Monocyte Conversion During Inflammation and Injury

Rachel M. Kratofil, Paul Kubes,* Justin F. Deniset*

Abstract—Monocytes are circulating leukocytes important in both innate and adaptive immunity, primarily functioning in
immune defense, inflammation, and tissue remodeling. There are 2 subsets of monocytes in mice (3 subsets in humans)
that are mobilized from the bone marrow and recruited to sites of inflammation, where they carry out their respective
functions in promoting inflammation or facilitating tissue repair. Our understanding of the fate of these monocyte subsets
at the site of inflammation is constantly evolving. This brief review highlights the plasticity of monocyte subsets and
their conversion during inflammation and injury.   (Arterioscler Thromb Vasc Biol. 2017;37:35-42. DOI: 10.1161/
ATVBAHA.116.308198.)
Key Words: infection ◼ inflammation ◼ innate immunity ◼ macrophages ◼ monocytes

Monocyte Populations in Mouse and Human
Monocytes are heterogeneous cells that circulate in the blood
and constitute a critical component of the innate immune
response to inflammation. In mice, 2 populations of mono-
cytes exist and can be discriminated by variable expression of
lymphocyte antigen 6C (Ly6C). Monocytes expressing high
levels of Ly6C (Ly6Chi monocytes) have proinflammatory and
antimicrobial functions and express high levels of C-C che-
mokine receptor 2 (CCR2) and low levels of CX3C chemo-
kine receptor 1 (CX3CR1; CCR2hiCX3CR1low).1 These Ly6Chi
Downloaded from http://ahajournals.org by on May 10, 2019
basis of the cell surface expression and cellular function, inter-
mediate monocytes exhibit both phagocytic function and anti-
inflammatory effects, as well as higher levels of intracellular
IL-1β and tumor necrosis factor (TNF)-α at steady state.7–9
The usage of new approaches such as lineage-tracing and
intravital microscopy has provided new understanding to the
origin, fate, and functions of these monocyte subsets in vivo.
In this brief review, we highlight the role of monocyte con-
version and plasticity in regulating myeloid cell development,
resolution of inflammation, and tissue homeostasis (Figure).

monocytes are able to transport antigens to the lymph node

and accumulate at sites of inflammation, where they can dif-
ferentiate into macrophages or dendritic cells (DCs) depend-
ing on the local cytokine environment.2,3 Ly6Clow monocytes,
also known as patrolling monocytes, survey the vasculature
by constantly crawling along the lumen of the vasculature and
are involved with early responses to inflammation and tissue
repair. These alternative monocytes express high levels of
CX3CR1 and low levels of CCR2 (CX3CR1hiCCR2low).
The dichotomy of monocyte subsets observed in mice is
also present in humans and is classified based on the expres-
sion levels of CD14, a component of the lipopolysaccharide
receptor complex, and CD16, the FCγRIII immunoglobulin
receptor.4 There are 3 subsets of monocytes that have been
described in humans: classical (CD14++CD16−), intermedi-
ate (CD14+CD16+), and nonclassical (CD14+CD16++) mono-
cytes.5–7 Human classical monocytes are analogous to murine
Ly6Chi monocytes, which are CCR2hiCX3CR1low, and human
nonclassical monocytes represent a subset similar to murine
Ly6Clow and CX3CR1hiCCR2low monocytes. The intermediate
monocyte subset in humans has been proposed as a monocyte
in transition from a classical to nonclassical monocyte. On the
Monocyte Development and Mobilization
Classical monocytes are derived from hematopoietic progeni-
tors in the bone marrow, specifically the common monocyte
progenitor, which differentiates from the macrophage DC
precursor.10 The development of blood monocytes from these
progenitors depends on the growth factor, macrophage colony-
stimulating factor (known as M-CSF or CD115).30 In mice
deficient in the colony-stimulating factor 1 receptor, the total
number of circulating monocytes is significantly reduced.31
Numerous transcription factors have been noted to regulate
monocyte development and differentiation, and their contribu-
tions have been extensively reviewed by Zhu et al.32 Importantly,
the C-C chemokine receptor 2 (CCR2) and its ligands, mono-
cyte chemoattractant protein-1 (or chemokine [C-C motif]
ligand [CCL2]) and monocyte chemoattractant protein-3
(CCL7), are required for the egress of classical Ly6Chi mono-
cytes from the bone marrow.11,30 The role of CCR2 in monocyte
egress from the bone marrow was identified in CCR2−/− mice.
These mice had fewer circulating monocytes in the blood and
increased retention in the bone marrow compared with a wild-
type mouse.11 The chemokines responsible for the activation of

Received on: July 19, 2016; final version accepted on: October 10, 2016.
From the Department of Microbiology, Immunology, and Infectious Diseases (R.M.K., P.K.) and Department of Physiology and Pharmacology (P.K.,
J.F.D.), Snyder Institute for Chronic Diseases, Cumming School of Medicine, University of Calgary, Canada.
*These authors are cosenior authors for this article.
Correspondence to Paul Kubes, PhD, Department of Physiology and Pharmacology, Snyder Institute for Chronic Diseases, Cumming School of Medicine,
University of Calgary, Calgary, AB T2N 4N1, Canada. E-mail pkubes@ucalgary.ca
© 2016 American Heart Association, Inc.
Arterioscler Thromb Vasc Biol is available at http://atvb.ahajournals.org DOI: 10.1161/ATVBAHA.116.308198

35
 36   Arterioscler Thromb Vasc Biol   January 2017
Nonstandard Abbreviations and Acronyms
CCL chemokine (C-C motif) ligand
CCR2 C-C chemokine receptor 2
CX3CR1 CX3C chemokine receptor 1
DC dendritic cell
IL interleukin
Ly6C lymphocyte antigen 6C
MI myocardial infarction
Nr4a1 nuclear receptor subfamily 4 group A member 1
TNF tumor necrosis factor
CCR2 and monocyte mobilization from the bone marrow are
CCL2 and CCL7, which can derive from multiple sources.11 A
recent study demonstrated that stromal cells in the bone mar-
row can also contribute to monocyte emigration by producing
CCL2 in response to circulating toll-like receptor ligands.33 In
addition, Zouggari et al34 showed that B cells, through the pro-
duction of CCL7, are able to mobilize monocytes out of the
bone marrow in response to acute myocardial infarction (MI).
