Neutrophil Biology

C Carmona-Rivera and MJ Kaplan, National Institutes of Health, Bethesda, MD, USA

Published by Elsevier Inc.

Introduction This article summarizes important components of neutro-

phil biology, including their development and microbicidal
Neutrophils are sentinel cells and are the first to arrive at sites of mechanisms and their potential pathogenic role.
infection to defend the host from invading microbes. The bone
marrow is the site of neutrophil production. Under conditions
of homeostasis, neutrophils can be found in the bone marrow,
lung, spleen, and liver. Typically, neutrophils have a very short Neutrophil Development
half-life and therefore are produced at a rate of 5–10
1010
cells day1 to maintain steady-state. Neutrophil production is Neutrophils originate in the bone marrow and enter the cir-
controlled by the synthesis of granulocyte-colony stimulating culation as terminally differentiated cells. Various complex
factor (G-CSF) in response to Interleukin-17A. During inflam- mechanisms and mediators are involved in the development of
mation, the longevity of neutrophils increases to ensure their neutrophil precursors and in their terminal differentiation.
presence at sites of damage. Neutrophils that die by apoptosis at Among them are cytokines, growth factors, microRNAs, and
sites of inflammation are removed by phagocytic cells such as transcriptional regulation. Balance between the transcription
macrophages and dendritic cells. Neutrophils are non-mitotic factors C/EBPα (CCAAT/enhancer binding protein α) and PU.1
cells and display a multilobulated nucleus, hence the alternative determine granulocytopoiesis commitment (Reddy et al., 2002;
name of polymorphonuclear cells (PMNs) (Figure 1). In cir- Dahl et al., 2003; Laslo et al., 2006). C/EBPα regulates granu-
culation, neutrophils remain in a resting state, which ensures locytic differentiation by competing with the transcription fac-
their potent toxic arsenals are not accidentally released, dam- tor NFI-A to modulate miR223 expression (Fazi et al., 2005).
aging the host. They are recruited from the circulation to the Other important factors include Gfi-1 (growth factor in-
tissue in response to microbes or other inflammatory stimuli. In dependent-1) and C/EBPε (Borregaard, 2010). Gfi-1 represses
tissue, they employ different mechanisms to damage pathogens. genes involved in proliferation of stem cells and monocyte-
As such, significant decreases in neutrophils can lead to in- promoting transcription factors (Hock et al., 2004; Zarebski
creased risk for severe infections (Klein, 2011), while tight et al., 2008). Neutrophils arise from hematopoietic stem cells
regulation of these cells is crucial to prevent tissue injury. (HSC) and follow a regulated differentiation process as follows:

LL-37 Myosin-9 DNA

10 µm

10 µm

10 µm
10 µm

Figure 1 Neutrophil morphology. Neutrophils have distinct multilobulated nucleus with multiple granules covering their cytoplasm. Neutrophils
isolated from a healthy volunteer express the granular protein LL-37 and cytosolic myosin-9. DNA was co-stained using Hoechst. Scale bar 10 mm.

