SCIENCE IMMUNOLOGY | REVIEW
IMMUNE REGULATION
The circadian immune system
Chen Wang1†, Lydia Kay Lutes1†, Coline Barnoud1, Christoph Scheiermann1,2*
The immune system is highly time-of-day dependent. Pioneering studies in the 1960s were the first to identify
immune responses to be under a circadian control. Only in the last decade, however, have the molecular factors
governing circadian immune rhythms been identified. These studies have revealed a highly complex picture
of the interconnectivity of rhythmicity within immune cells with that of their environment. Here, we provide a
global overview of the circadian immune system, focusing on recent advances in the rapidly expanding field of
circadian immunology.
INTRODUCTION
The circadian clock uses rhythmic environmental cues, such as light
and food intake, to establish approximately 24-hour rhythms in the
vast majority of physiological processes. These biological cycles
prepare organisms better to encounter regular, recurring events in
their environment. The immune system is no exception. It is highly
circadian-regulated and exhibits 24-hour rhythmicity both at steady
state and during activation.
Immune rhythms were first found in the innate immune system
in 1960 (1), and 10 years later, components of the adaptive immune
system were shown to be similarly time-of-day dependent (2). More
than half a century later, we now know that immune cellularity, mi-
gration, and function are all regulated by the circadian clock. The
number of circulating leukocytes oscillates in the blood of mam-
mals, peaking during the behavioral rest phase in nocturnal mice
(day) (3) and diurnal humans (night) (Box 1) (4). Circadian clocks
also modulate leukocyte migration across the body, effectively
gating the number of leukocytes at specific sites throughout the day
(3, 5–7). In addition, leukocyte effector functions are time-of-day
dependent as is the capacity of activated immune cells to proliferate
and produce cytokines (8–11). It is highly likely that these immune
rhythms evolved to defend against diurnal peaks of pathogen en-
counter, such as those occurring due to increased social interactions
or transmitted via food intake (12) that occur predominantly during
the behavioral active period.
Although the ability to detect and adapt to recurring environ-
mental rhythms exists in virtually all species, the molecular manner
and complexity of how circadian rhythms are maintained vary be-
tween organisms (13, 14). In mammals, the core circadian clock
machinery consists of a highly interconnected system of transcrip-
tion factors, forming several transcription-translation feedback loops
that activate and inhibit one another to drive 24-hour rhythms at
the molecular level (15). This network is often referred to as the core
clock and consists of the transcription factors BMAL1 [brain and
muscle aryl hydrocarbon receptor nuclear translocator (ARNT)–
like 1; encoded by Arntl], CLOCK (circadian locomotor output
cycles kaput), PER1/2/3 (period circadian protein homologs),
and CRY1/2 (cryptochromes 1/2) (15, 16). At the protein level,
BMAL1 and CLOCK form a heterodimeric transcription factor, which
binds to enhancer (E)–box regions of target genes, including those
1
Department of Pathology and Immunology, Faculty of Medicine, University of
Geneva, Geneva, Switzerland. 2Biomedical Center (BMC), Institute for Cardiovascular
Physiology and Pathophysiology, Walter Brendel Center for Experimental Medicine
(WBex), Ludwig-Maximilians-Universität München, Planegg-Martinsried, Germany.
*Corresponding author. Email: christoph.scheiermann@unige.ch
†These authors contributed equally to this work.
Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022
Copyright © 2022
The Authors, some
rights reserved;
exclusive licensee
American Association
for the Advancement
of Science. No claim
to original U.S.
Government Works
in the gene locus of the negative regulators Per1/2/3 and Cry1/2,
inducing their expression (15). Once expressed, PER and CRY also
form a heterodimeric transcription factor complex that inhibits the
Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023
expression of BMAL1 and CLOCK until concentrations of the re-
pressors are low enough to restart the cycle (15–17). In addition, the
REV-ERB nuclear receptor / [nuclear receptor subfamily 1
group D (NR1D); encoded by Nr1d1/2] and the retinoic acid receptor
(RAR)–related orphan receptors (RORs) make up a secondary feed-
back loop that stabilizes the core molecular clock (15, 16). However,
it is important to note that circadian rhythms can also occur in cells
that lack a nucleus, where rhythms are orchestrated by oscillations
in the oxidation of peroxiredoxin proteins (18).
Most circadian studies in the immune system have focused on
the role of BMAL1, because it is the only clock gene whose sole ab-
sence abrogates rhythmicity (17). Studies that disrupt the molecular
clock through removal of BMAL1 must therefore differentiate be-
tween effects mediated specifically by BMAL1 and those mediated
by the function of the clock in general, because the latter should
be corroborated with additional clock mutants whenever possible.
Box 1. Circadian terminologies.
Circadian rhythm: From the Latin circa diem (“about a day”), it refers to
approximately 24-hour cycles that are perpetuated by recurring
environmental cues, such as light and food intake, which drive daily
oscillations in the most physiological processes. Circadian rhythms can be
synchronized and are endogenous rhythms.
Diurnal rhythm: A daily oscillation. It may or may not be a circadian rhythm.
Diurnal: Refers to a species that is active mainly during the daytime, e.g.,
Homo sapiens.
Nocturnal: Refers to a species that is active mainly during the nighttime,
e.g., Mus musculus.
Zeitgeber time: Zeitgeber, from the German “time giver,” is an external
cue (such as light or food intake) that synchronizes the circadian clock of
an organism to its environment. Zeitgeber time (ZT) is the time in hours
after light onset, for example, lights on is ZT0 and lights off is ZT12 in a
12-hour light/12-hour dark cycle.
Circadian time (CT): A measure of subjective time used when organisms
are isolated from Zeitgebers of the environment (for example, in constant
darkness). CT0 represents the start of the subjective day, and CT12
represents the start of the subjective night.
Synchronization: The process where Zeitgeber signals reset an organism
or cells in culture to the same time or phase.
Suprachiasmatic nucleus (SCN): A brain region located within the anterior
hypothalamus that receives environmental light cues and constitutes the
central clock.
1 of 15
SCIENCE IMMUNOLOGY | REVIEW
Deficiency in various clock components, such as Bmal1 by itself
or double knockouts of Cry1 and Cry2 or Per1 and Per2, has been
associated with a variety of altered inflammatory phenotypes,
demonstrating the critical role of the circadian clock in the proper
function of the mammalian immune system (Table 1) (16). In this
Review, we will discuss how circadian regulation of leukocyte cel-
lularity, trafficking, and effector activity affects the function of
the immune system.
INNATE IMMUNITY
Response to bacterial components
Lipopolysaccharides (LPS) are found in the outer membrane of
Gram-negative bacteria. LPS bind to Toll-like receptor 4 (TLR4)
and initiate an inflammatory signaling cascade. This has been the most
widely used model of inflammation to assess circadian rhythms in
innate immune responses. It was first used in the 1960s, showing
that mice injected with LPS in the evening exhibited a notably
increased mortality compared with those challenged in the morning
(Box 2) (1). A few years later, studies reported similar results using
Gram-positive Streptococcus pneumoniae (19, 20). Many genes in
the LPS-induced signaling cascade are rhythmically expressed in
peritoneal macrophages at steady state, and splenocytes exhibit
rhythmic tumor necrosis factor– (TNF-) and interleukin-6 (IL-6)
production after LPS challenge (21). Mice treated with LPS at Zeitgeber
time 11 [ZT11; i.e., time (in hours) after light onset in a 12-hour
light:12-hour dark (LD) environment, “evening”] exhibited signifi-
cantly higher serum TNF- levels and suffered increased mortality
than those treated at ZT19 (“night”) (Box 1) (22). TNF receptor 1
(TNFR1)–deficient mice exhibited no time-of-day differences in
mortality rate, indicating that the temporal differences observed are
TNF- dependent (22). These data demonstrate that the innate im-
mune response to LPS is under circadian control.
Disrupting the circadian rhythm of mice affects the immune
response to LPS challenge. Mice that were desynchronized from
their normal circadian environment—by shifting them from the
usual 24-hour (12-hour:12-hour LD) cycle to a shorter 20-hour
(10-hour:10-hour LD) cycle—exhibited delayed recovery of loco-
motion and weight loss after LPS challenge compared with un-
disrupted mice (23). In addition, mice moved from a 12-hour:12-hour
LD schedule to constant darkness (DD; Box 1) exhibited a threefold
increased susceptibility to LPS compared with mice kept in rhythmic
LD conditions (24). Although cytokines such as TNF-, monocyte
chemoattractant protein-1 (MCP-1), IL-18, and IL-10 showed a dis-
tinct time-of-day oscillation in the peripheral blood of mice in LD
conditions after LPS challenge, the phase of this oscillation was
changed in mice kept in DD conditions (24). Interestingly, these
oscillations and the differences in survival could not be fully abro-
gated in Lyz2cre:Bmal1flox or Lyz2cre:Clockflox mice (24), which target
the circadian clock in the myeloid cell lineage [mostly granulocytes
and some macrophage subsets, as well as a fraction of monocytes (25)].
This indicated that light may exert an anti-inflammatory influence
on LPS responses in mice and that rhythmicity of this pheno­menon is
at least partially independent of myeloid cells.
Using a genome-wide screening approach, a recent study inves-
tigated the circadian transcriptome in murine lungs, showing that
genes related to immune function in steady state were enriched in
the middle of the day (26). However, upon challenge with LPS
during the evening, gene expression was reprogrammed, exhibiting
Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022
Box 2. Key circadian immune concepts.
Lethality to strong immune stimuli, such as in the form of high doses of
LPS, bacteria, or viruses, exhibits a circadian rhythm, peaking around the
onset of the behavioral activity phase.
Cytokine secretion from stimulated macrophages is circadian.
The numbers of leukocytes in blood are circadian, peaking during the
behavioral rest phase in both mice and humans. This is driven by circadian
rhythms in leukocyte trafficking. The mobilization of leukocytes from the
bone marrow into blood, the drainage of leukocytes from tissues, and the
homing of leukocytes to tissues are all rhythmic in a circadian manner.
Because these oscillations peak at different times of day, blood leukocyte
numbers exhibit an overall circadian oscillation.
Circadian rhythms in the host and microbiota interact with each other to
maintain metabolic homeostasis of the host. Host rhythms influence the
intestinal barrier and microbiota adhesion to the intestinal wall.
Microbiota produce metabolites in a time-of-day–dependent manner,
thus regulating the host rhythm.
Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023
a shift to a new set of genes and immunological processes. In addi-
tion, LPS challenge shortened the median period length of the circadian
rhythm in the lung from 24 hours (steady state) to 19.6 hours. Flow
cytometry analyses also detected oscillations in B cell and Gr1+
myeloid cell counts in the lungs of naïve mice, with a peak occurring
in the morning and a trough in the evening (26). Steady-state
alveolar macrophages presented an opposite and lower amplitude
rhythm, peaking in the evening. In contrast, LPS-treated animals
exhibited a dampened amplitude of oscillating B cell numbers,
whereas myeloperoxidase-positive (MPO+) granulocytes presented
circadian-­like variation (26). These data demonstrate the strong
interaction between a rhythmic environment and the ensuing im-
mune responses.
Circadian oscillation of granulocytes in the lung after LPS stim-
ulation is completely abrogated in whole-body Bmal1-deficient mice
(26). However, it is still unclear which cell types require Bmal1 to
induce this rhythmic granulocyte response. Mice with Bmal1-­deficient
myeloid cell subsets (Lyz2cre:Bmal1flox) still showed rhythmic re-
sponses to LPS challenge in the lung (27), indicating a myeloid-­
independent mechanism [analogous to the observation discussed
above (24)]. However, removing BMAL1 expression in bronchiolar
epithelial cells (using Ccspcre:Bmal1flox) ablated their diurnal secre-
tion of C-X-C motif chemokine ligand 5 (CXCL5) in response to
LPS treatment (27). Bronchiolar epithelial cells require BMAL1
expression to respond to oscillating glucocorticoid (GC) levels
(discussed further below), and their rhythmic production of CXCL5
is dependent on cell-intrinsic expression of the GC receptor (GR)
(28). Removing GR from the airway epithelium, however, still did
not abolish circadian oscillations in granulocyte recruitment or
rhythmic expression of TNF- and IL-6 in the lung after LPS stim-
ulation (28). Thus, although bronchiolar epithelial cell–intrinsic
expression of BMAL1 and the GR is sufficient to explain oscillating
levels of CXCL5, they do not drive the circadian control of neutro-
philic infiltration in response to inhaled LPS, indicating that there
are additional circadian mechanisms involved.
Apart from BMAL1 and CLOCK, the clock component REV-ERB
has also been implicated in controlling pulmonary LPS-induced
inflammation (29). Although wild-type mice exhibit increased neu-
trophil lung infiltration in the morning (ZT0) versus the evening
2 of 15
SCIENCE IMMUNOLOGY | REVIEW
Table 1. Alteration of circadian clock components in mouse strains and cell lines and their physiological relevance. NP-KLH, 4-hydroxy-3-
nitrophenylacetyl hapten conjugated to KLH (keyhole limpet hemocyanin); NP-CGG, chicken gamma globulin; KO, knockout; iPSCs, induced pluripotent stem
cells; DKO, double KO.
Cell type Mouse strain/cells
Monocyte/
Lyz2cre:Bmal1flox
macrophage
Lyz2cre:Bmal1flox
Lyz2cre:Bmal1flox Cx3cr1cre:Bmal1flox
Lyz2cre:Bmal1flox
Lyz2cre:Bmal1flox
Lyz2cre:Bmal1flox Rev-erb−/− full KO
Rev-erb−/− full KO
Rev-erb−/− full KO
Pharmacological target REV-ERB
Rev-erb−/− full KO
Rev-erb−/− full KO
Lyz2cre:Bmal1flox
Lyz2cre:Bmal1flox
REV-ERB/ agonist (SR9009,
GSK2667) administration
Microglia Rev-erb−/− full KO
Clec9cre:Bmal1flox
DC
Cdh5creERT2:Bmal1flox
Rev-erb−/− full KO Rev-erb−/− full
KO
Cd11ccre:Bmal1flox
Bmal1 full KO
Neutrophil Lyz2cre:Bmal1flox
hMRP8cre:Bmal1flox
continued on next page
Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022
Experimental model
LPS-induced inflammation
M1-stimulated macrophages
S. pneumoniae
L. monocytogenes
LPS-stimulated BMDMs
LPS-induced inflammation
LPS-induced inflammation
LPS-induced inflammation
LPS-induced inflammation
Induced fulminant hepatitis
DSS-induced colitis
TNF-–induced inflammation
EAE
Macrophages derived from
human iPSCs infected by HIV
LPS-induced inflammation
DC crawl in
BMDC from DKO mice
T. muris parasitic helminth
infection
DC-OVA vaccination
LPS-induced inflammation
Cell-intrinsic deletion in
neutrophils
Findings Ref
Time-dependent, endotoxic shock–induced mortality in
mice (peak at ZT8) is independent of Bmal1 expression (24)
in macrophages, as well as LD cycles.
Bmal1–Hif-1a regulates mitochondrial metabolism in
inflammatory macrophages in the scenario of
(32)
LPS-stimulated BMDMs, macrophages from cell-specific
knockout mice, and B16-F10–challenged mice.
Bmal1 deletion enhances macrophage motility and
(30)
phagocytosis, causing increased antibacterial immunity.
Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023
Deletion of Bmal1 abrogates diurnal variations in the
numbers of Ly6Chi monocytes in blood, spleen, and (43)
bone marrow during L. monocytogenes infection.
Relative activity of the NRF2 transcription factor is
(37)
significantly decreased in Bmal1−/− macrophages.
Circadian-dependent variation in cytokine response to
(40)
LPS is abrogated in Lyz2cre:Bmal1−/− and Rev-erb−/− mice.
REV-ERB in the murine macrophage suppresses Ccl2
(41)
expression induced by LPS challenge.
The time-of-day variation in pulmonary neutrophilia is
(29)
abrogated in Rev-erb−/− mice.
REV-ERB inhibits IL-1 production through the NF-B/
(34)
NLRP3 pathway.
REV-ERB regulates diurnal expression and activation of
NLRP3 inflammasome, attenuating NLRP3-driven (35)
inflammation.
REV-ERB represses p65 transcription and indirectly
(38)
represses Nlrp3 via the NF-B pathway.
Loss of Bmal1 abrogates rhythmic recruitment of
(84)
myeloid cells to arteries and veins.
Myeloid cell deficiency in Bmal1 abrogates time-of-day
(74)
differences in disease severity of EAE.
Pharmacological activation of REV-ERB/ inhibits HIV
(108)
promoter activity, impeding viral replication.
Diurnal microglial immunoreactivity in the
(42)
hippocampus is lost after Rev-erb deletion.
Increased DC migration into skin lymphatic vessels occurs
(48)
at ZT7, which is largely dependent on rhythms in CCL21.
BMDCs from KO mice showed enhanced expression of
(50)
CD86, MHCII, and proinflammatory cytokines.
DC clock regulates time-of-day–dependent efficiency of
worm expulsion by polarizing the immune system (111)
toward a TH2 response.
Impaired BMDCs migration to spleen after intravenous
(49)
injection
Similar neutrophils and G-CSF amount in mice with
(27)
Bmal1-sufficient and Bmal1-deficient myeloid cells.
Neutrophil Bmal1 controls the expression of Cxcl2 in
neutrophils, regulating neutrophil aging that peaks (44)
during the day.
3 of 15
SCIENCE IMMUNOLOGY | REVIEW

