Pflugers Arch - Eur J Physiol

DOI 10.1007/s00424-017-1945-7

INVITED REVIEW

Physiological roles of macrophages

Siamon Gordon 1,2 & Luisa Martinez-Pomares 3

Received: 20 January 2017 / Accepted: 29 January 2017

# The Author(s) 2017. This article is published with open access at Springerlink.com

Abstract Macrophages are present in mammals from Introduction

midgestation, contributing to physiologic homeostasis
throughout life. Macrophages arise from yolk sac and foetal BThe constancy of the internal environment is the condition
liver progenitors during embryonic development in the mouse for full and independent life: the mechanism that makes it
and persist in different organs as heterogeneous, self-renewing possible is that which assured that maintenance, within the
tissue-resident populations. Bone marrow-derived blood internal environment, conditions necessary for the life of these
monocytes are recruited after birth to replenish tissue- elements^ (Claude Bernard).
resident populations and to meet further demands during in- The concept of homeostasis as a physiological principle, a
flammation, infection and metabolic perturbations. term proposed by the neuroscientist Walter Cannon (1871–
Macrophages of mixed origin and different locations vary in 1945) and based on the work of Claude Bernard (1813–
replication and turnover, but are all active in mRNA and pro- 1878), has influenced the thinking of many macrophage in-
tein synthesis, fulfilling organ-specific and systemic trophic vestigators. Elie Metchnikoff (1845–1916), a comparative zo-
functions, in addition to host defence. In this review, we em- ologist who initially was intrigued by intracellular digestion in
phasise selected properties and non-immune functions of tis- invertebrates, became an experimental immunologist when he
sue macrophages which contribute to physiologic appreciated the role of phagocytes in tissue orthobiosis and
homeostasis. inflammation [67]. Hans Selye (1907–1982) drew attention to
the non-specific neuro-endocrine adaptations of the body to
biological stress; however, the contributions of the mononu-
Keywords Macrophages . Homeostasis . Tissue clear phagocyte system to the mechanisms of homeostasis and
heterogeneity . Physiology . Receptors . Phagocytosis stress responses have not achieved full recognition. In this
review, we consider topics related to the role of macrophages
in development, metabolism and tissue-specific functions in
different organs, to illustrate their remarkable ability to recog-
nise and respond to the changing needs of the body. A general
overview of the main topic covered is provided in Fig. 1.
* Siamon Gordon Blood monocytes and tissue macrophages display many
siamon.gordon@path.ox.ac.uk properties in common, but they adapt readily upon entering
* Luisa Martinez-Pomares different tissue microenvironments, where they contribute to
luisa.m@nottingham.ac.uk the general needs as well as specialised functions of individual
organs. Reciprocal interactions between neighbouring cells
1
Graduate Institute of Biomedical Sciences, College of Medicine, and tissue macrophages are mediated by close contacts as well
Chang Gung University, Taoyuan City 33302, Taiwan as secretory products which act at close range or distally via
2
Sir William Dunn School of Pathology, Oxford University, the circulation. Although primarily known as professional
Oxford, UK phagocytes [29], which engulf dying cells during develop-
3
School of Life Sciences, University of Nottingham, Nottingham, UK ment and subsequently during adult life, it has not been
 Pflugers Arch - Eur J Physiol

Fig. 1 Physiological roles of macrophages and macrophages patterns; (b) receptors for apoptotic cells; (c) cytokine receptors such as
heterogeneity. Phenotypic heterogeneity of tissue macrophages is IFN-γ and IL-4Rα that promote M1/M2-like activation, respectively; (d)
triggered by differences in cellular origin (self-replicated cells of receptors for neurotransmitters as well as (e) receptors that regulate
embryonic origin vs monocyte-derived), tissue environment and macrophage activation in situ such as the inhibitory receptors SIRP1-α
adaptation to the physiological requirements of the organism (shown on and CD200R. Integrins (f) mediate interaction with extracellular matrix.
the left). Macrophage heterogeneity underpins the vast range of Macrophages produce numerous molecules that contribute to tissue
physiological roles performed of these cells (shown on the right). remodelling and inflammation and mediate clearance of apoptotic
Phenotypic heterogeneity is consolidated by tissue-specific Bchromatin bodies, cell debris and soluble compounds such as collagen and
landscapes^ in macrophages in response to environmental signals. lysosomal hydrolases. High phagocytic and endocytic activities as well
Macrophages display a wide range of Bsensor systems^ in the form of as motility are central to the ability of macrophages to support tissue-
(a) cell surface and intracellular pattern recognition receptors that detect dependent functions. RNS reactive nitrogen species, ROS reactive
microbe-associated molecular patterns and damage-associated molecular oxygen species

generally appreciated that macrophage activities extend be- receptors to mediate phagocytic clearance of particulates, in-
yond immunology and host defence. They express a broad cluding micro-organisms and apoptotic cells, and for endocy-
repertoire of surface and intracellular receptors and produce tosis of soluble ligands. In addition, they are able to sense and
a large variety of regulated bioactive molecules; together with respond to the presence of microbial constituents, including
their ability to enter all tissues, in the steady state and on RNA and DNA, within their cytoplasm, activating intracellu-
demand, they constitute a dispersed organ system, able to lar inflammasomes [32], caspase processing and release of IL-
influence and respond to every other system in the body. 1β. A plethora of other cytokines, enzymes, growth factors,
lipid-derived and low molecular metabolite products can also
be secreted by macrophages in response to phagocytic and
Cellular characteristics of the macrophage lineage: various stimuli and, in turn, act on macrophage receptors by
relevance to physiology autocrine mechanisms. Plasma membrane receptors co-
operate in formation of a phagocytic synapse, excluding phos-
To set the stage for subsequent discussion, we briefly summa- phatases such as CD45, and allow receptor kinases to initiate
rise some general properties of macrophages relevant to their signalling pathways and cytoskeletal assembly, to bring about
functions in vivo. The cells originate from progenitors in the ingestion of particles [21]. A sequence of dynamic interac-
embryo and postnatally, as discussed further below, and are tions with vesicular membranes derived from the endoplasmic
seeded throughout the body as relatively long lived, terminally reticulum and Golgi apparatus accompanies phagosome mat-
differentiated haematopoietic cells. They are motile and dis- uration, fusion with primary lysosomes, acidification and di-
play a range of membrane receptors to recognise and respond gestion [31, 39, 71]. Macrophages can internalise its entire
to a large repertoire of host-derived and foreign ligands [29]. surface membrane in 20 min, but recycle components equally
These include adhesion molecules, opsonic and non-opsonic rapidly to maintain membrane homeostasis [51]. Heterophagy
Pflugers Arch - Eur J Physiol