750 Encyclopedia of Cell Biology, Volume 3 doi:10.1016/B978-0-12-394447-4.30112-2

 Cellular Immunology: Innate Immunity: Neutrophil Biology 751

Myeloblast of the cargo proteins contained in these granules include

defensins, bactericidal/permeability-increasing protein
Myeloblasts have a diameter of 10–20 mm, are derived from
(BPI), neutrophil elastase, myeloperoxidase (MPO),
HSCs and are normally found in the bone marrow. Morpho-
proteinase 3 (PR3), and cathepsin G (Faurschou and
logically, they are characterized by a large round to oval nu-
Borregaard, 2003).
cleus with a small basophilic cytoplasm with no evident
2. Secondary (specific) granules are synthesized during the
granules. During the myeloblast stage, levels of C/EBPα grad-
myelocyte stage and have an approximate diameter of
ually decrease (Bjerregaard et al., 2003). The appearance of
0.1 mm. They are characterized by the presence of the
granules marks the transition from myeloblast to promyelo-
glycoprotein lactoferrin and contain antimicrobial proteins
cyte and C/EBPε regulates the expression of granular proteins
such as neutrophil gelatinase-associated lipocalin (NGAL),
(Yamanaka et al., 1997).
human cationic antimicrobial protein (hCAP-18), and
lysozyme (Faurschou and Borregaard, 2003).
Promyelocyte 3. Tertiary (gelatinase) granules appear during the band stage
(Borregaard et al., 1995; Borregaard and Cowland, 1997).
The morphology of the promyelocyte is similar to that of the They are smaller than specific granules and contain gelati-
myeloblast. Cells are larger and nuclei are round to oval with nases such as leukolysin (matrix metalloproteinase (MMP)-
presence of nucleoli and diffuse chromatin distribution. The 25), MMP-8, and MMP-9.
endoplasmic reticulum (ER) is more prominent and azur-
ophilic granules (primary granules) accumulate in increasing Neutrophils also contain secretory vesicles that are syn-
numbers during this stage. These cells express CD33, CD13, thesized during the last stage of maturation. Protein content is
and CD15 on cell surface and they differentiate into dictated by the transcriptional activity during the granule for-
myelocytes. mation or by ‘targeting by timing of biosynthesis’ (Le Cabec
et al., 1996; Borregaard, 2010).

Myelocyte
During this stage, secondary granules become evident in the Neutrophil Migration into Sites of Inflammation
cytoplasm. Nuclei are round to oval and eccentric, chromatin
is coarse, and nucleoli are small. Formation of primary gran- Once mature, neutrophils are released from the bone marrow
ules decreases, while secondary granules are small and nu- in a tightly chemokine-regulated fashion (Geering and Simon,
merous. CD14 and CD11b become present as surface markers 2011). Chemokine receptors CXCR4 and CXCR2 play a critical
during this stage. role during neutrophil release from the bone marrow.
Retention of mature neutrophils in the bone marrow is asso-
ciated with mutations in CXCR4 or deletion of CXCR2
Metamyelocyte (Hernandez et al., 2003; Eash et al., 2009). Conversely,
Metamyelocytes have a ‘horseshoe-shape’ nucleus without deletion of CXCR4 receptor causes increased release of mature
nucleoli. Chromatin is denser and cytoplasm is filled with neutrophils to the circulation (Eash et al., 2009). Neutrophils
primary, secondary, and tertiary granules (Quesenberry and enter the circulation in a resting state and can become primed
Levitt, 1979; Lajtha et al., 1953). ER becomes thinly dispersed. by danger signals including bacterial products, cytokines and
chemokines. Primed neutrophils are mobilized to the sites of
infection or inflammation where they transmigrate into the
Band (Juvenile) tissues through interactions with the vascular endothelium.
This process involves three steps:
Bands are characterized by further condensation of the chro-
matin, with a ‘sausage-like’ or band conformation. The nucleus
is divided into two or more lobules connected by filamentous Tethering and Rolling
strands of heterochromatin. CD10 and CD16 markers dis-
tinguish bands from more immature forms. Upregulation of endothelial cell adhesion molecules, such as
selectins, initiates neutrophil tethering. This upregulation oc-
curs through mobilization of P-selectins from Weibel–Palade
Mature Neutrophils bodies and by upregulation of de novo synthesis of E-selectins.
Selectins are translocated to the apical membrane of the
The nucleus becomes more segmented (three to four lobules)
endothelial cell, where they transiently bind their respective
indicating full maturation of the neutrophil, which exits the
ligands P-selectin glycoprotein (PSGL-1) and E-selectin ligand-
bone marrow and enters the circulation (Bainton et al., 1971).
1 (ESL-1), expressed by activated neutrophils. These inter-
Granules are sequentially and continuously produced
actions activate intracellular cascades that include Src family
during granulocytic differentiation through vesicle budding
kinases Hck, Fgr, and Lyn that phosphorylate cytosolic pro-
from the Golgi apparatus. These compartments are classified
teins and trigger activation of syk (spleen tysorine kinase).
into three distinct subsets:
Activated syk triggers cytoskeletal rearrangement in the neu-
1. Primary (azurophilic) granules appear during the promye- trophil leading to activation of kinases that result in integrin
locyte stage. They are also known as peroxidase-positive activation (Yago et al., 2010; Ley et al., 2007; Woodfin et al.,
granules and measure around 0.3 mm in diameter. Some 2010; Barreiro and Sanchez-Madrid, 2009).
752 Cellular Immunology: Innate Immunity: Neutrophil Biology

