Neutrophil Function - From Mechanisms To Disease

IY30CH19-Zychlinsky ARI 17 February 2012 13:38
ANNUAL
REVIEWS Further Neutrophil Function:
Click here for quick links to
Annual Reviews content online,
including:
From Mechanisms to Disease
Access provided by Benemerita Universidad Autonoma de Puebla on 10/01/21. For personal use only.
• Other articles in this volume

• Top cited articles Borko Amulic, Christel Cazalet,
• Top downloaded articles
Garret L. Hayes, Kathleen D. Metzler,
Annu. Rev. Immunol. 2012.30:459-489. Downloaded from www.annualreviews.org
• Our comprehensive search
and Arturo Zychlinsky∗

Department of Cellular Microbiology, Max Planck Institute for Infection Biology,
Charitéplatz 1, 10117 Berlin, Germany; email: amulic@mpiib-berlin.mpg.de,
cazalet@mpiib-berlin.mpg.de, hayes@mpiib-berlin.mpg.de, metzler@mpiib-berlin.mpg.de,
zychlinsky@mpiib-berlin.mpg.de
Annu. Rev. Immunol. 2012. 30:459–89 Keywords

First published online as a Review in Advance on inflammation, antimicrobial, granule, phagocytosis, NET
January 3, 2012
The Annual Review of Immunology is online at Abstract

immunol.annualreviews.org
Neutrophils are the most abundant white blood cells in circulation,
This article’s doi:
and patients with congenital neutrophil deficiencies suffer from severe
10.1146/annurev-immunol-020711-074942
infections that are often fatal, underscoring the importance of these
Copyright c 2012 by Annual Reviews.
cells in immune defense. In spite of neutrophils’ relevance in immunity,
All rights reserved
research on these cells has been hampered by their experimentally in-
0732-0582/12/0423-0459$20.00
tractable nature. Here, we present a survey of basic neutrophil biology,
∗
All authors contributed equally to the work and with an emphasis on examples that highlight the function of neutrophils
are listed alphabetically.
not only as professional killers, but also as instructors of the immune
system in the context of infection and inflammatory disease. We focus
on emerging issues in the field of neutrophil biology, address questions
in this area that remain unanswered, and critically examine the experi-
mental basis for common assumptions found in neutrophil literature.
459
INTRODUCTION early and enthusiastic evolutionary biologist in-

terested in the phagocytic capacity of cells.
In the late nineteenth century, Paul Ehrlich,
Metchnikoff demonstrated that injury of
dissatisfied with what he considered an in-
starfish embryos resulted in recruitment of
excusable disinterest in the white blood cell,
phagocytic cells to the site of injury (3). He
began to utilize newly developed cell-staining
theorized (correctly) that these cells migrate to
techniques to examine subpopulations of leuko-
injured sites and participate in microbe diges-
cytes. His experimentation led to a new appreci-
tion. Remarkably, this prescient view of neu-
ation for the heterogeneity of white blood cells
trophil action still aptly summarizes, more than
and to the discovery of several novel leukocyte
a century later, the basic role of neutrophils
subpopulations. Ehrlich named one of these
in immunity. The uniquely lobulated nucleus
newly discovered cell types, characterized by a
of the neutrophil also inspired Metchnikoff to

“polymorphous nucleus” and a tendency to re-
rename these cells: He called them polymor-
tain neutral dyes, the “neutrophil” (1) (see also
phonuclear leukocytes (or PMNs), a title that

the sidebar, A Natural History of Neutrophils).
still enjoys frequent use and that is used inter-
The function of neutrophils was initially
changeably with neutrophil throughout this re-
shrouded in considerable mystery; their con-
view. Together with two other developmentally
spicuous presence during infections led several
related cell types, the eosinophils and basophils
researchers to arrive hastily at a rather ironic
(also discovered by Ehrlich), PMNs form the
conclusion: They surmised that neutrophils
granulocyte family of white blood cells, a fam-
promote infection, serving as cellular shuttles
ily whose hallmark is the presence of “granules,”
for bacteria (2). Their actual function, that of
unique storage structures important in antimi-
antimicrobial actors in the immune response,
crobial functions (see section on Granules and
was eventually demonstrated conclusively by a
Degranulation, below).
contemporary of Ehrlich, Elie Metchnikoff, an
Neutrophils were discovered at the dawn
of the immunological sciences; consequently,
elucidation of their role in the immune re-
sponse has been an ongoing process stretching
A NATURAL HISTORY OF NEUTROPHILS over more than a century. We now know that
they are key components of the innate immune
Phagocytes are ancient cells that evolved to allow multicellular response and vital in immune function; unfor-
organisms to thrive in the face of constant competition with mi- tunately, their importance has often been over-
crobes for resources. Metchnikoff ’s seminal theory of cellular shadowed by breakthroughs in the study of the
immunity was based on comparative embryology and observa- adaptive immune response (4). Admittedly, this
tions of phagocytes in various simple organisms, including the mi- situation is exacerbated by neutrophils’ notori-
croscopic crustacean Daphnia. Remarkably, even the slime mold ous experimental intractability: They exhibit a
Dictyostelium discoideum has phagocytic cells that protect it from short life span and are terminally differentiated,
infection (200). The short-lived neutrophil with a lobulated nu- preventing growth in tissue culture. The stan-
cleus and granule-packed cytoplasm is a more recent evolutionary dard tools of molecular biology, such as trans-
adaptation. In insects, phagocytes are long lived and have round fection and RNA interference, are of little use
nuclei. They do, however, produce hydrogen peroxide and carry when applied to these cells, and immortalized
distinct classes of granules (201). Bony fish and frogs have bona “neutrophil-like” cell lines rarely reflect the
fide neutrophils that are functionally similar to mammalian ones functional diversification of neutrophils. Fur-
(202, 203). In both zebrafish and rodents, neutrophils are less thermore, neutrophil-like cells studied in the
abundant than in humans, comprising only 15–20% of immune isolation of a culture dish most certainly do not
cells. In chimpanzees, neutrophils account for more than 50% of mimic the complex biological reality in tissues
the differential blood count (204). or circulation. Conclusions from in vitro stud-
ies should, therefore, be carefully interpreted.
460 Amulic et al.

Unfortunately, in vivo studies of neutrophil infection. Indeed, the number of neutrophils

function also raise concerns. Mouse neu- drastically increases during infection and some
trophils, the preferred model for in vivo diseases. Interestingly, neutrophils circulate
studies, differ in important aspects from their for only approximately 6–8 h and are among
human equivalents. This is perhaps best the shortest-lived cells in the human body.
exemplified by the differences in the respective Although the reason for this short life is unclear,
antimicrobial repertoires and the numbers of it may ensure neutrophil integrity; this hypoth-
PMNs in circulation (30% versus 70% in mice esis is bolstered by observations that apoptosis
and humans, respectively). prevents the release of noxious molecules.
Despite these difficulties, no picture of the Still, the question of why evolution opted for
immune response can be complete without eliminating neutrophils quickly as opposed
a comprehensive understanding of the neu- to reducing leakage of their dangerous cargo

trophil and its functions. The extensive nature remains an unanswered and intriguing mystery.
of neutrophil research, however, precludes a Mature neutrophils emerge from the bone
comprehensive review of the subject matter. marrow intent on pursuing one simple, yet
In this review, we intend to provide a survey essential, question: Has host integrity been
of basic neutrophil biology and function, while compromised by potentially harmful invaders?
emphasizing recent advances in neutrophil re- Should the answer prove to be “yes,” the
search and providing a critical assessment of neutrophil must swiftly enact a carefully
some current reports on PMN action. choreographed process to locate, attack, and
Our survey of the neutrophil begins in destroy the potential threat. At its disposal is
adult bone marrow where, under the in- an impressive arsenal of antimicrobial weapons
struction of growth factors and cytokines, that are deadly, indiscriminate, and brutish in
pluripotent hematopoietic cells differentiate their application. Although effective in their
into myeloblasts, a developmental cell type destructive capacity, these weapons can prove
committed to becoming granulocytes. As these to be just as dangerous to the host cells as to
precursor cells mature to neutrophils, they syn- their intended targets, the microbial invaders.
thesize proteins that are sorted into different Therefore, their deployment must be executed
granules (5). Traditionally, granules have been with exquisite precision and timing, at locations
subdivided into three different classes based where they are both contained and effective.
on their resident cargo molecules: azurophilic, How then does the neutrophil locate and
specific, and gelatinase granules. Although this identify infections? How does it transition
subdivision is practical, these designations are at the correct time and place from an in-
largely artificial. Granules are formed through a active cellular bystander to a fully activated
continuous process; vesicles bud from the Golgi microbial killing machine? This transition
apparatus and fuse, producing granular struc- process, during which the neutrophil inte-
tures. The content of these structures is dic- grates a complex barrage of environmental
tated by the transcriptional program active at cues and translates them into specific actions,
the time of their formation. As the maturing is known as neutrophil “activation.” As it
neutrophil sequentially alters its transcriptional pursues microbes, the neutrophil will enact an
profile, granule content changes, resulting in a impressive multitude of cellular mechanisms:
continuum of granule species with overlapping It will mobilize secretory vesicles and granules,
cargoes (6). identify chemotactic gradients and traverse
The release of neutrophils from the bone them through destruction and reorganization
marrow is tightly regulated in healthy in- of the actin skeleton, penetrate the endothelial
dividuals: Chemokines control the passage barrier and navigate a course through the
of PMNs into circulation and maintain a basement membrane, and begin transcription
pool of cells ready for release in case of of cytokines for recruitment of new immune
www.annualreviews.org • Neutrophil Functions 461

cells. Ultimately, upon arriving at the infection family kinases, Syk, phosphoinositide 3-kinase
site, it will seek the insulting pathogens and (PI3K), and p38 mitogen-activated protein
unleash its extensive arsenal of antimicrobial kinase (11–13). This cascade initiates a number
Selectins:
transmembrane weapons. The initiation of these processes oc- of changes in neutrophil biology and sets the
glycoproteins that curs in the bloodstream, where the neutrophil stage for integrin activation and firm adhesion.
mediate cell adhesion acts as a monitor for host distress, patrolling After selectin-mediated rolling, neutrophils
via binding to sugar vessels and vigilantly seeking out indications of enter a “firm adhesion” state mediated by the
moieties
an incipient inflammatory response. β2 integrin family of proteins (LFA-1 and
Integrins: Mac-1 proteins on the neutrophil); firm adhe-
transmembrane
sion is characterized by the arrest of neutrophil
receptors that mediate NEUTROPHIL ACTIVATION
attachment to the rolling in preparation for transendothelial
extracellular matrix, as At inflammatory sites, bacterial-derived and migration (13, 14). As the neutrophil rolls
well as direct cell-cell host-produced inflammatory signals are along the endothelium, interaction with
interaction and
abundant; these compounds stimulate the selectins, chemoattractants, cytokines, and

signaling
endothelial cells near the inflammatory site. bacterial products results in activation and
Oxidative/respiratory These stimulants, such as the bacterial-derived clustering of the β2 integrins on the surface of
burst: a rapid increase
lipopolysaccharide (LPS) and fMLP, as well the neutrophil (15, 16). The β2 integrins then
in oxygen
consumption upon as the classical chemoattractants and cytokines engage their endothelial ligands, members of
neutrophil activation tumor necrosis factor (TNF)-α, interleukin the ICAM-1 immunoglobulin superfamily,
due to production of (IL)-1β, and IL-17, prompt endothelial cells to resulting in arrest of neutrophil rolling and
ROS by the NADPH produce adhesion molecules on their luminal firm adhesion. This integrin engagement, as
oxidase
side: the P-selectins, E-selectins, and several well as continuing input from inflammatory
members of the integrin superfamily, the chemoattractants and cytokines, prepares the
ICAMs (5). As neutrophils traverse the circu- neutrophil for its final chemotactic pursuit: The
latory system, they continuously and randomly cell spreads, producing a leading-edge lamel-
probe the vessel wall; the postcapillary venules, lipodium where chemokine and phagocytic
where flow dynamics and the constricted space receptors are concentrated, the cytoskeleton is
are particularly amenable to increased random rebuilt and targeted toward movement along
probing, are often the best-suited location chemotactic gradients, and initiation of the
for neutrophils to encounter the stimulated neutrophil oxidative burst begins (17, 18).
endothelial cells (7, 8). Now firmly adhered, the neutrophil must
On the surface of neutrophils, two constitu- negotiate a path through the endothelium into
tively expressed proteins are critical for recog- the underlying tissue. In a process dependent
nition of the endothelial inflammatory signals: on β2 integrins and ICAMs, neutrophils
the glycoprotein P-selectin glycoprotein crawl along the vessel wall until a preferred
ligand-1 (PSGL-1) and L-selectin (9, 10). Upon site of transmigration is reached (19–21).
random contact with the endothelium, these Upon arrival at an endothelial cell junction, a
molecules engage the P- and E-selectins of complex interaction between (a) the neutrophil
endothelial cells, resulting in selectin-mediated integrins and their endothelial partners and
tethering of neutrophils to the vessel wall. (b) neutrophil surface proteins and various
This is followed by a characteristic “rolling” of endothelial junction molecules results in trans-
neutrophils along the endothelium. It is here migration through the endothelial junction
that the complex activation cascade begins (13). Once through the endothelial lining,
and the neutrophil commitment to microbial the neutrophil must navigate the basement
killing commences. What changes occur in the membrane, a protein mesh consisting largely
neutrophil at this early time point? The engage- of laminins and collagen type IV. Speculation
ment of PSGL-1 and L-selectin on neutrophils abounds that granule proteases assist in this
activates a variety of kinases, including Src migration by digesting the protein mesh
462 Amulic et al.