refers to uptake of extracellular cargo; autophagy, the seques- complex activation [19]. The stability or reversibility of phe-
tration of effete cytoplasm and intracellular organelles, is well notype in vivo needs a great deal of further study, at popula-
developed in macrophages [16]; genetic deficiency of lyso- tion and the single cell level. In addition, further studies on
somal digestion or components of the autophagy pathway can genetic and epigenetic mechanisms will contribute to a more
result in storage diseases [59] or chronic granulomatous in- sophisticated understanding of tissue-specific and generic
flammation [68], respectively. macrophage phenotypic heterogeneity.
The uptake and killing of microbes and opsonised cellular
targets by oxygen and nitrogen-derived radicals are mediated
by assembly and activation of an NADPH oxidase and induc- Macrophage heterogeneity under physiological
ible nitric oxide synthetase, respectively [23, 43]. The inges- conditions
tion of apoptotic targets, a major physiologic function of
tissue-resident and inflammatory macrophages, is mediated The immune system comprises a complex network of cellular
by a range of redundant receptors, using exposed phosphatidyl and soluble components that collaborate to respond to chal-
serine and other cues as ligands [2]. Apoptotic cell uptake lenges, both infectious and non-infectious, identified as poten-
inhibits the inflammatory response associated with uptake of tial threats to the organism. Accordingly, immune constituents
necrotic cells or microbes; anti-inflammatory products include are widely distributed throughout tissues with specialised lym-
TGF-β, IL-10 and PGE2 [38]. The importance of phagoptotic phoid organs, such as splenic white pulp and lymph nodes,
killing during phagocytosis has been emphasised by the stud- mostly providing unique sites for the induction of specific
ies of Guy Brown and colleagues [7]. Finally, macrophages acquired immunity.
can ingest membrane-enveloped nuclei or intracellular vesi- To date, newcomers to the field are still surprised to learn
cles released from living cells or undergo cell fusion, to form that healthy tissues incorporate immune cells, in particular,
multinucleated giant cells [35]. macrophages which are cells commonly associated with pro-
tection against infection and as mediators of tissue damage.
Macrophages, unlike neutrophils, are integral components of
Phenotypic adaptation of macrophages tissues; they contribute to organ development and mainte-
nance of homeostasis through mechanisms similar to those
Macrophage gene expression has been intensively studied in exploited during infection to detect pathogens and trigger in-
recent years and modules of genes regulated by a range of flammation. Widespread distribution of macrophages is ac-
cytokines and other stimuli characterised in tissue-resident companied by substantial tissue-specific phenotypic and gene
and inflammatory macrophages. Both pro- and anti- expression profile differences related to the specific environ-
inflammatory pathways can be induced in macrophage popu- mental cues in each organ that foster tissue-specific macro-
lations, mediated by plasma membrane or cytoplasmic recep- phage function [25]. Differences in gene expression are
tors. IFN-γ, for example, induces a characteristic profile of underpinned by the existence of distinct Bchromatin
genes and proteins, often termed Bclassical or M1 activation^ landscapes^ in each tissue macrophage population which are
[47]; prototypic markers include upregulation of MHCII anti- determined in large extent by the tissue environment [44].
gens, pro-inflammatory cytokines and induction of i-NOS, Furthermore, fully differentiated macrophages may retain the
whereas CD206 expression is selectively downregulated. By capacity to change their gene expression profile when exposed
contrast, interleukin-4 and interleukin-13, binding to a com- to a new tissue environment [44], which highlights the poten-
mon IL-4 receptor alpha chain, induce an Balternative, M2^ tial ability of macrophages to undergo reprogramming even
phenotype, characterised by upregulation of CD206, after differentiation. Phenotypically, there are instances of tis-
transglutaminase 2, arginase and, in the mouse, chitinase- sue adaptation in macrophages involving fine tuning of cellu-
like molecules such as YM1 [48]. An IL-13-specific receptor lar activation thresholds. In mucosal tissues such as the respi-
pathway has also been demonstrated by induction of macro- ratory and gastrointestinal tracts, macrophages display a
phage homokaryon formation [62]. Phenotypic changes in higher threshold of activation as a means to minimise
macrophages are underpinned by metabolic reprogramming. inflammation-mediated organ damage in response to repeated
M1 macrophages utilise Warburg metabolism and M2 macro- exposure to foreign material, including microbial products. In
phage oxidative metabolism [55]. The M1/M2 terminology the case of alveolar macrophages, exposure to IL-10, surfac-
arose as counterpart to Th1 and Th2 categories of adaptive tant proteins A and D and close interaction with respiratory
immune responses, associated with the above and related cy- epithelium through the inhibitory receptor CD200R and ανβ-
tokines. It is thought that instead of bimodal polarisation, a 6 tethered TGF-β, act as effective brakes of macrophage ac-
more complex phenotypic spectrum is more likely, especially tivation minimising the risk of unwanted inflammation [40].
in vivo [50, 52]. In truth, there is overlap with other cytokine- In the gut, macrophages are hyporesponsive to stimulation
induced stimuli, e.g. after IL-10 stimulation or immune [65]. Nevertheless, gut macrophages can trigger inflammation
 Pflugers Arch - Eur J Physiol

with one mechanism involving the detection of tissue damage Ly6Chigh monocytes are recruited to tissues under steady
through activation of the NLRC4 inflammasome which in- state and inflammatory conditions. In the absence of inflam-
duces processing of pro-IL-1β constitutively expressed by mation, Ly6Chigh monocytes sample the tissues and transport
intestinal macrophages [20]. This mechanism ensures detec- material to draining lymph nodes without differentiating into
tion of pathogens that cause tissue damage through the pro- macrophages or dendritic cells [18]. Under inflammatory con-
duction of virulence factors [6, 70]. Macrophages in internal ditions, Ly6Chigh monocytes differentiate into macrophages
organs, less exposed to microbial constituents, present a lower and can give rise to tissue macrophages with capacity for
activation threshold and are considered to be responsible for self-renewal. This process is highly dependent on the tissue-
the induction of systemic inflammation in response to blood- depletion strategy used and the organ under consideration
borne pathogens, which can have deadly consequences in the [18].
case of sepsis [30].