Firm Adhesion Hayashi et al., 2003). Upon stimulation, TLRs trigger a cascade
of intracellular events that induce inflammatory gene ex-
Neutrophil rolling along the endothelium induces conforma-
pression regulated by NF-kB elements (Tamassia et al., 2007).
tional changes in integrin adhesion of very late antigen-4
Microbes are internalized by neutrophils through phago-
(VLA-4; α4β1-integrin, CD49d/CD29b), lymphocyte function-
cytosis. The phagosome fuses with lysosomes to form the
associated antigen-1 (LFA-1; αLβ2-integrin, CD11a/CD18), and
phagolysosome, where the microbe is subsequently killed by
macrophage antigen-1 (MAC-1, αMβ2-integrin, CD11b/CD18)
generation of reactive oxygen species (ROS). ROS production
that bind to adhesion molecules on the endothelial membrane
is initiated by a nicotinamide adenine dinucleotide phos-
such as intercellular adhesion molecule (ICAM)-1 and -2,
phate-(NADPH)-oxidase complex-dependent process called
vascular cell adhesion molecule-1 (VCAM-1), and mucosal
oxidative burst. During this event, oxygen is converted into
vascular cell adhesion molecule-1 (MADCAM-1). LFA-1 is the
superoxide and transformed into hydrogen peroxide by
key molecule that mediates the transition from rolling to ad-
superoxide dismutases. Subsequently, MPO is released into
hesion (Phillipson et al., 2006). These interactions trigger
the phagolysosome through fusion with azurophilic granules,
strong adhesion of the neutrophil to the endothelium,
where it catalyzes the conversion of hydrogen peroxide and
allowing for transmigration.
chloride into hypochlorous acid (Klebanoff et al., 2013).