In-depth lineage-tracing experiments from Yona et al35
suggest that Ly6Chi monocytes are the precursors to Ly6Clow
monocytes in the blood and in the bone marrow during the
steady state. This concept is further supported by adoptive
transfer experiments, where fluorescently labeled Ly6Chi
monocytes become Ly6Clow monocytes in the blood and bone
marrow of the recipient mouse.15,36 However, there is also evi-
dence supporting the possibility of Ly6Clow monocytes origi-
Downloaded from http://ahajournals.org by on May 10, 2019
nating from an independent precursor. Carlin et al37 have noted
that CCR2−/− mice do not display a reduction in patrolling
Ly6Clow monocytes, despite reduced circulating Ly6Chi mono-
cytes. Furthermore, the disruption of factors that control proin-
flammatory monocyte development only moderately reduces
circulating counts of Ly6Clow monocytes when compared with
Ly6Chi monocytes.38,39 A possible explanation for these find-
ings is that patrolling monocytes can extend their lifespan to
compensate for the reduction in Ly6Chi precursors.35
It is known that the survival of Ly6Clow monocytes in
the bone marrow depends on the orphan nuclear receptor
Nuclear Receptor Subfamily 4 Group A Member 1 (Nr4a1).12
By analyzing the monocyte populations in the bone mar-
row of Nr4a1−/− mice, Ly6Clow monocytes arrested in the S
phase of the cell cycle and underwent apoptosis. Not only is
Nr4a1 essential for Ly6Clow monocyte survival, this transcrip-
tion factor is also involved in the differentiation of Ly6Clow
monocytes from Ly6Chi monocytes in the blood and in the
tissues. Adoptive transfer studies by Hanna et al12 showed
that Nr4a1−/− Ly6Chi monocytes do not convert to Ly6Clow
monocytes in the bone marrow, blood, and tissues. In addi-
tion, other groups have confirmed that Ly6Chi monocytes do
not convert in peripheral tissues of mice that received Nr4a1−/−
bone marrow.15,37 Nr4a1 is important for controlling the fate of
alternative Ly6Clow monocytes, and CX3CR1 also contributes
to the survival of these patrolling monocytes.12 However, it is
likely that in addition to Nr4a1 and CX3CR1 there are addi-
tional factors that drive the differentiation and survival of this
monocyte subset.
Monocyte Recruitment During Inflammation
The recruitment of monocytes to sites of inflammation is
critical for host defense. During inflammation, monocytes
circulate through the blood and extravasate into inflamed tis-
sues after the general paradigm of the leukocyte recruitment
cascade, involving rolling, adhesion, and transmigration.40
Whether it be an infection or sterile injury, monocyte recruit-
ment is a key feature.
Proinflammatory monocyte recruitment under various
inflammatory conditions has been predominantly dependent
on CCR2. However, as mentioned previously, CCR2 and
monocyte chemoattractant protein-1/monocyte chemoat-
tractant protein-3 are critical for the mobilization of mono-
cytes from the bone marrow into the bloodstream, as well as
maintaining blood monocyte homeostasis.11,41 Some studies
have attempted to study these events independently. Adoptive
transfer of monocytes from wild-type and CCR2−/− mice can
bypass efflux from the bone marrow and focus on the role of
monocyte recruitment from the blood into tissues. Serbina and
Pamer41 examined the recruitment of monocytes during infec-
tion with L. monocytogenes and in response to thioglycollate-
induced peritonitis, and they found that adoptively transferred
monocytes from CCR2−/− mice were still able to migrate from
the blood into the spleen. Alternatively, Tsou et al11 reported
that CCR2−/− mice did have a recruitment defect during thio-
glycollate-induced peritonitis. These authors argued that the
discrepancy among studies was that CCR2 was important
for early recruitment (24 hours) but not late recruitment (60
hours), as studied by Serbina and Pamer.41
In addition to the bone marrow, there exists a reservoir of
splenic monocytes within the subcapsular red pulp that can be
recruited to sites of inflammation.42 siRNA-mediated knock-
down of CCR2 in splenic Ly6Chi monocytes prevented their
recruitment to sites of inflammation.43 This study highlights
the requirement of CCR2 for recruitment to a site of injury
independently of bone marrow egress.
In some inflammatory diseases, monocytes exhibit differ-
ent recruitment mechanisms that can involve other chemokine
receptors and CCR2. In atherosclerosis, both inflammatory
and patrolling monocytes are involved in the disease progres-
sion.44,45 Ly6Chi monocytes were found to accumulate within
atherosclerotic plaques more efficiently than Ly6Clow mono-
cytes. Recruitment of Ly6Clow monocytes into plaques was
not dependent on CX3CR1 but partially dependent on CCR5.
Interestingly, Ly6Chi monocytes required CX3CR1 and
CCR2 and CCR5 to infiltrate into plaques.44 siRNA knock-
down of CCR2 reduced inflammatory monocyte recruitment
to atherosclerotic plaques and attenuated its disease progres-
sion in mice.43
In contrast to classical monocytes that flow in the circula-
tion and adhere at sites of endothelial activation, patrolling
monocytes are in constant contact with the endothelium under
steady-state conditions. Adhesion of these patrolling mono-
cytes to the endothelium depended on the β2 integrin LFA-1
(lymphocyte function-associated antigen-1; CD11a/CD18)
but not Mac-1 (macrophage-1 antigen; CD11b/CD18).13 Their
patrolling behavior was argued to be because of low expres-
sion of these adhesion molecules.13 Because integrins are
 Kratofil et al   Monocyte Fate and Function In Vivo   37
Downloaded from http://ahajournals.org by on May 10, 2019

Figure. Monocyte fate during sterile injury, infection, and their contribution to the replenishment of tissue-resident macrophage popula-
tions in vivo. In the bone marrow, proinflammatory monocytes are derived from the common monocyte progenitor (cMoP) that differ-
entiates from the macrophage dendritic cell precursor (MDP).10 Proinflammatory monocytes emigrate out of the bone marrow in a C-C
chemokine receptor 2 (CCR2)–dependent manner.11 The development of Ly6Clow patrolling monocytes from proinflammatory monocytes
in the bone marrow and blood depends on the transcription factor Nr4a1.12 In the blood, Ly6Clow patrolling monocytes are in constant
contact with the endothelium to provide immune surveillance to surrounding tissues.13 CCR2-dependent recruitment of proinflamma-
tory monocytes to inflammatory sites is critical for the resolution of inflammation. During sterile injury, proinflammatory monocytes can
differentiate into a Ly6Clow patrolling/repair monocyte14 or Ly6Clow alternative macrophage.15 Infection with Listeria monocytogenes or
Toxplasma gondii induces monocyte conversion from proinflammatory monocytes to tumor necrosis factor (TNF)-α/inducible nitric oxide
synthase–producing dendritic cells (TipDCs), which enhance bacterial clearance.16,17 The conversion of Ly6Chi monocytes to Ly6Clow mac-
rophages during a S. aureus skin infection is thought to occur in vivo.18,19 In addition to self-proliferation of embryonically derived macro-
phages, proinflammatory monocytes also contribute to the repopulation of resident macrophages in various tissues in the steady state
and after inflammation.20–29

found in different affinity states, this suggests that patrolling
could reflect an intermediate affinity state, whereas high affin-
ity would mediate firm adhesion. Furthermore, the CX3CR1
ligand fractalkine (CX3CL1) is expressed on endothelial cells
and contributes to the adhesion of patrolling monocytes to the
endothelium.13 Patrolling monocytes within the vasculature
provided immune surveillance to the surrounding tissues, as
these monocytes were able to extravasate rapidly into tissues
after the sterile inflammation, addition of irritants, or infection
with Listeria monocytogenes (L. monocytogenes).13
The role of Ly6Clow Nr4a1-dependent monocytes has
been further characterized by Carlin et al37 as accessory
cells of the endothelium, where they are able to respond to
a local toll-like receptor7 danger signal and orchestrate the
focal necrosis of endothelial cells by recruiting neutrophils.