subsequent to degranulation; however, conclu- point of high chemoattractant concentration,

sive experimental evidence for this is lacking. where no discernible gradient exists, the
Once the endothelial barrier has been neutrophil halts and begins the final release of
traversed, the neutrophil finds itself in a its antimicrobial arsenal; the neutrophil is now
much different inflammatory milieu: Here, the fully in an antimicrobial attack state.
environment is awash in a soup of chemoat- The complex signaling cascade leading to
tractants and inflammatory stimulants, both final neutrophil activation has several facets
host derived and of pathogenic origin. These worthy of note. The movement to ever-higher
compounds will now be the primary dictators concentrations of chemoattractant is key in
of neutrophil behavior and assume respon- this process, as individual chemoattractants
sibility for initiating the concluding steps of may have very different effects on neutrophil
neutrophil activation. In the interstitial space, physiology at different concentrations, a

the neutrophil follows chemotactic gradients phenomenon exemplified by one of the key
toward the invading microbes, pursuing host- neutrophil-recruiting chemokines and ac-
produced cytokines (e.g., IL-8) and, in parallel, tivators, IL-8. At low concentrations, IL-8
pathogen-derived chemoattractants (e.g., stimulates L-selectin shedding and increased
fMLP). During this process, these chemoat- expression of β2 integrins; slightly higher
tractants bind to their respective neutrophil concentrations result in initiation of the
receptors (often G protein–coupled receptors, oxidative burst. At the highest concentrations,
as is the case with the fMLP receptor FPR1 or IL-8 induces degranulation of neutrophils (27).
the chemokine receptors), which initiate a sig- In addition, many chemoattractant molecules
naling cascade dominated by the MAPK/ERK exert a “priming” effect. That is, alone they
pathway (22, 23). Downstream molecules stimulate the oxidative response only mildly,
prompt assembly of the oxidative burst ma- but they dramatically enhance the subsequent
chinery, a hallmark of neutrophil activation. response to other stimuli. A notable example of
Furthermore, the stimulation of FPR1 triggers this phenomenon is the strong priming effect
the release of ATP, whose autocrine action of LPS on the fMLP response (28). In this case,
through activation of purinergic receptors is exposure of the neutrophil to LPS induces
critical for the initiation of effective functional assembly of the NADPH oxidase machinery on
responses in neutrophils (24). Concomitantly, the membrane; fMLP stimulation then induces
a family of molecules, the pattern-recognition activation of this machinery (29). In contrast to
receptors, is activated through recognition of receptor priming, another critical feature of the
specific nonself patterns present on many mi- stimulation process is the desensitization to pre-
crobes (25). Perhaps the best-known example viously encountered ligands. Stimulation of the
of this family is the Toll-like receptors (TLRs); neutrophil by a chemoattractant often results
they are responsible for recognizing a number in endocytosis of the corresponding receptor,
of pathogen-derived compounds, collectively thus leading to a desensitization of the neu-
called pathogen-associated molecular patterns trophil to repeated stimulation with the same
(PAMPs), including LPS (TLR4), bacterial molecule (30, 31). The rich and varied input
lipopeptides (TLR2), flagellin (TLR5), and received by a neutrophil during this final leg of
DNA (TLR9). In neutrophils, all but one the activation process is complex, and the exact
of these receptors (TLR3) are constitutively effects of priming, desensitization, and signal-
expressed, and their stimulation contributes ing are incompletely understood. Regardless,
to further activation, e.g., induction of the the end result of this signaling cacophony is
oxidative burst (25, 26). As the neutrophil nears unambiguous: The neutrophil begins to imple-
its target, continued activation by chemoattrac- ment its regime of microbial killing, executing
tants further stimulates the oxidative response programs of phagocytosis, degranulation, and
and degranulation. Upon finally reaching a NETosis (i.e., the process of setting neutrophil

extracellular traps) (see the section on Neu- parsed by the complex neutrophilic signaling
trophils and the Elimination of Microbes, mechanisms, a process that gradually leads
below). to complete activation and culminates in the
The initiation of these microbicidal actions premiere killing functions of phagocytosis,
indicates the final stage of the neutrophil’s degranulation, and NETosis. It is, therefore,
journey through the activation process. How- more insightful to view neutrophil activation
ever, a prominent question remains largely as a continuum of processes, priming steps,
unanswered by the preceding exposition: What and signal cascades with varying effects and
exactly is meant by the (admittedly ambiguous) outcomes, all focused on the realization of
phrase “neutrophil activation”? A quick scan one goal: the transition of naive, circulating
of the literature presents the inexperienced neutrophils to their microbe-eliminating,
reader with a sometimes rather conflicting (and tissue-resident counterparts (Figure 1).
overwhelming) view of neutrophil activation.
In fact, one could be (erroneously) led to

believe that neutrophil activation refers only to
NEUTROPHILS AND THE
direct stimulation of the oxidative burst, as this
ELIMINATION OF MICROBES
has been the canonical in vitro activation assay The basic instruction set of the activated
for decades. This is, however, an oversimpli- neutrophil is both effective and ruthless in
fied view of a complex process. The myriad its simplicity: (1) kill microbes, (2) do no
interactions that occur during a neutrophil’s harm to the host, and (3) when in doubt, see
journey toward an inflammatory site must be rule 1. To fulfill this antimicrobial agenda,
a Capture b Rolling c Firm adhesion
Neutrophil
Integrin
P-selectin and ICAM
PSGL-1, E-selectin
L-selectin
Phagocytosis
Endothelial cell Degranulation
Cytokine secretion
NETs
Figure 1
Neutrophil recruitment to sites of inflammation. The circulating neutrophil must recognize signs of
inflammation and migrate to areas where its antimicrobial arsenal is needed for the elimination of infection.
(a) Close to the inflammatory sites, stimulated endothelial cells expose a class of molecules, the selectins,
which serve to capture circulating neutrophils and tether them to the endothelium. (b) Selectin-mediated
rolling along chemoattractant gradients then ensues, followed by (c) integrin-mediated firm adhesion.
Subsequently, the neutrophil traverses through the endothelium and arrives at the site of inflammation.
Here, the neutrophil releases cytokines that recruit other immune cells, and it begins to implement its
antimicrobial agenda. Among the processes employed are engulfment of microbes via receptor-mediated
phagocytosis, release of granular antimicrobial molecules through degranulation, and formation of
neutrophil extracellular traps (NETs).
464 Amulic et al.

neutrophils possess an array of toxic weapons needs of neutrophils. Granules are, however,
that are carefully regulated through controlled far more than just latent repository organelles
mechanisms. These antimicrobial weapons for dangerous substances; they are active and in-
Inflammation:
vary considerably in their methods of action dispensable participants in almost all neutrophil recruitment and
and thus reflect the neutrophil’s attempt to activities during inflammation. activation of immune
exploit any and all weaknesses that microbes As mentioned above, there are three cells upon infection or
might present during the course of infection. fundamental types of granules in neutrophils injury; when
uncontrolled it leads to
An understanding of these weapons, their (Figure 2). Azurophilic granules (also known
tissue damage
action, and their method of release is critical as peroxidase-positive or primary granules) are
to understanding neutrophil function. the largest, measuring approximately 0.3 μM
in diameter, and are the first formed during
neutrophil maturation. They are named for

Granules and Degranulation their ability to take up the basic dye azure A and
The neutrophil must safely transport a plethora contain myeloperoxidase (MPO), an enzyme
of dangerous substances through the blood- critical in the oxidative burst (32, 33). Other
stream and then correctly deploy them at the cargo of this granule class include the defensins,
appropriate time. Therefore, it comes as no lysozyme, bactericidal/permeability-increasing
surprise that a specialty storage organelle has protein (BPI), and a number of serine proteases:
evolved in neutrophils: the granule. Expect- neutrophil elastase (NE), proteinase 3 (PR3),
edly, these structures are replete with specifi- and cathepsin G (CG) (34). As such, these
cally tuned mechanics that address the unique granules are brimming with antimicrobial
Primary Secondary Tertiary Secretory

Granule type (azurophilic) (specific) (gelatinase) vesicles
Stage of Myeloblast Promyelocyte Myelocyte Metamyelocyte Band cell

formation PMN
Degranulation
propensity
Characteristic Lysozyme Complement receptor 1

proteins
Myeloperoxidase Lactoferrin FcγRIII
Elastase Gelatinase
Defensin
Other Cathepsin G, PR3, Gp91phox/p22phox, Gp91phox/p22phox, Gp91phox/p22phox,

proteins BPI, azurocidin, CD11b, collagenase, CD11b, MMP25, CD11b, MMP25, C1q-R,
sialidase, hCAP18, NGAL, B12BP, arginase-1, FPR, alkaline
β-glucuronidase SLPI, haptoglobin, β2-microglobulin, phosphatase, CD10,
pentraxin 3, CRISP3 CD13, CD14,
oroscomucoid, plasma proteins
β2-microglobulin,
heparanase, CRISP3
Figure 2
Neutrophil granules. Neutrophil granules carry a rich variety of antimicrobials and signaling molecules. They are typically divided into
three types (primary or azurophilic, secondary or specific, and tertiary or gelatinase). Additionally, structures called secretory vesicles
are also considered to be a granule subset. Considerable overlap exists in the cargo of the different granules, and their contents seem
determined by the timepoint during hematopoiesis at which they are produced (5). Granules also differ in their ability to mobilize, with
secretory vesicles being the first to fuse with the plasma membrane and the azurophilic granules demonstrating the least degranulation
propensity.

compounds and function as a primary reposi- granule subset has been traditionally associated
tory for the molecular weaponry of neutrophils. with a particular stage of neutrophil activation.
The second class of granules, the specific (or After neutrophils contact the endothelium,
secondary) granules, are smaller (0.1 μM stimulation through selectins and chemoattrac-
diameter), do not contain MPO, and are char- tants induces mobilization of secretory vesi-
acterized by the presence of the glycoprotein cles, whose membranes are rich in key factors
lactoferrin. These granules are formed after necessary for continued activation of the neu-
azurophilic granules; they also contain a wide trophil, including, among others, the β2 inte-
range of antimicrobial compounds including grins, complement and fMLP receptors, as well
NGAL, hCAP-18, and lysozyme (33, 35). The as the FcγRIII receptor CD16 (5, 38, 39, 42).
third class, the gelatinase (tertiary) granules, are Fusion of the secretory vesicles with the plasma
also MPO-negative, are smaller than specific membrane exposes these components to the ex-
granules, and contain few antimicrobials, ternal environment. This results in the transi-
but they serve as a storage location for a tion to firm adhesion, mediated by β2 integrin
number of metalloproteases, such as gelatinase interaction with the endothelium. As they pro-
and leukolysin. These granules are also the ceed through the endothelium, neutrophils are
last population of granules formed during exposed to further activation signals that initiate
neutrophil maturation (5). Finally, a fourth set mobilization of gelatinase granules, thereby re-
of structures, the secretory vesicles, are also leasing metalloproteases. The activity of these
commonly considered part of the neutrophil proteases may help neutrophils traverse the
granule family. In contrast to the classical basement membrane, although this has not
granules, these do not bud from the Golgi, been conclusively demonstrated (43, 44).
but instead are formed through endocytosis At the inflammatory site, complete acti-
in the end stages of neutrophil maturation vation of the neutrophil ensues, prompting
(36). Consequently, their cargo consists pre- initiation of the oxidative burst and mobiliza-
dominantly of plasma-derived proteins such as tion of the azurophilic and specific granules.
albumin. The membrane of secretory vesicles These granules either fuse with the phagosome
serves as a reservoir for a number of important (see section on Phagocytosis, below), con-
membrane-bound molecules employed during tributing to the antimicrobial activities of this
neutrophil migration. compartment, or fuse with the plasma mem-
As a neutrophil proceeds through activation, brane, releasing their potent antimicrobials
granules are mobilized and fuse with either the into the tissue. The fusion of specific granules
plasma membrane or the phagosome, releasing with the plasma or phagosomal membrane is of
their contents into the respective environment. particular importance for the oxidative burst,
In both cases, the membrane of the granule as flavocytochrome b558, a component of the
becomes a permanent part of the target mem- NADPH oxidase machinery, resides in the
brane, thus altering its molecular composition specific granule membrane (45). This fusion
(6). The different classes of granules demon- permits assembly of the NADPH oxidase com-
strate varying propensities for mobilization in plex and allows reactive oxygen species (ROS)
response to inflammatory signals: Azurophilic production both inside the phagolysosome and
granules are the most difficult to mobilize, fol- outside of the cell. Degranulation of primary
lowed by specific granules, gelatinase granules, and secondary granules contributes to the
and finally, secretory vesicles (37–41). The creation of an antimicrobial milieu at the in-
underlying mechanisms for this differential flammatory site and produces an environment
mobilization are not entirely understood, al- inhospitable to invading pathogens.
though regulation of intracellular calcium levels The release of granular proteins during de-
appears to play a salient role (32, 39). Because granulation presents the astute observer with
of this varying mobilization propensity, each a tempting proposition: Could these granular
466 Amulic et al.