Transmigration
Degranulation
Neutrophils transmigrate into the affected tissues through the
junctions between endothelial cells (paracellular transmigra- During this process, neutrophil granules translocate to the
tion) assisted by surface ligands ICAM-2, platelet endothelial phagosomal or plasma membrane, where they dock and fuse
cell adhesion molecule-1 (PECAM-1), and proteins of the with the membrane to release their contents. The release of
junctional adhesion molecule (JAM) family (Woodfin et al., granular components is tightly regulated mechanism that leads
2009). Endothelial PECAM-1 interacts with GPI-CD177 (NB1- to exocytosis. Granules' exocytosis requires increases in intra-
antigen) to facilitate transmigration (Sachs et al., 2007). Trans- cellular Ca2 þ and ATP and GTP hydrolysis. Actin cytoskeleton
cellular transmigration accounts for approximately 20% of the reorganization is also required. Granule released from neu-
transmigration mechanisms and occurs when neutrophils trophils depends on activation of intracellular signaling
penetrate individual endothelial cells under conditions of high pathways, including β-arrestins, the Rho guanosine tripho-
ICAM-1 expression and density (Yang et al., 2006). SNARE sphatase Rac2, soluble NSF attachment protein (SNAP) re-
proteins are essential in trafficking and fusion of intracellular ceptors, the src family of tyrosine kinases, and the tyrosine
organelles and exocytosis of granules to guide neutrophils phosphatase MEG2 (Lacy, 2006). Molecules such as neutro-
across the endothelial cell body (Carman et al., 2007). Once phil elastase, cathepsin G, and lysozymes degrade virulence
neutrophils migrate through the endothelium, they are ex- factors and the bacterial cell wall. Antibacterial proteins such
posed to a gradient of chemoattractants such as N-for- as BPI, cathelicidin, and defensins not only have the capacity
mylmethionyl-leucyl-phenylalanine (fMLP) and complement to kill microbes but also can damage the host’s tissues.
5a (C5a). At the sites of infection, neutrophils, along with
complement protein and immunoglobulin, recognize and
bind opsonized bacteria, and execute the various antimicrobial NETs
mechanisms including phagocytosis, degranulation, and neu- During NET formation, large strands of decondensed DNA in
trophil extracellular trap (NET) formation. During these pro- complex with granular proteins are extruded from the cell
cesses, released neutrophil molecules can also damage host (Brinkmann et al., 2004). The precise molecular mechanisms
tissue. Inflammation develops in response to pathogen- leading to NET formation are not yet fully elucidated. Various
associated molecular patterns (PAMPs, such as lipopoly- stimuli can trigger NET formation, including receptor-medi-
saccharide (LPS), flagellin, and nucleic acids) released into the ated signals (e.g., through TLR engagement) or through cal-
tissue during infection, and also in response to damage- cium channel formation through ionophores. These signals
associated molecular pattern molecules (DAMPs, such as heat- initiate a cascade of events, where calcium is released from its
shock proteins, high-mobility group box 1 (HMGB-1), ATP, reservoirs by activation of PKC. This leads to assembly of
and DNA) that are generated in response to sterile injury. NADPH oxidase and ROS generation (Matute et al., 2009).
H2O2 triggers activation and dissociation of the complex
named ‘azurosome,’ which is formed by neutrophil elastase,
Neutrophil Defense Mechanisms MPO, azurocidin, cathepsin G, eosinophil cationinc protein,
defensin-1, lysozyme, and lactoferrin, leading to translocation
Neutrophils can eliminate microbes by various intracellular of neutrophil elastase and MPO to the nucleus (Metzler et al.,
and extracellular mechanisms. 2014). Once in the nucleus, elastase partially cleaves histones
to promote chromatin decondensation (Papayannopoulos
et al., 2010; Metzler et al., 2014). Peptidylarginine deamini-
Phagocytosis
nase 4 (PAD4) is activated by PKCζ and translocates into the
Neutrophils sense invaders through Toll-like receptors (TLRs) nucleus where it citrullinates histones (Brinkmann et al.,
and other pattern-recognition receptors. Human neutrophils 2004; Neeli and Radic, 2013). This phenomenon appears to
express almost all TLRs, except TLR3 (Janke et al., 2009; be crucial for chromatin decondensation leading to NET
 Cellular Immunology: Innate Immunity: Neutrophil Biology 753

Unstimulated LPS

Lamin B Vesicles DNA

Figure 2 Cellular events during NETosis. NET formation is characterized by disintegration of the nuclear envelope followed by DNA
decondensation. Neutrophils from a healthy volunteer were stimulated with LPS for 30 min. A resident nuclear membrane protein, lamin B was
stained in red and vesicles were stained in green. Nuclear material was counterstained with Hoechst. Arrows indicate localization of lamin B before
and after stimulation with LPS.