Cell type Mouse strain/cells Experimental model Findings Ref

T cell deficiency in Bmal1 abrogates rhythmic,
cell-intrinsic CCR7 and S1PR1 expression, rhythmic T cell
T cell Cd4cre:Bmal1flox Steady state and EAE (66)
homing to LN, and differences in disease severity in an
EAE model.
Reduced T cell IFN-, IL-2, and TNF- production during
Cd4cre:Bmal1flox L. monocytogenes infection (56)
L. monocytogenes infection.
Abrogation of time-of-day–dependent expansion of
E8Icre:Bmal1flox DC-OVA vaccination OVA-specific T cells in response to vaccination performed (49)
at CT6 versus CT18
REV-ERB/ agonist (SR9009, Jurkat and primary human T Pharmacological activation of REV-ERBs inhibits HIV
(108)
GSK2667) administration cell lines infected by HIV promoter activity, impeding viral replication in vitro.

Rev-erb−/− full KO Rev-erb−/−/−/−

Perturbed light cycles Oscillations in TH17 differentiation are disrupted. (57)
DKO
Loss of rhythmic T cell cellularity in blood, lymph, and
Adrb2−/− full KO Integrin blocking (70)

Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023

peripheral LN.
Increased activation of the T cell CCR7 and CXCR4
Adrb2−/− full KO 2-AR agonist stimulation (68)
signaling pathways. Increased T cell retention in the LN.
Reduced IL-7Ra expression during the active phase, loss
Cd4cre:Nr3c1flox Steady state of rhythmic T cell CXCR4 expression, and loss of (69)
rhythmic T cell cellularity in blood and LN.
Clock19 In vitro anti-CD3 stimulation Loss of rhythmic T cell proliferative capacity. (10)
T cell deficiency in Bmal1 does not abrogate rhythmic
Cd4cre:Bmal1flox Steady state (87)
CXCR4 expression on Tregs in the spleen or inguinal LN.
Human peripheral blood Reduced peripheral B cell numbers, defective B cell
B cell Bmal1+/−, Bmal1−/− full KO (64)
lymphocytes immunization development, and IgG1 production.
Increased IgG and IgM production. Spontaneous
Cry1−/–Cry2−/− full DKO NP-KLH immunization (65)
development of an autoimmune phenotype.
flox
Cd19cre:Bmal1 Steady state Loss of rhythmic homing to LN. (66)
Loss of rhythmic B cell cellularity in blood, lymph, and
Adrb2−/− full KO Integrin blocking (70)
peripheral LN.
Loss of diurnal variation in IgM and IgG1 antibody
Adrb2−/− full KO NP-CGG immunization (70)
production.
Increased B cell retention in the LN and increased
Adrb2−/− full KO 2-AR agonist stimulation (68)
activation of the CCR7 and CXCR4 signaling pathways.
flox
Gut ILC3s Vav1cre:Bmal1 Mixed bone marrow chimera Reduced ILC3 cellularity in the gut. (95)
Reduced ILC3 cellularity in the gut and the remaining
Rorccre:Bmal1flox Steady state (96)
ILC3s are IL-17+ and IL-22+.
+
Loss of REV-ERB increases the percentage of IL-17 and
Steady-state and C. difficile
Rev-erb−/− full KO IL-22+ gut ILC3s and increases bacterial translocation to (97)
infection
mesenteric LNs during C. difficile infection.
Donor KO-derived ILC3s cells have reduced cellularity in
Il7rcre:Bmal1flox Mixed bone marrow chimera (90)
the gut compared with control Bmal1flox donor cells.
Reduced expression of ILC3 gut-homing molecules
Steady state and C. rodentium altered microbiota. After infection with C. rodentium, KO
Rorccre:Bmal1flox (95)
infection mice exhibit increased clinical score, bacterial burden,
and bacterial translocation.

(ZT12), these rhythms are lost in REV-ERB–deficient mice. This
phenomenon was shown to be driven by REV-ERB expression in
immune cells using bone marrow chimeric mice. Wild-type mice that
received REV-ERB–deficient bone marrow lost the time-of-day
difference in neutrophil infiltration, whereas those receiving REV-­
ERB–sufficient bone marrow retained rhythms after LPS stimula-
tion. Thus, these results indicate that leukocyte REV-ERB is one of
the factors that control rhythmic neutrophil infiltration into the
lungs after LPS challenge.
Macrophages and monocytes
Among immune cell types, the circadian rhythms in macro-
phages and monocytes have been the most studied to date (Table 1).
In an S. pneumoniae infection model, the phagocytic activity of

Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022 4 of 15

SCIENCE IMMUNOLOGY | REVIEW
macrophages was shown to be regulated by BMAL1 (30). Deletion
of Bmal1 in both Lyz2cre:Bmal1flox and Cx3cr1cre:Bmal1flox mice
showed a strong protective effect, rescuing animals from severe
bacteremia and weight loss. Further in vivo and ex vivo experi-
ments determined phagocytic function to be enhanced in Bmal1-­
deficient macrophages because Bmal1 deletion in macrophages
alters the actin cytoskeletal organization and enhances overall anti-
bacterial function (30). These data suggest that BMAL1 is a strong
regulator of macrophage morphology and motility.
An additional key element affecting the inflammatory status of
macrophages appears to be the circadian regulation of cellular me-
tabolism (31). A recent study showed BMAL1 signaling to regulate
mitochondrial metabolism in M1 macrophages upon stimulation
with interferon- (IFN-) and LPS (32). Bmal1-deficient macro-
phages exhibited dramatically reduced oxygen consumption; on the
other hand, succinate dehydrogenase–mediated production of reac-
tive oxygen species was enhanced compared with control cells. This
indicates that BMAL1 promotes mitochondrial function. Mechanis-
tically, these effects may be mediated by a posttranscriptional meta-
bolic regulation (33). Comprehensive profiling of the transcriptome
and proteome of bone marrow–derived macrophages (BMDMs)
showed that only 15% of the oscillatory proteome was found to have
an oscillating mRNA, indicating that most of the proteome oscillates
because of posttranscriptional regulation (33). Further analy-
sis showed that proteins involved in degradation and translation,
such as tRNA-aminoacylation proteins and ubiquitin ligase, are
rhythmically enriched, which could explain the phenomenon.
Moreover, tricarboxylic acid–related proteins and oxygen con-
sumption rate exhibit circadian oscillations, resulting in rhythms of
adenosine triphosphate production (33). Together, these data show
that the clock gene Bmal1 can regulate the metabolic function of
macrophages.
Recent studies have focused on the connections between clock
components, particularly REV-ERB, and inflammasome pathways
(Table 1) (34–37). In mouse and human macrophages, NLR family
pyrin domain containing 3 (NLRP3, a major component of the in-
flammasome) is expressed in a diurnal manner under the control of
REV-ERB (35). NLRP3 modulates the expression and secretion of IL-1
and IL-18 in the peritoneum, resulting in time-of-day–dependent
disease severity changes in a mouse model of fulminant hepatitis (38).
Mechanistically, chromatin immunoprecipitation (ChIP) assays
showed that REV-ERB can directly bind to the Nlrp3 promoter re-
gion in BMDMs (35, 38). Pharmacological activation of REV-ERB
inhibited the NLRP3 inflammasome and reduced the severity of fulmi-
nant hepatitis (35, 38). REV-ERB was further shown to repress the
function of the nuclear factor B (NF-B)/NLRP3 axis, and REV-ERB–­
deficient mice exhibited a more activated NLRP3 inflammasome (38).
Enhanced inflammasome activity rendered these mice more sensitive
to experimental colitis, demonstrating critical interactions between
circadian rhythms and inflammasome regulation. These studies
show the strong anti-inflammatory functions of REV-ERB, in
addition to its role in the circadian clock.
On the basis of this important anti-inflammatory role, REV-ERB
is currently used as target in therapeutic studies. One study found
that the REV-ERB agonist GSK4112 can inhibit the expression of
NLRP3 and reduce the production of IL-1 in an LPS-induced in-
flammation model (34). Another study showed that an NF-B–
driven long noncoding RNA (lncRNA), Lnc-UC, can promote the
expression of REV-ERB and, in a negative feedback loop, can
Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022
inactivate NF-B and the NLRP3 inflammasome in macrophages
(39). Furthermore, earlier studies reported that REV-ERB can also
regulate the expression of the cytokine IL-6 and the CC motif
chemokine ligand 2 (CCL2) in macrophages (40, 41), as well
as CCL2 in microglial cells (42). These data implicate REV-ERB
as an important mediator of anti-inflammatory roles of the clock
machinery.
With respect to monocytes, in a sterile thioglycollate peritonitis
model and after infection with Listeria monocytogenes, the recruit-
ment of Ly6Chigh inflammatory monocytes to the inflamed site was
demonstrated to exhibit a diurnal variation, regulated by the myeloid
cell–intrinsic clock (43). Mice infected at ZT8 (“afternoon”) showed
better control of L. monocytogenes bacterial spread compared with
ZT0 (“morning”) infected mice, with increased levels of the cyto-
kines and chemokines IL-1, TNF-, IFN-, and CCL2. Mice with
Bmal1-deficient myeloid cell subsets (Lyz2cre:Bmal1flox) were more
Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023
susceptible to L. monocytogenes, showing reduced survival and an in-
crease in plasma of the same proinflammatory cytokines and chemo-
kines (43). Deletion of Bmal1 also impaired the diurnal oscillation
of Ly6Chigh monocyte numbers in blood, spleen, and bone marrow,
as well as in the peritoneum. Furthermore, ChIP analysis showed
that the BMAL1-CLOCK heterodimer binds directly to the E-box of
Ccl2, controlling its rhythmic expression (43). This work provided
evidence for a crucial role of Bmal1 in inflammatory monocytes.
In summary, macrophages and monocytes exhibit a strong circadian
rhythm, particularly with respect to the circadian components BMAL1
and REV-ERB, which are implicated in controlling metabolism,
function, and cytokine production in these cells.
Neutrophils
The number of neutrophils in blood oscillates with time of day, and
their phenotype exhibits a diurnal change that has been associated
with aging. Both phenomena are controlled by Bmal1 in a cell-­
autonomous manner, as demonstrated with the use of an hMRP8Cre:
Bmal1flox mouse line that targets neutrophils (44). ChIP assays
demonstrated that BMAL1 can directly bind to the E-box in the pro-
moter region of Cxcl2, which governs neutrophil aging in a CXCL2-
C-X-C motif chemokine receptor 2 (CXCR2)–dependent manner.
In contrast, CXCR4 acts as an antagonist to CXCR2-induced aging,
favoring neutrophil clearance and protection of the cardiovascu-
lar system (44). Surprisingly, although neutrophils only represent
a minor fraction of leukocytes in nonhematopoietic tissues under
steady-state conditions, they were reported to be important regula-
tors of the hematopoietic stem cell niche and to alter clock gene ex-
pression in various organs (5, 45). For example, neutrophils were
shown to infiltrate the mouse liver in a time-of-day–dependent man-
ner, which altered the hepatocyte clock because of neutrophil elastase
(NE) secretion (46). Neutrophils peaked in the liver during the light
phase, where they secreted elastase and activated c-Jun N-terminal
kinase (JNK) signaling in hepatocytes, ultimately triggering Bmal1
expression. Intriguingly, this phenomenon may also relate to the
human liver, as a strong correlation between activation of JNK, NE
levels, and BMAL1 expression was observed in human patient sam-
ples (46). In other tissues, neutrophil infiltration was shown to alter the
transcriptional profile of the whole organ (47). Neutrophil infiltration of
the intestine, for instance, suppressed the expression of IL-23 in
resident macrophages, thereby reducing the concentration of granu-
locyte colony-stimulating factor (G-CSF) in blood plasma (47). These
studies demonstrate the strong influence of circadian rhythms on
5 of 15
SCIENCE IMMUNOLOGY | REVIEW