components also serve as signaling molecules whereas others may be redundant. One of the
for subsequent inflammatory cell recruitment? challenges in understanding the neutrophil’s
Recent studies have provided experimental evi- antimicrobial mechanisms is to study their
dence suggesting this does seem to be the case: function during concerted action and in con-
Granule proteins from neutrophils, including ditions that mimic an infection site. Therefore,
PR3 and azurocidin, can induce monocyte re- testing the relevance of antimicrobials in vivo
cruitment. Furthermore, neutrophil granule is essential. This is, however, particularly chal-
proteins may increase macrophage bacterial lenging; ablation of a single antimicrobial gene
clearance by enhancing phagocytosis (46). This may only subtly affect immune defense. In ad-
could be advantageous in situations in which the dition, much biochemical identification of neu-
extracellular concentration of released granule trophil antimicrobials has been performed in
proteins is insufficient to exert extensive micro- rabbits and humans, species with abundant neu-
bicidal effects. In such cases, the granule pro- trophils. Mice, which are genetically tractable,
teins would instead operate as signaling and re- have neutrophils that function differently from
cruitment factors (see section on Neutrophils those of other species. Indeed, as already men-
in Immune Cell Cross Talk, below). tioned, mice lack the genes for some antimicro-
By necessity, most data on neutrophil bials identified in humans. Interestingly, there
degranulation and its effects on neutrophil ac- are few clinically relevant innate immune de-
tivity have been acquired through biochemical ficiencies that directly link antimicrobial activ-
approaches performed exclusively in vitro. A ity with a particular mutation. Thus, with few
pertinent question therefore presents itself: Is exceptions, evidence for clinical or biological
this process truly relevant during the in vivo relevance of these molecules is still lacking.
inflammatory response? The data here are There are three main types of antimicro-
sparse, and understandably so: Historically, the bials: (a) cationic peptides and proteins that
possibilities for such an in vivo observation have bind to microbial membranes, (b) enzymes,
been restrained by technical limitations. Most and (c) proteins that deprive microorganisms
evidence for in vivo degranulation relies on of essential nutrients. Here we present an
observation of increased levels of extracellular overview of this rich field of investigation.
granular proteins at inflammatory sites. Even There are more than 800 antimicrobial
so, release of granular components could occur peptides described in nature, some of them
primarily through other means, most notably highly conserved throughout evolution (47).
through formation of neutrophil extracellular These peptides are often charged, a feature that
traps, cell damage, or cell lysis. With the probably promotes their initial interaction with
advent of intravital microscopy techniques, microbial surfaces. Under artificial conditions,
direct observation of the degranulation process many of these peptides disrupt the membrane
in vivo may soon be realized. integrity. Because in vitro tests are often exe-
cuted at high antimicrobial concentrations to
obtain maximal microbial killing in the shortest
Antimicrobial Proteins possible time, it is unclear whether this disrup-
Neutrophils produce a plethora of peptides and tion reflects their mechanism of action under
proteins that directly or indirectly kill microbes physiological conditions. Alternatively, some
(Table 1). Many of these antimicrobials were antimicrobials are thought to disrupt essential
identified through biochemical fractionation of microbial functions, such as DNA replication,
neutrophil extracts, and their in vitro activity transcription, or production of energy. Little
is easily demonstrated in optimized conditions; is known about antimicrobial concentrations
nonetheless, showing in vivo relevance is chal- achieved at inflammatory sites or in the phago-
lenging. The diversity of antimicrobials sug- some. This information, as well as information
gests that some of them evolved to act together, about the synergistic interactions of different

Table 1 Mechanism of action of neutrophil antimicrobial proteins

Antimicrobial peptide Antimicrobial mechanisma
Cationic antimicrobial peptides
α-defensins (HNP-1, HNP-2, Permeabilize membrane bilayers containing negatively charged
HNP-3, HNP-4) phospholipids
Inhibit DNA, RNA as well as protein biosynthesis
Inhibition of bacterial cell wall synthesis
LL-37 Transmembrane pore-forming
BPI Increase bacterial permeability and hydrolysis of bacterial
phospholipids by binding to LPS
Histones Unknown mechanism
Proteolytic enzymes
Lysozyme Degrades bacterial cell wall
Proteinase 3 (PR3) Mechanism independent of a proteolytic activity by binding to the

bacterial membrane
Neutrophil elastase (NE), Cleaves bacterial virulence factors and outer membrane
cathepsin G (CG) proteins
Mechanism independent of a proteolytic activity by binding to
the bacterial membrane
Azurocidin Mechanism independent of a proteolytic activity by binding to the
bacterial membrane
Metal chelator proteins
Lactoferrin Alters bacterial growth by binding to iron, an essential bacterial
nutrient
Binds to the lipid A part of LPS, causing a release of LPS from
the cell wall and an increase in membrane permeability
Calprotectin Alters bacterial growth by sequestering manganese and zinc
a
Only direct actions of neutrophil antimicrobial proteins on microbes are listed in the table.
antimicrobials, is essential for designing appro- from larger proteins, and in addition to their
priate in vitro conditions to probe mechanisms antimicrobial activity, they may potentiate
of action. DNA activation of dendritic cells (DCs) (50).
The neutrophil cationic antimicrobial Neutrophils also contain a number of
peptides include defensins and cathelicidins. full-length cationic antimicrobial proteins,
Neutrophils mostly produce α-defensins, a including BPI and histones. BPI is cationic
protein family whose members possess multi- and binds LPS avidly, much like its structural
ple disulfide bonds and whose structures may cousin the LPS binding protein. BPI binding to
change under physiological conditions and LPS results in increased bacterial permeability
increase their activity (48). A surprising num- and hydrolysis of bacterial phospholipids; cell
ber of functions are assigned to defensins, but death then follows (51). Interestingly, histones
none have been validated in vivo. Interestingly, are extremely effective antimicrobials and
inhibition of bacterial cell wall synthesis (49) were one of the first antimicrobials described
was recently shown at low concentrations that (52). The significance of histones (and of the
may be more similar to those present at inflam- peptides derived from them) as microbials
matory sites. Cathelicidins, including the well- remains to be demonstrated in vivo (53).
studied LL-37, are proteolytically processed Given their dual role as an architectural
468 Amulic et al.

scaffold for DNA and as antimicrobials, their Reactive Oxygen Species

in vivo significance is particularly difficult to Upon activation, neutrophils produce ROS in
demonstrate. a process called the respiratory burst. It is mis- Chronic
The second class of neutrophil antimi- leading to think of ROS as a single entity be- granulomatous
crobials encompasses a broad assortment of cause they differ in their stability, reactivity, and disease (CGD):
proteolytic enzymes that participate in microbe permeability to membranes (62). However, all
caused by mutations
destruction. Lysozyme destroys the bacterial rendering the
ROS can modify and damage other molecules, NADPH oxidase
wall, making it an obvious antimicrobial, as properties exploited by the host cell for signal- nonfunctional,
shown in mice deficient in this enzyme (54). ing and antimicrobial action. characterized by
Surprisingly, this occurred independently of its The NADPH oxidase complex assembles susceptibility to
enzymatic activity (55). Neutrophils also con- on the phagosomal and plasma membranes
infection and
tain several serine proteases (including PR3, autoinflammation

and begins the reactive oxygen cascade by
CG, and NE, collectively known as the serpro- reducing molecular oxygen to superoxide.
cidins) that exhibit differing specificities. They Downstream of superoxide, many potential
are tightly regulated intra- and extracellularly reactions can occur (for details, see References
by serpins, indicating that their activity is 62–64). Superoxide, though not a strong
deployed under specific conditions. NE cleaves oxidant, rapidly dismutates, forming hydrogen
enterobacterial virulence factors with high peroxide. Superoxide can also react with nitric
specificity (56), indicating the possibility of the oxide, which is produced at high levels at
coevolution of microbial virulence factors and inflammatory sites, to form peroxynitrite, a
antimicrobial effectors. Of further interest, NE strong oxidant. Upon degranulation into the
mutations in humans, but not genetic ablation phagosome, MPO can react with hydrogen
of this enzyme in mice, result in neutropenia. peroxide to produce various reactive species,
This can be rescued by the administration of including hypohalous acids. Hypochlorous
recombinant granulocyte macrophage colony- acid, thought to be the major product of MPO
stimulating factor (GM-CSF); however, these in the phagosome, is more reactive than su-
patients still exhibit significant susceptibility peroxide and is antimicrobial in vitro. Thus, it
to infections. Mice deficient in NE or CG is assumed to have direct antimicrobial effects
are highly susceptible to bacterial and fungal in the phagosome. However, a theoretical
infections (57, 58). Another protein, azuro- model of the phagosome suggests that most of
cidin, is a member of the same family but lacks the hypochlorous acid produced would react
protease activity. Unexpectedly, it still kills with host proteins before reaching the bac-
microbes, suggesting that these proteins may terium. This model predicts that chloramines,
all have antimicrobial activity independent produced when hypochlorous acid reacts
of proteolysis, perhaps as a result of their with amine groups, may be the most relevant
cationicity. These serine proteases also play a antimicrobial actors in the phagosome (65).
salient role in autoimmunity (see discussion in ROS are clearly important for neutrophil
section on Autoimmunity, below) (59). antimicrobial activity: Neutrophils from
The final class of neutrophil antimicrobials chronic granulomatous disease (CGD) patients
consists of a number of proteins that chelate kill microbes poorly, making these patients
essential metals from microbes and possibly susceptible to many infections. Interestingly,
impact bacterial growth. Two of these chela- CGD patients can control catalase-negative
tors are lactoferrin, first identified in milk, bacteria, which produce, but do not degrade,
which binds preferentially to iron, and cal- their own hydrogen peroxide, thus providing
protectin (also called S100A and many other a substrate for reactions downstream in the
names), which sequesters zinc (60) and results in reactive oxygen cascade (66). NADPH ox-
“nutritional immunity” (61). idase is also implicated in the regulation of

inflammation, which explains why CGD (73, 74). Furthermore, superoxide generation
patients often suffer from autoinflammatory leads to an ionic influx into the phagosome to
diseases (67). compensate for charge; this may activate gran-
Paradoxically, although MPO is required ule proteases by releasing them from their pu-
for neutrophil microbicidal activity in vitro, tative matrix (75). There is controversy around
MPO-deficient individuals do not have striking which ions and which channel are responsible
clinical manifestations (68, 69). Some MPO- for charge compensation, but this theory of
deficient individuals suffer from frequent or se- protease activation is certainly intriguing (69).
vere infections, especially with Candida species, Studies of ROS are hampered by various
and a few have been mistaken for CGD patients. technical issues. Ideally, a probe for ROS
However, most MPO-deficient individuals in should be specific, targetable to particular
the developed world have apparently normal intracellular compartments, and capable of
immunity. The mild effects of MPO deficiency being used in vivo. Traditional probes for
suggest that MPO’s products are not essential ROS do not meet these specifications; in
for antimicrobial action. Indeed, in the absence addition, the probes often become radical
of MPO, other reactive species (e.g., superox- species (76). One promising new approach
ide, hydrogen peroxide, hydroperoxyl radical, for ROS detection that meets these criteria is
peroxynitrite) can still be produced in the the use of redox-sensitive fluorescent protein-
neutrophil phagosome; hydroperoxyl radical is based probes, such as roGFP and HyPer
predicted to be present at antimicrobial concen- (76). Other methods that can be used in vivo
trations (65). However, there may be a broader include transcription profiling of superoxide
reason for this discrepancy. Modern technolo- or hydrogen peroxide–sensitive genes as well
gies can distinguish between individuals who as the detection of relatively stable products of
are partially and completely MPO deficient, reactive oxygen using mass spectrometry (76).
and partial MPO deficiency does not correlate
with pathology (70). Residual activity of MPO
may be sufficient for antimicrobial activity: In Phagocytosis
the case of CGD, even 1% of normal NADPH Phagocytosis is the major mechanism to re-
oxidase activity leads to an improved prognosis move pathogens and cell debris. It is an active,
(71). Epidemiological studies distinguishing receptor-mediated process during which a par-
the degrees of MPO deficiency and their ticle is internalized by the cell membrane into
correlation with clinical manifestations may be a vacuole called the phagosome. As with other
necessary to understand the function of MPO. phagocytes, the mechanistic details of internal-
In addition to direct antimicrobial action, ization depend on the type of interaction be-
ROS can modify host molecules. Because tween the neutrophil and the microorganism.
these species are highly reactive, they are often Interaction can be direct, through recognition
thought to be too nonspecific to be involved in of PAMPs by pattern-recognition receptors, or
signaling. However, specificity can be achieved opsonin mediated. The latter mechanism is bet-
on the submolecular level, by cellular redox ter characterized and includes two prototypical
buffering systems and by limited diffusion of examples: FcγR-mediated phagocytosis, which
ROS owing to their short half-lives (72). A relies on the formation of pseudopod extensions
well-studied example of ROS in signaling is for engulfment of IgG-opsonized particles, and
the reversible regulation of various targets complement receptor-mediated phagocytosis,
(including phosphatases, metalloproteinases, which does not require membrane extensions
and caspases) by direct oxidation of cysteine or pseudopods (77).
residues. In addition, neutrophil granule After engulfment, the nascent phagosome
proteases can be regulated by oxidative inacti- is relatively benign to microorganisms, acquir-
vation of their inhibitors or by direct oxidation ing its lethal properties only after a drastic
470 Amulic et al.

maturation process. Our understanding of importance of phagocytosis in the innate

this process is largely based on studies in immune defense.
macrophages, and although these are certainly
Autophagy: a process
instructive, essential differences exist in neu- in which cellular
trophils. Macrophage phagocytosis follows an Neutrophil Extracellular Traps
contents are degraded
endocytic maturation pathway: In neutrophils, Upon stimulation, neutrophils can undergo in lysosomes,
phagosome maturation happens upon fusion of NETosis, an active form of cell death that especially in
conditions of nutrient
granules to the phagosome, whereby delivery leads to release of decondensed chromatin into
scarcity and infection
of antimicrobial molecules into the phagoso- the extracellular space (86, 87). The fibrous
mal lumen occurs. Simultaneously, assembly structures termed NETs contain histones as
of the NADPH oxidase on the phagosomal well as antimicrobial granular and cytoplasmic
membrane allows ROS production, and jointly, proteins (88). NETs trap many types of mi-
these two mechanisms create an environment crobes ex vivo and have been found in various
toxic to most pathogens. Neutrophil phago- disease models in vivo; they are thought to
somal pH regulation also differs significantly kill microbes by exposing them to high local
from that observed in macrophages. While the concentrations of antimicrobials (89).
macrophage phagosome gradually acidifies, The mechanism of NET formation is not
neutrophil phagosomal pH is initially alkaline completely understood. The reactive oxygen
(78) and remains neutral for prolonged periods pathway is involved, as NADPH oxidase and
of time (79). The maintenance of this alkaline MPO are required for NET formation in re-
pH is essential for the activation of the major sponse to chemical and biological stimuli (87,
serine proteases NE and CG, and it is sustained 90, 91). Nitric oxide donors can induce NETs
via NADPH oxidase activity, despite contin- via a mechanism that also requires ROS (90), a
uing fusion of acidic granules. Key events of finding that awaits genetic confirmation. All ac-
the maturation process are described in more tivators of NET formation tested so far require
detail in Reference 80. ROS production. S. aureus may be an exception,
Not all pathogens succumb to the hostile although those experiments were done using
environment of the phagosome. In fact, some pharmacological inhibitors, not cells deficient
have evolved strategies to survive inside neu- in ROS production (92). Upstream of NADPH
trophils. These strategies include interfering oxidase, the Raf-MEK-ERK pathway is impli-
with engulfment, modulating phagosome cated in NET formation (93), but further along
maturation, and creating a more hospitable in the process, NE translocates from the gran-
intraphagosomal environment. The polysac- ules to the nucleus and degrades histones, lead-
charide capsule expressed by Staphylococcus ing to chromatin decondensation (94). Histone
aureus confers antiphagocytic properties (81). citrullination may also play a role in NET for-
Helicobacter pylori can disrupt targeting of mation, although this has not been confirmed
NADPH oxidase to the phagosome so that in primary human neutrophils (95–97). Au-
superoxide anions accumulate extracellularly tophagy is also thought to be required for NET
rather than in the phagosome (82). Francisella formation, but this has so far been shown only
tularensis prevents triggering of the oxidative using a nonspecific inhibitor of autophagy (98).
burst and also inhibits ROS production in The majority of research on NETs has been
response to other stimuli (83). Finally, other conducted ex vivo. Ideally, to test the relevance
pathogens, such as Salmonella typhimurium and of NETs, a “NETs knockout” organism should
Streptococcus pyogenes, can efficiently block gran- be generated to investigate its response to
ule fusion with the phagosome (84, 85). The pathogens. Unfortunately, it is not possible to
variety of mechanisms evolved by intracellular eliminate the main components of NETs—
pathogens to resist killing and enable survival DNA and histones—from an infection model.
within the phagosome further emphasizes the Moreover, the factors that are important for