formation. The nuclear membrane disintegrates and decon- of neutrophils have been identified through expression of
densed DNA mixes with granules and cytosolic proteins specific markers, gene expression, and functional aspects in
(Figure 2). This complex of nuclear material and cytoplasmic healthy and pathological conditions. Some examples are
and nuclear proteins is extruded to the extracellular space as mentioned below:
NETs (Brinkmann et al., 2004) that serve to trap and immo-
bilize microbes and represent a lytic mechanism of neutrophil Olfactomedin 4-Expressing Neutrophils
cell death. The presence of antimicrobial proteins
(e.g., histones, defensins, elastase, proteinase 3, cathepsin G, Olfactomedin 4 (OLFM4) is a matrix glycoprotein expressed
lactoferrin, and MPO) in these structures supports their during the myelocyte and metamyelocyte stages of maturation,
microbicidal role toward various pathogens, including bac- and levels of this protein decline as neutrophils mature
teria, viruses, fungi, and parasites (Urban et al., 2009; (Clemmensen et al., 2012). OLFM4 is expressed in approxi-
McCormick et al., 2010). Neutrophils can also release mito- mately 25% of peripheral blood neutrophils and its presence is
chondrial DNA in NETs, without undergoing a cell death not regulated at the transcriptional level, suggesting microRNA
program (Yousefi et al., 2009, 2008) in response to LPS and (miRNA) regulation (Clemmensen et al., 2012). OLFM4 is a
other stimuli. In addition, Staphylococcus aureus can trigger NET granular protein that co-localizes with NGAL (Clemmensen
formation in a rapid, non-lytic, and oxidant-independent et al., 2012) and is externalized in NETs (Welin et al., 2013).
mechanism called vital NETosis (Pilsczek et al., 2010). During While the exact role of this neutrophil subset remains to be
this event, the plasma membrane remains intact. Vesicles determined, OLFM4 may have immunoregulatory roles (Liu
containing DNA and proteins are pinched off from the nuclear et al., 2013).
membrane and fuse to the plasma membrane, releasing NETs.
Vital NETosis has been also documented in response to TLR4- CD177-Expressing Neutrophils
mediated platelets binding to neutrophils (Clark et al., 2007). The glycoprotein CD177 (NB1) is a glycosyl-phosphatidyli-
nositol-anchored receptor that is differentially expressed in
circulating neutrophils (Stroncek, 2007; Hu et al., 2009).
Neutrophil Subsets Neutrophils that express CD177 demonstrate higher affinity to
PECAM-1. CD177 associates with membrane PR3 promoting
Given their short half-life, neutrophils have been difficult to infiltration into tissues (Figure 3). CD177-positive neutrophils
study and characterize in detail. While considered a homo- are increased in the autoimmune vasculitis granulomatosis
genous leukocyte subset for many years, emerging evidence with polyangiitis (Hu et al., 2009). It also appears that these
suggests that these cells carry significant heterogeneity and cells synthesize higher levels of IL-17. More information is
plasticity with various degrees of phagocytic potential, protein required regarding their role in induction of tissue damage and
synthesis, and oxidative metabolism. Indeed, various neutro- vascular inflammation.
phil subsets in response to physiological and pathological
conditions have been described (Gallin, 1984; Beyrau et al.,
Low-Density Granulocytes
2012). It is still unclear if neutrophil subsets represent truly
distinct lineages or develop from a single neutrophil precursor Low-density granulocytes (LDGs) are a distinct proin-
in response to the microenvironment. Up to this date, subsets flammatory subset of neutrophils identified in the
754 Cellular Immunology: Innate Immunity: Neutrophil Biology

volunteers after intravenous LPS administration. Regulatory

neutrophils are CD62Ldim/CD11cbright, demonstrate hyper-
PR3 DNA segmented nucleus and are capable of producing large quan-
tities of H2O2. Suppression of T cells requires formation of an
immunological synapse through neutrophil αMβ2-integrin
(MAC-1) and large amounts of H2O2 (Pillay et al., 2012). It
remains to be better understood whether specific neutrophil
subsets serve different roles in modulation of adaptive
immunity.