Intravital microscopy revealed that these patrolling monocytes
are enriched in the skin and kidney microvasculature in the
steady state, and during toll-like receptor7–mediated inflam-
mation, these monocytes are retained within the kidney capil-
laries.37 Importantly, this intravascular retention of patrolling
monocytes during inflammation was independent of CCR2,
suggesting a specialized function for the Ly6Clow monocyte
subset. Recently, Hanna et al46 demonstrated that patrolling
monocytes are enriched in the lung microvasculature and are
able to reduce tumor metastasis to the lung, which was depen-
dent on Nr4a1 and CX3CR1. Although these monocytes did
 38   Arterioscler Thromb Vasc Biol   January 2017
not directly kill tumor cells, they increased natural killer cell
recruitment and activation in the lung which contributed to
tumor cell killing.
Monocyte Fate and Function In Vivo During
Inflammation
On recruitment to a site of inflammation, monocytes were
generally thought to differentiate into macrophages while
maintaining the same inflammatory phenotype. The extensive
description of monocyte/macrophage plasticity in vitro has
prompted the re-evaluation of this concept to include the pos-
sibility of monocyte, and macrophages can functionally adapt
locally with the changing inflammatory environment. It has
only been in recent years that conclusive evidence of mono-
cyte conversion in vivo and the mechanisms that regulate this
process have been demonstrated in the context of sterile injury
and infection (Figure).
Sterile Injury
Monocyte subsets have distinct functions within the blood and
tissue during steady state and during inflammation. The tradi-
tional view on monocyte function is that monocytes are pre-
cursors to macrophages and DCs that extravasate into tissues
and differentiate into professional antigen-presenting cells,
where they are then able to resolve inflammation and facilitate
wound repair.1,3,30,47,48 Depletion of monocytes/macrophages
at different stages of inflammation has highlighted a critical
role for these cells in mediating resolution of the response.49–51
Recently, it has been shown that Ly6Clow wound macrophages
Downloaded from http://ahajournals.org by on May 10, 2019
arose from circulating Ly6Chi monocytes recruited to a sponge
implantation (sterile wound) in the skin.3 Fluorescent mic-
roparticle labeling of monocyte subsets in vivo demonstrated
that Ly6Chi monocytes infiltrated into the skin and slowly tran-
sitioned into repair macrophages expressing F4/80 and Mer
tyrosine kinase. Cytokine profiling at the site of inflammation
revealed that early on (day 1) Ly6Chi monocytes expressed
higher levels of IL-β and TNF-α, whereas over the course
of wound healing (day 14) Ly6Clow macrophages expressed
TGF-β (transforming growth factor-beta) and VEGF (vascular
endothelial growth factor), congruent with a more reparative
phenotype. The possibility of alternative Ly6Clow monocytes
being present in this tissue was not examined. A similar
dynamic has been described after MI. This biphasic response
is characterized by the accumulation of Ly6Chi monocytes in
the first phase to clear cellular debris followed by the arrival
of Ly6Clow monocytes/macrophages in the reparative phase
that contributes to collagen deposition and scar formation.15,52
These Ly6Chi monocytes depend on CCR2 to be recruited to
the MI tissue, where they differentiate into Ly6ClowNr4a1hi
macrophages. In mice lacking Nr4a1 on hematopoietic cells,
the inflammatory response during MI is increased, and wound
healing of the infarcted myocardium is compromised. This
study demonstrates that Nr4a1 modulates both inflammatory
and reparative phases of MI.15
A new paradigm has emerged that challenges the current
dogma of monocyte to macrophage differentiation within a tis-
sue environment. In a study by Jakubzick et al,2 using parabio-
sis and bromodeoxyuridine pulse-chase analysis, monocytes
were able to be tracked through steady-state nonlymphoid
organs and lymph nodes and monitored for their expression
of monocyte, macrophage, or DC markers. It was found that
monocytes were able to traffic through tissues and emigrate to
lymph nodes while retaining their monocyte markers and not
differentiating into macrophages or DCs.2 This concept was
further demonstrated during a sterile injury in the liver. Using
intravital microscopy, Dal-Secco et al14 demonstrated that
CCR2-expressing proinflammatory monocytes transitioned
into CX3CR1-expressing reparative monocytes at the site of
injury. These monocytes did not express macrophage or DC
markers, and their phenotypic switch was driven by the cyto-
kines IL-4 and IL-10. A local reprogramming of monocytes at
the site of inflammation or injury may be a more efficient way
to resolve homeostasis in tissues rather than relying on addi-
tional recruitment of repair monocytes or macrophages subse-
quent to the first wave of proinflammatory cells. Moreover, it
allows for monocyte transition as the local environment heals.
The switch from inflammatory to alternative monocyte also
may drive the healing process. Indeed, delaying the switch in
monocyte phenotype also prevented proper wound healing.14
Further studies are needed to determine whether this mono-
cyte–monocyte conversion phenomenon is limited to the liver
or applies to other tissues as well.
Infection
Infection with L. monocytogenes induces a robust innate
response in the liver and spleen, where Ly6Chi monocyte
recruitment is an integral component of this defense. To dem-
onstrate this role, antibody blockade of CD11b or deficiency
of CCR2 enhanced susceptibility to infection by preventing
monocyte recruitment to infected tissues.53,54 During infection
with L. monocytogenes, Ly6Chi monocytes emigrate from the
bone marrow in a CCR2-dependent manner. These monocytes
traffic through the blood to sites of infection, where they dif-
ferentiate into a CD11c+, TNF-α/inducible nitric oxide syn-
thase–producing DC which enhance bacterial clearance.16,41,55
Shi et al56 revealed that inflammatory monocytes, and not neu-
trophils, are essential for bacterial clearance during the early
innate and late adaptive phases of the immune response to L.
monocytogenes.
T. gondii is an obligate intracellular parasite that infects
immunologically privileged sites such as the central nervous
system and muscle tissue, where it establishes a life-long
chronic infection.57,58 A critical component of innate defense
against T. gondii infection is Gr1+Ly6C+ monocytes, which
produce nitric oxide and TNF-α. Monocytes are recruited in a
CCR2-dependent manner in response to T. gondii, and macro-
phage migration inhibitory factor mediates immunity against
T. gondii by promoting activation and conversion of Ly6Chi
monocytes into TNF-α/inducible nitric oxide synthase–pro-
ducing DCs during murine toxoplasmosis.17
A leading cause of skin and soft-tissue infections is because
of Staphylococcus aureus infections, which can lead to life-
threatening conditions, such as sepsis and endocarditis.18,59
During a localized S. aureus infection in the ear, Ly6Chi mono-
cytes get recruited to the infectious site but do not contrib-
ute to the initiation or clearance of infection.18 Inflammatory
 Kratofil et al   Monocyte Fate and Function In Vivo   39
monocytes were predominantly responsible for the expansion
of dermal macrophages during S. aureus infection, although it
was not demonstrated whether these Ly6Chi monocytes differ-
entiated into these dermal macrophages.18 A S. aureus biofilm
has been shown to skew macrophages toward an alternatively
activated M2 phenotype,19 thereby suppressing the immune
system and subsequently allowing S. aureus to persist as a
biofilm. Interestingly, S. aureus biofilms resulted in an exag-
gerated macrophage infiltrate when compared with a nonbio-
film abscess infection,19 indicating that these newly recruited
macrophages are presumably monocyte derived.