NET formation, such as NADPH oxidase, establishing the correct environmental condi-
MPO, and NE, are also critical for other an- tions to launch the adaptive immune response.
timicrobial neutrophil functions. For now, the The cytokines released by PMNs are often
Cystic fibrosis:
caused by defects in evidence for the relevance of NETs is indirect. synthesized de novo. Although neutrophils
the CFTR ion On the one hand, bacteria that express DNases transcribe little after leaving the bone marrow,
transporter, as virulence factors disseminate more efficiently once activated, these cells undergo a tran-
characterized by thick, in the host, which may point to evolutionary scriptional burst that results in the synthesis
sticky mucus and
pressure to avoid entrapment by NETs (99, of signaling molecules (110, 111). Compared
decreases in lung and
digestive function 100). In addition, a persistent Aspergillus with other immune cells (e.g., macrophages),
infection in a CGD patient was cleared after neutrophils typically produce lower amounts
gene therapy, which restored NADPH oxidase of cytokines per cell, but they are so abundant
activity, NET formation, and NET-mediated at inflammatory sites that their contribution
but not phagocytosis-mediated killing by the to total cytokine levels is significant (4). Fur-
patient’s neutrophils ex vivo (101). On the other thermore, neutrophil-secreted proteases can
hand, the immune system has redundant mech- modulate signaling networks in vivo through
anisms to fight infection, and it may be that cytokine processing (112).
NETs are especially important under certain The initial neutrophil cytokine response is
conditions, such as during infections with large an appeal for immunological reinforcement.
pathogens that are not readily phagocytosed. The most abundantly produced cytokine, IL-8,
NETs can also have detrimental effects on primarily serves to recruit other neutrophils
the host. Because NETs expose self molecules (113). Similarly, neutrophil-derived proinflam-
extracellularly, they lead to autoimmunity: matory IL-1β and TNF-α induce other cells
NETs have been implicated in systemic to produce neutrophil chemoattractants (114,
lupus erythematosus (SLE), an autoimmune 115) (for a comprehensive list of cytokines
disease characterized by the formation of produced by neutrophils, please see References
autoantibodies, often against chromatin and 115, 116). In addition to cytokines, neutrophils
neutrophil components (102–106) (see section release other signaling mediators, including
on Autoimmunity, below). Platelet-induced granule contents (117), lipids (118), and ROS
NETs, formed during sepsis, are associated such as hydrogen peroxide (119). They also
with hepatotoxicity due to tissue damage communicate via cell-cell contact (120). Here
(107). Platelets also bind to NETs, raising the we provide examples of how neutrophils
possibility that NETs nucleate blood clots in interact with other cells to shape the immune
the context of deep vein thrombosis (108). response (see Figure 3).
NETs have also been observed in the airway
fluids of cystic fibrosis patients, where they
may increase the viscosity of the sputum and Monocytes and Macrophages
decrease lung function (109). As they respond to infection or injury,
neutrophils and their relatives in the mono-
cyte/macrophage lineage coordinate their
NEUTROPHILS IN IMMUNE activities, leading to alternating waves of re-
CELL CROSS TALK cruitment of these two cell types. Macrophages
Neutrophils participate in the communica- and patrolling monocytes are among the initial
tion networks that form the foundations of detectors of PAMPs and endogenous activators,
immunity, issuing instructions to practically the danger-associated molecular patterns (121),
all other immune cells. As one of the first cell and these cells work to summon large numbers
types to arrive at sites of infection, neutrophils of neutrophils to the inflammatory locus. The
secrete cytokines and chemokines critical in the influx of neutrophils is followed closely by the
unfolding of the inflammatory response and in arrival of monocytes, suggesting a causal link
472 Amulic et al.

Tissue
T cell
Activation and CD8+ Lymph node
IFN-γ
differentiation DC T cell
Crosspriming
ROS?
T cell Arginase? Neutrophil
IFN-γ Activation DC
DC
NK cell
IL-12 DC Antigen
presentation
CD4+
Macrophage Neutrophil T cell Th1
Activation Neutrophil
Bacteria
Neutrophil Monocyte DC
Blood
Figure 3
Neutrophil communication with other immune cells. Neutrophils interact with a variety of cell types. They are important both for
recruitment of monocytes and dendritic cells (DCs) to infected tissues and for enhancement of macrophage and DC activity. In
contrast, in the lymph nodes, neutrophils impede DC function by inhibiting antigen presentation to CD4+ cells. Neutrophils also
interact with the adaptive arm of the immune system: They can act as antigen-presenting cells by cross-presenting antigen to CD8+ T
cells; they also secrete IL-12, which activates T cells. T cells, in turn, activate neutrophils by secreting IFN-γ. Finally, neutrophils,
DCs and natural killer (NK) cells colocalize and enhance each other’s activity via receptor-receptor interactions and soluble mediators.
behind these temporal dynamics. Indeed, neu- microbicidal activity (129). The circuitous
trophils recruit monocytes via several different nature of the cross talk of these two cell types
mechanisms. They express classical monocyte becomes obvious during inflammation abate-
chemoattractants such as CCL2 (MCP-1) ment: Monocytes, recruited by neutrophils
(122), CCL3 (MIP-1α) (123), CCL20 (MIP- and differentiated into macrophages, repress
3α), and CCL19 (MIP-3β) (124). Additionally, further neutrophil chemotaxis and ensure
and perhaps more unexpectedly, neutrophils the appropriate removal of their postmortem
use granule proteins to induce extravasation remains (see section on Neutrophils and
of monocytes in vivo, as shown for LL-37, Resolution of Inflammation, below).
azurocidin (HBP/CAP37), and CG (125–127).
Monocyte recruitment is also affected indirectly
by neutrophils: via upregulation of endothelial Dendritic Cells
adhesion factors, increase of transendothelial Neutrophils can also recruit and activate
permeability, enhancement of production of DCs in vivo. This was recently illustrated
chemoattractants by other cell types, and mod- in a mouse model of Leishmaniasis, where
ulation of the activities of these chemokines subcutaneous inoculation of Leishmania major
via proteolytic processing (reviewed in 128). triggered a massive and rapid infiltration of
In addition to recruitment, neutrophils mod- neutrophils (130). These cells secrete the
ulate monocyte and macrophage cytokine chemokine CCL3, recruiting DCs to the
production (128), directly enhancing their site of inoculation and initiating a protective

Th1 response (131). Interestingly, activated performed in vitro, and their interpretation is
neutrophils can induce the maturation of DCs frustratingly difficult owing to the question-
in vitro through specific receptor-receptor able purity of cell preparations. Recently, it
DC-SIGN:
dendritic cell–specific interactions between Mac-1 and DC-SIGN, was shown that neutrophils, NK cells, and DCs
intercellular adhesion leading to local secretion of TNF-α (120). interact in a ménage à trois involving both
molecule-3-grabbing In this case, the reduced levels of cytokine cytokine signaling and direct cell-cell contact
nonintegrin production foster specificity, as only proximal (137, 138). In one report, infection of mice
Granulocyte DCs receive the maturation signal. A similar with Legionella pneumophila triggered produc-
receptor 1 (Gr1): activation model was earlier proposed for Tox- tion of IFN-γ by NK cells; this was dependent
the anti-Gr1 antibody
oplasma gondii (132). Neutrophil-activated DCs on both PMN-derived IL-18 and DC-derived
RB6-8C5 reacts with
both Ly6G (specific produce the proinflammatory cytokine IL-12 IL-12 (137). Similarly, human neutrophils, NK
for neutrophils) and and induce proliferation of T cells (120, 132). cells, and DCs colocalize at inflammatory sites,
Ly6C (present on However, some of these experiments should and a positive feedback loop has been proposed
many immune cell
be interpreted cautiously because they are on the basis of in vitro data. In this scheme, neu-
types)
based on the injection of the anti-Gr1 antibody trophils interact with a specific subset of DCs,
Th17 cells: subset of (RB6), which depletes neutrophils but may also (via CD18-ICAM-1 interactions), prompting
T helper cells that
result in depletion of many other cell types in the DCs to produce IL-12p70, which in turn
produce IL-17,
important in mice. The anti-Ly6G monoclonal antibody is stimulates IFN-γ production by NK cells and
inflammation and more specific and hence a better reagent for this further activates neutrophils. Simultaneously,
implicated in type of experiment (133). The crucial role of neutrophils also activate NK cells by direct con-
autoimmunity neutrophils in DC activation was recently contact (139). Additional in vitro interactions be-
firmed using anti-Ly6G antibody depletion: In tween neutrophils and NK cells are extensively
Mycobacterium tuberculosis infection, timely traf- reviewed in Reference 138.
ficking of DCs to lymph nodes and activation of
CD4+ T cells were both dependent on PMNs.
Furthermore, this study demonstrated that Lymphocytes
DCs presented bacterial antigens when they A surprising finding in recent years is the exten-
ingested infected neutrophils just as efficiently sive cross talk between cells located at opposite
as they did via direct uptake of Mycobacterium ends of the immune spectrum. Previously
(134). In sharp contrast to the above findings, thought to belong to isolated compartments,
a separate study using an immunization model neutrophils and T cells shape and impact
showed that neutrophils recruited to lymph each other’s functions, both qualitatively and
nodes compete for antigen with DCs and quantitatively (140). Neutrophils affect T cell
macrophages and that these neutrophils inhibit function indirectly via DCs, as outlined above,
their interactions with T cells (135). It is possi- but can also influence T cell function directly.
ble that neutrophils have site-specific effects on PMNs secrete IL-12, which may be crucial for
DCs and can be stimulatory at peripheral sites Th1 cell differentiation (141, 142). They also
and inhibitory in the lymph nodes. Neutrophils express several T cell chemoattractants (116)
exhibit fascinating and somewhat enigmatic be- as well as B cell development and maturation
havior in the lymph nodes, where they engage factors (143, 144). Cytokine communication
in swarming activity in response to parasitic occurs in both directions: For instance, IFN-γ,
infection (136). The functions and mechanistic which is secreted by T cells, prolongs neu-
details of these swarms are unknown and trophil life span, induces gene expression, and
represent questions of immense interest. increases phagocytic capacity (145). The T
helper 17 (Th17) cell subset secretes IL-17,
Natural Killer Cells a key cytokine in the control of neutrophil
Studies of interactions between neutrophil and dynamics, which acts by upregulating expres-
natural killer (NK) cells have historically been sion of CXCL8 (IL-8), G-CSF, and TNF-α
474 Amulic et al.

by epithelial, endothelial, and stromal cells Although some collateral damage to host
(146). Collectively, these Th17-associated tissues is inevitable during infection, neu-
cytokines increase granulopoeisis as well as the trophils must be removed before they have
Ulcerative colitis: a
recruitment and life span of neutrophils. serious, detrimental effects on inflamed tissues. type of inflammatory
Neutrophils potentially have suppressive ef- Resolution of inflammation is an active process bowel disease
fects on T cells via two proposed mechanisms: that limits further leukocyte infiltration and characterized by ulcers
(a) L-arginine depletion by release of arginase, removes apoptotic cells from inflamed sites. and tissue erosion in
the colon and rectum
which inhibits T cell responses in vitro (147), This process is essential for maintenance of
and (b) hydrogen peroxide–mediated suppres- tissue homeostasis and, if impeded, leads to
sion, as proposed in a cancer model (119) (see “nonresolving inflammation,” a problematic
section on Cancer, below). Direct evidence of condition that contributes to many diseases.
such interactions in vivo is still missing.

Interestingly, neutrophils influence CD8+
T cell responses by cross-presenting exogenous

antigens in vivo. Using mice in which profes- Apoptosis and Clearance
sional antigen-presenting cells do not express Apoptosis is a central aspect of inflammation
functional MHC class I, Beauvillain et al. (148) resolution. Once neutrophils have executed
showed that antigen-pulsed neutrophils can their antimicrobial agenda, they die via a built-
induce differentiation of cytotoxic T cells. in cell-death program. However, not only does
These striking findings imply that neutrophils apoptosis reduce the number of neutrophils
have characteristics of antigen-presenting cells. present, it also produces signals that abro-
Neutrophils also appear capable of expressing gate further neutrophil recruitment. Phagocy-
MHC class II and costimulatory molecules tosis of apoptotic neutrophils also reprograms
under inflammatory conditions (149–151), macrophages to adopt an anti-inflammatory
and they can present antigen to CD4+ T cells phenotype.
in vitro (152–154). However, the functional Neutrophil death is influenced by inflamma-
significance for protective immunity remains tory mediators such as GM-CSF and LPS and
unclear, especially in light of the finding that by environmental conditions such as hypoxia,
mouse neutrophils that migrate to the lymph all of which prolong neutrophil survival. The
node have a negative effect on CD4 responses signaling networks that regulate survival have
in an immunization system (135). In humans, also been well characterized. These networks
there are large variations in the ability of also control the expression of known antiapo-
donors to express MHC class II (149, 151), ptotic (Mcl-1 and A1) or proapoptotic proteins
suggesting concomitant variations in the ability (Bad, Bax, Bak, and Bid), and they also activate
to activate T cells, a finding that could have caspases (for an extensive review, see Reference
implications for susceptibility to autoimmune 155). Given that neutrophils are terminally
diseases. Therefore, neutrophil modulation of differentiated, it is unexpected that molecules
adaptive immunity seems to be highly complex controlling cell proliferation regulate survival.
and is only now starting to be unraveled. Proposed to have prosurvival effects, one such
protein is survivin. It is expressed more highly
in immature neutrophils than in mature ones,
NEUTROPHILS AND but its expression can be restored in mature
RESOLUTION OF cells by inflammatory signals such as G-CSF or
INFLAMMATION GM-CSF. In line with these findings, survivin
The lethal cargo of neutrophils is not only is also highly expressed in neutrophils at sites
destructive toward invading microbes, but of inflammation, such as cystic fibrosis sputum,
also harmful to host cells. Thus, neutrophil appendix infiltrates, and intestines of patients
deployment must be tightly controlled. with ulcerative colitis (156).