Role of macrophages in development

Origin of tissue macrophages
Osteoclasts and bone remodelling
In addition to tissue-specific environmental cues, tissue-
specific characteristics in macrophages can be linked in part Bone remodelling occurs throughout life and is influenced by
to on site differentiation of macrophages from embryonic pre- physical activity, diet and hormones. Bone generation, pro-
cursors during development and maintenance of macrophage moted by cells of mesenchymal origin termed osteoblasts, is
numbers in tissues through self-renewal [26, 63] with imprint- balanced by the action of cells of haematopoietic origin
ing of macrophage transcriptional programmes by tissue fac- termed osteoclasts that mediate bone reabsorption. Bone for-
tors of particular importance [26]. Indeed, the original idea of mation and degradation need to be balanced as disequilibrium
monocytes as precursors of tissue macrophages that gave rise would lead to osteoporosis (loss of bone mass) or
to the concept of Bthe mononuclear phagocyte system^ pro- osteopetrosis (excess bone mineralisation). Further, bone re-
posed by van Furth et al. 1974 has been questioned by recent modelling controls Ca2+ fluxes into and from the extracellular
studies demonstrating that in many anatomical sites, tissue fluid which is required for the maintenance of the levels of
macrophages originate from precursors derived from the yolk calcium in blood [4]. Osteoclasts belong to the myeloid line-
sac during early embryogenesis with a second wave of embry- age, are M-CSF (CSF-1)-dependent and are multinucleated.
onic precursors derived from foetal liver. This embryonic or- Osteoclastogenesis is dependent on receptor activator of
igin contrasts with the bone marrow origin for most immune NFκB ligands (RANKL). Osteoclasts attach to the bone sur-
cells [18, 26, 33]. Definitive haematopoiesis takes place in the face and assume a polarised morphology. The area attached to
bone marrow after birth. These findings were accompanied by the bone is sealed, bone is degraded by the release of acid and
the identification in the mouse of two populations of circulat- acidic collagenolytic enzymes and released material is secret-
ing monocytes: classical Ly6chi monocytes that originate di- ed at the opposite side of the cell through transcytosis [4].
rectly from Ly6c + precursors and non-classical Ly6clow Exchange of signals between osteoblasts and osteoclasts en-
monocytes that derive from Ly6chigh monocytes through the sures that bone remodelling is regulated [64]. For instance,
action of the transcription factor Nr4a1 [18]. Further, mono- osteoblast apoptosis upon bone damage promotes osteoclast
cytes are now perceived to be not just transitional cells on their recruitment to the bone surface and osteoblast-derived
way to tissues; some authors refer to circulating Ly6Clo mono- RANKL is essential for osteoclast differentiation. IL-33 pro-
cytes as Bblood macrophages^ involved in the maintenance of duced by osteoblasts inhibits osteoclast formation. Similarly,
the integrity of the endothelial barrier [33]. Ly6Chigh mono- osteoclasts influence osteoblast function through the produc-
cytes contribute to replenishment of tissue macrophages with tion of cardiotrophin-1 and semaphorin 4D (CD100), which
the rate of replenishment varying among anatomical sites. In promote and inhibit bone formation, respectively [64]. As a
the steady state, monocyte-derived tissue macrophages are consequence of lack of bone resorption, bone marrow
almost non-existent in the case of brain as microglia are de- haematopoiesis does not occur due to lack of its anatomical
rived from yolk sac precursors. Similar scenarios have been niche. Csf1-deficient animals develop osteopetrosis and sur-
observed in the case of Langerhans cells in the epidermis, vive to adulthood because of extramedullary haematopoiesis
alveolar macrophages in the lung and Kupffer cells in the liver. in the spleen and liver and generation of osteoclasts takes
Monocyte-derived resident macrophages are more prevalent place by compensatory expression of VEGF. Osteoclast gen-
in the heart and pancreas (which display a slow replacement eration enables the establishment of bone marrow
rate) and gut and dermis (which display a higher replacement haematopoiesis by enabling the creation of the haematopoietic
rate) [26, 63]. niche [54].
Pflugers Arch - Eur J Physiol

Remodelling deficiencies in the absence of macrophages Macrophages and lung homeostasis