Genetic Defects That Affect Neutrophil Numbers and/

or Function

While genetic diseases that affect neutrophil numbers and

function are rare, they provide additional understanding into
the molecular mechanisms by which these cells are crucial in
host defense. A few are mentioned below:

Severe Congenital Neutropenia

Figure 3 Neutrophil heterogeneity. Emerging evidence supports the Severe congenital neutropenia (SCN) results from mutations
notion of heterogeneity among leukocytes. A subset of neutrophils in various neutrophil genes including neutrophil elastase
expressing membrane-bound PR3 can be found in neutrophil (ELANE), HAX-1, and GfI-1. Neutrophil formation is arrested
preparations from healthy volunteers. PR3 is stained in green and at the promyelocytic stage, patients display very low neutro-
DNA in blue. Scale bar 10 mm. phil counts and develop life-threatening infections.

mononuclear cell fraction of patients with systemic lupus Leukocyte Adhesion Disorders
erythematosus (SLE) (Hacbarth and Kajdacsy-Balla, 1986;
Leukocyte adhesion disorders (LAD) develop due to defects in
Denny et al., 2010) and other autoimmune conditions such as
neutrophil cell adhesion to the endothelium or the extra-
psoriasis (Lin et al., 2011). LDGs synthesize higher levels of
cellular matrix. Mutations in genes encoding for specific
proinflammatory cytokines (Denny et al., 2010) and are
integrins (such as LAD-I and LAD-II diseases) are characterized
primed to form significantly higher numbers of NETs
by severe infections due to defects in neutrophil recruitment.
(Villanueva et al., 2011). LDG-NETs contain higher concen-
trations of metalloproteinases, such as MMP-9 that trigger
endothelial damage (Carmona-Rivera et al., in press). Mol- Chronic Granulomatous Disease
ecules externalized by LDG-NETs may serve as modified
sources of autoantigens in SLE (Carmona-Rivera and Kaplan, Chronic granulomatous disease (CGD) is the most common
2013; Knight et al., 2012). of the genetic neutrophil function disorders. It is a hetero-
geneous group of hereditary diseases characterized by an in-
ability of phagocytic cells to kill engulfed organisms due to
Proangiogenic MMP-9 Delivering Neutrophils defects in ROS generation through NAPDH oxidase complex
(Figure 4). This is due to defects in subunits of phagocyte
Vascular endothelial growth factor (VEGF)-A facilitates the NADPH oxidase (Kuhns et al., 2010; Bianchi et al., 2009).
recruitment of a subset of neutrophils that express high levels Patients suffer from recurrent, life-threatening infections,
of MMP-9 to a site of hypoxia. The CD11b þ /Gr-1 þ /CXCR4 þ underlying the important role of ROS production in host
leukocyte subset has been reported to be involved in the defense.
revascularization of transplanted pancreatic islets. They are
more primed and prone to respond to a hypoxic stimulus,
expressing more CXCR4 and containing more MMP-9 with Mutations in the Gene Encoding MPO
proangiogenic capabilities (Christoffersson et al., 2012). Mutations in the gene encoding MPO have been described in
patients that suffer from recurrent Candida infections. One out
of 3000 individuals may harbor mutations that affect MPO
T-Cell Regulatory Neutrophils production, activity or trafficking (Winterbourn et al., 2006).
Recent evidence indicates that neutrophils can also have im-
portant regulatory roles in adaptive immune responses
Papillon–Lefèvre syndrome
through effects on T and B lymphocyte proliferation, acti-
vation, and differentiation. A subset of human neutrophils Papillon–Lefèvre syndrome (PLS) is an autosomal recessive
that can suppress T cells response was identified in healthy disorder caused by a deficiency in cathepsin C, an enzyme that
 Cellular Immunology: Innate Immunity: Neutrophil Biology 755

NADPH oxidase CGD

Cath-G
MPO
PLS
ELA

Phagolysosome

SGD
G
el
at
Sp
ec

Azu

C/EBP

Figure 4 Genetic diseases that affect neutrophil function. Genetic alterations that affect neutrophil function can trigger life-threatening infections.
Patients with mutation in subunits of the NADPH oxidase develop CGD. PLS is characterized by deficiency of proteinase activity present in
azurophilic granules. Lack of secondary and tertiary granules is found in patients with mutation in the transcription factor C/EBPε that develop
neutrophil-SGD. Azu, azurophilic granules; Cath G, Cathepsin G; ELA, elastase; Gelat, gelatinase granules; MPO, myeloperoxidase; Spec, specific
granules.