Similarly, cyclin-dependent kinases func- their vicinity (epithelial cells, endothelial cells,
tion as prosurvival factors in neutrophils. fibroblasts, platelets, and leukocytes) and par-
Pharmacological inhibition of these cell cycle ticipate in the transcellular biosynthesis of lipid
Wegener’s
granulomatosis: regulators induce caspase-dependent apoptosis mediators with anti-inflammatory and prore-
vasculitis affecting the and block life-span extension by survival factors solving activities, such as lipoxins, resolvins, and
lungs, nose, and (157). More recently, prosurvival effects were protectins. A major lipid mediator class switch
kidneys; inflammation also attributed to proliferating cell nuclear thus exists, governed by temporally regulated
leads to reduced blood
antigen (PCNA). This factor usually resides expression of different lipoxygenases and the
flow, tissue
destruction, and in the nucleus, where it is involved in DNA mobilization of different fatty acid substrates.
damage of vital organs replication, but in neutrophils, it associates The different biosynthesis pathways of prore-
Prostaglandins and with procaspases in the cytosol and is thought solving lipid mediators have been reviewed in
leukotrienes: lipids to prevent their activation. During apoptosis, detail elsewhere (118). Interestingly, microor-
synthesized by PCNA is targeted for proteosomal degradation, ganisms are also a source of lipid precursors
cyclooxygenases and
which correlates with an increase in caspase-3 that can be used by neutrophils for resolvin
5-lipoxygenase,
and caspase-8 activities. This mechanism is rel- synthesis. Thus, microbes also likely participate
respectively, in the
arachidonic acid evant in Wegener’s granulomatosis and sepsis, in synthesis of mediators with proresolving
pathway; have where stabilization of PCNA is associated with functions at the site of infection (159, 160).
proinflammatory resistance of neutrophils to apoptosis (158). How do lipid mediators contribute to
functions including Equally important for the resolution of in- the termination of inflammation? Lipoxins,
leukocyte recruitment
flammation is the proper removal of apoptotic resolvins, and protectins exert cell-type specific
cells. This relies on the release of “find-me” effects, promoting monocyte/macrophage
signals at early stages of cell death, which at- recruitment and activation while inhibiting
tract phagocytes. Likewise, distinct “eat me” neutrophil functions. The inhibitory effect
signals are required for specific recognition of extends to all essential steps of neutrophil
apoptotic cells. Ingestion of apoptotic cells by responses: migration, adhesion, and activation.
macrophages drives the production of the anti- All three lipid mediators reduce neutrophil
inflammatory cytokines tumor growth factor recruitment, a process that involves the lipoxin-
(TGF)-β and IL-10 (155). Failure to clear these A4 receptor and the leukotriene B4 receptor
apoptotic cells, by contrast, results in secondary (BLT1) (161–167). Ariel et al. (168) also pro-
necrosis and release of products that generate posed an interesting mechanism of action for
proinflammatory signals (Figure 4). lipoxins, resolvins, and protectins in clearing in-
flammatory sites. They showed that neutrophil
exposure to these lipids increases expression
Lipid Mediator Class Switch of CCR5 on the surface of late apoptotic neu-
Soluble mediators play a crucial role in the trophils, leading to efficient sequestration of the
resolution of inflammation. In neutrophils, chemoattractants CCL3 and CCL5. The se-
a particularly prominent role is assumed by questration of these chemokines means they are
lipid mediators. The successful progression unavailable to recruit neutrophils to inflamed
of inflammation appears to hinge on a shift sites (168) (Figure 4). This mechanism com-
in the composition of secreted lipids. At early plements other anti-inflammatory processes
stages of inflammation, neutrophils synthesize in which chemokines are inactivated by neu-
proinflammatory lipid mediators, such as trophil proteases. Of these lipids, lipoxins are
prostaglandins and leukotrienes. These are the most completely understood. In addition to
derived from arachidonate precursor molecules neutrophil recruitment, lipoxins can inhibit the
and are synthesized through the cyclooxy- shedding of L-selectin and the upregulation of
genase and lipoxygenase pathways. During β2 integrins in response to proinflammatory
the later stages of the inflammatory response, stimuli, thereby reducing adhesion of neu-
neutrophils interact with various cell types in trophils to endothelial cells (169, 170). Finally,
476 Amulic et al.

Initiation Resolution
of inflammation Leukotrienes Prostaglandins TNF-α Lipoxins Resolvins Protectins IL-10 TGF-β of inflammation
Platelets
Monocyte
Lipoxins
Neutrophil
Lipoxin
Resolvins
Protectins
Macrophage
Apoptotic
neutrophil Chemokines CCR5
IL-10
Leukotrienes TGF-β
Prostaglandins PGE-2
Chemokines Chemokine clearance
Microorganisms ?
TNF-α NETotic
IL-6 neutrophil Macrophage
Figure 4
From inflammation to homeostasis: neutrophil apoptosis and lipid mediator class switching in the resolution of inflammation. At the
site of infection, resident macrophages initiate an inflammatory response, secreting proinflammatory cytokines and chemokines that
alert the immune system and promote neutrophil recruitment. In the early stages of inflammation, microbes trigger the production of
proinflammatory lipid mediators, such as leukotrienes and prostaglandins, which also recruit neutrophils. As inflammation progresses, a
switch occurs, and anti-inflammatory lipid mediators such as lipoxins, resolvins, and protectins are produced. Notably, interaction of
neutrophils with platelets induces the production of lipoxins. Anti-inflammatory lipid mediators initiate the resolution of inflammation
by blocking neutrophil and promoting monocyte recruitment. Monocytes differentiated into macrophages ingest apoptotic neutrophils,
driving the production of the anti-inflammatory cytokines tumor growth factor (TGF)-β and IL-10 and prostaglandin-E2 (PGE-2),
which drive the lipid mediator class switch. Proresolving lipid mediators also promote the expression of CCR5 on the surface of
apoptotic neutrophils, providing a means of scavenging chemokines. Chemokine clearance upon phagocytosis of apoptotic neutrophils
by macrophages further contributes to the reduction of neutrophil infiltration and the return to tissue homeostasis. The contribution of
macrophages to the clearance of NETotic neutrophils, and how this could impact inflammation resolution, is currently unknown. A
timeline of the inflammation process from initiation to resolution is summarized in the upper part of the figure.
lipoxins also impact neutrophil activation by Disorders Associated with

inhibiting ROS and peroxynitrite production, Nonresolved Inflammation
NF-κB activation, and IL-8 expression (170).
The failure of neutrophils to apoptose or mal-
In addition to directly impacting neu-
functions in the removal of their apoptotic re-
trophil functions, lipid mediators promote
mains result in chronic inflammation. These
nonphlogistic (noninflammatory) phagocyto-
conditions lead to the accumulation of cyto-
sis of apoptotic neutrophils by monocytes
toxic substances and are associated with severe Chronic obstructive
and macrophages. In the presence of anti-
pathologies, including cystic fibrosis, chronic pulmonary disease
inflammatory lipids, engulfment of apoptotic (COPD): lung disease
obstructive pulmonary disease (COPD), and
neutrophils is not accompanied by the release of caused by noxious
rheumatoid arthritis (RA). The severity of in-
proinflammatory mediators, as typically occurs particles or gas, e.g.,
flammation often directly correlates with poor
during macrophage activation. Instead, produc- tobacco smoking;
clinical outcome. inflammation leads to
tion of the anti-inflammatory cytokines TGF-β
COPD is a major cause of death in indus- lung obstruction
and IL-10 is increased (163, 171).
trialized nations, where smoking is a prime

instigator of this disease. A chronic neutrophil the previous section. It is, however, unknown
infiltration in the lungs of COPD patients whether all neutrophils are capable of adapting
promotes tissue damage and organ dysfunc- to the changing chemoattractant environment
Rheumatoid arthritis
(RA): chronic tion. One of the key molecules controlling or if different subsets of neutrophils are suc-
inflammatory disease the inflammatory response in the lung is cessively involved. The relevance of this model
that affects many leukotriene A4 hydrolase (LTA4H). This in human disease remains to be established,
tissues and organs but enzyme has two opposing activities. First, its although the clinical similarities between this
primarily synovial
hydrolase activity converts leukotriene A4 into mouse model and human RA are encouraging.
joints; severe
inflammation causes leukotriene B4, a potent neutrophil chemoat-
deformity tractant and proinflammatory agent. Second, NEUTROPHILS IN DISEASE
LTA4H is an aminopeptidase that inactivates
Neutrophils are prominent players in the innate

a specific neutrophil chemoattractant, the
immune response and the clearance of infec-
proline-glycine-proline tripeptide (PGP), thus
tion, a subject addressed in several prominent
conferring the enzyme with anti-inflammatory

reviews. However, neutrophil action can also
properties. Interestingly, tobacco smoke selec-
support disease progression in other illnesses.
tively inhibits only the aminopeptidase activity
A host of autoimmune disorders belong to this
of LTA4H, promoting the accumulation of
category. In addition, certain malignant cancers
both leukotriene B4 and PGP. This in turn
are also prime examples of illnesses in which
promotes neutrophil recruitment and fuels
neutrophils play a salient role.
chronic lung inflammation (172).
Another prime example of a disease linked to
nonresolving inflammation is RA. Neutrophils Cancer
are the most abundant leukocytes present in the The link between cancer and inflammation
synovial fluid of RA patients, and their role in was noted as early as 1863 by Rudolf Virchow
pathogenesis has been demonstrated in several (177). Since then, it has been proposed that
animal models. These models primarily used neutrophil-derived ROS have the potential to
neutrophil depletion or adoptive transfer of initiate tumor formation by genotoxic stress
wild-type neutrophils in leukotriene-deficient and induction of genomic instability. Although
mice (173–175). In one model, synthesis this has been demonstrated in vitro (178, 179),
of leukotriene B4 by neutrophils in joints convincing evidence for PMN-mediated DNA
is essential for disease development (174). mutagenesis in vivo is still lacking. Neutrophils
Leukotriene B4 can act in an autocrine manner do, however, impact cancer progression.
via the neutrophil receptor BLT1 to promote They are abundant in tumors and influence
the recruitment of a first wave of neutrophils tumor development through several secreted
into the joint. Later, the recruitment of a mediators, including cytokines, ROS, and
second wave of neutrophils is independent of matrix-degrading proteases (reviewed in Ref-
this leukotriene B4–BLT1 pathway. At this erence 180). The majority of findings support
stage, immune complexes are essential for a “protumor” and “antihost” effect of these
stimulating infiltrating neutrophils to deliver cells; clinical studies indicate that neutrophil
IL-1β into the joint. This in turn induces the infiltration of tumors is associated with poorer
production of chemokines by synovial tissue prognosis (181, 182). Indeed, some cancers
cells and sustains neutrophil recruitment (175, seem to actively recruit neutrophils through
176). These studies exemplify the complex production of IL-8 (183). In agreement with
regulation cascades involving lipids, cytokines, this, antibody depletion of neutrophils reduces
and chemokines that orchestrate neutrophil tumor growth (184). The protumor function
recruitment in chronic inflammation. They of neutrophils operates at multiple levels,
also demonstrate the cross talk between neu- including production of angiogenic factors
trophils and other immune cells discussed in (185), enhancement of metastasis (186), and
478 Amulic et al.