neutrophils, which appear to play much more prominent roles than was shown to promote TH17 differentiation. It does this by inhibit-
previously anticipated in various organs even when their numbers ing the expression of nuclear factor, interleukin 3 regulated (NFIL3),
present in those tissues are low. which itself is a repressor of RORt (encoded by Rorc)—a positive
regulator of TH17 differentiation (57–63). Rhythmicity in these
Dendritic cells transcription factors modulates TH17 differentiation. Thus, TH17
Dendritic cells (DCs) are phagocytic antigen-­presenting cells (APCs) that development is favored during the mouse behavioral rest phase
directly link the innate immune system to the adaptive immune because of high NFIL3 activity during the active phase. Disrupting
system by collecting antigen at distal sites and presenting it to circadian rhythms through the use of a jetlag model increased the
cells of the adaptive immune system in immune organs. A recent frequency of TH17 cells and made mice more susceptible to dextran
study found that the migration of mouse DCs into afferent lym- sulfate sodium (DSS)–induced colitis (57). The numbers of TH1
phatic vessels of the skin occurs in a rhythmic manner, with a cells in the intestine were also reduced in Clock mutant mice, sug-
peak around ZT7 (“day”) and a trough at ZT19 (night) (Fig. 1) (48). gesting that the circadian clock also plays a role in intestinal TH1
This rhythmic migration is driven by diurnal expression of several differentiation in addition to TH17 cells (57). Because these tran-
adhesion molecules on lymphatic endothelial cells (LECs) such as scription factors mediating peripheral T cell differentiation are also
CCL21, lymphatic vessel endothelial receptor 1 (LYVE-1), CD99, clock genes, fine-tuning T cell phenotypes in peripheral sites may
and junctional adhesion molecule A (JAM-A), as well as DC-­ be affected by the circadian rhythm. However, future studies are

Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023

expressed CCR7. Synchronization of bone marrow–derived DCs required to assess a potential time-of-day–dependent role for RORt
(BMDCs) and lineage-­specific ablation of BMAL1 function in conven- and ROR in stabilizing TH17 commitment and to address whether
tional DCs (Clec9acre:Bmal1flox), LECs (Prox1creERT2:Bmal1flox), or T cell–specific ablation of BMAL1 changes the capacity for TH17
endothelial cells (Cdh5creERT2:Bmal1flox) implicated both DC-­ differentiation in a circadian manner.
autonomous and LEC circadian clocks in these rhythms (Box 1). In contrast to T cells, B cell differentiation appears to be depen-
ChIP analyses indicated that BMAL1 directly controls the expression dent on the molecular clock machinery. Complete BMAL1-deficient
of Ccl21 and Ccr7, as well as the adhesion molecule Lyve1 (48). mice exhibit a partial block in B cell development, as demonstrated
Another study using BMDCs as APCs demonstrated that expan- by a significantly reduced percentage of B220high, mature B cells and
sion of antigen-­specific CD8 T cells is time-of-day dependent a significantly increased percentage of B220low out of the total bone
and partially driven by the DC intrinsic clock, given that dele- marrow cells (64). This effect was demonstrated via adoptive transfer
tion of Bmal1 in BMDCs impaired T cell migration to the spleen experiments to be non–B cell autonomous and to involve the bone
after adoptive transfer (49). An ex vivo study using REV-ERB– and marrow stromal cell compartment (64). This is in line with data from
REV-ERB–deficient BMDCs showed that loss of either REV-ERB mice with a B cell–intrinsic deletion of BMAL1 that exhibit normal
or REV-ERB enhanced the expression of the maturation markers B cell development and function (56). In contrast, mice exhibiting
CD86 and major histocompatibility complex II (MHCII), as well as double deficiency for the BMAL1-repressive clock components Cry1
proinflammatory cytokines such as Il1b, Il6, and Il12b (50). Inversely, and Cry2 show increased serum immunoglobulin G (IgG) levels, in-
the REV-ERB–specific agonist SR9009 inhibited the expression of creased phosphorylation of the proximal B cell receptor signaling
maturation markers and proinflammatory cytokines in BMDCs. pathway, and an overall autoimmune phenotype (65). These results
Consistent with the data in macrophages (34–37), REV-ERB pres- indicate a non–cell-autonomous role for the circadian clock in B cell
ents anti-inflammatory function in DCs, a phenotype that appears development and differentiation.
to be independent of its role within the clock. In summary, these
data demonstrate that DC functions are circadian in nature and Lymphocyte trafficking
can be altered by targeting the clock machinery. Circadian rhythms govern the rhythmic shuttling of lymphocytes
between blood, lymph (both peaking during the behavioral rest
phase), and tissues (peaking around the onset of the active phase)
ADAPTIVE IMMUNITY (3, 66–71). Thus far, three mechanisms have been identified to ex-
Given the longer time frame compared with innate immune re- plain these daily migration patterns, which likely co-govern the
sponses, it is surprising that adaptive immunity exhibits circadian overall phenotype: BMAL1 activity in both lymphocytes and endo-
rhythmicity as well, even weeks after an initial challenge (Table 1) thelial cells (3, 66), the sympathetic nervous system via activation of
(7, 16, 51–55). Here, we provide a summary of the major findings the 2–adrenergic receptor (AR) (70), and GCs (69), the latter two
and highlight the most recent developments. of which are discussed in the next section.
BMAL1 controls lymphocyte trafficking via cell-intrinsic and
Development of lymphocytes cell-extrinsic means. Cell-intrinsic expression of BMAL1 in T cells
Current data indicate that T cell development in the thymus is inde- and B cells governs rhythmic cellularity in lymph nodes (LNs) by
pendent of the circadian clock because T cells with an intrinsic controlling circadian oscillations in the expression of the LN-­
deletion of BMAL1 exhibit no defects in thymic development (56). homing factor CCR7 and the egress factor S1pr1 in lymphocytes
In addition, the frequency of CD4+ T helper 17 (TH17) cells in the (66). Endothelial BMAL1 expression is required for the circadian
intestinal lamina propria (LP) is unchanged in these mice, which protein expression of the adhesion molecules intercellular adhe-
suggested that intestinal TH17 differentiation is also not directed by sion molecule–1 (ICAM-1) and vascular cell adhesion molecule–1
BMAL1 (56). However, several clock genes are involved in or even (VCAM-1) in endothelial cells, which guide rhythmic lymphocyte
master regulators of TH17 differentiation, including Nr1d1 and the homing into tissues (3,  66,  72). Without oscillatory expression of
ROR family of transcription factors (Rorc and Ror in particular). these adhesion receptors, rhythmic tissue recruitment is lost
Using whole-body knockout mice, REV-ERB (encoded by Nr1d1) (3, 66, 72). Thus, BMAL1 is extrinsically (through endothelial cells)

Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022 6 of 15

 SCIENCE IMMUNOLOGY | REVIEW

in the day (ZT2) showed increased IL-2

production compared with infection at
night (ZT14) (56). Together, these data
indicate that, although adaptive immune
responses are clearly rhythmic, different
routes of immunization may be re-
sponsible for some of these observed
time shifts in peak response times.
Oscillations in T cell responses to
antigen stimulations are due, at least in
part, to rhythmic changes in the de-
gree to which naïve T cells are poised
to respond to stimulation throughout
the circadian cycle. The temporal dif-
ferences in T cell vaccination responses
correlated with oscillations in the mRNA

Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023

expression of T cell receptor (TCR) sig-
naling pathway regulators (49). Positive
regulators of T cell activation (ZAP70,
AKT, and phospholipase C–) peaked
during the day, the time at which
OVA-loaded DC vaccination yielded
the highest T cell responses. In contrast,
negative regulators of TCR signaling
[SHP1 (small heterodimer partner 1),
CSK (C-terminal Src kinase), and PTEN
phosphatase and tensin homolog] peaked
at night (49). These data suggest that
naïve T cells are poised to respond to
antigens in a time-of-day–dependent
Fig. 1. DCs migrate into skin lymphatics in a circadian manner. DC migration peaks during the daytime (left) and
manner and, together with rhythmic DC
troughs during the nighttime (right) in mice. This oscillation is driven by rhythmic secretion of CCL21 by LECs and the trafficking and function, contribute to the
rhythmic expression CCR7 on DCs and adhesion molecules LYVE-1, CD99, and JAM-A on LECs (48). All of these mole- oscillations in vaccination responses.
cules show increased expression during the daytime (left) and decreased expression at nighttime (right). These pro- B cells demonstrate rhythmic effec-
teins are directly regulated by LEC or DC-intrinsic mechanisms, respectively, where BMAL1 binds to promoter regions tor responses through cell-intrinsic, clock-­
in their genes. driven TLR9 protein expression (73).
When mice were stimulated with OVA-
CpG (a TLR9 agonist) at peak TLR9
and intrinsically essential for normal lymphocyte circulation between expression on LN B cells [occurring around ZT19 (night)], they ex-
blood, lymph, and tissues. hibited increased cytokine production and proliferation compared
with stimulation during times of lower TLR9 expression (73). Thus,
Effector functions these correlative data indicate that, similar to T cells, B cells may also
Adaptive immune cells also display rhythmic effector functions. be poised to exhibit strong effector responses in a circadian manner.
T cells exhibit clock-dependent, rhythmic proliferation (10) and Rhythmic effector function can also affect the severity of auto-
show oscillatory IFN- production after in vitro stimulation (11). In immune diseases. Mice lacking BMAL1 in T cells show reduced severity
addition, several studies have demonstrated that the strongest adap- scores after induction of experimental autoimmune encephalomyeli-
tive immune response occurs if antigen is encountered in the skin tis (EAE; an animal model of multiple sclerosis) (66). Furthermore, in
when LNs are near their peak cellularity, which, in mice, occurs be- the skin, it was shown that rhythmic mRNA expression of the IL-23
tween late afternoon and late night (Table 1) (66, 69, 70, 73, 74). In receptor (IL-23R) in T cells correlated with imiquimod (IMQ)–­
contrast, mice immunized intravenously with ovalbumin (OVA)– induced psoriasis severity in murine skin (75). IL-23 is released in
CREDIT: A. MASTIN/SCIENCE IMMUNOLOLGY

loaded DCs produce more OVA-specific T cells in the spleen when psoriatic lesions by DCs and macrophages, which stimulates IL-23R–
immunized in the day (ZT6) versus at night (ZT18) (10). Mice vac- expressing cells to release IL-22 and IL-17, further exacerbating the
cinated with OVA-loaded DCs during the day have a significantly disease (76). ChIP assays demonstrated that CLOCK binds to the
increased percentage of OVA-specific, IFN-producing effector promoter region of IL-23R in T cells (75). On the other hand, mice
T cells in the spleen 7 days after vaccination compared with vaccina- with a loss-of-function mutation in Clock (Clock 19) lost rhythmic
tion during the night (CT18) (49). These mice further exhibit a sig- IL-23R expression on T cells, displayed reduced IL-22 expression,
nificantly lower bacterial load when challenged with a lethal dose of and exhibited significantly reduced psoriasis severity compared
L. monocytogenes 7 days after DC-OVA vaccination (49), corrobo- with wild-type mice (75). REV-ERB has also been shown to con-
rating previous observations where L. monocytogenes–infected mice trol inflammatory cytokine production in T cells in the context

Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022 7 of 15

SCIENCE IMMUNOLOGY | REVIEW
of IMQ-induced psoriasis (77). REV-ERB–deficient mice exhibited
significantly increased numbers of IL-17+ T cells. In contrast, stim-
ulating REV-ERB with the synthetic agonist SR9009 suppressed
IL-17 production by T cells and reduces the severity of IMQ-­
induced psoriasis (77). These data point to a crucial role for clock tran-
scription factors in autoimmune diseases.
There is evidence that long-term disruption of an organism’s
rhythmic environment affects adaptive immune cell function. Aged
mice who undergo chronic jetlag (CJL) display a reduced life span
(78), and extended CJL (~85 weeks) starting from a young age exac-
erbates immune senescence and mortality (79). CJL resulted in
significantly increased numbers of PD-1 +CD44 +CD4 + T cells,
PD-1+CD44+CD153+CD4+ T cells, T follicular helper cells (Tfh), and
T regulatory cells (Tregs) in the spleen—all phenotypes associated
with T cell senescence (79). In addition, this phenomenon was ac-
companied by significantly higher percentages and numbers of
germinal center B cells (B220+CD19+CD95+GL7+) (79). A second
study used high-throughput RNA sequencing data from the Cancer
Genome Atlas to show that disrupted circadian rhythms in tumors
correlate with increased tumor-associated T cell anergy in human
cancers (80). Across 716 samples and 29 cancer types, negative reg-
ulators of the circadian clock were down-regulated, whereas the
positive regulator ARNTL2 was up-regulated in most types of can-
cers investigated, suggesting that tumors lose oscillatory mRNA
expression of circadian clock genes (80). Furthermore, high expres-
sion of clock genes was positively correlated with levels of T cell
anergy markers in tumor samples, such as programmed cell death
protein 1 (PD-1) and cytotoxic T-lymphocyte associated protein 4
(CTLA-4) (80). These findings indicate a link between dysregulated
clock gene expression and T cell anergy in tumors, indicating that
adaptive immune cell senescence is influenced by circadian rhythms
in aging and in the context of cancer.
NEUROENDOCRINE CONTROL OF THE IMMUNE SYSTEM
The nervous system directly conveys external, circadian signals to
an organism’s immune system, setting leukocytes in tune with
rhythms in the environment. The adrenergic branch of the sympa-
thetic nervous system is under circadian control and uses the cate-
cholamines epinephrine and norepinephrine as neurotransmitters
to convey circadian oscillations (81). The central circadian clock,
situated in the suprachiasmatic nucleus (SCN) of the hypothalamus
(Box 1), synchronizes the rhythmic synthesis and secretion of cate-
cholamines, which ultimately results in the peripheral control of
immune cell trafficking through AR signaling (52).
One of the first evidence of adrenergic control over hematopoietic
cell migration was uncovered by the mechanistic demonstration that
the circadian clock regulates hematopoietic stem and progenitor cell
(HSPC) trafficking (82). The circadian clock synchronizes the local
secretion of norepinephrine that activates the 3-AR on bone marrow
stromal cells. Adrenergic signaling then results in the degradation of
the transcription factor S1P and the down-regulation of the bone mar-
row retention factors VCAM-1 and CXCL12 as well as endothelial
selectins. These changes ultimately lead to the oscillatory mobilization
of HSPCs from the bone marrow into the blood (82). In subsequent
years, these data were extended to demonstrate that a similar process
occurs in the recruitment of HSPCs to the bone marrow as well as in
myeloid cell recruitment to skeletal muscle (72). These rhythms in
homing are driven by circadian oscillations of VCAM-1, P-selectin,
Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022
and E-selectin expression in bone marrow sinusoids, as well as
ICAM-1 and CCL2 expression in skeletal muscle endothelial cells
both at the mRNA and at the protein level (72). Removal of nerve-­
derived adrenergic signals via surgical, chemical, or genetic denerva-
tion abrogates the circadian differences in leukocyte migration to the
bone marrow, the muscle, and the LN (68, 70, 72). An additional study
provided mechanistic insight into how adrenergic signaling regulates
two distinct peaks of HSPC activity, in the blood and bone marrow,
across the 24-hour period (83). The first peak, driven by the onset of
light, increases norepinephrine levels. This triggers HSPC prolifera-
tion and differentiation that, together with increased vascular perme-
ability, lead to blood replenishment. The onset of darkness induces a
second peak in HSPC activity via increased TNF- secretion, which
mediates HSPC maintenance (83). These data provide insights into
how adrenergic signaling can account for two opposite events, name-
ly, rhythmic egress and homing of hematopoietic cells to and from the
Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023
bone marrow. Local adrenergic signaling also plays a critical role in
oscillatory inflammatory leukocyte recruitment to arteries and veins
(72, 84). In a TNF-–induced inflammation model, it was demon-
strated that 2-AR signaling regulates the rhythmic expression of
adhesion molecules within both vascular beds. This has functional
implications for a time-of-day–dependent predisposition to intra-
vascular, thrombotic events, involving rhythmic adhesion of leuko-
cytes and platelets to endothelial cells (84). These findings show
that leukocyte trafficking is heavily controlled by circadian oscil-
lations in the sympathetic nervous system.
2-AR signaling is also a crucial regulator of lymphocyte effector
functions after time-of-day–dependent immunizations. Immune re-
actions were observed to be stronger when immunization was per-
formed at ZT17 (night; coinciding with enhanced adrenergic tone)
than at ZT5 (day), a phenotype that was abrogated in 2-AR–deficient
animals (70). Mice immunized at night displayed higher IgG1 and
IgM-specific antibody titers as well as increased antigen-specific
germinal center B cells and Tfh cells than day-immunized animals.
These results thus suggest that adrenergic signaling contributes to a
diurnal variation in the adaptive immune response, particularly with
respect to controlling the numbers of lymphocytes in LNs.
Together with the sympathetic nervous system, the parasympa-
thetic nervous system has been shown to regulate circadian trafficking
of HSPCs and leukocytes (85). At night, cholinergic signals from the
parasympathetic nervous system are antagonizing sympathetic-­
derived noradrenergic signals. This has been suggested to repress 3-
AR signaling at this time and suggested to result in reduced bone
marrow egress of both HSPCs and leukocytes, whereas the 2-AR–
mediated bone marrow homing capacity of cells is retained. During
daytime, disinhibition of noradrenergic tone triggers egress of the cells
from the bone marrow. Coordination between these two branches of
the nervous system has, thus, been suggested to result in diurnal oscil-
lations of HSPCs and leukocyte egress from the bone marrow (85).
The central nervous system also coordinates circadian GC re-
lease from the adrenal cortex in a time-of-day–dependent manner.
GC levels peak at the beginning of the active phase in both mice and
humans and act by binding to the GR, promoting its translocation
to the nucleus where this complex acts as a transcription factor. GCs
are best known for their immunosuppressive roles; however, they
are more accurately described as immune modulators that play
important immunostimulatory and inhibitory roles in addition to
regulating circadian lymphocyte migration (53, 86). One example
of this is the GC-regulated expression of the interleukin 7 receptor
8 of 15
SCIENCE IMMUNOLOGY | REVIEW
alpha (IL-7R) on T cells, which is required for normal T cell
proliferation, differentiation, and survival. GC binding to the GR in
T cells promotes the expression of IL-7R protein by binding to GR
elements (GREs) in the Il7r enhancer (69). Removing GR or the
GREs in the Il7r locus in T cells ablates the normally observed
rhythmic IL-7R expression, and mice lacking T cell–intrinsic GR
or Il7r GREs lose circadian rhythmicity in T cell cellularity in the
blood and lymphoid organs (69). This is due to loss of rhythmic,
IL-7–driven CXCR4 expression, and mice lacking T cell–intrinsic
GR or Il7r GREs exhibit a similar phenotype to CXCR4-deficient
T cells (69). Because CXCR4 is the receptor for CXCL12 and crucial
for driving leukocyte trafficking, interrupting this axis disrupts
circadian oscillations in leukocyte migration (3). There is additional
evidence that GC-induced CXCR4 expression preferentially recruits
Tregs specifically during the active phase to inflamed joints in a mouse
model for arthritis (87). Although Tregs show no overt oscillations in
circadian clock genes (87), during both homeostasis and chronic
inflammation, these cells exhibit circadian CXCR4 expression that
matches the rhythmic oscillations of serum GCs (87). These data
reveal a critical role for GCs in guiding the circadian distribution of
lymphocytes throughout the body.
GUT IMMUNITY
There is a significant amount of evidence connecting circadian
rhythms to the function of the digestive system, gut microbiota, and
intestinal immunity. The timing of food intake exerts a strong
influence on peripheral clocks and causes fluctuations in microbiota,
gut cell motility, and nutrient absorption (88, 89). Food intake also
triggers rhythmic release of vasoactive intestinal peptide (VIP) from
enteric neurons in the intestine (90, 91). VIP binds to G protein–
coupled receptor VIP receptor 2 (VIPR2) on immune cells, such as
intestinal innate lymphoid cells group 2 (ILC2s) and ILC3s, and in-
fluences their function in the gut (88, 90, 91). Although the impact
of food intake and microbiome activity on the circadian rhythm of
the organism has been recently reviewed (88, 89), here we will focus
on how the circadian rhythm directs immune cells in the gut, par-
ticularly ILC3s (Fig. 2 and Table 1).
Gut tolerance
During periods of food intake (generally the behavioral active phase
of an organism), the gut is poised to absorb a maximum amount of
nutrients, which also facilitates microbiota to traverse the intestinal
epithelial barrier. Recently, a study revealed that MHCII protein ex-
pression on murine small intestine epithelial cells (SIECs) is rhyth-
mic, is Bmal1 dependent, and peaks in the resting phase (92). This
was paired with rhythmic permeability of the small intestine (SI),
which, in contrast, peaks during the active phase (92). Rhythmic
MHCII expression on SIECs, microbiota composition, and feeding
rhythms all contributed to circadian SI permeability, suggesting that
rhythmic SI permeability is due to oscillations in microbiota detec-
tion (92). Indeed, rhythmic MHCII expression on SIECs could be
induced in germ-free mice after colonization with mouse segmented
filamentous bacteria (SFB), but not with rat SFB, which is unable to
attach to murine SIECs (92). SIECs lacking CDC42 expression, which
is essential for SFB to attach to the gut epithelium, also lost diurnal
MHCII expression (92). Furthermore, abrogation of MHCII ex-
pression in SIECs (Villin-cre:I-Abflox) reduced the numbers of CD4+
intraepithelial T cell lymphocytes in the gut and exacerbated the
Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022
severity of a Crohn’s disease model in mice (92). These data
demonstrate that there is a delicate circadian balance between pro-
moting gut permeability for nutrient absorbance during feeding
hours and increasing intestinal immune surveillance of microbiota
during the resting hours, which allows organisms to anticipate
nutrient uptake while maintaining homeostasis.
Gut microbiota
In recent years, the gut microbiota has been shown to be highly in-
fluenced by host circadian rhythm and rhythmic food intake. Gut
bacteria rhythmically adhere to the intestinal wall and exhibit time-
of-day–dependent changes in species composition. The microbiota
also has oscillatory production of metabolites that rhythmically
affect host rhythms, both locally in the gut and in relatively distant
sites such as the liver (93). Changes in diet affect the gut microbiome
and can weaken immune responses against bacteria or even alter im-
Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023
mune cell migration patterns in the blood and other tissues (88).
The circadian immune response against Salmonella Typhimurium
has recently been demonstrated to be driven by oscillations in feed-
ing behavior and the activity of the gut microbiota. Mice pretreated
with streptomycin are more susceptible to oral Salmonella Typh-
imurium when infected early during the day (ZT4; higher bacterial
count in the colon and mesenteric LNs) compared with the night
(ZT16) (94). In contrast, nonantibiotic-treated mice infected with
Salmonella Typhimurium orally at ZT12 exhibit an increased bacte-
rial burden in the SI 24 hours later and suffer from significantly in-
creased mortality compared with those infected at ZT0 (morning)
(89). This demonstrates that preserving microbial diversity dramatically
changes infection outcome. This is due to food intake–driven oscil-
lations in the protein expression of antimicrobial peptides (AMPs)
(89). Food intake peaks early in the active phase and promotes SFB
to associate with SIECs. This triggers rhythmic activation of an innate
immune circuit where a MyD88-dependent response in CD11c+
myeloid cells activates ILC3s to secrete IL-22, leading to signal trans-
ducer and activator of transcription 3 (STAT3) activation in SIECs
and circadian synthesis of AMPs (Reg3g, Lcn2, and S100A8) (89).
These oscillations are completely ablated in fasting mice or in animals
lacking SFB. They are also dependent on the expression of MyD88 in
CD11c+ cells (Cd11ccre:Myd88flox) and STAT3 in SIECs (Villincre:
Stat3 flox), as well as the presence of gut ILC3s (89). Because
Salmonella Typhi­murium is resistant to Lcn2, it gains a competitive
advantage in the early active phase (ZT12), outcompeting the healthy
microbiota that is susceptible to Lcn2 (89). These results demonstrate
how the host circadian rhythm affects microbial rhythms that, in turn,
guide rhythmic host immunity.
Regulation of ILC3s
In 2019, three papers were published exploring the role of the circa-
dian clock in ILC3 homeostasis (Table 1) (95–98). All showed that
intestinal ILC3s exhibit high clock gene mRNA expression com-
pared with other immune and ILC cell subsets in the SI-LP, which
is in line with the critical role that Rorc (a clock gene that encodes
the transcription factor RORγt) and Nfil3 (a clock-associated
gene) play as defining transcription factors in ILC3 development
(90, 95–97, 99–101). ILC3s also exhibit circadian oscillations of the
key transcription factors ROR and RUNX1, which are involved
in ILC3 differentiation, as well as the ILC3 signature cytokines IL-17
and IL-22 (90, 96, 97). ILC3s receive their circadian cues primarily
from light via the SCN, and removal of this brain region by
9 of 15
 SCIENCE IMMUNOLOGY | REVIEW

A LN to the intestine, because these cells

display reduced surface protein ex-
pression of the gut-homing molecules
47, CCR9, and CXCR4 (95). However,
SI-LP ILC3s with an impaired molec-
ular clock exhibited increased frequency
of IL-17+ and IL-22+ cells and increased
expression of the proapoptotic Bcl-2–
like 11 protein BIM (96, 97). Together,
these data show gut ILC3s to be a highly
rhythmic immune cell population that
is synchronized by light.
Intriguingly, the effect of ablation of
B the circadian clock varies for different
ILC3 subsets. Although NKp46+ ILC3s
exhibited reduced numbers and in-

Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023

creased IL-22 protein expression, double-­
negative (DN; NKp46−CCR6−) and CCR6+
ILC3s exhibited increased numbers and
IL-17A protein expression when the
circadian clock was disrupted by loss of
REV-ERB. In line with this, REV-ERB–
deficient mice showed an altered chro-
matin landscape in the Il17a locus (97).
This suggests that lack of REV-ERB
C opens up ROR response elements (RORE)
binding sites at that site for additional
transcription factor binding and ulti-
mately increased IL-17 expression in DN
and CCR6+ ILC3s (97).
BMAL1 deficiency also impairs the
relationship between gut ILC3s and in-
testinal bacteria. The circadian oscillations
of microbiota populations are altered in
mice with BMAL1-deficient ILC3s, in-
dicating that the activated phenotype of
ILC3s lacking Bmal1 may be connected
Fig. 2. Circadian rhythms control the number and function of ILC3s in the SI. (A) During the active phase, ILC3s to the intestinal microbiota (95). In ad-
are present in low numbers and exhibit a nonactivated phenotype (low IL-17 and IL-22 expression) in the small intes- dition, when mice with BMAL1-deficient
tinal LP (96, 97). This is partially due to increased feeding-dependent VIP secretion from enteric VIP neurons during ILC3s are given antibiotics for 2 weeks,
the active phase (90, 91), which likely represses IL-22 expression by binding VIPR2 on ILC3s (91). Because IL-22 signals clearing much of the intestinal micro-
epithelial cells to express AMPs, low IL-22 levels allow microbes to thrive during the active/feeding phase, increasing biota, the frequency of IL-17+ and IL-22+
nutrient absorption (91). (B) During the rest phase, there are many ILC3s in the LP (96). IL-17 expression is increased ILC3s decreases and the total number
in ILC3s at this time (96, 97), and reduced food intake causes low VIP levels, promoting IL-22 expression by ILC3s (91).
of ILC3s in the intestine is restored to
These activated ILC3s promote antimicrobial functions and increase barrier maintenance, thus reducing nutrient
the level of ILC3-BMAL1–sufficient
absorption in the SI. (C) In the absence of a circadian rhythm, ILC3 numbers in the gut are dramatically reduced
(90, 95–97), but those that remain have an activated phenotype (96, 97). Mice with clock gene–deficient ILC3s also
animals (96). In agreement with this,
exhibit increased bacterial translocation during infection (95, 97), which may cause the high levels of IL-17 and IL-22 when mice with BMAL1-deficient ILC3s
on the LP ILC3s in these mice. were infected with Clostridium difficile
or Citrobacter rodentium, their ILC3s
electrolytic lesion or genetic ablation of Bmal1 from the forebrain were hyperresponsive and they had more bacterial translocation
CREDIT: A. MASTIN/SCIENCE IMMUNOLOLGY