Replenishment of Tissue-Resident
Macrophages by Monocytes In Vivo
Tissue-resident macrophages are critical for organ homeosta-
sis and innate immune defenses during inflammation.60 Local
cues called enhancer landscapes have been shown to regulate
macrophage function and their microenvironment within the
tissue to maintain homeostasis and influence the outcome of
inflammatory responses.20 It has been previously thought that
all macrophages are derived from monocytes; however, fate-
mapping studies have established that many tissue-resident
macrophages such as microglia, Kupffer cells, Langerhans
cells, and alveolar macrophages are embryonically derived
and can self-maintain through local proliferation.21–23,60
It is important to understand how monocytes contrib-
ute to the replenishment of tissue-resident populations in
the steady state and after inflammation. In the steady state,
monocytes are noted to constantly replenish tissue-resident
Downloaded from http://ahajournals.org by on May 10, 2019
macrophages in the intestine24–26 and the skin.61 As well in
the steady-state dermis, an extravascular pool of Ly6Chi
monocytes continuously generates monocyte-derived DCs.62
Whereas, in other tissues such as the heart,63,64 pancreas,65
and aorta,63,66 bone marrow–derived monocytes contribute to
only a small proportion or subset of resident macrophages.
After stimulation or tissue injury, monocyte-derived cells are
able to replace embryonically derived macrophages.15,27,63,64
Alternatively, monocytes can also differentiate into new
transient67 or long-term resident66 macrophage populations.
Theurl et al67 identified a novel mechanism in which transient
monocyte-derived, ferroportin 1-expressing macrophages
mediate iron recycling. Importantly, these macrophages
accumulate during mouse models of hemolytic anemia and
sickle cell disease, and preventing their recruitment can lead
to kidney and liver damage. During atherosclerotic develop-
ment, Ly6Chi monocytes infiltrate into the plaque and persist
long term within this environment through local prolifera-
tion. This has challenged the concept that proinflammatory
monocytes are constantly recruited during atherosclerosis to
maintain macrophage load within the lesion. Collectively,
these mechanisms contribute to a heterogeneous popula-
tion of tissue macrophages derived from different origins.
Whether their origin dictates their function in these organs
is not well understood. In the liver, monocyte-derived mac-
rophages are able to obtain a similar transcriptional profile as
embryonically derived Kupffer cells following disease, main-
tain bacterial phagocytic function, and acquire the capacity
to self-renew.27–29,68 Additional work is needed to evaluate
this functional differentiation in other tissue environments
especially in organs with more restricted regenerative capac-
ity after injury such as the brain and the heart.
Monocyte Conversion in Humans
The identification of the intermediate monocyte population,
which has features of both classical and nonclassical mono-
cytes,8,69,70 has contributed to the concept that circulating
monocyte subsets in humans are developmentally related.
Mirroring the model of Ly6Chi monocytes serving as the pre-
cursor cell for patrolling Ly6Clow monocytes in the mouse sys-
tem,35 mobilized classical monocytes from the bone marrow
are proposed to mature into nonclassical monocytes (through
an intermediate subset) in human blood. Gene expression
profiling of these subsets has demonstrated increased expres-
sion of genes associated with maturation going from classical
monocytes to intermediate monocytes and nonclassical mono-
cytes.8 Similar to monocyte development in mice, M-CSF
is noted to play a role in this differentiation in humans.
Stimulation with this factor mediates sequential increase of
intermediate monocytes followed by nonclassical monocytes
over time,71 possibly through induction of CD16 expression.72
Further, blockade of M-CSF results in decreased levels of
both intermediate and nonclassical populations in rheumatoid
arthritis patients,73 demonstrating the ability to modulate this
process during an inflammatory state.
Numerous studies have noted increases in circulating
CD16+ monocytes in cardiovascular disease, trauma, sep-
sis, and autoimmunity,72,74–79 suggesting that this maturation/
polarization response may be an important mechanism in
regulating immune responses and in some cases contributing
to disease pathogenesis. In fact, different monocyte subset
levels have been correlated with poor clinical outcomes,79–83
although causal relationships have not yet been established.
Kinetic studies have provided circumstantial evidence for
this monocyte conversion during the course of an inflammatory
response. After MI, Tsujioka et al78 described sequential mobi-
lization of both CD14++CD16− and CD14+CD16+ monocytes
peaking at day 3 and 5 post infarct, respectively. The peak lev-
els of these populations coincide with their respective appear-
ances in the heart tissue during the inflammatory (12 hours
to 5 days) and proliferative phase (5–12 days) after injury as
evaluated in an independent study.84 This would suggest that
monocyte conversion could occur in both the blood and locally
at the injury site, consistent with results obtained in mouse
models of MI.15,52 In addition to the heart, local polarization
of monocytes has also been suggested in the liver injury repair
after resection surgery.76 This study noted an acute increase
in circulating intermediate monocytes at day 1 after surgery
and their subsequent accumulation at the injury site at day 3.
Interestingly, a concurrent enhancement of angiogenic mono-
cytes (CD14++TIE2+) previously associated with intermediate
monocytes70 was detected at the local surgical site.76 It is unclear
whether this resulted from preferential recruitment of this
subset from the circulation or local differentiation. Recently,
Shalova et al85 have demonstrated the ability of circulating
monocytes to functionally reprogram over the course of sepsis.
In this study, monocytes taken from patients with active sepsis
 40   Arterioscler Thromb Vasc Biol   January 2017
display immunosuppressive and reparative features, possibly
through induction of hypoxia-inducible factor 1. Importantly,
during the recovery stage, circulating monocytes from these
patients re-establish normal proinflammatory function on stim-
ulation similar to monocytes taken from healthy individuals.
Although expansion of CD16+ monocytes has been previously
noted during sepsis,74 monocytes used in this study were only
selected based on CD14+ expression. As such, it is not known
whether these changes are because of the enrichment of 1 sub-
set in particular or the reprogramming of all subsets.
Collectively, human studies to date have provided
important information on the characteristics, presence, and
potential function of monocyte subsets under both homeo-
static and inflammatory conditions. The challenge moving
forward is to develop new strategies to effectively track the
fate of specific subsets in the human context. This in turn
will help better evaluate the potential importance of mono-
cyte conversion during inflammation and identify targets for
therapeutic use.
Sources of Funding
This work is supported by Alberta Innovates Health Solutions
Postgraduate Fellowship (to J.F. Deniset) and Canadian Institutes of
Health Research and Heart and Stroke Foundation of Canada (to P.
Kubes).
Disclosures
None.
References
Downloaded from http://ahajournals.org by on May 10, 2019
1. Geissmann F, Jung S, Littman DR. Blood monocytes consist of two princi-
pal subsets with distinct migratory properties. Immunity. 2003;19:71–82.
doi: 10.1016/S1074-7613(03)00174-2.
2. Jakubzick C, Gautier EL, Gibbings SL, et al. Minimal differentiation
of classical monocytes as they survey steady-state tissues and transport
antigen to lymph nodes. Immunity. 2013;39:599–610. doi: 10.1016/j.
immuni.2013.08.007.