suppression of the antitumor immune response human renal cell carcinoma, MDSCs have
(119, 187). Using the anti-Ly6G antibody, identical morphology and express the same sur-
Fridlender and colleagues (187) depleted neu- face markers as do activated neutrophils (190,
Acute-phase
trophils and confirmed their tumorigenic role. 191). MDSCs inhibit T cell proliferation by proteins: secreted by
Moreover, the study showed that neutrophils in limiting L-arginine availability via arginase and liver, concentration in
the tumor microenvironment could, under cer- NOS activities, both of which use this amino plasma changes by
tain circumstances, be induced to target their acid as a substrate (189, 191, 192). Furthermore, 25% or more during
inflammation
cytotoxic arsenal at tumor cells, whose growth MDSCs are strong producers of ROS, which
they usually help to fuel. Pharmacological suppresses T cell responses (119, 192). Inter-
inhibition of TGF-β signaling led tumor- fering with the release of MDSCs or using drug
associated neutrophils to assume a heightened interventions to polarize neutrophil responses
proinflammatory state, causing a reduction in in the tumor microenvironment could repre-

tumor growth. These alternatively activated sent novel therapeutic strategies against cancer.
neutrophils underwent a complete reversal in

their effect on CD8+ T cells, serving to activate
rather than suppress these cells. Differential Autoimmunity
neutrophil responses were also demonstrated in Deregulated neutrophil cell death and/or
a melanoma study. In this instance, increased clearance often accompanies autoimmune syn-
systemic levels of the acute-phase protein dromes (193–195) and may play a major role
serum amyloid A (SAA-1) induced neutrophils in disease pathogenesis, given that release of
to secrete the anti-inflammatory cytokine IL- proteolytic and cytotoxic molecules from neu-
10, which also inhibited T cell responses. Cross trophils can trigger organ damage. Neutrophil
talk with invariant NKT cells could counter products act as both targets and mediators of
this response, restoring a proinflammatory autoimmunity. MPO and PR3 are the main tar-
activation status (188). Thus, investigation of gets of antineutrophil cytoplasmic antibodies
neutrophils in cancer has revealed considerable (ANCA), autoantibodies directed against anti-
plasticity in their responses. Although little gens present in the cytoplasm of neutrophils.
evidence currently supports the existence of Wegener’s granulomatosis is consistently as-
different populations, it is likely that neutrophil sociated with the presence of ANCA. Further-
responses are more flexible and less stereotyped more, the extent of organ damage in patients
than previously thought. with Wegener’s granulomatosis correlates with
Another major mechanism of tumor escape the PMN infiltrate rather than with traditional
from immune control has recently been autoimmunity parameters such as T cell acti-
attributed to a heterogeneous category of im- vation or autoantibody titers (196). Likewise,
mature myeloid cells, called myeloid-derived ANCA bind MPO and PR3 expressed on the
suppressor cells (MDSCs) (189). In healthy surface of activated neutrophils, promoting
individuals, MDSCs are found in the bone degranulation and release of chemoattractants
marrow, where they differentiate into mature and ROS, which together lead to a vicious
neutrophils and monocytes. In cancer and cycle of tissue damage and inflammation. Early
some autoimmune and infectious diseases, reports also suggest that, in an inflammatory en-
differentiation is partially blocked, leading to vironment, ANCA accelerate ROS-dependent
accumulation of these precursors, which act as neutrophil apoptosis, suggesting a feed-forward
powerful suppressors of T cell functions. MD- cycle culminating in organ damage (194, 195).
SCs have characteristics of neutrophils, and in SLE is another chronic autoimmune disease
mice, they are typically detected using the neu- affecting multiple tissues and organs. Autoan-
trophil surface markers CD11b+ and Gr-1+ , tibodies produced in SLE are predominantly
although they consist of variable proportions either ANCA or directed against chromatin.
of monocytic and granulocytic cells (189). In Although neutrophils had long been suspected

to be causative agents, their role in SLE patho- to produce IFN-α, a central cytokine in SLE
genesis remained elusive. The recent discovery pathogenesis (103, 104). However, it remains to
of a link between SLE and NET formation be determined if DCs can present NET com-
Vasculitis:
inflammation of blood has helped to shed light on this quandary. ponents or if they contribute to autoreactive B
vessels It was proposed that TNF-α and IFN-α cell activation. It is also possible that NETs are
prime cells for NET formation in response to involved in other autoimmune diseases. Should
anti-PR3, antiribonucleoprotein, anti-HNP, this prove to be the case, understanding the
or anti-LL-37 autoantibodies (103, 104, 106). role of NETs may provide critical insights into
Thus, high levels of inflammatory cytokines in the role of microbial infections as a trigger of
autoimmune patients are believed to sensitize autoimmunity.
neutrophils to NETosis, whereas autoanti-
bodies may trigger a switch from apoptosis to

NETosis. Additional evidence suggesting a CONCLUDING REMARKS
role for NETs in autoimmune pathology was Neutrophils are specialized phagocytes that
obtained when NETs were identified in renal arose as an evolutionary adaptation in verte-
and/or skin biopsies from patients with SLE brates to coordinate and execute one of the most
and small vessel vasculitis (103–106). Several fundamental physiological responses: inflam-
studies have reported the presence of a particu- mation. They are endowed with antimicrobial
lar subset of neutrophils in PBMC preparations mechanisms that make them the preeminent
from pediatric and adult SLE patients. These microbe exterminators of the immune system.
low-density granulocytes display phenotypic In addition to this important role, PMNs also
characteristics of immature neutrophils with network with many other immune cells and
nonsegmented nuclei and higher expression help regulate the initiation of specific T and
of MPO, NE, and defensin-3, and they may B cell immunity. However, neutrophils do not
be related to the MDSCs discussed previously always act in ways beneficial to the host: Uncon-
(see section on Cancer, above) (197, 198). trolled neutrophil responses can exacerbate and
An increased capacity to form NETs and a even cause autoimmune and inflammatory dis-
heightened cytotoxicity toward endothelial eases. Many challenges remain in understand-
cells could bestow them with pathogenic ing neutrophil function: Is there specialization
properties in lupus (105). among PMNs? Are they more plastic than we
Because NETs appear to be formed during suspect? How do they make decisions before
autoimmune disease, their timely removal may deploying their armamentaria? How do they
be an essential mechanism for maintaining kill microbes? How specific are their instruc-
tissue homeostasis. Human serum contains the tions to other cells? Answering these questions
nuclease DNase I, which degrades NETs in will better define neutrophils’ role in defense
vitro. Notably, a familial form of SLE is linked and disease and will provide a rational path for
to a mutation in DNase I (199). Furthermore, pursuing new therapies. Moreover, neutrophils
in a cohort of SLE patients, 36% exhibited can potentially provide insights into several
either elevated titers of autoantibodies directed unique aspects of basic cell biology. Their strik-
against NET components or inhibitors of ingly short life spans make them excellent mod-
DNase I, both of which may protect NETs els for investigating cell death, whereas their
from degradation. Most notably, impaired reliance on ROS as biochemical effectors may
NET degradation correlates with development reveal novel ways for relaying intracellular
of lupus nephritis, one of the most severe signals. The uniquely lobulated neutrophil
manifestations of SLE (102). nucleus is a feat of higher-order nuclear
Can it be that NETs play a general role architecture that is just beginning to yield
in modulation of autoimmune responses? We its secrets. In short, exciting times await the
know that NETs induce plasmacytoid DCs humble neutrophil.
480 Amulic et al.

DISCLOSURE STATEMENT
The authors are not aware of any affiliations, memberships, funding, or financial holdings that
might be perceived as affecting the objectivity of this review.
ACKNOWLEDGMENTS
We thank Diane Schad for assistance with graphic design and Cornelia Heinz for administrative
help. G.H. is an Alexander von Humboldt Foundation Scholar, and B.A. is supported by an EMBO
Long-Term Fellowship.
LITERATURE CITED
1. Ehrlich P. 1880. Methodologische Beiträge zur Physiologie und Pathologie der verschiedenen Formen
der Leukocyten. Z. Klin. Med. 1:553–60

2. Waller A. 1846. Microscopic observation on the perforation of capillaries by the corpuscles of blood and
the origin of mucus and pus globules. Lond. Edinburgh Philos. Mag. J. Sci. 2:271–80
3. Metchnikoff E. 1893. Lecon sur la pathologie comparee de inflammation. Ann. Inst. Pasteur 7:348–57
4. Nathan C. 2006. Neutrophils and immunity: challenges and opportunities. Nat. Rev. Immunol. 6:173–82
5. Borregaard N. 2010. Neutrophils, from marrow to microbes. Immunity 33:657–70
6. Borregaard N, Cowland JB. 1997. Granules of the human neutrophilic polymorphonuclear leukocyte.
Blood 89:3503–21
7. Finger EB, Puri KD, Alon R, Lawrence MB, von Andrian UH, Springer TA. 1996. Adhesion through
L-selectin requires a threshold hydrodynamic shear. Nature 379:266–69
8. Lawrence MB, Kansas GS, Kunkel EJ, Ley K. 1997. Threshold levels of fluid shear promote leukocyte
adhesion through selectins (CD62L,P,E). J. Cell Biol. 136:717–27
9. Kansas GS. 1996. Selectins and their ligands: current concepts and controversies. Blood 88:3259–87
10. McEver RP, Cummings RD. 1997. Role of PSGL-1 binding to selectins in leukocyte recruitment.
J. Clin. Investig. 100:S97–103
11. Yago T, Shao B, Miner JJ, Yao L, Klopocki AG, et al. 2010. E-selectin engages PSGL-1 and CD44
through a common signaling pathway to induce integrin αLβ2-mediated slow leukocyte rolling. Blood
116:485–94
12. Mueller H, Stadtmann A, Van Aken H, Hirsch E, Wang D, et al. 2010. Tyrosine kinase Btk regulates
E-selectin-mediated integrin activation and neutrophil recruitment by controlling phospholipase C
(PLC) γ2 and PI3Kγ pathways. Blood 115:3118–27
13. Ley K, Laudanna C, Cybulsky MI, Nourshargh S. 2007. Getting to the site of inflammation: the leukocyte
adhesion cascade updated. Nat. Rev. Immunol. 7:678–89
14. Campbell JJ, Hedrick J, Zlotnik A, Siani MA, Thompson DA, Butcher EC. 1998. Chemokines and the
arrest of lymphocytes rolling under flow conditions. Science 279:381–84
15. Constantin G, Majeed M, Giagulli C, Piccio L, Kim JY, et al. 2000. Chemokines trigger immediate β2
integrin affinity and mobility changes: differential regulation and roles in lymphocyte arrest under flow.
Immunity 13:759–69
16. Liu S, Kiosses WB, Rose DM, Slepak M, Salgia R, et al. 2002. A fragment of paxillin binds the α4
integrin cytoplasmic domain (tail) and selectively inhibits α4-mediated cell migration. J. Biol. Chem.
277:20887–94
17. Goldfinger LE, Han J, Kiosses WB, Howe AK, Ginsberg MH. 2003. Spatial restriction of α4 inte-
grin phosphorylation regulates lamellipodial stability and α4β1-dependent cell migration. J. Cell Biol.
162:731–41
18. Xu J, Wang F, Van Keymeulen A, Herzmark P, Straight A, et al. 2003. Divergent signals and cytoskeletal
assemblies regulate self-organizing polarity in neutrophils. Cell 114:201–14
19. Schenkel AR, Chew TW, Muller WA. 2004. Platelet endothelial cell adhesion molecule deficiency or
blockade significantly reduces leukocyte emigration in a majority of mouse strains. J. Immunol. 173:6403–
8

20. Phillipson M, Heit B, Colarusso P, Liu L, Ballantyne CM, Kubes P. 2006. Intraluminal crawling of
neutrophils to emigration sites: a molecularly distinct process from adhesion in the recruitment cascade.
J. Exp. Med. 203:2569–75
21. Wong CH, Heit B, Kubes P. 2010. Molecular regulators of leucocyte chemotaxis during inflammation.
Cardiovasc. Res. 86:183–91
22. Zarbock A, Ley K. 2008. Mechanisms and consequences of neutrophil interaction with the endothelium.
Am. J. Pathol. 172:1–7
23. Selvatici R, Falzarano S, Mollica A, Spisani S. 2006. Signal transduction pathways triggered by selective
formylpeptide analogues in human neutrophils. Eur. J. Pharmacol. 534:1–11
24. Chen Y, Yao Y, Sumi Y, Li A, To UK, et al. 2010. Purinergic signaling: a fundamental mechanism in
neutrophil activation. Sci. Signal. 3:ra45
25. Sabroe I, Dower SK, Whyte MK. 2005. The role of Toll-like receptors in the regulation of neutrophil
migration, activation, and apoptosis. Clin. Infect. Dis. 41(Suppl. 7):S421–26

26. Parker LC, Whyte MK, Dower SK, Sabroe I. 2005. The expression and roles of Toll-like receptors in
the biology of the human neutrophil. J. Leukoc. Biol. 77:886–92

27. Ley K. 2002. Integration of inflammatory signals by rolling neutrophils. Immunol. Rev. 186:8–18
28. Guthrie LA, McPhail LC, Henson PM, Johnston RB Jr. 1984. Priming of neutrophils for enhanced
release of oxygen metabolites by bacterial lipopolysaccharide. Evidence for increased activity of the
superoxide-producing enzyme. J. Exp. Med. 160:1656–71
29. El-Benna J, Dang PM, Gougerot-Pocidalo MA. 2008. Priming of the neutrophil NADPH oxidase
activation: role of p47phox phosphorylation and NOX2 mobilization to the plasma membrane. Semin.
Immunopathol. 30:279–89
30. Didsbury JR, Uhing RJ, Tomhave E, Gerard C, Gerard N, Snyderman R. 1991. Receptor class desen-
sitization of leukocyte chemoattractant receptors. Proc. Natl. Acad. Sci. USA 88:11564–68
31. Claing A, Laporte SA, Caron MG, Lefkowitz RJ. 2002. Endocytosis of G protein–coupled receptors:
roles of G protein–coupled receptor kinases and β-arrestin proteins. Prog. Neurobiol. 66:61–79
32. Nusse O, Lindau M. 1988. The dynamics of exocytosis in human neutrophils. J. Cell Biol. 107:2117–23
33. Lacy P. 2005. The role of Rho GTPases and SNAREs in mediator release from granulocytes. Pharmacol.
Ther. 107:358–76
34. Faurschou M, Borregaard N. 2003. Neutrophil granules and secretory vesicles in inflammation. Microbes
Infect. 5:1317–27
35. Faurschou M, Borregaard N. 2003. Neutrophil granules and secretory vesicles in inflammation. Microbes
Infect. 5:1317–27
36. Borregaard N, Sorensen OE, Theilgaard-Monch K. 2007. Neutrophil granules: a library of innate
immunity proteins. Trends Immunol. 28:340–45
37. Borregaard N, Kjeldsen L, Lollike K, Sengelov H. 1992. Ca2+ -dependent translocation of cytosolic
proteins to isolated granule subpopulations and plasma membrane from human neutrophils. FEBS Lett.
304:195–97
38. Borregaard N, Kjeldsen L, Sengelov H, Diamond MS, Springer TA, et al. 1994. Changes in subcellular
localization and surface expression of L-selectin, alkaline phosphatase, and Mac-1 in human neutrophils
during stimulation with inflammatory mediators. J. Leukoc. Biol. 56:80–87
39. Sengelov H, Kjeldsen L, Borregaard N. 1993. Control of exocytosis in early neutrophil activation.
J. Immunol. 150:1535–43
40. Kjeldsen L, Bjerrum OW, Askaa J, Borregaard N. 1992. Subcellular localization and release of human
neutrophil gelatinase, confirming the existence of separate gelatinase-containing granules. Biochem. J.
287(Pt. 2):603–10
41. Kjeldsen L, Bainton DF, Sengelov H, Borregaard N. 1993. Structural and functional heterogene-
ity among peroxidase-negative granules in human neutrophils: identification of a distinct gelatinase-
containing granule subset by combined immunocytochemistry and subcellular fractionation. Blood
82:3183–91
42. Faurschou M, Sorensen OE, Johnsen AH, Askaa J, Borregaard N. 2002. Defensin-rich granules of human
neutrophils: characterization of secretory properties. Biochim. Biophys. Acta Mol. Cell Res. 1591:29–35
482 Amulic et al.