processes various serine proteases crucial in antimicrobial perpetuation of chronic inflammatory/autoimmune diseases.
defense (Figure 4). The immunodeficiency that develops in A few examples are mentioned below:
these patients in mild and the mutation affects mostly mature
neutrophils (De Haar et al., 2006; Sorensen et al., 2014).
Vasculitis
Vasculitis is a group of diseases characterized by inflammation
Neutrophil-Specific Granule Deficiency
of the blood vessels. Small vessels vasculitis (SVV) is a subtype
Neutrophil-specific granule deficiency (SGD) is a rare con- of this disease where patients synthesize anti-neutrophil cyto-
genital disease where neutrophils display atypical bilobed plasmic antibodies (ANCA) recognizing PR3 or MPO. Both
nuclei and lack expression of secondary and tertiary granular activated neutrophils and ANCAs have been implicated in the
proteins with multiple defects in antimicrobial strategies and development of vasculitis. MPO and PR3 are externalized dur-
severe bacterial infections (Figure 4). Mutations in the tran- ing NET formation and autoantibodies against PR3 induce
scription factor C/EBPε are the primary genetic defects NETosis (Kessenbrock et al., 2009). Dysregulated neutrophil cell
(Gombart et al., 2001). death has been proposed to play a pathogenic role in SSV.

Rheumatoid Arthritis
Neutrophils in Systemic Autoimmune Diseases
Rheumatoid Arthritis (RA) is an autoimmune disease that re-
While neutrophils are key players in the innate response to sults in destructive erosive inflammation of synovial joints
infections, they can also contribute to the development and (De Rycke et al., 2004). Neutrophils are the most abundant
756 Cellular Immunology: Innate Immunity: Neutrophil Biology

