IRF-3 degradation. Cell Mol Biol (Noisy-le-grand)
2006;52:29–41.
171. Shi H-X, et al. Positive regulation of interferon regulatory factor
3 activation by Herc5 via ISG15 modification. Mol Cell Biol.
2010;30:2424–36.
172. Prinarakis E, Chantzoura E, Thanos D, Spyrou G. S-glutath-
ionylation of IRF3 regulates IRF3-CBP interaction and activa-
tion of the IFN[beta] pathway. EMBO J. 2008;27:865–75.
123
Singapore Immunology Network: SIgN (2012) 53:25–40
173. Xu H-G, Ren W, Lu C, Zhou G-P. Characterization of the
human IRF-3 promoter and its regulation by the transcription
factor E2F1. Mol Biol Rep. 2010;37:3073–80.
174. Xu H-G, Ren W, Zou L, Wang Y, Jin R, Zhou G-P. Direct
repression of the human IRF-3 promoter by E2F1. Immunoge-
netics. 2011;63:189–96.
175. Lim C-A, et al. Genome-wide mapping of RELA(p65) binding
identifies E2F1 as a transcriptional activator recruited by NF-jB
upon TLR4 activation. Mol Cell. 2007;27:622–35.
176. Li C, Ma L, Chen X. Interferon regulatory factor 3-CL, an
isoform of IRF3, antagonizes activity of IRF3. Cell Mol
Immunol. 2011;8:67–74.
177. Karpova AY, Ronco LV, Howley PM. Functional character-
ization of interferon regulatory factor 3a (IRF-3a), an alternative
splice isoform of IRF-3. Mol Cell Biol. 2001;21:4169–76.
178. Marozin S, Altomonte J, Stadler F, Thasler WE, Schmid RM,
Ebert O. Inhibition of the IFN-beta response in hepatocellular
carcinoma by alternative spliced isoform of IFN regulatory
factor-3. Mol Ther. 2008;16:1789–97.
179. Goh FG, Thomson SJP, Krausgruber T, Lanfrancotti A, Copley
RR, Udalova IA. Beyond the enhanceosome: cluster of novel jB
sites downstream of the human IFN-b gene is essential for
lipopolysaccharide-induced gene activation. Blood. 2010;116:
5580–8.
180. Yount JS, Gitlin L, Moran TM, Lopez CB. MDA5 participates
in the detection of paramyxovirus infection and is essential for
the early activation of dendritic cells in response to Sendai virus
defective interfering particles. J Immunol. 2008;180:4910–8.
181. Honda K, et al. IRF-7 is the master regulator of type-I inter-
feron-dependent immune responses. Nature. 2005;434:772–7.
182. Carrigan SO, et al. IFN regulatory factor 3 contributes to the
host response during Pseudomonas aeruginosa lung infection in
mice. J Immunol. 2010;185:3602–9.