43. Delclaux C, Delacourt C, D’Ortho MP, Boyer V, Lafuma C, Harf A. 1996. Role of gelatinase B and
elastase in human polymorphonuclear neutrophil migration across basement membrane. Am. J. Respir.
Cell Mol. Biol. 14:288–95
44. Singer II, Scott S, Kawka DW, Kazazis DM. 1989. Adhesomes: specific granules containing receptors
for laminin, C3bi/fibrinogen, fibronectin, and vitronectin in human polymorphonuclear leukocytes and
monocytes. J. Cell Biol. 109:3169–82
45. Jesaitis AJ, Buescher ES, Harrison D, Quinn MT, Parkos CA, et al. 1990. Ultrastructural localization
of cytochrome b in the membranes of resting and phagocytosing human granulocytes. J. Clin. Investig.
85:821–35
46. Soehnlein O, Zernecke A, Weber C. 2009. Neutrophils launch monocyte extravasation by release of
granule proteins. Thromb. Haemost. 102:198–205
47. Brogden KA. 2005. Antimicrobial peptides: pore formers or metabolic inhibitors in bacteria? Nat. Rev.
Microbiol. 3:238–50
48. Schroeder BO, Wu Z, Nuding S, Groscurth S, Marcinowski M, et al. 2011. Reduction of disulphide
bonds unmasks potent antimicrobial activity of human β-defensin 1. Nature 469:419–23
49. Schneider T, Kruse T, Wimmer R, Wiedemann I, Sass V, et al. 2010. Plectasin, a fungal defensin, targets
the bacterial cell wall precursor Lipid II. Science 328:1168–72
50. Lande R, Gregorio J, Facchinetti V, Chatterjee B, Wang YH, et al. 2007. Plasmacytoid dendritic cells
sense self-DNA coupled with antimicrobial peptide. Nature 449:564–69
51. Canny G, Levy O. 2008. Bactericidal/permeability-increasing protein (BPI) and BPI homologs at mu-
cosal sites. Trends. Immunol. 29:541–47
52. Hirsch JG. 1958. Bactericidal action of histone. J. Exp. Med. 108:925–44
53. Park CB, Yi KS, Matsuzaki K, Kim MS, Kim SC. 2000. Structure-activity analysis of buforin II, a histone
H2A-derived antimicrobial peptide: the proline hinge is responsible for the cell-penetrating ability of
buforin II. Proc. Natl. Acad. Sci. USA 97:8245–50
54. Markart P, Korfhagen TR, Weaver TE, Akinbi HT. 2004. Mouse lysozyme M is important in pulmonary
host defense against Klebsiella pneumoniae infection. Am. J. Respir. Crit. Care Med. 169:454–58
55. Nash JA, Ballard TN, Weaver TE, Akinbi HT. 2006. The peptidoglycan-degrading property of lysozyme
is not required for bactericidal activity in vivo. J. Immunol. 177:519–26
56. Weinrauch Y, Drujan D, Shapiro SD, Weiss J, Zychlinsky A. 2002. Neutrophil elastase targets virulence
factors of enterobacteria. Nature 417:91–94
57. Belaaouaj A, McCarthy R, Baumann M, Gao Z, Ley TJ, et al. 1998. Mice lacking neutrophil elastase
reveal impaired host defense against gram negative bacterial sepsis. Nat. Med. 4:615–18
58. Tkalcevic J, Novelli M, Phylactides M, Iredale JP, Segal AW, Roes J. 2000. Impaired immunity and
enhanced resistance to endotoxin in the absence of neutrophil elastase and cathepsin G. Immunity 12:201–
10
59. Campanelli D, Detmers PA, Nathan CF, Gabay JE. 1990. Azurocidin and a homologous serine protease
from neutrophils. Differential antimicrobial and proteolytic properties. J. Clin. Investig. 85:904–15
60. Corbin BD, Seeley EH, Raab A, Feldmann J, Miller MR, et al. 2008. Metal chelation and inhibition of
bacterial growth in tissue abscesses. Science 319:962–65
61. Weinberg ED. 1975. Nutritional immunity. Host’s attempt to withhold iron from microbial invaders.
J. Am. Med. Assoc. 231:39–41
62. Hampton MB, Kettle AJ, Winterbourn CC. 1998. Inside the neutrophil phagosome: oxidants, myeloper-
oxidase, and bacterial killing. Blood 92:3007–17
63. Bogdan C, Rollinghoff M, Diefenbach A. 2000. Reactive oxygen and reactive nitrogen intermediates in
innate and specific immunity. Curr. Opin. Immunol. 12:64–76
64. Winterbourn CC. 2008. Reconciling the chemistry and biology of reactive oxygen species. Nat. Chem.
Biol. 4:278–86
65. Winterbourn CC, Hampton MB, Livesey JH, Kettle AJ. 2006. Modeling the reactions of superoxide
and myeloperoxidase in the neutrophil phagosome: implications for microbial killing. J. Biol. Chem.
281:39860–69
66. Segal BH, Leto TL, Gallin JI, Malech HL, Holland SM. 2000. Genetic, biochemical, and clinical features
of chronic granulomatous disease. Medicine 79:170–200

67. Seger RA. 2010. Chronic granulomatous disease: recent advances in pathophysiology and treatment.
Neth. J. Med. 68:334–40
68. Klebanoff SJ. 2005. Myeloperoxidase: friend and foe. J. Leukoc. Biol. 77:598–625
69. Nauseef WM. 2007. How human neutrophils kill and degrade microbes: an integrated view. Immunol.
Rev. 219:88–102
70. Kutter D. 1998. Prevalence of myeloperoxidase deficiency: population studies using Bayer-Technicon
automated hematology. J. Mol. Med. 76:669–75
71. Kuhns DB, Alvord WG, Heller T, Feld JJ, Pike KM, et al. 2010. Residual NADPH oxidase and survival
in chronic granulomatous disease. N. Engl. J. Med. 363:2600–10
72. Nathan C. 2003. Specificity of a third kind: reactive oxygen and nitrogen intermediates in cell signaling.
J. Clin. Investig. 111:769–78
73. Johnson D, Travis J. 1979. Oxidative inactivation of human α-1-proteinase inhibitor - further evidence
for methionine at the reactive center. J. Biol. Chem. 254:4022–26

74. Shao BH, Belaaouaj A, Verlinde C, Fu XY, Heinecke JW. 2005. Methionine sulfoxide and proteolytic
cleavage contribute to the inactivation of cathepsin G by hypochlorous acid—an oxidative mechanism
for regulation of serine proteinases by myeloperoxidase. J. Biol. Chem. 280:29311–21

75. Reeves EP, Lu H, Lortat-Jacob H, Messina CGM, Bolsover S, et al. 2002. Killing activity of neutrophils
is mediated through activation of proteases by K+ flux. Nature 416:291–97
76. Murphy MP, Holmgren A, Larsson NG, Halliwell B, Chang CJ, et al. 2011. Unraveling the biological
roles of reactive oxygen species. Cell Metab. 13:361–66
77. Underhill DM, Ozinsky A. 2002. Phagocytosis of microbes: complexity in action. Annu. Rev. Immunol.
20:825–52
78. Segal AW, Geisow M, Garcia R, Harper A, Miller R. 1981. The respiratory burst of phagocytic cells is
associated with a rise in vacuolar pH. Nature 290:406–9
79. Jankowski A, Scott CC, Grinstein S. 2002. Determinants of the phagosomal pH in neutrophils. J. Biol.
Chem. 277:6059–66
80. Lee WL, Harrison RE, Grinstein S. 2003. Phagocytosis by neutrophils. Microbes Infect. 5:1299–306
81. Luong TT, Lee CY. 2002. Overproduction of type 8 capsular polysaccharide augments Staphylococcus
aureus virulence. Infect. Immun. 70:3389–95
82. Allen LA, Beecher BR, Lynch JT, Rohner OV, Wittine LM. 2005. Helicobacter pylori disrupts NADPH
oxidase targeting in human neutrophils to induce extracellular superoxide release. J. Immunol. 174:3658–
67
83. McCaffrey RL, Schwartz JT, Lindemann SR, Moreland JG, Buchan BW, et al. 2010. Multiple mecha-
nisms of NADPH oxidase inhibition by type A and type B Francisella tularensis. J. Leukoc. Biol. 88:791–805
84. Joiner KA, Ganz T, Albert J, Rotrosen D. 1989. The opsonizing ligand on Salmonella typhimurium
influences incorporation of specific, but not azurophil, granule constituents into neutrophil phagosomes.
J. Cell Biol. 109:2771–82
85. Staali L, Bauer S, Morgelin M, Bjorck L, Tapper H. 2006. Streptococcus pyogenes bacteria modulate mem-
brane traffic in human neutrophils and selectively inhibit azurophilic granule fusion with phagosomes.
Cell. Microbiol. 8:690–703
86. Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, et al. 2004. Neutrophil extracellular
traps kill bacteria. Science 303:1532–35
87. Fuchs TA, Abed U, Goosmann C, Hurwitz R, Schulze I, et al. 2007. Novel cell death program leads to
neutrophil extracellular traps. J. Cell Biol. 176:231–41
88. Urban CF, Ermert D, Schmid M, Abu-Abed U, Goosmann C, et al. 2009. Neutrophil extracellular traps
contain calprotectin, a cytosolic protein complex involved in host defense against Candida albicans. PLoS
Pathog. 5:e1000639
89. Papayannopoulos V, Zychlinsky A. 2009. NETs: a new strategy for using old weapons. Trends Immunol.
30:513–21
90. Patel S, Kumar S, Jyoti A, Srinag BS, Keshari RS, et al. 2010. Nitric oxide donors release extracellular
traps from human neutrophils by augmenting free radical generation. Nitric Oxide 22:226–34
91. Metzler KD, Fuchs TA, Nauseef WM, Reumaux D, Roesler J, et al. 2011. Myeloperoxidase is required
for neutrophil extracellular trap formation: implications for innate immunity. Blood 117:953–59
484 Amulic et al.

92. Pilsczek FH, Salina D, Poon KKH, Fahey C, Yipp BG, et al. 2010. A novel mechanism of rapid nuclear
neutrophil extracellular trap formation in response to Staphylococcus aureus. J. Immunol. 185:7413–25
93. Hakkim A, Fuchs TA, Martinez NE, Hess S, Prinz H, et al. 2011. Activation of the Raf-MEK-ERK
pathway is required for neutrophil extracellular trap formation. Nat. Chem. Biol. 7:75–77
94. Papayannopoulos V, Metzler KD, Hakkim A, Zychlinsky A. 2010. Neutrophil elastase and myeloperox-
idase regulate the formation of neutrophil extracellular traps. J. Cell Biol. 191:677–91
95. Neeli I, Khan SN, Radic M. 2008. Histone deimination as a response to inflammatory stimuli in neu-
trophils. J. Immunol. 180:1895–902
96. Wang Y, Li M, Stadler S, Correll S, Li P, et al. 2009. Histone hypercitrullination mediates chromatin
decondensation and neutrophil extracellular trap formation. J. Cell Biol. 184:205–13
97. Li P, Li M, Lindberg MR, Kennett MJ, Xiong N, Wang Y. 2010. PAD4 is essential for antibacterial
innate immunity mediated by neutrophil extracellular traps. J. Exp. Med. 207:1853–62
98. Remijsen Q, Vanden Berghe T, Wirawan E, Asselbergh B, Parthoens E, et al. 2011. Neutrophil extra-
cellular trap cell death requires both autophagy and superoxide generation. Cell Res. 21:290–304
99. Beiter K, Wartha F, Albiger B, Normark S, Zychlinsky A, Henriques-Normark B. 2006. An endonuclease

allows Streptococcus pneumoniae to escape from neutrophil extracellular traps. Curr. Biol. 16:401–7
100. Buchanan JT, Simpson AJ, Aziz RK, Liu GY, Kristian SA, et al. 2006. DNase expression allows the
pathogen group A Streptococcus to escape killing in neutrophil extracellular traps. Curr. Biol. 16:396–400
101. Bianchi M, Hakkim A, Brinkmann V, Siler U, Seger RA, et al. 2009. Restoration of NET formation by
gene therapy in CGD controls aspergillosis. Blood 114:2619–22
102. Hakkim A, Furnrohr BG, Amann K, Laube B, Abed UA, et al. 2010. Impairment of neutrophil extracel-
lular trap degradation is associated with lupus nephritis. Proc. Natl. Acad. Sci. USA 107:9813–18
103. Garcia-Romo GS, Caielli S, Vega B, Connolly J, Allantaz F, et al. 2011. Netting neutrophils are major
inducers of type I IFN production in pediatric systemic lupus erythematosus. Sci. Transl. Med. 3:73ra20
104. Lande R, Ganguly D, Facchinetti V, Frasca L, Conrad C, et al. 2011. Neutrophils activate plasmacytoid
dendritic cells by releasing self-DNA-peptide complexes in systemic lupus erythematosus. Sci. Transl.
Med. 3:73ra19
105. Villanueva E, Yalavarthi S, Berthier CC, Hodgin JB, Khandpur R, et al. 2011. Netting neutrophils
induce endothelial damage, infiltrate tissues, and expose immunostimulatory molecules in systemic lupus
erythematosus. J. Immunol. 187:538–52
106. Kessenbrock K, Krumbholz M, Schonermarck U, Back W, Gross WL, et al. 2009. Netting neutrophils
in autoimmune small-vessel vasculitis. Nat. Med. 15:623–25
107. Clark SR, Ma AC, Tavener SA, McDonald B, Goodarzi Z, et al. 2007. Platelet TLR4 activates neutrophil
extracellular traps to ensnare bacteria in septic blood. Nat. Med. 13:463–69
108. Fuchs TA, Brill A, Duerschmied D, Schatzberg D, Monestier M, et al. 2010. Extracellular DNA traps
promote thrombosis. Proc. Natl. Acad. Sci. USA 107:15880–85
109. Marcos V, Zhou Z, Yildirim AO, Bohla A, Hector A, et al. 2010. CXCR2 mediates NADPH oxidase-
independent neutrophil extracellular trap formation in cystic fibrosis airway inflammation. Nat. Med.
16:1018–23
110. Subrahmanyam YV, Yamaga S, Prashar Y, Lee HH, Hoe NP, et al. 2001. RNA expression patterns
change dramatically in human neutrophils exposed to bacteria. Blood 97:2457–68
111. Kobayashi SD, Voyich JM, Buhl CL, Stahl RM, DeLeo FR. 2002. Global changes in gene expression
by human polymorphonuclear leukocytes during receptor-mediated phagocytosis: Cell fate is regulated
at the level of gene expression. Proc. Natl. Acad. Sci. USA 99:6901–6
112. Meyer-Hoffert U, Wiedow O. 2010. Neutrophil serine proteases: mediators of innate immune responses.
Curr. Opin. Hematol. 18:19–24
113. Scapini P, Lapinet-Vera JA, Gasperini S, Calzetti F, Bazzoni F, Cassatella MA. 2000. The neutrophil as
a cellular source of chemokines. Immunol. Rev. 177:195–203
114. Sica A, Matsushima K, Van Damme J, Wang JM, Polentarutti N, et al. 1990. IL-1 transcriptionally
activates the neutrophil chemotactic factor/IL-8 gene in endothelial cells. Immunology 69:548–53
115. Kasama T, Miwa Y, Isozaki T, Odai T, Adachi M, Kunkel SL. 2005. Neutrophil-derived cytokines:
potential therapeutic targets in inflammation. Curr. Drug Targets Inflamm. Allergy 4:273–9