3. Crane MJ, Daley JM, van Houtte O, Brancato SK, Henry WL Jr, Albina
JE. The monocyte to macrophage transition in the murine sterile wound.
PLoS One. 2014;9:e86660. doi: 10.1371/journal.pone.0086660.
4. Passlick B, Flieger D, Ziegler-Heitbrock HW. Identification and charac-
terization of a novel monocyte subpopulation in human peripheral blood.
Blood. 1989;74:2527–2534.
5. Hulsmans M, Sam F, Nahrendorf M. Monocyte and macrophage contribu-
tions to cardiac remodeling. J Mol Cell Cardiol. 2016;93:149–155. doi:
10.1016/j.yjmcc.2015.11.015.
6. Shi C, Pamer EG. Monocyte recruitment during infection and inflamma-
tion. Nat Rev Immunol. 2011;11:762–774. doi: 10.1038/nri3070.
7. Glezeva N, Horgan S, Baugh JA. Monocyte and macrophage subsets
along the continuum to heart failure: misguided heroes or targetable
villains? J Mol Cell Cardiol. 2015;89(pt B):136–145. doi: 10.1016/j.
yjmcc.2015.10.029.
8. Wong KL, Tai JJ, Wong WC, Han H, Sem X, Yeap WH, Kourilsky P,
Wong SC. Gene expression profiling reveals the defining features of the
classical, intermediate, and nonclassical human monocyte subsets. Blood.
2011;118:e16–e31. doi: 10.1182/blood-2010-12-326355.
9. Mukherjee R, Kanti Barman P, Kumar Thatoi P, Tripathy R, Kumar Das
B, Ravindran B. Non-classical monocytes display inflammatory fea-
tures: validation in sepsis and systemic lupus erythematosus. Sci Rep.
2015;5:13886. doi: 10.1038/srep13886.
10. Hettinger J, Richards DM, Hansson J, Barra MM, Joschko AC, Krijgsveld
J, Feuerer M. Origin of monocytes and macrophages in a committed pro-
genitor. Nat Immunol. 2013;14:821–830. doi: 10.1038/ni.2638.
11. Tsou CL, Peters W, Si Y, Slaymaker S, Aslanian AM, Weisberg SP, Mack
M, Charo IF. Critical roles for CCR2 and MCP-3 in monocyte mobili-
zation from bone marrow and recruitment to inflammatory sites. J Clin
Invest. 2007;117:902–909. doi: 10.1172/JCI29919.
12. Hanna RN, Carlin LM, Hubbeling HG, Nackiewicz D, Green AM, Punt
JA, Geissmann F, Hedrick CC. The transcription factor NR4A1 (Nur77)
controls bone marrow differentiation and the survival of Ly6C- mono-
cytes. Nat Immunol. 2011;12:778–785. doi: 10.1038/ni.2063.
13. Auffray C, Fogg D, Garfa M, Elain G, Join-Lambert O, Kayal S, Sarnacki
S, Cumano A, Lauvau G, Geissmann F. Monitoring of blood vessels and
tissues by a population of monocytes with patrolling behavior. Science.
2007;317:666–670. doi: 10.1126/science.1142883.
14. Dal-Secco D, Wang J, Zeng Z, Kolaczkowska E, Wong CH, Petri B,
Ransohoff RM, Charo IF, Jenne CN, Kubes P. A dynamic spectrum of
monocytes arising from the in situ reprogramming of CCR2+ monocytes
at a site of sterile injury. J Exp Med. 2015;212:447–456. doi: 10.1084/
jem.20141539.
15. Hilgendorf I, Gerhardt LM, Tan TC, Winter C, Holderried TA, Chousterman
BG, Iwamoto Y, Liao R, Zirlik A, Scherer-Crosbie M, Hedrick CC, Libby
P, Nahrendorf M, Weissleder R, Swirski FK. Ly-6Chigh monocytes
depend on Nr4a1 to balance both inflammatory and reparative phases in
the infarcted myocardium. Circ Res. 2014;114:1611–1622. doi: 10.1161/
CIRCRESAHA.114.303204.
16. Shi C, Velázquez P, Hohl TM, Leiner I, Dustin ML, Pamer EG. Monocyte
trafficking to hepatic sites of bacterial infection is chemokine indepen-
dent and directed by focal intercellular adhesion molecule-1 expression. J
Immunol. 2010;184:6266–6274. doi: 10.4049/jimmunol.0904160.
17. Ruiz-Rosado Jde D, Olguín JE, Juárez-Avelar I, Saavedra R, Terrazas
LI, Robledo-Avila FH, Vazquez-Mendoza A, Fernández J, Satoskar AR,
Partida-Sánchez S, Rodriguez-Sosa M. MIF promotes classical activation
and conversion of inflammatory Ly6C(high) monocytes into TipDCs dur-
ing murine toxoplasmosis. Mediators Inflamm. 2016;2016:9101762. doi:
10.1155/2016/9101762.
18. Feuerstein R, Seidl M, Prinz M, Henneke P. MyD88 in macrophages is
critical for abscess resolution in staphylococcal skin infection. J Immunol.
2015;194:2735–2745. doi: 10.4049/jimmunol.1402566.
19. Thurlow LR, Hanke ML, Fritz T, Angle A, Aldrich A, Williams SH,
Engebretsen IL, Bayles KW, Horswill AR, Kielian T. Staphylococcus
aureus biofilms prevent macrophage phagocytosis and attenuate inflam-
mation in vivo. J Immunol. 2011;186:6585–6596. doi: 10.4049/
jimmunol.1002794.
20. Lavin Y, Winter D, Blecher-Gonen R, David E, Keren-Shaul H, Merad
M, Jung S, Amit I. Tissue-resident macrophage enhancer landscapes are
shaped by the local microenvironment. Cell. 2014;159:1312–1326. doi:
10.1016/j.cell.2014.11.018.
21. Gomez Perdiguero E, Klapproth K, Schulz C, Busch K, Azzoni E, Crozet L,
Garner H, Trouillet C, de Bruijn MF, Geissmann F, Rodewald HR. Tissue-
resident macrophages originate from yolk-sac-derived erythro-myeloid
progenitors. Nature. 2015;518:547–551. doi: 10.1038/nature13989.
22. Ginhoux F, Greter M, Leboeuf M, Nandi S, See P, Gokhan S, Mehler MF,
Conway SJ, Ng LG, Stanley ER, Samokhvalov IM, Merad M. Fate map-
ping analysis reveals that adult microglia derive from primitive macro-
phages. Science. 2010;330:841–845. doi: 10.1126/science.1194637.
23. Schulz C, Gomez Perdiguero E, Chorro L, Szabo-Rogers H, Cagnard
N, Kierdorf K, Prinz M, Wu B, Jacobsen SE, Pollard JW, Frampton J,
Liu KJ, Geissmann F. A lineage of myeloid cells independent of Myb
and hematopoietic stem cells. Science. 2012;336:86–90. doi: 10.1126/
science.1219179.