have been observed in mammary glands, kidney and pancreas,
which suggest an important role in morphogenesis; this can be Alveoli are lined by the alveolar epithelium which is mainly
due to the contribution of macrophages to extracellular matrix composed of type I and type II alveolar epithelial cells. Type I
remodelling and a role in maintenance of the viability and epithelial cells cover 90% of the alveoli surface while type II
function of stem cells [73], as observed in mammary glands epithelial cells are less abundant and mediate the synthesis,
and liver. secretion and recycling of lung surfactant [5, 46]. Alveolar
surfactant is composed of lipids and proteins and keeps alveoli
Macrophages and erythropoiesis open by reducing surface tension during breathing. Alveolar
macrophages reside in the alveoli and establish close interac-
Definitive erythropoiesis occurs in the bone marrow; in par- tions with alveolar epithelial cells, some of which involve
ticular, terminal erythropoiesis takes place in erythroblastic interactions such as CD200R-CD200 and CD47-SIPRα as
islands where tight clusters can be found between erythroid inhibitory receptors. Surfactant proteins A and D also contrib-
precursors with a central macrophage [8]. Molecules implicat- ute to inhibition of cellular activation in alveolar macrophages
ed in the interaction between erythroid precursors and macro- by engaging SIRPα [37]. Alveolar macrophages are derived
phages include the erythroblast macrophage protein (Emp), from embryonically derived foetal monocytes soon after birth
the haemoglobin-haptoglobin receptor CD163, VLA-4 on through a process highly dependent on GM-CSF [18]. They
erythroblasts and VCAM-1 on macrophages, and ICAM-4 replicate in situ and, thus, are independent from monocyte
on erythroblasts and αV integrin on macrophages. input in most instances [18]. Alveolar macrophages mediate
Erythroblastic islands have also been found in areas of defin- 20–30% of surfactant lipid metabolism which explains the
itive haematopoiesis such as foetal liver and splenic red pulp. development of pulmonary alveolar proteinosis in GM-CSF-
Macrophages have been shown to promote erythropoiesis deficient mice and in mice lacking the ABC transporters
through the release of ferritin by exocytosis, which is (ABC-A1, ABCG1 or peroxisome proliferator-activating re-
internalised by erythroblasts where iron is released through ceptor gamma (PPAR- ) which display inefficient surfactant
acidification and proteolysis [15]; factors associated with catabolism in alveolar macrophages. These same genes are
chronic inflammation such as IFN- , TNF-α, TGF-β and IL- involved in cholesterol metabolism and transport by circulat-
6 inhibit erythropoiesis [15]. Erythropoiesis is completely de- ing monocytes and their deficiency has been linked to the
pendent on the phagocytic activity of macrophages. development of atherosclerosis [46].
Macrophages phagocytose extruded erythrocyte nuclei which
are released during the final phase of erythrocyte differentia-
tion [15]. This is possible through the maintenance of adhe- Macrophages and metabolism
sion molecules such as Emp and β1 integrin and exposure of
phosphatidyl serine at the surface of the membrane-enveloped Macrophages and metabolic syndrome
nuclei [15].
The involvement of macrophages in metabolic homeostasis
Macrophages and brain development was highlighted by major changes in macrophage numbers
(10–15 to 45–60% of stromal cells) and phenotype (shift from
Microglia constitute the resident macrophage population in alternative activation to classical activation) in white adipose
the brain and spinal cord and are yolk sac-derived. In addition tissue in obese insulin-resistant animals and humans [9, 17,
to responding to tissue injury, ramified microglia have been 28, 49, 57]. These changes have also been observed in other
shown to contribute to synaptic maturation during brain de- metabolic tissues, liver and muscle, which implies a strong
velopment [58]. This process is termed Bsynaptic pruning^ functional link between macrophage phenotype and parame-
and involves the clearance of immature or defective neuronal ters of tissue homeostasis [49]. This is in agreement with mac-
synapses [1]; microglia dysfunction is now considered a major rophages contributing actively to maintenance of tissue ho-
cause of neuropsychiatric or neurological disorders as exem- meostasis, but also acting as sensors of tissue dysfunction
plified by the link between mutations in the gene encoding and promoters of inflammation in the absence of infection.
CSF1R to hereditary diffuse leukoencephalopathy with spher- In the case of the metabolic syndrome, the central role of
oids [1]. Loss of function mutations in TREM-2, a receptor macrophages derives in part from the negative impact of in-
expressed by microglia in the CNS, have been associated with flammation on signalling events downstream of the insulin
increased risk of developing late-onset Alzheimer’s disease receptor, making the cells within metabolic tissues resistant
[75]. Microglia protect against extension of amyloid fibrils to the action of insulin [49]. This effect in turn will lead to
by wrapping around amyloid-β plaques [11] and TREM-2 maladaptation in glucose metabolism, with increased insulin
deficiency has been shown to disrupt this process [75]. production by islet cells in the pancreas required to
 Pflugers Arch - Eur J Physiol

compensate for this inflammation-related effect and establish- CD47 and thrombospondin-1 which results in increased up-
ment of a pre-diabetic state [49]. In healthy white adipose take by red pulp macrophages [15]. Splenic macrophages
tissue, insulin sensitivity in adipocytes is maintained through have also been implicated in the removal of intracellular in-
the action of resident macrophages that express the anti- clusions in erythrocytes through a process that leaves the
inflammatory cytokine IL-10 acting as insulin sensitiser. erythrocyte intact. These inclusion bodies are thought to be
These adipose-tissue macrophages have a phenotype consis- caused by oxidative damage [15].
tent with alternative activation resulting from exposure to IL-4
and IL-13. PPAR- expression by macrophages is essential for
control of alternative activation and improves insulin resis- Macrophages and tissue regeneration
tance [56]; eosinophils have been identified as the source of
IL-4 in adipose tissue and lack of eosinophils has been asso- Self-contained inflammation with preservation of organ func-
ciated with activation of adipose tissue macrophages and a tion depends on multifaceted interactions between macro-
tendency to develop insulin resistance in response to obesity phages and recruited immune cells, particularly neutrophils
[72]. Changes in macrophage phenotype in white adipose tis- and monocytes that ensure timely resolution through changes
sue after chronic increase in energy uptake occur in response in the activation state of macrophages (from a M1 to a M2-like
to metabolic stress in the adipocytes; this leads to the recruit- phenotype). These constitute a Blipid mediator class switch^
ment of inflammatory monocytes by CCL2, CCL5 and CCL8 which shifts the production of lipid mediators from pro-
[73]. These inflammatory monocytes differentiate into classi- inflammatory (for instance the eicosanoids prostaglandin E2,
cally activated macrophages, engulf dying adipocytes and fur- and leukotriene B4 that promote vasodilatation and chemo-
ther promote cellular activation. Cytokine production by these taxis) to pro-resolving lipids (such as lipoxins, resolvins,
activated macrophages is considered major contributors to protectins and maresins), production of proteins such as
local and systemic insulin resistance. In the case of hepatic annexin A1, gaseous mediators (carbon monoxide), chemo-
insulin resistance, Kupffer cells regulate the oxidation of fatty kine inactivation and release of acetylcholine by the vagus
acids in hepatocytes; hepatic steatosis develops if Kupffer nerve [34]. Resident macrophages dominate the induction of
cells are unable to adopt an alternative activation phenotype inflammatory responses in response to injury in the presence
because of loss of PPAR-δ in myeloid cells [73]. and absence of infection through activation of receptors by
DAMPs and MAMPs. After stimulation, macrophages pro-
Macrophages and iron recycling duce an array of cytokines such as TNF-α, IL-1β and IL-6
and chemokines as well as pro-inflammatory eicosanoids de-
Resident bone marrow macrophages and red pulp macro- rived from arachidonic acid, an omega-6 fatty acid.
phages are essential for the process of erythrophagocytosis Vasodilation and edema are followed by recruitment of neu-
to recycle iron. Senescent or damaged erythrocytes are phago- trophils which are the first cells that enter the inflamed site.
cytosed and digested to extract heme [22]. Upon uptake, heme Neutrophil recruitment is favoured by the rapid exposure of P-
is degraded through the action of heme oxygenase (HO; selectin (CD62P) on activated endothelial cells followed by
predominantly HO-1) to release iron into the cytoplasm. engagement of β2 integrins by the adhesion molecules
Cytoplasmic iron is exported by ferroportin located at the ICAM-1 and ICAM-2, culminating in the process of diapede-
plasma membrane [22, 33]. Iron flux from macrophages ex- sis. Cues for directional cell migration are provided by the
ceeds iron adsorption through diet and iron stored in hepato- formation of chemokine gradients that guide the migrating
cytes [22]. Intriguingly, heme itself has been shown to induce neutrophil to the site of injury. Resolution of inflammation is
the development of F4/80hi, VCAMhi iron recycling macro- now viewed as an active process and its de-regulation can
phages in bone marrow and spleen through induction of the hamper restoration of tissue function with lack of resolution
transcription factor Spi-c [33]. Red pulp macrophages also leading to chronic inflammation and scar formation and fibro-
express Nramp1 and the haptoglobin-haemoglobin receptor sis. The collaborative nature of the resolution process is illus-
(CD163) which further emphasises their important role in sys- trated by the requirement for transcellular metabolic events
temic iron haemostasis [22]. There are several hypotheses between leukocytes and epithelial cells and platelets for the
regarding the signals recognised on aged erythrocytes by mac- synthesis of lipoxins, the first described pro-resolving lipids.
rophages for their removal. These signals include the exposure Because of their potential for tissue damage, it appears coun-
of the universal apoptotic marker phosphatidyl serine in se- terintuitive to consider neutrophils’ essential partners for the
nescent erythrocytes, in spite of the fact that erythrocytes lack resolution of inflammation. However, neutrophils promote
a cell nucleus and mitochondria and do not undergo canonical resolution by enhancing monocyte recruitment by inducing
apoptotic process [15]. Other possibilities include the loss of changes in endothelial cells, for instance through shedding
CD47 expression, i.e. lack of an inhibitory signal, and confor- of IL-6Rα which complexes with IL-6. The IL-6Rα-IL-6
mational changes in CD47 that enable an interaction between complex engages gp130 on endothelial cells and induces the
Pflugers Arch - Eur J Physiol