(using Camk2acre:Bmal1flox mice) completely abrogated rhyth- from the gut to the mesenteric LN or spleen, respectively (95, 97).
micity in ILC3s (95). In contrast, ILC3s appear to be only mildly In addition, mice with BMAL1-deficient ILC3s showed overall
influenced by microbial or rhythmic feeding cues (95), which is sur- reduced survival during C. rodentium infection compared with mice
prising, given that food intake strongly influences gut bacterial with BMAL1-sufficient ILC3s (95, 97). Mice with BMAL1-deficient
composition. Ablating the circadian clock in Rorc-expressing cells ILC3s also exhibited more severe gut inflammation during infection.
(using Rorccre:Bmal1flox mice), which includes gut ILC3s, causes a re- Furthermore, human patients with inflammatory bowel disease
duction in the frequency and number of ILC3s in the intestine, but exhibit ILC3 populations with dysregulated clock genes, compared
not the spleen, blood, or lung (90, 95–97). This is likely due to an with that of control patients (96). Together, these data make a
inability of clock-deficient ILC3s to migrate from the mesenteric strong case that circadian regulation of ILC3s is crucial for

Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022 10 of 15

SCIENCE IMMUNOLOGY | REVIEW
maintaining gut homeostasis with the microbiota and managing
gut infection.
Although the number of ILC3s in the gut appears to be con-
trolled by the central clock and light, the function of ILC3s may be
influenced by food-induced expression of VIP. In early 2020, two
papers showed that ILC3s express the VIPR2, which binds VIP (90, 91).
ILC3s and VIP-expressing neurons colocalize in the intestine, and
food consumption, which induces VIP production, changes the ex-
pression of IL-22 by ILC3s (90,  91). Although one paper showed
that in vitro and in vivo VIPR2 stimulation caused reduced IL-22
protein expression in ILC3s (90), the other demonstrated the
opposite phenomenon, namely, increased IL-22 (91). Both groups
argued that the VIP-induced changes they found were important
for maintaining gut homeostasis during feeding, either for reducing
inflammation via high IL-22 levels (90) or for supporting efficient
nutrient absorption and balance with the microbiota via low IL-22
levels (91). Despite the differences in the role for VIPR2 stimula-
tion, IL-22 production was shown to be rhythmic in ILC3s, and this
rhythmicity could not be entirely abolished by removing cell-intrinsic
BMAL1 (90). It is clear from these findings that there is an import-
ant connection between circadian rhythm, VIP production, and
ILC3 homeostasis. However, more research is necessary to elucidate
the role of this neuroimmune axis in the gut.
IMMUNE RESPONSES TO PATHOGENS
Although, initially, most studies focused on sterile inflammation to
study the influence of circadian rhythms, the importance of circadian
rhythms during pathogen infection has emerged as a key factor in
disease progression. This has become even more obvious in the con-
text of the severe acute respiratory syndrome coronavirus 2 (SARS-
CoV-2) pandemic.
Many facets of SARS-CoV-2 infection and subsequent human
immune responses are time-of-day dependent. SARS-CoV-2 viral
uptake and replication in cultured human monocytes were found to
be regulated in a circadian manner, with higher entry and multipli-
cation of the virus at specific times compared with other times (102).
Furthermore, the pattern of secreted cytokines was dependent on
the time of virus encounter. The highest virus load correlated with
higher IL-6, IL-1, and IL-10 protein levels, suggesting that circadian
rhythms affect both viral infection and the host immune response.
In addition, SARS-CoV-2 entry and replication in human Calu-3
lung epithelial cells was shown in vitro to be under circadian regu-
lation of both BMAL1 and REV-ERBs (103). The circadian clock
components repress angiotensin-converting enzyme 2, the primary
entry receptor of SARS-CoV-2 in human epithelial cells, leading to
the limitation of virus entry and replication. These data make it clear
that the circadian oscillations play an important role in viral cell
invasion and the human immune response to the virus.
Besides SARS-CoV-2, circadian rhythms play important roles in
other viral infections (Table 1). Viral replication and dissemination
of both herpes and influenza A viruses depend on the time of infec-
tion, with mice infected at ZT0 (morning) with a luciferase-expressing
herpes virus (M3:luc MuHV4) exhibiting 10-fold higher viral repli-
cation than mice infected at ZT10 (evening) (104). However, this
time-of-day difference is abrogated in complete Bmal1−/− mice with
levels of viral infection being higher than in wild-type animals.
Similar results were obtained for respiratory syncytial virus (RSV)
and parainfluenza virus type 3, with higher viral replication of
Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022
both viruses in Bmal1−/− cells and a more severe RSV infection in
Bmal1−/− mice (105). The importance of BMAL1 in antiviral re-
sponse was also highlighted in the context of Sendai virus (SeV)
infection (106). Bmal1−/− mice were more susceptible to SeV infec-
tion because of a defective control of viral replication, resulting in
higher cytokine expression and an overall increased lung inflam-
mation. In the context of hepatitis B infection, REV-ERB plays a
direct role in controlling the expression of NTCP (sodium taurocho-
late cotransporting polypeptide), the primary receptor used by
hepatitis B virus (HBV) to enter cells (107). Pharmacological activa-
tion of REV-ERBs with the agonist SR9009 resulted in the inhibi-
tion of HBV entry into cells, whereas BMAL1, in contrast, was
shown to bind to HBV DNA and enhance viral promoter activity.
REV-ERBs were additionally shown to regulate HIV-1 replication
through the modulation of HIV-1 long-terminal repeat promoter
activity (108). Furthermore, susceptibility to influenza infection
Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023
is time-of-day dependent in mice, with animals infected at ZT11
(evening) experiencing a more severe disease and a higher mortality
compared with animals infected at ZT23 (morning) (109). This is
despite the fact that mice infected at ZT8 (afternoon) feature more
CD8+IFN+ T cells compared with those infected at ZT20 (night)
(66). Together, these results highlight the importance of the circadian
clock components in antiviral response.
Bacterial infections are also affected by circadian mechanisms.
As mentioned above, mice were shown to be more susceptible to oral
Salmonella Typhimurium infection when gavaged in the morning
compared with the evening (94). This increased susceptibility resulted
in greater colonization of the colon by Salmonella Typhimurium
and a greater proinflammatory response. Time-of-day–dependent
susceptibility to bacterial infection was also shown in the context of
pulmonary S. pneumoniae, with mice infected in the morning
(ZT0) experiencing higher lung and blood bacterial burden 48 hours
after infection than mice infected in the evening (ZT12) (27).
Another example comes from the genital pathogen Chlamydia
muridarum, responsible for genital tract infection and causing
infertility (110). Mice infected at ZT3 (morning) had higher
C. muridarum infectivity compared with mice infected at ZT15
(evening). This was accompanied by increased pathological lesions
and higher protein levels of CXCL1, IL-10, IL-1, and IFN- 1 week
after infection (110). Collectively, these results emphasize time-of-
day–dependent mechanisms in antibacterial defense.
Pathogenic infection with parasites has also been shown to be
clock-regulated. Expulsion of the parasite Trichuris muris is con-
trolled by the DC clock (111). Mice infected with this parasite in the
morning showed a better clearance efficiency because of a stronger
TH2 immune response in the morning compared with the evening.
In contrast, mice with BMAL1-­deficient DCs (Cd11ccre:BMAL1flox)
no longer showed this time-of-day difference. The importance of
circadian clocks in immune cells has also been demonstrated in
the context of Leishmania major infection (112). L. major infection
performed in the morning led to lower parasite burden and foot-
pad swelling compared with infection in the afternoon. This
was paralleled by a rhythmic infiltration of neutrophils and
macrophages to the site of infection, due to a rhythmic chemo-
kine profile. The implication of the circadian clock in these phe-
nomena was confirmed by generating BMDMs from Bmal1−/− mice
in vitro and in immune cells in vivo (using bone marrow chimera
from Bmal1−/− Rag2–deficient donors), which led to the ablation
of all the described oscillations (112). Together, these findings
11 of 15
 SCIENCE IMMUNOLOGY | REVIEW

Vaccination responses in humans also

exhibit circadian rhythmicity (Fig. 3). In
a cohort of 298 participants (>65 years
old), morning vaccination with an in-
fluenza vaccine was shown to induce a
higher antibody response compared with
afternoon vaccination (113). Another
influenza vaccination study suggested
that timing of sample collection could
explain the observed differences in
antibody titers between morning- and
afternoon-vaccinated participants (114).
Furthermore, vaccination with the anti-
tuberculosis agent Bacillus Calmette-
Guérin (BCG) in the morning was able
to better induce a nonspecific, trained

Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023

immunity response compared with later
times in the day (115). In view of the
current SARS-CoV-2 pandemic, adminis-
tration of a protein-based, inactivated
SARS-CoV-2 vaccine (BBIBP-CorV,
Sinopharm) in the morning was demon-
strated to enhance vaccine efficacy (116).
Participants that received morning doses
had a stronger immune response to the
vaccination, with a twofold higher level
of neutralizing antibodies compared
with the afternoon vaccination cohort.
Morning vaccination also yielded stronger
Fig. 3. Benefits of morning vaccination compared with afternoon/evening vaccination in humans. Immune
B cell and Tfh cell responses as well as
response to vaccination is time-of-day dependent with patients vaccinated in the morning experiencing a better
higher frequencies of monocytes and
response to vaccines compared with patients vaccinated in the afternoon/evening. This improved immune response
was observed for three vaccines—BCG (115), influenza (113), and SARS-CoV-2 (116)—and results in (i) greater anti-
DCs. Participants vaccinated in the
body response (113, 116), (ii) enhanced short-term immunity with higher cytokine production (IFN-, TNF-, and IL-1) morning additionally exhibited increased
(115, 116), and (iii) increased long-term immunity with increased frequencies of memory B cells (116). Mono, mono- long-term immunity and had higher
cytes; Mem B, memory B cell. percentages of spike- and RBD-specific
memory B cells (116,  117). A similar
show that oscillations in parasite infectivity and clearance are SARS-CoV-2 vaccination study performed on a U.K. cohort of health
dependent on immune cell clocks. care workers vaccinated with either Pfizer mRNA vaccine or AstraZeneca
adenoviral vaccine found that anti–spike protein responses were higher
in people who received their shot later in the day, suggesting that peak
IMMUNE RESPONSES TO VACCINATION times may vary between different vaccines or vaccination regimes
Given the importance of circadian rhythms in regulating many aspects (118). Overall, these data indicate that circadian rhythms play a role
of immune responses, it follows that circadian rhythms could be used in human vaccination responses and that those circadian rhythms
advantageously in targeted therapies such as vaccination protocols could be harnessed to optimize vaccination strategies.
(Table  1) (55). A pioneering work in mice demonstrated that
vaccination with OVA at ZT19 (night), using the TLR9 ligand
CpG-oligodeoxynucleotides as adjuvant, resulted in increased CONCLUDING REMARKS
antigen-induced lymphocyte proliferation and increased IFN- The recent years have provided substantial new arguments that
production compared with ZT7 (day) vaccination (73). This phe- rhythmicity significantly affects immune responses. In particular,
nomenon was correlated with a rhythmic expression of TLR9 on several recent advances in gut immunity have demonstrated gut-­
CREDIT: A. MASTIN/SCIENCE IMMUNOLOLGY

macrophages and B cells. Circadian vaccination responses were fur- associated immunity, principally ILC3s, to exhibit time-of-day
ther observed in the CD8 T cell response to an intravenously injected differences. Because Nr1d1 and Rorc play important roles in both
DC vaccine (49). This was largely mediated by the CD8 T cell–­ circadian rhythms and as master regulators of TH17 and ILC3
intrinsic clock and circadian modulation of the TCR signaling path- development, this indicates a strong circadian control of the develop-
way, which enhanced the activation and proliferation potential of ment of these subsets. It will be important to distinguish between
T cells during the day. These studies demonstrate that circadian the circadian and developmental roles of these transcription factors
changes in the steady-state expression of immune molecules and in the future.
activation potential of immune cells can have a profound impact on Clinical data indicate time-of-day differences in long-term immune
the strength of vaccination responses. responses in humans, indicating that chronobiological considerations

Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022 12 of 15

SCIENCE IMMUNOLOGY | REVIEW
are finding their way into the clinic. Although most of these datasets
are still assessed retrospectively for potential time-of-day effects, a
few studies are now already incorporating circadian aspect into the
study design for prospective trials. Although a lot of ground has
been made, these advances have also yielded additional unanswered
questions. One question is the relevance for circadian rhythmicity
in vaccination responses. Do the observed differences in antibody
titers provide a better outcome after challenge? Assessing this in an
experimental and clinical setting will be essential. Furthermore, where
is rhythmicity in adaptive immunity defined and how is it maintained?
It may be that the very initial response at the site of challenge
defines how the ensuing response unfolds in a circadian manner. It
is currently completely unclear how circadian rhythmicity in im-
mune responses seen months after the initial stimulus is maintained
by the body, and why these differences do not wane or even become
bigger. An answer may lie in the importance of circadian rhythms
in balancing physiology, maintaining the correct level of the immune
responses. Studies showing that lack of circadian rhythmicity can
lead to exaggerated responses support this notion. Given the recent
major developments in this relatively new field, the coming years
are sure to provide answers to these exciting questions.
REFERENCES AND NOTES
1. F. Halberg, E. A. Johnson, B. W. Brown, J. J. Bittner, Susceptibility rhythm to E. coli
endotoxin and bioassay. Proc. Soc. Exp. Biol. Med. 103, 142–144 (1960).
2. G. Fernandes, F. Halberg, E. J. Yunis, R. A. Good, Circadian rhythmic plaque-forming cell
response of spleens from mice immunized with SRBC. J. Immunol. 117, 962–966 (1976).
3. W. He, S. Holtkamp, S. M. Hergenhan, K. Kraus, A. de Juan, J. Weber, P. Bradfield,
J. M. P. Grenier, J. Pelletier, D. Druzd, C. S. Chen, L. M. Ince, S. Bierschenk, R. Pick,
M. Sperandio, M. Aurrand-Lions, C. Scheiermann, Circadian expression of migratory
factors establishes lineage-specific signatures that guide the homing of leukocyte
subsets to tissues. Immunity 49, 1175–1190.e7 (2018).
4. C. Wyse, G. O'Malley, A. N. Coogan, S. McConkey, D. J. Smith, Seasonal and daytime
variation in multiple immune parameters in humans: Evidence from 329,261 participants
of the UK Biobank cohort. iScience 24, 102255 (2021).
5. I. Cossio, D. Lucas, A. Hidalgo, Neutrophils as regulators of the hematopoietic niche. Blood
133, 2140–2148 (2019).
6. M. Baxter, D. W. Ray, Circadian rhythms in innate immunity and stress responses.
Immunology 161, 261–267 (2020).
7. R. Pick, W. He, C. S. Chen, C. Scheiermann, Time-of-day-dependent trafficking
and function of leukocyte subsets. Trends Immunol. 40, 524–537 (2019).
8. S. Wang, F. Li, Y. Lin, B. Wu, Targeting REV-ERB for therapeutic purposes: Promises
and challenges. Theranostics 10, 4168–4182 (2020).
9. G. A. Timmons, J. R. O'Siorain, O. D. Kennedy, A. M. Curtis, J. O. Early, Innate rhythms: Clocks
at the center of monocyte and macrophage function. Front. Immunol. 11, 1743 (2020).
10. E. E. Fortier, J. Rooney, H. Dardente, M. P. Hardy, N. Labrecque, N. Cermakian, Circadian
variation of the response of T cells to antigen. J. Immunol. 187, 6291–6300 (2011).
11. T. Bollinger, A. Leutz, A. Leliavski, L. Skrum, J. Kovac, L. Bonacina, C. Benedict, T. Lange,
J. Westermann, H. Oster, W. Solbach, Circadian clocks in mouse and human CD4+ T cells.
PLOS ONE 6, e29801 (2011).
12. E. Challet, The circadian regulation of food intake. Nat. Rev. Endocrinol. 15, 393–405 (2019).
13. K. Wager-Smith, S. A. Kay, Circadian rhythm genetics: From flies to mice to humans.
Nat. Genet. 26, 23–27 (2000).
14. D. Stoleru, Y. Peng, P. Nawathean, M. Rosbash, A resetting signal between Drosophila
pacemakers synchronizes morning and evening activity. Nature 438, 238–242 (2005).
15. J. S. Takahashi, Transcriptional architecture of the mammalian circadian clock. Nat. Rev.
Genet. 18, 164–179 (2017).
16. C. Scheiermann, J. Gibbs, L. Ince, A. Loudon, Clocking in to immunity. Nat. Rev. Immunol.
18, 423–437 (2018).
17. M. K. Bunger, L. D. Wilsbacher, S. M. Moran, C. Clendenin, L. A. Radcliffe, J. B. Hogenesch,
M. C. Simon, J. S. Takahashi, C. A. Bradfield, Mop3 is an essential component of the master
circadian pacemaker in mammals. Cell 103, 1009–1017 (2000).
18. J. S. O'Neill, G. van Ooijen, L. E. Dixon, C. Troein, F. Corellou, F. Y. Bouget, A. B. Reddy, A. J. Millar,
Circadian rhythms persist without transcription in a eukaryote. Nature 469, 554–558 (2011).
19. R. D. Feigin, V. H. San Joaquin, M. W. Haymond, R. G. Wyatt, Daily periodicity
of susceptibility of mice to pneumococcal infection. Nature 224, 379–380 (1969).
Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022
20. P. G. Shackelford, R. D. Feigin, Periodicity of susceptibility to pneumococcal infection:
Influence of light and adrenocortical secretions. Science 182, 285–287 (1973).
21. M. Keller, J. Mazuch, U. Abraham, G. D. Eom, E. D. Herzog, H. D. Volk, A. Kramer, B. Maier,
A circadian clock in macrophages controls inflammatory immune responses. Proc. Natl.
Acad. Sci. U.S.A. 106, 21407–21412 (2009).
22. M. L. Mul Fedele, I. Aiello, C. S. Caldart, D. A. Golombek, L. Marpegan, N. Paladino,
Differential thermoregulatory and inflammatory patterns in the circadian response
to LPS-induced septic shock. Front. Cell. Infect. Microbiol. 10, 100 (2020).
23. G. L. Pearson, M. Savenkova, J. J. Barnwell, I. N. Karatsoreos, Circadian desynchronization
alters metabolic and immune responses following lipopolysaccharide inoculation in male
mice. Brain Behav. Immun. 88, 220–229 (2020).
24. V. Lang, S. Ferencik, B. Ananthasubramaniam, A. Kramer, B. Maier, Susceptibility rhythm
to bacterial endotoxin in myeloid clock-knockout mice. eLife 10, e62469 (2021).
25. C. L. Abram, G. L. Roberge, Y. Hu, C. A. Lowell, Comparative analysis of the efficiency
and specificity of myeloid-Cre deleting strains using ROSA-EYFP reporter mice.
J. Immunol. Methods 408, 89–100 (2014).
26. J. A. Haspel, S. Chettimada, R. S. Shaik, J. H. Chu, B. A. Raby, M. Cernadas, V. Carey,
V. Process, G. M. Hunninghake, E. Ifedigbo, J. A. Lederer, J. Englert, A. Pelton, A. Coronata,
L. E. Fredenburgh, A. M. Choi, Circadian rhythm reprogramming during lung
Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023
inflammation. Nat. Commun. 5, 4753 (2014).
27. J. Gibbs, L. Ince, L. Matthews, J. Mei, T. Bell, N. Yang, B. Saer, N. Begley, T. Poolman,
M. Pariollaud, S. Farrow, F. DeMayo, T. Hussell, G. S. Worthen, D. Ray, A. Loudon, An
epithelial circadian clock controls pulmonary inflammation and glucocorticoid action.
Nat. Med. 20, 919–926 (2014).
28. L. M. Ince, Z. Zhang, S. Beesley, R. M. Vonslow, B. R. Saer, L. C. Matthews, N. Begley,
J. E. Gibbs, D. W. Ray, A. S. I. Loudon, Circadian variation in pulmonary inflammatory
responses is independent of rhythmic glucocorticoid signaling in airway epithelial cells.
FASEB J. 33, 126–139 (2019).
29. M. Pariollaud, J. E. Gibbs, T. W. Hopwood, S. Brown, N. Begley, R. Vonslow, T. Poolman,
B. Guo, B. Saer, D. H. Jones, J. P. Tellam, S. Bresciani, N. C. Tomkinson, J. Wojno-Picon,
A. W. Cooper, D. A. Daniels, R. P. Trump, D. Grant, W. Zuercher, T. M. Willson,
A. S. MacDonald, B. Bolognese, P. L. Podolin, Y. Sanchez, A. S. Loudon, D. W. Ray, Circadian
clock component REV-ERB controls homeostatic regulation of pulmonary inflammation.
J. Clin. Invest. 128, 2281–2296 (2018).
30. G. B. Kitchen, P. S. Cunningham, T. M. Poolman, M. Iqbal, R. Maidstone, M. Baxter,
J. Bagnall, N. Begley, B. Saer, T. Hussell, L. C. Matthews, D. H. Dockrell, H. J. Durrington,
J. E. Gibbs, J. F. Blaikley, A. S. Loudon, D. W. Ray, The clock gene Bmal1 inhibits
macrophage motility, phagocytosis, and impairs defense against pneumonia. Proc. Natl.
Acad. Sci. U.S.A. 117, 1543–1551 (2020).
31. D. Guan, M. A. Lazar, Interconnections between circadian clocks and metabolism. J. Clin.
Invest. 131, e148278 (2021).
32. R. K. Alexander, Y. H. Liou, N. H. Knudsen, K. A. Starost, C. Xu, A. L. Hyde, S. Liu, D. Jacobi,
N. S. Liao, C. H. Lee, Bmal1 integrates mitochondrial metabolism and macrophage
activation. eLife 9, e54090 (2020).
33. E. J. Collins, M. P. Cervantes-Silva, G. A. Timmons, J. R. O'Siorain, A. M. Curtis, J. M. Hurley,
Post-transcriptional circadian regulation in macrophages organizes temporally distinct
immunometabolic states. Genome Res. 31, 171–185 (2021).
34. D. Yu, X. Fang, Y. Xu, H. Xiao, T. Huang, Y. Zhang, Y. Ge, Y. Li, L. Zong, J. Gao, Rev-erb can
regulate the NF-B/NALP3 pathway to modulate lipopolysaccharide-induced acute lung
injury and inflammation. Int. Immunopharmacol. 73, 312–320 (2019).
35. B. Pourcet, M. Zecchin, L. Ferri, J. Beauchamp, S. Sitaula, C. Billon, S. Delhaye, J. Vanhoutte,
A. Mayeuf-Louchart, Q. Thorel, J. T. Haas, J. Eeckhoute, D. Dombrowicz, C. Duhem,
A. Boulinguiez, S. Lancel, Y. Sebti, T. P. Burris, B. Staels, H. M. Duez, Nuclear receptor
subfamily 1 group d member 1 regulates circadian activity of NLRP3 inflammasome to reduce
the severity of fulminant hepatitis in mice. Gastroenterology 154, 1449–1464.e20 (2018).
36. B. Pourcet, H. Duez, Circadian control of inflammasome pathways: Implications
for circadian medicine. Front. Immunol. 11, 1630 (2020).
37. J. O. Early, D. Menon, C. A. Wyse, M. P. Cervantes-Silva, Z. Zaslona, R. G. Carroll,
E. M. Palsson-McDermott, S. Angiari, D. G. Ryan, S. E. Corcoran, G. Timmons, S. S. Geiger,
D. J. Fitzpatrick, D. O'Connell, R. J. Xavier, K. Hokamp, L. A. J. O'Neill, A. M. Curtis, Circadian
clock protein BMAL1 regulates IL-1 in macrophages via NRF2. Proc. Natl. Acad. Sci. U.S.A.
115, E8460–E8468 (2018).
38. S. Wang, Y. Lin, X. Yuan, F. Li, L. Guo, B. Wu, REV-ERB integrates colon clock with
experimental colitis through regulation of NF-B/NLRP3 axis. Nat. Commun. 9, 4246 (2018).
39. S. Wang, Y. Lin, F. Li, Z. Qin, Z. Zhou, L. Gao, Z. Yang, Z. Wang, B. Wu, An NF-B-driven
lncRNA orchestrates colitis and circadian clock. Sci. Adv. 6, eabb5202 (2020).
40. J. E. Gibbs, J. Blaikley, S. Beesley, L. Matthews, K. D. Simpson, S. H. Boyce, S. N. Farrow,
K. J. Else, D. Singh, D. W. Ray, A. S. Loudon, The nuclear receptor REV-ERB mediates
circadian regulation of innate immunity through selective regulation of inflammatory
cytokines. Proc. Natl. Acad. Sci. U.S.A. 109, 582–587 (2012).
13 of 15
SCIENCE IMMUNOLOGY | REVIEW
41. S. Sato, T. Sakurai, J. Ogasawara, M. Takahashi, T. Izawa, K. Imaizumi, N. Taniguchi, H. Ohno,
T. Kizaki, A circadian clock gene, Rev-erb, modulates the inflammatory function of macrophages
through the negative regulation of Ccl2 expression. J. Immunol. 192, 407–417 (2014).
42. P. Griffin, J. M. Dimitry, P. W. Sheehan, B. V. Lananna, C. Guo, M. L. Robinette, M. E. Hayes,
M. R. Cedeno, C. J. Nadarajah, L. A. Ezerskiy, M. Colonna, J. Zhang, A. Q. Bauer, T. P. Burris,
E. S. Musiek, Circadian clock protein Rev-erb regulates neuroinflammation. Proc. Natl.