24. Rivollier A, He J, Kole A, Valatas V, Kelsall BL. Inflammation switches
the differentiation program of Ly6Chi monocytes from antiinflammatory
macrophages to inflammatory dendritic cells in the colon. J Exp Med.
2012;209:139–155. doi: 10.1084/jem.20101387.
25. Tamoutounour S, Henri S, Lelouard H, et al. CD64 distinguishes macro-
phages from dendritic cells in the gut and reveals the Th1-inducing role
of mesenteric lymph node macrophages during colitis. Eur J Immunol.
2012;42:3150–3166. doi: 10.1002/eji.201242847.
26. Bain CC, Scott CL, Uronen-Hansson H, Gudjonsson S, Jansson O, Grip
O, Guilliams M, Malissen B, Agace WW, Mowat AM. Resident and
pro-inflammatory macrophages in the colon represent alternative con-
text-dependent fates of the same Ly6Chi monocyte precursors. Mucosal
Immunol. 2013;6:498–510. doi: 10.1038/mi.2012.89.
27. Movita D, van de Garde MD, Biesta P, Kreefft K, Haagmans B, Zuniga E,
Herschke F, De Jonghe S, Janssen HL, Gama L, Boonstra A, Vanwolleghem
T. Inflammatory monocytes recruited to the liver within 24 hours after
virus-induced inflammation resemble Kupffer cells but are functionally
distinct. J Virol. 2015;89:4809–4817. doi: 10.1128/JVI.03733-14.
28. Scott CL, Zheng F, De Baetselier P, Martens L, Saeys Y, De Prijck S,
Lippens S, Abels C, Schoonooghe S, Raes G, Devoogdt N, Lambrecht
BN, Beschin A, Guilliams M. Bone marrow-derived monocytes give rise
 Kratofil et al   Monocyte Fate and Function In Vivo   41
to self-renewing and fully differentiated Kupffer cells. Nat Commun.
2016;7:10321. doi: 10.1038/ncomms10321.
29. Elsegood CL, Chan CW, Degli-Esposti MA, Wikstrom ME, Domenichini
A, Lazarus K, van Rooijen N, Ganss R, Olynyk JK, Yeoh GC. Kupffer
cell-monocyte communication is essential for initiating murine liver
progenitor cell-mediated liver regeneration. Hepatology. 2015;62:1272–
1284. doi: 10.1002/hep.27977.
30. Auffray C, Sieweke MH, Geissmann F. Blood monocytes: development,
heterogeneity, and relationship with dendritic cells. Annu Rev Immunol.
2009;27:669–692. doi: 10.1146/annurev.immunol.021908.132557.
31. Dai XM, Ryan GR, Hapel AJ, Dominguez MG, Russell RG, Kapp
S, Sylvestre V, Stanley ER. Targeted disruption of the mouse colony-
stimulating factor 1 receptor gene results in osteopetrosis, mononuclear
phagocyte deficiency, increased primitive progenitor cell frequencies,
and reproductive defects. Blood. 2002;99:111–120. doi: 10.1182/blood.
V99.1.111.
32. Zhu YP, Thomas GD, Hedrick CC. 2014 Jeffrey M. Hoeg Award Lecture:
Transcriptional Control of Monocyte Development. Arterioscler Thromb
Vasc Biol. 2016;36:1722–1733. doi: 10.1161/ATVBAHA.116.304054.
33. Shi C, Jia T, Mendez-Ferrer S, Hohl TM, Serbina NV, Lipuma L, Leiner
I, Li MO, Frenette PS, Pamer EG. Bone marrow mesenchymal stem and
progenitor cells induce monocyte emigration in response to circulating
toll-like receptor ligands. Immunity. 2011;34:590–601. doi: 10.1016/j.
immuni.2011.02.016.
34. Zouggari Y, Ait-Oufella H, Bonnin P, et al. B lymphocytes trigger mono-
cyte mobilization and impair heart function after acute myocardial infarc-
tion. Nat Med. 2013;19:1273–1280. doi: 10.1038/nm.3284.
35. Yona S, Kim KW, Wolf Y, Mildner A, Varol D, Breker M, Strauss-Ayali
D, Viukov S, Guilliams M, Misharin A, Hume DA, Perlman H, Malissen
B, Zelzer E, Jung S. Fate mapping reveals origins and dynamics of mono-
cytes and tissue macrophages under homeostasis. Immunity. 2013;38:79–
91. doi: 10.1016/j.immuni.2012.12.001.
36. Varol C, Landsman L, Fogg DK, Greenshtein L, Gildor B, Margalit R,
Kalchenko V, Geissmann F, Jung S. Monocytes give rise to mucosal, but
not splenic, conventional dendritic cells. J Exp Med. 2007;204:171–180.
doi: 10.1084/jem.20061011.
37. Carlin LM, Stamatiades EG, Auffray C, Hanna RN, Glover L, Vizcay-
Downloaded from http://ahajournals.org by on May 10, 2019
Barrena G, Hedrick CC, Cook HT, Diebold S, Geissmann F. Nr4a1-
dependent Ly6C(low) monocytes monitor endothelial cells and orchestrate
their disposal. Cell. 2013;153:362–375. doi: 10.1016/j.cell.2013.03.010.
38. Alder JK, Georgantas RW III, Hildreth RL, Kaplan IM, Morisot S, Yu X,
McDevitt M, Civin CI. Kruppel-like factor 4 is essential for inflammatory
monocyte differentiation in vivo. J Immunol. 2008;180:5645–5652. doi:
10.4049/jimmunol.180.8.5645.
39. Kurotaki D, Osato N, Nishiyama A, Yamamoto M, Ban T, Sato H,
Nakabayashi J, Umehara M, Miyake N, Matsumoto N, Nakazawa M,
Ozato K, Tamura T. Essential role of the IRF8-KLF4 transcription factor
cascade in murine monocyte differentiation. Blood. 2013;121:1839–1849.
doi: 10.1182/blood-2012-06-437863.
40. Ley K, Laudanna C, Cybulsky MI, Nourshargh S. Getting to the site of
inflammation: the leukocyte adhesion cascade updated. Nat Rev Immunol.
2007;7:678–689. doi: 10.1038/nri2156.
41. Serbina NV, Pamer EG. Monocyte emigration from bone marrow dur-
ing bacterial infection requires signals mediated by chemokine receptor
CCR2. Nat Immunol. 2006;7:311–317. doi: 10.1038/ni1309.
42. Swirski FK, Nahrendorf M, Etzrodt M, Wildgruber M, Cortez-Retamozo
V, Panizzi P, Figueiredo JL, Kohler RH, Chudnovskiy A, Waterman P,
Aikawa E, Mempel TR, Libby P, Weissleder R, Pittet MJ. Identification of
splenic reservoir monocytes and their deployment to inflammatory sites.
Science. 2009;325:612–616. doi: 10.1126/science.1175202.
43. Leuschner F, Dutta P, Gorbatov R, et al. Therapeutic siRNA silencing in
inflammatory monocytes in mice. Nat Biotechnol. 2011;29:1005–1010.
doi: 10.1038/nbt.1989.
44. Tacke F, Alvarez D, Kaplan TJ, Jakubzick C, Spanbroek R, Llodra J, Garin
A, Liu J, Mack M, van Rooijen N, Lira SA, Habenicht AJ, Randolph GJ.