production of E-selectin (CD62E) and VCAM1, thus promot- have been found to produce Wnt7b which promotes repair of
ing monocyte recruitment. In addition, apoptotic neutrophils basement membrane [10].
change macrophage activation as uptake of apoptotic neutro- Scar formation during wound healing can compromise or-
phils is non-phlogistic, inducing IL-10, TGF-β and lipoxin gan function after injury and multiple reports have linked Th2
synthesis by macrophages [66]. These changes are associated immunity to the regulation of tissue fibrosis, although the
with the sensing of an Binflammation threshold^ established molecular processes of this effect are poorly understood [24,
early during the induction phase and highly dependent on the 74]. Knipper et al. [42] recently revealed an important cross-
nature and extent of the original insult. Molecular mechanisms talk between macrophages and fibroblasts during wound
underpinning this concept include the fact that prostaglandin healing in mouse injured skin, required to control the forma-
E2 induces changes in the lipoxygenase activity of neutrophils tion of collagen fibrils. This effect is dependent on the expres-
from 5-lipoxygenase to 15-lipoxygenase, required for synthe- sion of IL-4-Rα by myeloid cells which, among others, in-
sis of lipoxins [34]. During cardiac damage, inflammatory duces the production of Relm-α, a well-characterised marker
neutrophils and monocytes release oncostatin-M that induces of macrophage activation in response to IL-4. IL-4Rα signal-
the release by cardiomyocytes of Reg3β. Reg3β promotes the ling during wound healing promotes the expression of
recruitment of reparative macrophages which favours the Relm-α by macrophages, and Relm-α, in turn, induces the
elimination of neutrophils [26]. In addition, macrophages con- synthesis of the collagen-modifying enzyme lysyl hydroxy-
trol the neutrophil lifespan through the release of TNF-α, Fas lase 2 by fibroblasts, leading to fibrosis [42]. In line with the
ligand (FasL) and TRAIL [34]. major influence of the tissue environment on macrophage bi-
The fate of the recruited monocytes and their contribution ology, in liver and muscle, Th2 immunity promotes tissue
to repair depends on the organ and extent of damage. In the regeneration through direct action on parenchymal cells [27,
heart, recruited monocytes give rise to pro-inflammatory mac- 36]; this suggests that macrophage contribution to wound
rophages, while resident macrophages mediate tissue repair. healing is largely dependent on the intrinsic regenerative ca-
In the spinal cord, both the pro-inflammatory and repair mac- pacity of the tissue and, likely, the nature of the insult.
rophages are monocyte-derived. While the pro-inflammatory
macrophages derive from monocytes recruited through the
CCR2-CCL2 pathway, repair-promoting macrophages de- Macrophages and thermogenesis
rived from monocytes are recruited through a pathway involv-
ing VLA-4, VCAM-1 and CD73 [26]. In other scenarios, Brown fat, termed brown adipose tissue, generates heat
recruited monocytes have been shown to convert from an (thermogenesis) and plays an important role in maintaining
initial pro-inflammatory (M1) phenotype to a healing, pro- body temperature. In contrast to white adipocytes, brown ad-
resolving (M2) phenotype [14, 60] consistent with plasticity ipocytes contain multilocular fat droplets, a large number of
of myeloid cells. This scenario has been observed in kidney mitochondria, and express high levels of UCP1 (uncoupling
[45] and muscle [3]. protein 1). UCP1 is located in the inner membrane of the
A key aspect during tissue regeneration and repair is the mitochondria and uncouples electron transport to promote
cross-talk between macrophages and stem cells. For instance, heat instead of ATP in brown adipocytes. In humans, brown
in liver, macrophage depletion has been shown to reduce in- adipose tissue was thought to be restricted to neonates but
vasiveness of liver progenitor cells into the parenchyma which recent data have shown the presence of brown fat in adults
is required for differentiation into hepatocytes [10]. In addi- [12, 41, 69]. Adult human brown fat is dispersed in the
tion, macrophages can be induced to produce Wnt3a after supraclavicular, para-aortic and suprarenal regions, is cold-
uptake of hepatic debris. Wnt3a promotes expression of inducible and shares many characteristic with the cold-
Numb in liver progenitor cells which inhibits Notch and pro- inducible beige fat found in the subcutaneous white adipose
motes differentiation into hepatocytes instead of biliary cells tissue of rodents [61]. Beige fat refers to the presence of white
[10]. Macrophages contribute to muscle regeneration with the adipocytes functionally similar to brown adipocytes within
sequential change from an early M1-like phenotype into a white fat and promotion of beige fat formation is considered
M2-like phenotype, essential for this process. Disruption of as a potential therapeutic tool in the fight against obesity [41].
these changes in the phenotype of macrophages by blocking Macrophages have been shown to be essential for adaptation
early inflammation (treatment with IL-10 or blocking IFN- ) to cold exposure that leads to lipolysis in white fat and ther-
or late resolution (treatment with anti-IL-10) compromises mogenesis in brown fat. The effect of macrophages depends
muscle regeneration. Myogenic precursor cells are stimulated on the acquisition of an alternative activated phenotype driven
to migrate, grow and survive by macrophages with M1 cells by lL-4/IL-13 and Stat-6 and that leads to secretion of cate-
stimulating proliferation and inhibiting fusion and M2 macro- cholamines by these cells [53]. Indeed, IL-4 exposure of mac-
phages stimulating commitment of myogenic precursors and rophages induces production of the three enzymes required for
differentiation to form myotubes [10]. In kidney, macrophages catecholamine production: tyrosine hydroxylase, dopa
 Pflugers Arch - Eur J Physiol