Acad. Sci. U.S.A. 116, 5102–5107 (2019).
43. K. D. Nguyen, S. J. Fentress, Y. Qiu, K. Yun, J. S. Cox, A. Chawla, Circadian gene Bmal1 regulates
diurnal oscillations of Ly6C(hi) inflammatory monocytes. Science 341, 1483–1488 (2013).
44. J. M. Adrover, C. Del Fresno, G. Crainiciuc, M. I. Cuartero, M. Casanova-Acebes, L. A. Weiss,
H. Huerga-Encabo, C. Silvestre-Roig, J. Rossaint, I. Cossio, A. V. Lechuga-Vieco,
J. Garcia-Prieto, M. Gomez-Parrizas, J. A. Quintana, I. Ballesteros, S. Martin-Salamanca,
A. Aroca-Crevillen, S. Z. Chong, M. Evrard, K. Balabanian, J. Lopez, K. Bidzhekov, F. Bachelerie,
F. Abad-Santos, C. Munoz-Calleja, A. Zarbock, O. Soehnlein, C. Weber, L. G. Ng,
C. Lopez-Rodriguez, D. Sancho, M. A. Moro, B. Ibanez, A. Hidalgo, A neutrophil timer
coordinates immune defense and vascular protection. Immunity 50, 390–402.e10 (2019).
45. M. Casanova-Acebes, C. Pitaval, L. A. Weiss, C. Nombela-Arrieta, R. Chevre, N. A-González,
Y. Kunisaki, D. Zhang, N. van Rooijen, L. E. Silberstein, C. Weber, T. Nagasawa,
P. S. Frenette, A. Castrillo, A. Hidalgo, Rhythmic modulation of the hematopoietic niche
through neutrophil clearance. Cell 153, 1025–1035 (2013).
46. M. Crespo, B. Gonzalez-Teran, I. Nikolic, A. Mora, C. Folgueira, E. Rodriguez, L. Leiva-Vega,
A. Pintor-Chocano, M. Fernandez-Chacon, I. Ruiz-Garrido, B. Cicuendez, A. Tomas-Loba,
N. A-González, A. Caballero-Molano, D. Beiroa, L. Hernandez-Cosido, J. L. Torres,
N. J. Kennedy, R. J. Davis, R. Benedito, M. Marcos, R. Nogueiras, A. Hidalgo, N. Matesanz,
M. Leiva, G. Sabio, Neutrophil infiltration regulates clock-gene expression to organize
daily hepatic metabolism. Elife 9, e59258 (2020).
47. M. Casanova-Acebes, J. A. Nicolas-Avila, J. L. Li, S. Garcia-Silva, A. Balachander,
A. Rubio-Ponce, L. A. Weiss, J. M. Adrover, K. Burrows, N. A-González, I. Ballesteros, S. Devi,
J. A. Quintana, G. Crainiciuc, M. Leiva, M. Gunzer, C. Weber, T. Nagasawa, O. Soehnlein,
M. Merad, A. Mortha, L. G. Ng, H. Peinado, A. Hidalgo, Neutrophils instruct homeostatic
and pathological states in naive tissues. J. Exp. Med. 215, 2778–2795 (2018).
48. S. J. Holtkamp, L. M. Ince, C. Barnoud, M. T. Schmitt, F. Sinturel, V. Pilorz, R. Pick, S. Jemelin,
M. Muhlstadt, W. H. Boehncke, J. Weber, D. Laubender, J. Philippou-Massier, C. S. Chen,
L. Holtermann, D. Vestweber, M. Sperandio, B. U. Schraml, C. Halin, C. Dibner, H. Oster,
J. Renkawitz, C. Scheiermann, Circadian clocks guide dendritic cells into skin lymphatics.
Nat. Immunol. 22, 1375–1381 (2021).
49. C. C. Nobis, G. Dubeau Laramee, L. Kervezee, D. Maurice De Sousa, N. Labrecque,
N. Cermakian, The circadian clock of CD8 T cells modulates their early response
to vaccination and the rhythmicity of related signaling pathways. Proc. Natl. Acad. Sci.
U.S.A. 116, 20077–20086 (2019).
50. M. Amir, S. Campbell, T. M. Kamenecka, L. A. Solt, Pharmacological modulation
and genetic deletion of REV-ERB and REV-ERB regulates dendritic cell development.
Biochem. Biophys. Res. Commun. 527, 1000–1007 (2020).
51. A. Nakai, K. Suzuki, Adrenergic control of lymphocyte trafficking and adaptive immune
responses. Neurochem. Int. 130, 104320 (2019).
52. C. Scheiermann, Y. Kunisaki, P. S. Frenette, Circadian control of the immune system.
Nat. Rev. Immunol. 13, 190–198 (2013).
53. A. Shimba, K. Ikuta, Glucocorticoids regulate circadian rhythm of innate and adaptive
immunity. Front. Immunol. 11, 2143 (2020).
54. N. Labrecque, N. Cermakian, Circadian clocks in the immune system. J. Biol. Rhythms 30,
277–290 (2015).
55. N. Cermakian, S. K. Stegeman, K. Tekade, N. Labrecque, Circadian rhythms in adaptive
immunity and vaccination. Semin. Immunopathol. 44, 193–207 (2021).
56. S. Hemmers, A. Y. Rudensky, The cell-intrinsic circadian clock is dispensable
for lymphocyte differentiation and function. Cell Rep. 11, 1339–1349 (2015).
57. X. Yu, D. Rollins, K. A. Ruhn, J. J. Stubblefield, C. B. Green, M. Kashiwada, P. B. Rothman,
J. S. Takahashi, L. V. Hooper, TH17 cell differentiation is regulated by the circadian clock.
Science 342, 727–730 (2013).
58. C. Chang, C. S. Loo, X. Zhao, L. A. Solt, Y. Liang, S. P. Bapat, H. Cho, T. M. Kamenecka,
M. Leblanc, A. R. Atkins, R. T. Yu, M. Downes, T. P. Burris, R. M. Evans, Y. Zheng, The nuclear
receptor REV-ERB modulates Th17 cell-mediated autoimmune disease. Proc. Natl. Acad.
Sci. U.S.A. 116, 18528–18536 (2019).
59. M. Amir, S. Chaudhari, R. Wang, S. Campbell, S. A. Mosure, L. B. Chopp, Q. Lu, J. Shang,
O. B. Pelletier, Y. He, C. Doebelin, M. D. Cameron, D. J. Kojetin, T. M. Kamenecka, L. A. Solt,
REV-ERBalpha regulates TH17 cell development and autoimmunity. Cell Rep. 25,
3733–3749.e8 (2018).
60. M. Ciofani, A. Madar, C. Galan, M. Sellars, K. Mace, F. Pauli, A. Agarwal, W. Huang,
C. N. Parkhurst, M. Muratet, K. M. Newberry, S. Meadows, A. Greenfield, Y. Yang, P. Jain,
F. K. Kirigin, C. Birchmeier, E. F. Wagner, K. M. Murphy, R. M. Myers, R. Bonneau,
D. R. Littman, A validated regulatory network for Th17 cell specification. Cell 151,
289–303 (2012).
Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022
61. J. R. Huh, M. W. Leung, P. Huang, D. A. Ryan, M. R. Krout, R. R. Malapaka, J. Chow, N. Manel,
M. Ciofani, S. V. Kim, A. Cuesta, F. R. Santori, J. J. Lafaille, H. E. Xu, D. Y. Gin, F. Rastinejad,
D. R. Littman, Digoxin and its derivatives suppress TH17 cell differentiation by
antagonizing RORt activity. Nature 472, 486–490 (2011).
62. J.-Y. Lee, J. A. Hall, M. Pokrovskii, L. Kroehling, L. Wu, D. R. Littman, ROR enforces stability
of the T-helper-17 cell effector program. bioRxiv 2020.12.15.422921 [Preprint].
22 December 2020. https://doi.org/10.1101/2020.12.15.422921.
63. X. O. Yang, B. P. Pappu, R. Nurieva, A. Akimzhanov, H. S. Kang, Y. Chung, L. Ma, B. Shah,
A. D. Panopoulos, K. S. Schluns, S. S. Watowich, Q. Tian, A. M. Jetten, C. Dong, T helper 17
lineage differentiation is programmed by orphan nuclear receptors ROR alpha and ROR
gamma. Immunity 28, 29–39 (2008).
64. Y. Sun, Z. Yang, Z. Niu, J. Peng, Q. Li, W. Xiong, A. N. Langnas, M. Y. Ma, Y. Zhao, MOP3,
a component of the molecular clock, regulates the development of B cells. Immunology
119, 451–460 (2006).
65. Q. Cao, X. Zhao, J. Bai, S. Gery, H. Sun, D. C. Lin, Q. Chen, Z. Chen, L. Mack, H. Yang,
R. Deng, X. Shi, L. W. Chong, H. Cho, J. Xie, Q. Z. Li, M. Muschen, A. R. Atkins, C. Liddle,
R. T. Yu, S. Alkan, J. W. Said, Y. Zheng, M. Downes, R. M. Evans, H. P. Koeffler, Circadian
clock cryptochrome proteins regulate autoimmunity. Proc. Natl. Acad. Sci. U.S.A. 114,
12548–12553 (2017).
66. D. Druzd, O. Matveeva, L. Ince, U. Harrison, W. He, C. Schmal, H. Herzel, A. H. Tsang,
Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023
N. Kawakami, A. Leliavski, O. Uhl, L. Yao, L. E. Sander, C. S. Chen, K. Kraus, A. de Juan,
S. M. Hergenhan, M. Ehlers, B. Koletzko, R. Haas, W. Solbach, H. Oster, C. Scheiermann,
Lymphocyte circadian clocks control lymph node trafficking and adaptive immune
responses. Immunity 46, 120–132 (2017).
67. E. Haus, M. H. Smolensky, Biologic rhythms in the immune system. Chronobiol. Int. 16,
581–622 (1999).
68. A. Nakai, Y. Hayano, F. Furuta, M. Noda, K. Suzuki, Control of lymphocyte egress from
lymph nodes through 2-adrenergic receptors. J. Exp. Med. 211, 2583–2598 (2014).
69. A. Shimba, G. Cui, S. Tani-Ichi, M. Ogawa, S. Abe, F. Okazaki, S. Kitano, H. Miyachi,
H. Yamada, T. Hara, Y. Yoshikai, T. Nagasawa, G. Schutz, K. Ikuta, Glucocorticoids drive
diurnal oscillations in T cell distribution and responses by inducing interleukin-7 receptor
and CXCR4. Immunity 48, 286–298.e6 (2018).
70. K. Suzuki, Y. Hayano, A. Nakai, F. Furuta, M. Noda, Adrenergic control of the adaptive
immune response by diurnal lymphocyte recirculation through lymph nodes. J. Exp. Med.
213, 2567–2574 (2016).
71. S. D. House, S. Ruch, W. F. Koscienski III, C. W. Rocholl, R. L. Moldow, Effects
of the circadian rhythm of corticosteroids on leukocyte-endothelium interactions
in the AM and PM. Life Sci. 60, 2023–2034 (1997).
72. C. Scheiermann, Y. Kunisaki, D. Lucas, A. Chow, J. E. Jang, D. Zhang, D. Hashimoto,
M. Merad, P. S. Frenette, Adrenergic nerves govern circadian leukocyte recruitment
to tissues. Immunity 37, 290–301 (2012).
73. A. C. Silver, A. Arjona, W. E. Walker, E. Fikrig, The circadian clock controls toll-like receptor
9-mediated innate and adaptive immunity. Immunity 36, 251–261 (2012).
74. C. E. Sutton, C. M. Finlay, M. Raverdeau, J. O. Early, J. DeCourcey, Z. Zaslona, L. A. J. O'Neill,
K. H. G. Mills, A. M. Curtis, Loss of the molecular clock in myeloid cells exacerbates
T cell-mediated CNS autoimmune disease. Nat. Commun. 8, 1923 (2017).
75. N. Ando, Y. Nakamura, R. Aoki, K. Ishimaru, H. Ogawa, K. Okumura, S. Shibata, S. Shimada,
A. Nakao, Circadian gene clock regulates psoriasis-like skin inflammation in mice.
J. Invest. Dermatol. 135, 3001–3008 (2015).
76. A. Chiricozzi, R. Saraceno, M. S. Chimenti, E. Guttman-Yassky, J. G. Krueger, Role of IL-23
in the pathogenesis of psoriasis: A novel potential therapeutic target? Expert Opin. Ther.
Targets 18, 513–525 (2014).
77. S. Wang, M. Kozai, H. Mita, Z. Cai, M. A. Masum, O. Ichii, K. Takada, M. Inaba, REV-ERB
agonist suppresses IL-17 production in T cells and improves psoriatic dermatitis
in a mouse model. Biomed. Pharmacother. 144, 112283 (2021).
78. A. J. Davidson, M. T. Sellix, J. Daniel, S. Yamazaki, M. Menaker, G. D. Block, Chronic jet-lag
increases mortality in aged mice. Curr. Biol. 16, R914–R916 (2006).
79. H. Inokawa, Y. Umemura, A. Shimba, E. Kawakami, N. Koike, Y. Tsuchiya, M. Ohashi,
Y. Minami, G. Cui, T. Asahi, R. Ono, Y. Sasawaki, E. Konishi, S. H. Yoo, Z. Chen, S. Teramukai,
K. Ikuta, K. Yagita, Chronic circadian misalignment accelerates immune senescence
and abbreviates lifespan in mice. Sci. Rep. 10, 2569 (2020).
80. Y. Wu, B. Tao, T. Zhang, Y. Fan, R. Mao, Pan-cancer analysis reveals disrupted circadian
clock associates with T cell exhaustion. Front. Immunol. 10, 2451 (2019).
81. S. Leach, K. Suzuki, Adrenergic signaling in circadian control of immunity. Front. Immunol.
11, 1235 (2020).
82. S. Mendez-Ferrer, D. Lucas, M. Battista, P. S. Frenette, Haematopoietic stem cell release is
regulated by circadian oscillations. Nature 452, 442–447 (2008).
83. K. Golan, A. Kumari, O. Kollet, E. Khatib-Massalha, M. D. Subramaniam, Z. S. Ferreira,
F. Avemaria, S. Rzeszotek, A. Garcia-Garcia, S. Xie, E. Flores-Figueroa, S. Gur-Cohen,
T. Itkin, A. Ludin-Tal, H. Massalha, B. Bernshtein, A. K. Ciechanowicz, A. Brandis,
T. Mehlman, S. Bhattacharya, M. Bertagna, H. Cheng, E. Petrovich-Kopitman, T. Janus,
14 of 15
SCIENCE IMMUNOLOGY | REVIEW
N. Kaushansky, T. Cheng, I. Sagi, M. Z. Ratajczak, S. Mendez-Ferrer, J. E. Dick, R. P. Markus,
T. Lapidot, Daily onset of light and darkness differentially controls hematopoietic stem
cell differentiation and maintenance. Cell Stem Cell 23, 572–585.e7 (2018).
84. A. de Juan, L. M. Ince, R. Pick, C. S. Chen, F. Molica, G. Zuchtriegel, C. Wang, D. Zhang,
D. Druzd, M. E. T. Hessenauer, G. Pelli, I. Kolbe, H. Oster, C. Prophete, S. M. Hergenhan,