Monocyte subsets differentially employ CCR2, CCR5, and CX3CR1 to
accumulate within atherosclerotic plaques. J Clin Invest. 2007;117:185–
194. doi: 10.1172/JCI28549.
45. Swirski FK, Libby P, Aikawa E, Alcaide P, Luscinskas FW, Weissleder R,
Pittet MJ. Ly-6Chi monocytes dominate hypercholesterolemia-associated
monocytosis and give rise to macrophages in atheromata. J Clin Invest.
2007;117:195–205. doi: 10.1172/JCI29950.
46. Hanna RN, Cekic C, Sag D, et al. Patrolling monocytes control tumor
metastasis to the lung. Science. 2015;350:985–990. doi: 10.1126/science.
aac9407.
47. Brancato SK, Albina JE. Wound macrophages as key regulators of repair:
origin, phenotype, and function. Am J Pathol. 2011;178:19–25. doi:
10.1016/j.ajpath.2010.08.003.
48. Peng X, Zhang J, Xiao Z, Dong Y, Du J. CX3CL1-CX3CR1 interac-
tion increases the population of Ly6C(-)CX3CR1(hi) macrophages con-
tributing to unilateral ureteral obstruction-induced fibrosis. J Immunol.
2015;195:2797–2805. doi: 10.4049/jimmunol.1403209.
49. Duffield JS, Forbes SJ, Constandinou CM, Clay S, Partolina M, Vuthoori
S, Wu S, Lang R, Iredale JP. Selective depletion of macrophages reveals
distinct, opposing roles during liver injury and repair. J Clin Invest.
2005;115:56–65. doi: 10.1172/JCI22675.
50. Goren I, Allmann N, Yogev N, Schürmann C, Linke A, Holdener M,
Waisman A, Pfeilschifter J, Frank S. A transgenic mouse model of induc-
ible macrophage depletion: effects of diphtheria toxin-driven lysozyme
M-specific cell lineage ablation on wound inflammatory, angiogenic, and
contractive processes. Am J Pathol. 2009;175:132–147. doi: 10.2353/
ajpath.2009.081002.
51. Lucas T, Waisman A, Ranjan R, Roes J, Krieg T, Müller W, Roers A, Eming
SA. Differential roles of macrophages in diverse phases of skin repair. J
Immunol. 2010;184:3964–3977. doi: 10.4049/jimmunol.0903356.
52. Nahrendorf M, Swirski FK, Aikawa E, Stangenberg L, Wurdinger T,
Figueiredo JL, Libby P, Weissleder R, Pittet MJ. The healing myocardium
sequentially mobilizes two monocyte subsets with divergent and com-
plementary functions. J Exp Med. 2007;204:3037–3047. doi: 10.1084/
jem.20070885.
53. Rosen H, Gordon S, North RJ. Exacerbation of murine listeriosis by a
monoclonal antibody specific for the type 3 complement receptor of
myelomonocytic cells. Absence of monocytes at infective foci allows
Listeria to multiply in nonphagocytic cells. J Exp Med. 1989;170:27–37.
doi: 10.1084/jem.170.1.27.
54. Kurihara T, Warr G, Loy J, Bravo R. Defects in macrophage recruitment
and host defense in mice lacking the CCR2 chemokine receptor. J Exp
Med. 1997;186:1757–1762. doi: 10.1084/jem.186.10.1757.
55. Serbina NV, Salazar-Mather TP, Biron CA, Kuziel WA, Pamer EG.
TNF/iNOS-producing dendritic cells mediate innate immune defense
against bacterial infection. Immunity. 2003;19:59–70. doi: 10.1016/
S1074-7613(03)00171-7.
56. Shi C, Hohl TM, Leiner I, Equinda MJ, Fan X, Pamer EG. Ly6G+ neu-
trophils are dispensable for defense against systemic Listeria mono-
cytogenes infection. J Immunol. 2011;187:5293–5298. doi: 10.4049/
jimmunol.1101721.
57. Serbina NV, Jia T, Hohl TM, Pamer EG. Monocyte-mediated defense
against microbial pathogens. Annu Rev Immunol. 2008;26:421–452. doi:
10.1146/annurev.immunol.26.021607.090326.
58. Neal LM, Knoll LJ. Toxoplasma gondii profilin promotes recruitment of
Ly6Chi CCR2+ inflammatory monocytes that can confer resistance to bac-
terial infection. PLoS Pathog. 2014;10:e1004203. doi: 10.1371/journal.
ppat.1004203.
59. Thammavongsa V, Missiakas DM, Schneewind O. Staphylococcus aureus
degrades neutrophil extracellular traps to promote immune cell death.
Science. 2013;342:863–866. doi: 10.1126/science.1242255.
60. Ginhoux F, Guilliams M. Tissue-resident macrophage ontogeny and homeo-
stasis. Immunity. 2016;44:439–449. doi: 10.1016/j.immuni.2016.02.024.
61. Capucha T, Mizraji G, Segev H, et al. Distinct murine mucosal Langerhans
cell subsets develop from pre-dendritic cells and monocytes. Immunity.
2015;43:369–381. doi: 10.1016/j.immuni.2015.06.017.
62. Tamoutounour S, Guilliams M, Montanana Sanchis F, Liu H, Terhorst D,
Malosse C, Pollet E, Ardouin L, Luche H, Sanchez C, Dalod M, Malissen
B, Henri S. Origins and functional specialization of macrophages and
of conventional and monocyte-derived dendritic cells in mouse skin.
Immunity. 2013;39:925–938. doi: 10.1016/j.immuni.2013.10.004.
63. Epelman S, Lavine KJ, Beaudin AE, et al. Embryonic and adult-derived
resident cardiac macrophages are maintained through distinct mechanisms
at steady state and during inflammation. Immunity. 2014;40:91–104. doi:
10.1016/j.immuni.2013.11.019.
64. Heidt T, Courties G, Dutta P, Sager HB, Sebas M, Iwamoto Y, Sun Y, Da
Silva N, Panizzi P, van der Laan AM, van der Lahn AM, Swirski FK,
Weissleder R, Nahrendorf M. Differential contribution of monocytes to
heart macrophages in steady-state and after myocardial infarction. Circ
Res. 2014;115:284–295. doi: 10.1161/CIRCRESAHA.115.303567.
65. Calderon B, Carrero JA, Ferris ST, Sojka DK, Moore L, Epelman S,
Murphy KM, Yokoyama WM, Randolph GJ, Unanue ER. The pancreas
anatomy conditions the origin and properties of resident macrophages. J
Exp Med. 2015;212:1497–1512. doi: 10.1084/jem.20150496.
 42   Arterioscler Thromb Vasc Biol   January 2017

66. Robbins CS, Hilgendorf I, Weber GF, et al. Local proliferation domi-
nates lesional macrophage accumulation in atherosclerosis. Nat Med.
2013;19:1166–1172. doi: 10.1038/nm.3258.
67. Theurl I, Hilgendorf I, Nairz M, et al. On-demand erythrocyte disposal
and iron recycling requires transient macrophages in the liver. Nat Med.
2016;22:945–951. doi: 10.1038/nm.4146.