decarboxylase and dopamine β-hydroxylase [53]; the β3- 7. Brown GC, Vilalta A, Fricker M (2015) Phagoptosis—cell death by
phagocytosis—plays central roles in physiology, host defense and
adrenergic agonist CL-316246 was able to rescue the thermo-
pathology. Curr Mol Med 15:842–851
genic defect in animals unable to produce IL-4 and IL-13 [53]. 8. Chasis JA, Mohandas N (2008) Erythroblastic islands: niches for
In agreement with these findings in mice, oral administration erythropoiesis. Blood 112:470–478
of the β3-adrenergic receptor agonist mirabegron can stimu- 9. Chawla A, Nguyen KD, Goh YP (2011) Macrophage-mediated
late human brown fat thermogenesis in humans [13]. inflammation in metabolic disease. Nat Rev Immunol 11:738–749
10. Chazaud B (2014) Macrophages: supportive cells for tissue repair
Macrophages, together with eosinophils and Th2 cytokines, and regeneration. Immunobiology 219:172–178
have also been implicated in the remodelling of subcutaneous 11. Condello C, Yuan P, Schain A, Grutzendler J (2015) Microglia
white fat into beige fat in mice [61]. constitute a barrier that prevents neurotoxic protofibrillar Abeta42
hotspots around plaques. Nat Commun 6:6176
12. Cypess AM, Lehman S, Williams G, Tal I, Rodman D, Goldfine
AB, Kuo FC, Palmer EL, Tseng YH, Doria A, Kolodny GM, Kahn
CR (2009) Identification and importance of brown adipose tissue in
Conclusion and therapeutic opportunities adult humans. N Engl J Med 360:1509–1517
13. Cypess AM, Weiner LS, Roberts-Toler C, Franquet Elia E, Kessler
Macrophages are remarkable for their ability to sense their SH, Kahn PA, English J, Chatman K, Trauger SA, Doria A,
surroundings, produce a wide range of immune mediators Kolodny GM (2015) Activation of human brown adipose tissue
by a beta3-adrenergic receptor agonist. Cell Metab 21:33–38
alongside growth factors and opsonins, and their endocytic
14. Das A, Sinha M, Datta S, Abas M, Chaffee S, Sen CK, Roy S
and phagocytic activity. The recognition of macrophages as (2015) Monocyte and macrophage plasticity in tissue repair and
intrinsic components of organs, adapted to the particular tis- regeneration. Am J Pathol 185:2596–2606
sue environment and actively involved in homeostatic pro- 15. de Back DZ, Kostova EB, van Kraaij M, van den Berg TK, van
cesses, opens the opportunity for macrophage-centred ther- Bruggen R (2014) Of macrophages and red blood cells; a complex
love story. Front Physiol 5:9
apies that can be exploited to steer pathological processes 16. Deretic V (2016) Autophagy in leukocytes and other cells: mecha-
towards outcomes that favour maintenance of organ func- nisms, subsystem organization, selectivity, and links to innate im-
tion. Furthermore, the potential plasticity shown by these munity. J Leukoc Biol 100:969–978
cells [44] makes it possible to envisage the possibility of 17. Donath MY, Shoelson SE (2011) Type 2 diabetes as an inflamma-
tory disease. Nat Rev Immunol 11:98–107
using in vitro-differentiated cells to treat disease by adoptive
18. Epelman S, Lavine KJ, Randolph GJ (2014) Origin and functions of
transfer. tissue macrophages. Immunity 41:21–35
19. Fleming BD, Mosser DM (2011) Regulatory macrophages: setting
the threshold for therapy. Eur J Immunol 41:2498–2502
Open Access This article is distributed under the terms of the Creative 20. Franchi L, Kamada N, Nakamura Y, Burberry A, Kuffa P, Suzuki S,
Commons Attribution 4.0 International License (http:// Shaw MH, Kim YG, Nunez G (2012) NLRC4-driven production of
creativecommons.org/licenses/by/4.0/), which permits unrestricted use, IL-1beta discriminates between pathogenic and commensal bacteria
distribution, and reproduction in any medium, provided you give appro- and promotes host intestinal defense. Nat Immunol 13:449–456
priate credit to the original author(s) and the source, provide a link to the 21. Freeman SA, Goyette J, Furuya W, Woods EC, Bertozzi CR,
Creative Commons license, and indicate if changes were made. Bergmeier W, Hinz B, van der Merwe PA, Das R, Grinstein S
(2016) Integrins form an expanding diffusional barrier that coordi-
nates phagocytosis. Cell 164:128–140
22. Ganz T (2012) Macrophages and systemic iron homeostasis. J
References Innate Immun 4:446–453
23. Gaudet RG, Bradfield CJ, and MacMicking JD 2016 Evolution of
cell-autonomous effector mechanisms in macrophages versus non-
1. Amit I, Winter DR, Jung S (2016) The role of the local environment immune cells. Microbiol Spectr 4
and epigenetics in shaping macrophage identity and their effect on 24. Gause WC, Wynn TA, Allen JE (2013) Type 2 immunity and
tissue homeostasis. Nat Immunol 17:18–25 wound healing: evolutionary refinement of adaptive immunity by
2. Arandjelovic S, Ravichandran KS (2015) Phagocytosis of apoptotic helminths. Nat Rev Immunol 13:607–614
cells in homeostasis. Nat Immunol 16:907–917 25. Gautier EL, Shay T, Miller J, Greter M, Jakubzick C, Ivanov S,
3. Arnold L, Henry A, Poron F, Baba-Amer Y, van Rooijen N, Helft J, Chow A, Elpek KG, Gordonov S, Mazloom AR, Ma'ayan
Plonquet A, Gherardi RK, Chazaud B (2007) Inflammatory mono- A, Chua WJ, Hansen TH, Turley SJ, Merad M, Randolph GJ,
cytes recruited after skeletal muscle injury switch into Immunological GC (2012) Gene-expression profiles and transcrip-
antiinflammatory macrophages to support myogenesis. J Exp tional regulatory pathways that underlie the identity and diversity of
Med 204:1057–1069 mouse tissue macrophages. Nat Immunol 13:1118–1128
4. Bar-Shavit Z (2007) The osteoclast: a multinucleated, hematopoi- 26. Ginhoux F, Guilliams M (2016) Tissue-resident macrophage ontog-
etic-origin, bone-resorbing osteoimmune cell. J Cell Biochem 102: eny and homeostasis. Immunity 44:439–449
1130–1139 27. Goh YP, Henderson NC, Heredia JE, Red Eagle A, Odegaard JI,
5. Bhattacharya J, Westphalen K (2016) Macrophage-epithelial inter- Lehwald N, Nguyen KD, Sheppard D, Mukundan L, Locksley RM,
actions in pulmonary alveoli. Semin Immunopathol 38:461–469 Chawla A (2013) Eosinophils secrete IL-4 to facilitate liver regen-
6. Blander JM, Sander LE (2012) Beyond pattern recognition: five eration. Proc Natl Acad Sci U S A 110:9914–9919
immune checkpoints for scaling the microbial threat. Nat Rev 28. Gordon S (2007) Macrophage heterogeneity and tissue lipids. J
Immunol 12:215–225 Clin Invest 117:89–93
Pflugers Arch - Eur J Physiol