U. Albrecht, J. Ripperger, E. Montanez, C. A. Reichel, O. Soehnlein, B. R. Kwak,
P. S. Frenette, C. Scheiermann, Artery-associated sympathetic innervation drives
rhythmic vascular inflammation of arteries and veins. Circulation 140, 1100–1114 (2019).
85. A. Garcia-Garcia, C. Korn, M. Garcia-Fernandez, O. Domingues, J. Villadiego, D. Martin-Perez,
J. Isern, J. A. Bejarano-Garcia, J. Zimmer, J. A. Perez-Simon, J. J. Toledo-Aral, T. Michel,
M. S. Airaksinen, S. Mendez-Ferrer, Dual cholinergic signals regulate daily migration
of hematopoietic stem cells and leukocytes. Blood 133, 224–236 (2019).
86. R. Dumbell, O. Matveeva, H. Oster, Circadian clocks, stress, and immunity. Front.
Endocrinol. (Lausanne) 7, 37 (2016).
87. L. E. Hand, K. J. Gray, S. H. Dickson, D. A. Simpkins, D. W. Ray, J. E. Konkel, M. R. Hepworth,
J. E. Gibbs, Regulatory T cells confer a circadian signature on inflammatory arthritis.
Nat. Commun. 11, 1658 (2020).
88. D. Zheng, K. Ratiner, E. Elinav, Circadian influences of diet on the microbiome
and immunity. Trends Immunol. 41, 512–530 (2020).
89. J. F. Brooks II, C. L. Behrendt, K. A. Ruhn, S. Lee, P. Raj, J. S. Takahashi, L. V. Hooper, The
microbiota coordinates diurnal rhythms in innate immunity with the circadian clock. Cell
184, 4154–4167.e12 (2021).
90. C. Seillet, K. Luong, J. Tellier, N. Jacquelot, R. D. Shen, P. Hickey, V. C. Wimmer,
L. Whitehead, K. Rogers, G. K. Smyth, A. L. Garnham, M. E. Ritchie, G. T. Belz, The
neuropeptide VIP confers anticipatory mucosal immunity by regulating ILC3 activity. Nat.
Immunol. 21, 168–177 (2020).
91. J. Talbot, P. Hahn, L. Kroehling, H. Nguyen, D. Li, D. R. Littman, Feeding-dependent VIP
neuron-ILC3 circuit regulates the intestinal barrier. Nature 579, 575–580 (2020).
92. T. Tuganbaev, U. Mor, S. Bashiardes, T. Liwinski, S. P. Nobs, A. Leshem, M. Dori-Bachash,
C. A. Thaiss, E. Y. Pinker, K. Ratiner, L. Adlung, S. Federici, C. Kleimeyer, C. Moresi,
T. Yamada, Y. Cohen, X. Zhang, H. Massalha, E. Massasa, Y. Kuperman, P. A. Koni,
A. Harmelin, N. Gao, S. Itzkovitz, K. Honda, H. Shapiro, E. Elinav, Diet diurnally regulates
small intestinal microbiome-epithelial-immune homeostasis and enteritis. Cell 182,
1441–1459.e21 (2020).
93. C. A. Thaiss, M. Levy, T. Korem, L. Dohnalova, H. Shapiro, D. A. Jaitin, E. David, D. R. Winter,
M. Gury-BenAri, E. Tatirovsky, T. Tuganbaev, S. Federici, N. Zmora, D. Zeevi, M. Dori-Bachash,
M. Pevsner-Fischer, E. Kartvelishvily, A. Brandis, A. Harmelin, O. Shibolet, Z. Halpern,
K. Honda, I. Amit, E. Segal, E. Elinav, Microbiota diurnal rhythmicity programs host
transcriptome oscillations. Cell 167, 1495–1510.e12 (2016).
94. M. M. Bellet, E. Deriu, J. Z. Liu, B. Grimaldi, C. Blaschitz, M. Zeller, R. A. Edwards, S. Sahar,
S. Dandekar, P. Baldi, M. D. George, M. Raffatellu, P. Sassone-Corsi, Circadian clock regulates
the host response to Salmonella. Proc. Natl. Acad. Sci. U.S.A. 110, 9897–9902 (2013).
95. C. Godinho-Silva, R. G. Domingues, M. Rendas, B. Raposo, H. Ribeiro, J. A. da Silva,
A. Vieira, R. M. Costa, N. L. Barbosa-Morais, T. Carvalho, H. Veiga-Fernandes, Light-
entrained and brain-tuned circadian circuits regulate ILC3s and gut homeostasis. Nature
574, 254–258 (2019).
96. F. Teng, J. Goc, L. Zhou, C. Chu, M. A. Shah, G. Eberl, G. F. Sonnenberg, A circadian clock is
essential for homeostasis of group 3 innate lymphoid cells in the gut. Sci. Immunol. 4,
eaax1215 (2019).
97. Q. Wang, M. L. Robinette, C. Billon, P. L. Collins, J. K. Bando, J. L. Fachi, C. Secca, S. I. Porter,
A. Saini, S. Gilfillan, L. A. Solt, E. S. Musiek, E. M. Oltz, T. P. Burris, M. Colonna, Circadian
rhythm–dependent and circadian rhythm–independent impacts of the molecular clock
on type 3 innate lymphoid cells. Sci. Immunol. 4, eaay7501 (2019).
98. Q. Wang, M. Colonna, Keeping time in group 3 innate lymphoid cells. Nat. Rev. Immunol.
20, 720–726 (2020).
99. C. Seillet, L. C. Rankin, J. R. Groom, L. A. Mielke, J. Tellier, M. Chopin, N. D. Huntington,
G. T. Belz, S. Carotta, Nfil3 is required for the development of all innate lymphoid cell
subsets. J. Exp. Med. 211, 1733–1740 (2014).
100. T. L. Geiger, M. C. Abt, G. Gasteiger, M. A. Firth, M. H. O'Connor, C. D. Geary, T. E. O'Sullivan,
M. R. van den Brink, E. G. Pamer, A. M. Hanash, J. C. Sun, Nfil3 is crucial for development
of innate lymphoid cells and host protection against intestinal pathogens. J. Exp. Med.
211, 1723–1731 (2014).
101. W. Xu, R. G. Domingues, D. Fonseca-Pereira, M. Ferreira, H. Ribeiro, S. Lopez-Lastra,
Y. Motomura, L. Moreira-Santos, F. Bihl, V. Braud, B. Kee, H. Brady, M. C. Coles,
C. Vosshenrich, M. Kubo, J. P. Di Santo, H. Veiga-Fernandes, NFIL3 orchestrates
the emergence of common helper innate lymphoid cell precursors. Cell Rep. 10,
2043–2054 (2015).
102. A. B. Diallo, L. Gay, B. Coiffard, M. Leone, S. Mezouar, J. L. Mege, Daytime variation
in SARS-CoV-2 infection and cytokine production. Microb. Pathog. 158, 105067 (2021).
103. X. Zhuang, S. Tsukuda, F. Wrensch, P. A. C. Wing, M. Schilling, J. M. Harris, H. Borrmann,
S. B. Morgan, J. L. Cane, L. Mailly, N. Thakur, C. Conceicao, H. Sanghani, L. Heydmann,
Wang et al., Sci. Immunol. 7, eabm2465 (2022) 3 June 2022
C. Bach, A. Ashton, S. Walsh, T. K. Tan, L. Schimanski, K. A. Huang, C. Schuster, K. Watashi,
T. S. C. Hinks, A. Jagannath, S. R. Vausdevan, D. Bailey, T. F. Baumert, J. A. McKeating, The
circadian clock component BMAL1 regulates SARS-CoV-2 entry and replication in lung
epithelial cells. iScience 24, 103144 (2021).
104. R. S. Edgar, A. Stangherlin, A. D. Nagy, M. P. Nicoll, S. Efstathiou, J. S. O'Neill, A. B. Reddy,
Cell autonomous regulation of herpes and influenza virus infection by the circadian
clock. Proc. Natl. Acad. Sci. U.S.A. 113, 10085–10090 (2016).
105. T. Majumdar, J. Dhar, S. Patel, R. Kondratov, S. Barik, Circadian transcription factor
BMAL1 regulates innate immunity against select RNA viruses. Innate Immun. 23,
147–154 (2017).
106. A. Ehlers, W. Xie, E. Agapov, S. Brown, D. Steinberg, R. Tidwell, G. Sajol, R. Schutz,
R. Weaver, H. Yu, M. Castro, L. B. Bacharier, X. Wang, M. J. Holtzman, J. A. Haspel, BMAL1
links the circadian clock to viral airway pathology and asthma phenotypes. Mucosal
Immunol. 11, 97–111 (2018).
107. X. Zhuang, D. Forde, S. Tsukuda, V. D'Arienzo, L. Mailly, J. M. Harris, P. A. C. Wing,
H. Borrmann, M. Schilling, A. Magri, C. O. Rubio, R. J. Maidstone, M. Iqbal, M. Garzon,
R. Minisini, M. Pirisi, S. Butterworth, P. Balfe, D. W. Ray, K. Watashi, T. F. Baumert,
J. A. McKeating, Circadian control of hepatitis B virus replication. Nat. Commun. 12, 1658
(2021).
Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023
108. H. Borrmann, R. Davies, M. Dickinson, I. Pedroza-Pacheco, M. Schilling, A. Vaughan-Jackson,
A. Magri, W. James, P. Balfe, P. Borrow, J. A. McKeating, X. Zhuang, Pharmacological
activation of the circadian component REV-ERB inhibits HIV-1 replication. Sci. Rep. 10,
13271 (2020).
109. S. Sengupta, S. Y. Tang, J. C. Devine, S. T. Anderson, S. Nayak, S. L. Zhang, A. Valenzuela,
D. G. Fisher, G. R. Grant, C. B. Lopez, G. A. FitzGerald, Circadian control of lung
inflammation in influenza infection. Nat. Commun. 10, 4107 (2019).
110. S. R. Lundy, T. Ahmad, T. Simoneaux, I. Benyeogor, Y. Robinson, Z. George, D. Ellerson,
W. Kirlin, T. Omosun, F. O. Eko, C. M. Black, U. Blas-Machado, J. P. DeBruyne,
J. U. Igietseme, Q. He, Y. O. Omosun, Effect of time of day of infection on chlamydia
infectivity and pathogenesis. Sci. Rep. 9, 11405 (2019).
111. T. W. Hopwood, S. Hall, N. Begley, R. Forman, S. Brown, R. Vonslow, B. Saer, M. C. Little,
E. A. Murphy, R. J. Hurst, D. W. Ray, A. S. MacDonald, A. Brass, D. A. Bechtold, J. E. Gibbs,
A. S. Loudon, K. J. Else, The circadian regulator BMAL1 programmes responses to parasitic
worm infection via a dendritic cell clock. Sci. Rep. 8, 3782 (2018).
112. S. Kiessling, G. Dubeau-Laramee, H. Ohm, N. Labrecque, M. Olivier, N. Cermakian, The
circadian clock in immune cells controls the magnitude of Leishmania parasite infection.
Sci. Rep. 7, 10892 (2017).
113. J. E. Long, M. T. Drayson, A. E. Taylor, K. M. Toellner, J. M. Lord, A. C. Phillips, Morning
vaccination enhances antibody response over afternoon vaccination: A cluster-
randomised trial. Vaccine 34, 2679–2685 (2016).
114. R. K. Kurupati, A. Kossenkoff, S. Kannan, L. H. Haut, S. Doyle, X. Yin, K. E. Schmader, Q. Liu,
L. Showe, H. C. J. Ertl, The effect of timing of influenza vaccination and sample collection
on antibody titers and responses in the aged. Vaccine 35, 3700–3708 (2017).
115. L. C. J. de Bree, V. P. Mourits, V. A. Koeken, S. J. Moorlag, R. Janssen, L. Folkman,
D. Barreca, T. Krausgruber, V. Fife-Gernedl, B. Novakovic, R. J. Arts, H. Dijkstra,
H. Lemmers, C. Bock, L. A. Joosten, R. van Crevel, C. S. Benn, M. G. Netea, Circadian
rhythm influences induction of trained immunity by BCG vaccination. J. Clin. Invest. 130,
5603–5617 (2020).
116. H. Zhang, Y. Liu, D. Liu, Q. Zeng, L. Li, Q. Zhou, M. Li, J. Mei, N. Yang, S. Mo, Q. Liu, M. Liu,
S. Peng, H. Xiao, Time of day influences immune response to an inactivated vaccine
against SARS-CoV-2. Cell Res. 31, 1215–1217 (2021).
117. C. Barnoud, C. Wang, C. Scheiermann, Timing vaccination against SARS-CoV-2. Cell Res.
31, 1146–1147 (2021).
118. W. Wang, P. Balfe, D. W. Eyre, S. F. Lumley, D. O'Donnell, F. Warren, D. W. Crook, K. Jeffery,
P. C. Matthews, E. B. Klerman, J. A. McKeating, Time of day of vaccination affects
SARS-CoV-2 antibody responses in an observational study of health care workers.
J. Biol. Rhythms 37, 124–129 (2022).
Funding: Research in the laboratory is funded by the European Research Council (ERC CoG
101001233, CIRCADYN), the Swiss National Science Foundation (SNF; 310030_182417/1), the
Swiss Cancer League (KLS-4836-08-2019), the Geneva Cancer league (2106), the Fondation
privée des HUG (RC05-03), EU ITN (813284, INTEGRATA), and the Novartis Foundation for
Medical-Biological Research (20A019); C.W. is the recipient of a fellowship of the Chinese
Scholarship Council (CSC). Competing interests: The authors declare that they have no
competing interests.
Submitted 15 December 2021
Accepted 9 May 2022
Published 3 June 2022
10.1126/sciimmunol.abm2465
15 of 15
 The circadian immune system
Chen Wang, Lydia Kay Lutes, Coline Barnoud, and Christoph Scheiermann

Sci. Immunol., 7 (72), eabm2465.

DOI: 10.1126/sciimmunol.abm2465

Downloaded from https://www.science.org at Universidade de Sao Paulo on May 03, 2023

View the article online
https://www.science.org/doi/10.1126/sciimmunol.abm2465
Permissions
https://www.science.org/help/reprints-and-permissions

Use of this article is subject to the Terms of service

Science Immunology (ISSN ) is published by the American Association for the Advancement of Science. 1200 New York Avenue NW,
Washington, DC 20005. The title Science Immunology is a registered trademark of AAAS.
Copyright © 2022 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim
to original U.S. Government Works