68. David BA, Rezende RM, Antunes MM, et al. Combination of mass cytom-
etry and imaging analysis reveals origin, location, and functional repopu-
lation of liver myeloid cells in mice [published online ahead of print
August 25, 2016]. Gastroenterology. doi: 10.1053/j.gastro.2016.08.024.
69. Cros J, Cagnard N, Woollard K, Patey N, Zhang SY, Senechal B, Puel
A, Biswas SK, Moshous D, Picard C, Jais JP, D’Cruz D, Casanova JL,
Trouillet C, Geissmann F. Human CD14dim monocytes patrol and sense
nucleic acids and viruses via TLR7 and TLR8 receptors. Immunity.
2010;33:375–386. doi: 10.1016/j.immuni.2010.08.012.
70. Zawada AM, Rogacev KS, Rotter B, Winter P, Marell RR, Fliser D, Heine
GH. SuperSAGE evidence for CD14++CD16+ monocytes as a third mono-
cyte subset. Blood. 2011;118:e50–e61. doi: 10.1182/blood-2011-01-326827.
71. Weiner LM, Li W, Holmes M, Catalano RB, Dovnarsky M, Padavic K,
Alpaugh RK. Phase I trial of recombinant macrophage colony-stimulating
factor and recombinant gamma-interferon: toxicity, monocytosis, and
clinical effects. Cancer Res. 1994;54:4084–4090.
72. West SD, Goldberg D, Ziegler A, Krencicki M, Du Clos TW, Mold C.
Transforming growth factor-β, macrophage colony-stimulating factor and
C-reactive protein levels correlate with CD14(high)CD16+ monocyte
induction and activation in trauma patients. PLoS One. 2012;7:e52406.
doi: 10.1371/journal.pone.0052406.
73. Korkosz M, Bukowska-Strakova K, Sadis S, Grodzicki T, Siedlar
M. Monoclonal antibodies against macrophage colony-stimulating
factor diminish the number of circulating intermediate and non-
classical (CD14(++)CD16(+)/CD14(+)CD16(++)) monocytes in rheu-
matoid arthritis patient. Blood. 2012;119:5329–5330. doi: 10.1182/
blood-2012-02-412551.
74. Poehlmann H, Schefold JC, Zuckermann-Becker H, Volk HD, Meisel
C. Phenotype changes and impaired function of dendritic cell subsets
in patients with sepsis: a prospective observational analysis. Crit Care.
2009;13:R119. doi: 10.1186/cc7969.
Downloaded from http://ahajournals.org by on May 10, 2019
angiogenic potential are selectively induced by liver resection and accu-
mulate near the site of liver regeneration. BMC Immunol. 2014;15:50. doi:
10.1186/s12865-014-0050-3.
77. Tapp LD, Shantsila E, Wrigley BJ, Pamukcu B, Lip GY. The
CD14++CD16+ monocyte subset and monocyte-platelet interactions in
patients with ST-elevation myocardial infarction. J Thromb Haemost.
2012;10:1231–1241. doi: 10.1111/j.1538-7836.2011.04603.x.
78. Tsujioka H, Imanishi T, Ikejima H, et al. Impact of heterogeneity of human
peripheral blood monocyte subsets on myocardial salvage in patients with
primary acute myocardial infarction. J Am Coll Cardiol. 2009;54:130–
138. doi: 10.1016/j.jacc.2009.04.021.
79. Urra X, Villamor N, Amaro S, Gómez-Choco M, Obach V, Oleaga L,
Planas AM, Chamorro A. Monocyte subtypes predict clinical course and
prognosis in human stroke. J Cereb Blood Flow Metab. 2009;29:994–
1002. doi: 10.1038/jcbfm.2009.25.
80. Baeten D, Boots AM, Steenbakkers PG, Elewaut D, Bos E, Verheijden
GF, Berheijden G, Miltenburg AM, Rijnders AW, Veys EM, De
Keyser F. Human cartilage gp-39+,CD16+ monocytes in periph-
eral blood and synovium: correlation with joint destruction in
rheumatoid arthritis. Arthritis Rheum. 2000;43:1233–1243. doi:
10.1002/1529-0131(200006)43:6<1233::AID-ANR6>3.0.CO;2-9.
81. Kaito M, Araya S, Gondo Y, Fujita M, Minato N, Nakanishi M, Matsui
M. Relevance of distinct monocyte subsets to clinical course of isch-
emic stroke patients. PLoS One. 2013;8:e69409. doi: 10.1371/journal.
pone.0069409.
82. Rogacev KS, Cremers B, Zawada AM, et al. CD14++CD16+ mono-
cytes independently predict cardiovascular events: a cohort study of 951
patients referred for elective coronary angiography. J Am Coll Cardiol.
2012;60:1512–1520. doi: 10.1016/j.jacc.2012.07.019.
83. Rogacev KS, Zawada AM, Emrich I, Seiler S, Böhm M, Fliser D, Woollard
KJ, Heine GH. Lower Apo A-I and lower HDL-C levels are associated
with higher intermediate CD14++CD16+ monocyte counts that predict
cardiovascular events in chronic kidney disease. Arterioscler Thromb Vasc
Biol. 2014;34:2120–2127. doi: 10.1161/ATVBAHA.114.304172.
84. van der Laan AM, Ter Horst EN, Delewi R, Begieneman MP, Krijnen PA,
Hirsch A, Lavaei M, Nahrendorf M, Horrevoets AJ, Niessen HW, Piek JJ.
Monocyte subset accumulation in the human heart following acute myo-

75. Rossol M, Kraus S, Pierer M, Baerwald C, Wagner U. The CD14(bright)
CD16+ monocyte subset is expanded in rheumatoid arthritis and promotes
expansion of the Th17 cell population. Arthritis Rheum. 2012;64:671–
677. doi: 10.1002/art.33418.
76. Schauer D, Starlinger P, Zajc P, Alidzanovic L, Maier T, Buchberger
E, Pop L, Gruenberger B, Gruenberger T, Brostjan C. Monocytes with
cardial infarction and the role of the spleen as monocyte reservoir. Eur
Heart J. 2014;35:376–385. doi: 10.1093/eurheartj/eht331.
85. Shalova IN, Lim JY, Chittezhath M, et al. Human monocytes undergo
functional re-programming during sepsis mediated by hypoxia-
inducible factor-1α. Immunity. 2015;42:484–498. doi: 10.1016/j.
immuni.2015.02.001.

Highlights
• Phenotypically distinct monocyte subsets identified in both mice and humans have been suggested to form a part of a maturation continuum
under steady-state and inflammatory conditions. Their recruitment to sites of injury is mediated by both common and distinct mechanisms
depending on the inflammatory context.
• Recent studies have provided the first definitive in vivo evidence of monocyte/macrophage polarization within an inflammatory site. This con-
version mechanism importantly contributes to proper immune response resolution and tissue repair.
• Many tissue-resident macrophage populations are now known to arise from an embryonic origin. However, under inflammatory conditions,
proinflammatory monocytes can contribute to their replenishment forming a heterogeneous population of tissue macrophage within different
organs. Details on the cellular plasticity of monocytes in this context and the associated functional consequences have only started to emerge.