29. Gordon S (2016) Phagocytosis: the legacy of Metchnikoff. Cell 52. Murray PJ, Allen JE, Biswas SK, Fisher EA, Gilroy DW, Goerdt S,
166:1065–1068 Gordon S, Hamilton JA, Ivashkiv LB, Lawrence T, Locati M,
30. Gotts JE, Matthay MA (2016) Sepsis: pathophysiology and clinical Mantovani A, Martinez FO, Mege JL, Mosser DM, Natoli G,
management. BMJ 353:i1585 Saeij JP, Schultze JL, Shirey KA, Sica A, Suttles J, Udalova I,
31. Groves E, Dart AE, Covarelli V, Caron E (2008) Molecular mech- van Ginderachter JA, Vogel SN, Wynn TA (2014) Macrophage
anisms of phagocytic uptake in mammalian cells. Cell Mol Life Sci activation and polarization: nomenclature and experimental guide-
65:1957–1976 lines. Immunity 41:14–20
32. Guo H, Callaway JB, Ting JP (2015) Inflammasomes: mechanism 53. Nguyen KD, Qiu Y, Cui X, Goh YP, Mwangi J, David T,
of action, role in disease, and therapeutics. Nat Med 21:677–687 Mukundan L, Brombacher F, Locksley RM, Chawla A (2011)
33. Haldar M, Murphy KM (2014) Origin, development, and homeo- Alternatively activated macrophages produce catecholamines to
stasis of tissue-resident macrophages. Immunol Rev 262:25–35 sustain adaptive thermogenesis. Nature 480:104–108
34. Headland SE, Norling LV (2015) The resolution of inflammation: 54. Niida S, Kaku M, Amano H, Yoshida H, Kataoka H, Nishikawa S,
principles and challenges. Semin Immunol 27:149–160 Tanne K, Maeda N, Nishikawa S, Kodama H (1999) Vascular en-
35. Helming L, Winter J, Gordon S (2009) The scavenger receptor dothelial growth factor can substitute for macrophage colony-
CD36 plays a role in cytokine-induced macrophage fusion. J Cell stimulating factor in the support of osteoclastic bone resorption. J
Sci 122:453–459 Exp Med 190:293–298
36. Heredia JE, Mukundan L, Chen FM, Mueller AA, Deo RC, 55. O'Neill LA, Pearce EJ (2016) Immunometabolism governs dendrit-
Locksley RM, Rando TA, Chawla A (2013) Type 2 innate signals ic cell and macrophage function. J Exp Med 213:15–23
stimulate fibro/adipogenic progenitors to facilitate muscle regener- 56. Odegaard JI, Ricardo-Gonzalez RR, Goforth MH, Morel CR,
ation. Cell 153:376–388 Subramanian V, Mukundan L, Red Eagle A, Vats D, Brombacher
37. Hussell T, Bell TJ (2014) Alveolar macrophages: plasticity in a F, Ferrante AW, Chawla A (2007) Macrophage-specific
tissue-specific context. Nat Rev Immunol 14:81–93 PPARgamma controls alternative activation and improves insulin
38. Janssen WJ, Bratton DL, Jakubzick CV, and Henson PM 2016 resistance. Nature 447:1116–1120
Myeloid Cell Turnover and Clearance. Microbiol Spectr 4 57. Olefsky JM, Glass CK (2010) Macrophages, inflammation, and
39. Jaumouille V, and Grinstein S 2016 Molecular mechanisms of insulin resistance. Annu Rev Physiol 72:219–246
phagosome formation. Microbiol Spectr 4 58. Paolicelli RC, Bolasco G, Pagani F, Maggi L, Scianni M, Panzanelli
40. Kaur M, Bell T, Salek-Ardakani S, Hussell T (2015) Macrophage P, Giustetto M, Ferreira TA, Guiducci E, Dumas L, Ragozzino D,
adaptation in airway inflammatory resolution. Eur Respir Rev 24: Gross CT (2011) Synaptic pruning by microglia is necessary for
510–515 normal brain development. Science 333:1456–1458
41. Kim SH, Plutzky J (2016) Brown fat and browning for the treat- 59. Pastores GM, Hughes DA (2015) Non-neuronopathic lysosomal
ment of obesity and related metabolic disorders. Diabetes Metab J storage disorders: disease spectrum and treatments. Best Pract Res
40:12–21 Clin Endocrinol Metab 29:173–182
42. Knipper JA, Willenborg S, Brinckmann J, Bloch W, Maass T,
60. Porcheray F, Viaud S, Rimaniol AC, Leone C, Samah B,
Wagener R, Krieg T, Sutherland T, Munitz A, Rothenberg ME,
Dereuddre-Bosquet N, Dormont D, Gras G (2005) Macrophage
Niehoff A, Richardson R, Hammerschmidt M, Allen JE, Eming
activation switching: an asset for the resolution of inflammation.
SA (2015) Interleukin-4 receptor alpha signaling in myeloid cells
Clin Exp Immunol 142:481–489
controls collagen fibril assembly in skin repair. Immunity 43:803–
61. Qiu Y, Nguyen KD, Odegaard JI, Cui X, Tian X, Locksley RM,
816
Palmiter RD, Chawla A (2014) Eosinophils and type 2 cytokine
43. Lambeth JD (2004) NOX enzymes and the biology of reactive
signaling in macrophages orchestrate development of functional
oxygen. Nat Rev Immunol 4:181–189
beige fat. Cell 157:1292–1308
44. Lavin Y, Winter D, Blecher-Gonen R, David E, Keren-Shaul H,
Merad M, Jung S, Amit I (2014) Tissue-resident macrophage en- 62. Sheikh F, Dickensheets H, Pedras-Vasconcelos J, Ramalingam T,
hancer landscapes are shaped by the local microenvironment. Cell Helming L, Gordon S, Donnelly RP (2015) The interleukin-13
159:1312–1326 receptor-alpha1 chain is essential for induction of the alternative
45. Lee S, Huen S, Nishio H, Nishio S, Lee HK, Choi BS, Ruhrberg C, macrophage activation pathway by IL-13 but not IL-4. J Innate
Cantley LG (2011) Distinct macrophage phenotypes contribute to Immun 7:494–505
kidney injury and repair. J Am Soc Nephrol 22:317–326 63. Sieweke MH, Allen JE (2013) Beyond stem cells: self-renewal of
46. Lopez-Rodriguez E, Gay-Jordi G, Mucci A, Lachmann N, Serrano- differentiated macrophages. Science 342:1242974
Mollar A 2016 Lung surfactant metabolism: early in life, early in 64. Sims NA, Walsh NC (2012) Intercellular cross-talk among bone
disease and target in cell therapy. Cell Tissue Res cells: new factors and pathways. Curr Osteoporos Rep 10:109–117
47. Martinez FO, Gordon S (2014) The M1 and M2 paradigm of mac- 65. Smythies LE, Sellers M, Clements RH, Mosteller-Barnum M,
rophage activation: time for reassessment. F1000Prime Rep 6:13 Meng G, Benjamin WH, Orenstein JM, Smith PD (2005) Human
48. Martinez FO, Helming L, Milde R, Varin A, Melgert BN, Draijer C, intestinal macrophages display profound inflammatory anergy de-
Thomas B, Fabbri M, Crawshaw A, Ho LP, Ten Hacken NH, Cobos spite avid phagocytic and bacteriocidal activity. J Clin Invest 115:
Jimenez V, Kootstra NA, Hamann J, Greaves DR, Locati M, 66–75
Mantovani A, Gordon S (2013) Genetic programs expressed in 66. Soehnlein O, Lindbom L (2010) Phagocyte partnership during the
resting and IL-4 alternatively activated mouse and human macro- onset and resolution of inflammation. Nat Rev Immunol 10:427–
phages: similarities and differences. Blood 121:e57–e69 439
49. McNelis JC, Olefsky JM (2014) Macrophages, immunity, and met- 67. Teti G, Biondo C, and Beninati C 2016. The phagocyte,
abolic disease. Immunity 41:36–48 Metchnikoff, and the foundation of immunology. Microbiol
50. Mosser DM, Edwards JP (2008) Exploring the full spectrum of Spectr 4
macrophage activation. Nat Rev Immunol 8:958–969 68. van de Veerdonk FL, Dinarello CA (2014) Deficient autophagy
51. Muller WA, Steinman RM, Cohn ZA (1983) Membrane proteins of unravels the ROS paradox in chronic granulomatous disease.
the vacuolar system. III. Further studies on the composition and Autophagy 10:1141–1142
recycling of endocytic vacuole membrane in cultured macrophages. 69. van Marken Lichtenbelt WD, Vanhommerig JW, Smulders NM,
J Cell Biol 96:29–36 Drossaerts JM, Kemerink GJ, Bouvy ND, Schrauwen P, Teule GJ
 Pflugers Arch - Eur J Physiol