cells found in the synovial fluid of RA patients (Mohr et al., Bjerregaard, M.D., Jurlander, J., Klausen, P., Borregaard, N., Cowland, J.B., 2003.
1981) and they form NETs more readily than control neutro- The in vivo profile of transcription factors during neutrophil differentiation in
human bone marrow. Blood 101, 4322–4332.
phils (Khandpur et al., 2013). Various citrullinated proteins
Borregaard, N., 2010. Neutrophils, from marrow to microbes. Immunity 33,
released during NET formation are RA autoantigens (Khandpur 657–670.
et al., 2013; Pratesi et al., 2014). NETs induce a proin- Borregaard, N., Cowland, J.B., 1997. Granules of the human neutrophilic
flammatory phenotype in fibroblast-like synoviocytes, which polymorphonuclear leukocyte. Blood 89, 3503–3521.
may lead to enhanced articular damage (Khandpur et al., 2013). Borregaard, N., Sehested, M., Nielsen, B.S., Sengelov, H., Kjeldsen, L., 1995.
Biosynthesis of granule proteins in normal human bone marrow cells. Gelatinase
is a marker of terminal neutrophil differentiation. Blood 85, 812–817.
SLE Brandt, L., Hedberg, H., 1969. Impaired phagocytosis by peripheral blood
granulocytes in systemic lupus erythematosus. Scandinavian Journal of
SLE is a systemic autoimmune syndrome characterized by Haematology 6, 348–353.
Brinkmann, V., Reichard, U., Goosmann, C., et al., 2004. Neutrophil extracellular
the development of autoantibodies to nuclear components,
traps kill bacteria. Science 303, 1532–1535.
immune complex deposition, inflammation, and organ Carman, C.V., Sage, P.T., Sciuto, T.E., et al., 2007. Transcellular diapedesis is
damage (Tsokos, 2011). Neutrophils may contribute to loss of initiated by invasive podosomes. Immunity 26, 784–797.
tolerance and organ damage in SLE. Abnormalities in Carmona-Rivera, C., Kaplan, M.J., 2013. Low-density granulocytes: A distinct class
phagocytosis, activation, and NET formation have been de- of neutrophils in systemic autoimmunity. Seminars in Immunopathology 35,
455–463.
scribed in SLE neutrophils (Brandt and Hedberg, 1969; Carmona-Rivera, C., Zhao, W., Yalavarthi, S., Kaplan, M.J., in press. Neutrophil
Courtney et al., 1999; Hashimoto et al., 1982; Villanueva et al., extracellular traps induce endothelial dysfunction in systemic lupus
2011; Carmona-Rivera et al., in press). Dysregulated NET deg- erythematosus through the activation of matrix metalloproteinase-2. Annals of the
radation through inhibition of DNASE-I has also been described Rheumatic Diseases. doi: 10.1136/annrheumdis-2013-204837.
Christoffersson, G., Vagesjo, E., Vandooren, J., et al., 2012. VEGF-A recruits a
(Hakkim et al., 2010; Leffler et al., 2012). NETs also promote
proangiogenic MMP-9-delivering neutrophil subset that induces angiogenesis in
synthesis of cytokines considered key in lupus pathogenesis and transplanted hypoxic tissue. Blood 120, 4653–4662.
inhibition of NET formation decreases lupus phenotype in Clark, S.R., Ma, A.C., Tavener, S.A., et al., 2007. Platelet TLR4 activates neutrophil
animal models (Villanueva et al., 2011; Knight et al., 2013). extracellular traps to ensnare bacteria in septic blood. Nature Medicine 13,
463–469.
Clemmensen, S.N., Bohr, C.T., Rorvig, S., et al., 2012. Olfactomedin 4 defines
a subset of human neutrophils. Journal of Leukocyte Biology 91,
Conclusion 495–500.
Courtney, P.A., Crockard, A.D., Williamson, K., et al., 1999. Increased apoptotic
Neutrophils are specialized, terminally differentiated cells that peripheral blood neutrophils in systemic lupus erythematosus: Relations with
disease activity, antibodies to double stranded DNA, and neutropenia. Annals of
are endowed with crucial roles in host defense against mi-
the Rheumatic Diseases 58, 309–314.
crobes. Genetic or acquired deficiencies in neutrophil numbers Dahl, R., Walsh, J.C., Lancki, D., et al., 2003. Regulation of macrophage and
or function can be life-threatening and lead to severe in- neutrophil cell fates by the PU.1:C/EBPalpha ratio and granulocyte colony-
fections, while unchecked neutrophil responses or extended stimulating factor. Nature Immunology 4, 1029–1036.
neutrophil half-life may lead to chronic inflammatory and De Haar, S.F., Hiemstra, P.S., Van Steenbergen, M.T., Everts, V., Beertsen, W.,
2006. Role of polymorphonuclear leukocyte-derived serine proteinases in defense
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remain with regards to their specialization and plasticity and De Rycke, L., Peene, I., Hoffman, I.E., et al., 2004. Rheumatoid factor and
their interactions with other cells of the innate and adaptive anticitrullinated protein antibodies in rheumatoid arthritis: Diagnostic value,
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Denny, M.F., Yalavarthi, S., Zhao, W., et al., 2010. A distinct subset of
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See also: Cell Communication: Prototypic Integrative
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Processes: Wound Healing: An Orchestrated Process of Cell Cycle, Eash, K.J., Means, J.M., White, D.W., Link, D.C., 2009. CXCR4 is a key regulator of
Adhesion, and Signaling. Vertical Integration: Applications: A neutrophil release from the bone marrow under basal and stress granulopoiesis
Review on Various Mathematical Modeling Approaches for Wound conditions. Blood 113, 4711–4719.
Healing Faurschou, M., Borregaard, N., 2003. Neutrophil granules and secretory vesicles in
inflammation. Microbes and Infection 5, 1317–1327.
Fazi, F., Rosa, A., Fatica, A., et al., 2005. A minicircuitry comprised of microRNA-
223 and transcription factors NFI-A and C/EBPalpha regulates human
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