(2009) Cold-activated brown adipose tissue in healthy men. N Engl 73. Wynn TA, Chawla A, Pollard JW (2013) Macrophage biology in
J Med 360:1500–1508 development, homeostasis and disease. Nature 496:445–455
70. Vance RE, Isberg RR, Portnoy DA (2009) Patterns of pathogenesis: 74. Wynn TA, Ramalingam TR (2012) Mechanisms of fibrosis: thera-
discrimination of pathogenic and nonpathogenic microbes by the peutic translation for fibrotic disease. Nat Med 18:1028–1040
innate immune system. Cell Host Microbe 6:10–21 75. Yuan P, Condello C, Keene CD, Wang Y, Bird TD, Paul SM, Luo
71. Vieira OV, Botelho RJ, Grinstein S (2002) Phagosome maturation: W, Colonna M, Baddeley D, Grutzendler J (2016) TREM2
aging gracefully. Biochem J 366:689–704 haplodeficiency in mice and humans impairs the microglia barrier
72. Wu D, Molofsky AB, Liang HE, Ricardo-Gonzalez RR, Jouihan function leading to decreased amyloid compaction and severe axo-
HA, Bando JK, Chawla A, Locksley RM (2011) Eosinophils sus- nal dystrophy. Neuron 92:252–264
tain adipose alternatively activated macrophages associated with
glucose homeostasis. Science 332:243–247