116. Scapini P, Lapinet-Vera JA, Gasperini S, Calzetti F, Bazzoni F, Cassatella MA. 2000. The neutrophil as
a cellular source of chemokines. Immunol. Rev. 177:195–203
117. Yang D, de la Rosa G, Tewary P, Oppenheim JJ. 2009. Alarmins link neutrophils and dendritic cells.
Trends Immunol. 30:531–37
118. Serhan CN. 2007. Resolution phase of inflammation: novel endogenous anti-inflammatory and prore-
solving lipid mediators and pathways. Annu. Rev. Immunol. 25:101–37
119. Schmielau J, Finn OJ. 2001. Activated granulocytes and granulocyte-derived hydrogen peroxide are
the underlying mechanism of suppression of t-cell function in advanced cancer patients. Cancer Res.
61:4756–60
120. van Gisbergen KP, Sanchez-Hernandez M, Geijtenbeek TB, van Kooyk Y. 2005. Neutrophils mediate
immune modulation of dendritic cells through glycosylation-dependent interactions between Mac-1 and
DC-SIGN. J. Exp. Med. 201:1281–92
121. Cailhier JF, Partolina M, Vuthoori S, Wu S, Ko K, et al. 2005. Conditional macrophage ablation
demonstrates that resident macrophages initiate acute peritoneal inflammation. J. Immunol. 174:2336–42
122. Yoshimura T, Takahashi M. 2007. IFN-γ-mediated survival enables human neutrophils to produce
MCP-1/CCL2 in response to activation by TLR ligands. J. Immunol. 179:1942–49

123. Katsura T, Kobayashi K, Hosaka M, Sugihara S, Kasama T, et al. 1993. Desensitization of delayed-type
hypersensitivity in mice: suppressive environment. Med. Inflamm. 2:205–10
124. Scapini P, Laudanna C, Pinardi C, Allavena P, Mantovani A, et al. 2001. Neutrophils produce biologically
active macrophage inflammatory protein-3α (MIP-3α)/CCL20 and MIP-3β/CCL19. Eur. J. Immunol.
31:1981–88
125. Soehnlein O, Zernecke A, Eriksson EE, Rothfuchs AG, Pham CT, et al. 2008. Neutrophil secretion
products pave the way for inflammatory monocytes. Blood 112:1461–71
126. De Y, Chen Q, Schmidt AP, Anderson GM, Wang JM, et al. 2000. LL-37, the neutrophil granule-
and epithelial cell-derived cathelicidin, utilizes formyl peptide receptor-like 1 (FPRL1) as a receptor to
chemoattract human peripheral blood neutrophils, monocytes, and T cells. J. Exp. Med. 192:1069–74
127. Chertov O, Ueda H, Xu LL, Tani K, Murphy WJ, et al. 1997. Identification of human neutrophil-
derived cathepsin G and azurocidin/CAP37 as chemoattractants for mononuclear cells and neutrophils.
J. Exp. Med. 186:739–47
128. Soehnlein O, Weber C, Lindbom L. 2009. Neutrophil granule proteins tune monocytic cell function.
Trends Immunol. 30:538–46
129. Soehnlein O, Kai-Larsen Y, Frithiof R, Sorensen OE, Kenne E, et al. 2008. Neutrophil primary granule
proteins HBP and HNP1-3 boost bacterial phagocytosis by human and murine macrophages. J. Clin.
Investig. 118:3491–502
130. Peters NC, Egen JG, Secundino N, Debrabant A, Kimblin N, et al. 2008. In vivo imaging reveals an
essential role for neutrophils in Leishmaniasis transmitted by sand flies. Science 321:970–74
131. Charmoy M, Brunner-Agten S, Aebischer D, Auderset F, Launois P, et al. 2010. Neutrophil-derived
CCL3 is essential for the rapid recruitment of dendritic cells to the site of Leishmania major inoculation
in resistant mice. PLoS Pathog. 6:e1000755
132. Bennouna S, Bliss SK, Curiel TJ, Denkers EY. 2003. Cross-talk in the innate immune system: neutrophils
instruct recruitment and activation of dendritic cells during microbial infection. J. Immunol. 171:6052–58
133. Daley JM, Thomay AA, Connolly MD, Reichner JS, Albina JE. 2008. Use of Ly6G-specific monoclonal
antibody to deplete neutrophils in mice. J. Leukoc. Biol. 83:64–70
134. Blomgran R, Ernst JD. 2011. Lung neutrophils facilitate activation of naive antigen-specific CD4+
T cells during Mycobacterium tuberculosis infection. J. Immunol. 186:7110–19
135. Yang CW, Strong BS, Miller MJ, Unanue ER. 2010. Neutrophils influence the level of antigen presen-
tation during the immune response to protein antigens in adjuvants. J. Immunol. 185:2927–34
136. Chtanova T, Schaeffer M, Han SJ, van Dooren GG, Nollmann M, et al. 2008. Dynamics of neutrophil
migration in lymph nodes during infection. Immunity 29:487–96
137. Sporri R, Joller N, Hilbi H, Oxenius A. 2008. A novel role for neutrophils as critical activators of NK
cells. J. Immunol. 181:7121–30
138. Costantini C, Cassatella MA. 2011. The defensive alliance between neutrophils and NK cells as a novel
arm of innate immunity. J. Leukoc. Biol. 89:221–33
486 Amulic et al.

139. Costantini C, Calzetti F, Perbellini O, Micheletti A, Scarponi C, et al. 2011. Human neutrophils interact
with both 6-sulfo LacNAc+ DC and NK cells to amplify NK-derived IFNγ: role of CD18, ICAM-1,
and ICAM-3. Blood 117:1677–86
140. Muller I, Munder M, Kropf P, Hansch GM. 2009. Polymorphonuclear neutrophils and T lymphocytes:
strange bedfellows or brothers in arms? Trends Immunol. 30:522–30
141. Romani L, Mencacci A, Cenci E, Spaccapelo R, Del Sero G, et al. 1997. Neutrophil production of IL-12
and IL-10 in candidiasis and efficacy of IL-12 therapy in neutropenic mice. J. Immunol. 158:5349–56
142. Tateda K, Moore TA, Deng JC, Newstead MW, Zeng X, et al. 2001. Early recruitment of neutrophils
determines subsequent T1/T2 host responses in a murine model of Legionella pneumophila pneumonia.
J. Immunol. 166:3355–61
143. Scapini P, Bazzoni F, Cassatella MA. 2008. Regulation of B-cell-activating factor (BAFF)/B lymphocyte
stimulator (BLyS) expression in human neutrophils. Immunol. Lett. 116:1–6
144. Huard B, McKee T, Bosshard C, Durual S, Matthes T, et al. 2008. APRIL secreted by neutrophils
binds to heparan sulfate proteoglycans to create plasma cell niches in human mucosa. J. Clin. Investig.
118:2887–95
145. Ellis TN, Beaman BL. 2004. Interferon-γ activation of polymorphonuclear neutrophil function.
Immunology 112:2–12
146. Ouyang W, Kolls JK, Zheng Y. 2008. The biological functions of T helper 17 cell effector cytokines in
inflammation. Immunity 28:454–67
147. Munder M, Schneider H, Luckner C, Giese T, Langhans CD, et al. 2006. Suppression of T-cell functions
by human granulocyte arginase. Blood 108:1627–34
148. Beauvillain C, Delneste Y, Scotet M, Peres A, Gascan H, et al. 2007. Neutrophils efficiently cross-prime
naive T cells in vivo. Blood 110:2965–73
149. Gosselin EJ, Wardwell K, Rigby WF, Guyre PM. 1993. Induction of MHC class II on human poly-
morphonuclear neutrophils by granulocyte/macrophage colony-stimulating factor, IFN-γ, and IL-3.
J. Immunol. 151:1482–90
150. Mudzinski SP, Christian TP, Guo TL, Cirenza E, Hazlett KR, Gosselin EJ. 1995. Expression of HLA-
DR (major histocompatibility complex class II) on neutrophils from patients treated with granulocyte-
macrophage colony-stimulating factor for mobilization of stem cells. Blood 86:2452–53
151. Reinisch W, Lichtenberger C, Steger G, Tillinger W, Scheiner O, et al. 2003. Donor dependent,
interferon-γ induced HLA-DR expression on human neutrophils in vivo. Clin. Exp. Immunol. 133:476–
84
152. Fanger NA, Liu C, Guyre PM, Wardwell K, O’Neil J, et al. 1997. Activation of human T cells by major
histocompatability complex class II expressing neutrophils: proliferation in the presence of superantigen,
but not tetanus toxoid. Blood 89:4128–35
153. Radsak M, Iking-Konert C, Stegmaier S, Andrassy K, Hansch GM. 2000. Polymorphonuclear neu-
trophils as accessory cells for T-cell activation: major histocompatibility complex class II restricted
antigen-dependent induction of T-cell proliferation. Immunology 101:521–30
154. Culshaw S, Millington OR, Brewer JM, McInnes IB. 2008. Murine neutrophils present Class II restricted
antigen. Immunol. Lett. 118:49–54
155. Kennedy AD, DeLeo FR. 2009. Neutrophil apoptosis and the resolution of infection. Immunol. Res.
43:25–61
156. Altznauer F, Martinelli S, Yousefi S, Thurig C, Schmid I, et al. 2004. Inflammation-associated cell
cycle–independent block of apoptosis by survivin in terminally differentiated neutrophils. J. Exp. Med.
199:1343–54
157. Rossi AG, Sawatzky DA, Walker A, Ward C, Sheldrake TA, et al. 2006. Cyclin-dependent kinase
inhibitors enhance the resolution of inflammation by promoting inflammatory cell apoptosis. Nat. Med.
12:1056–64
158. Witko-Sarsat V, Mocek J, Bouayad D, Tamassia N, Ribeil JA, et al. 2010. Proliferating cell nuclear
antigen acts as a cytoplasmic platform controlling human neutrophil survival. J. Exp. Med. 207:2631–45
159. Arita M, Clish CB, Serhan CN. 2005. The contributions of aspirin and microbial oxygenase to the
biosynthesis of anti-inflammatory resolvins: novel oxygenase products from omega-3 polyunsaturated
fatty acids. Biochem. Biophys. Res. Commun. 338:149–57

160. Vance RE, Hong S, Gronert K, Serhan CN, Mekalanos JJ. 2004. The opportunistic pathogen Pseudomonas
aeruginosa carries a secretable arachidonate 15-lipoxygenase. Proc. Natl. Acad. Sci. USA 101:2135–39
161. Devchand PR, Arita M, Hong S, Bannenberg G, Moussignac RL, et al. 2003. Human ALX receptor
regulates neutrophil recruitment in transgenic mice: roles in inflammation and host defense. FASEB J.
17:652–59
162. Arita M, Ohira T, Sun YP, Elangovan S, Chiang N, Serhan CN. 2007. Resolvin E1 selectively interacts
with leukotriene B4 receptor BLT1 and ChemR23 to regulate inflammation. J. Immunol. 178:3912–17
163. Schwab JM, Chiang N, Arita M, Serhan CN. 2007. Resolvin E1 and protectin D1 activate inflammation-
resolution programmes. Nature 447:869–74
164. Spite M, Norling LV, Summers L, Yang R, Cooper D, et al. 2009. Resolvin D2 is a potent regulator of
leukocytes and controls microbial sepsis. Nature 461:1287–91
165. Xu ZZ, Zhang L, Liu T, Park JY, Berta T, et al. 2010. Resolvins RvE1 and RvD1 attenuate inflammatory
pain via central and peripheral actions. Nat. Med. 16:592–97

166. Krishnamoorthy S, Recchiuti A, Chiang N, Yacoubian S, Lee CH, et al. 2010. Resolvin D1 binds human
phagocytes with evidence for proresolving receptors. Proc. Natl. Acad. Sci. USA 107:1660–65
167. Oh SF, Pillai PS, Recchiuti A, Yang R, Serhan CN. 2011. Pro-resolving actions and stereoselective
biosynthesis of 18S E-series resolvins in human leukocytes and murine inflammation. J. Clin. Investig.
121:569–81
168. Ariel A, Fredman G, Sun YP, Kantarci A, Van Dyke TE, et al. 2006. Apoptotic neutrophils and T cells
sequester chemokines during immune response resolution through modulation of CCR5 expression.
Nat. Immunol. 7:1209–16
169. Filep JG, Zouki C, Petasis NA, Hachicha M, Serhan CN. 1999. Anti-inflammatory actions of lipoxin
A(4) stable analogs are demonstrable in human whole blood: modulation of leukocyte adhesion molecules
and inhibition of neutrophil-endothelial interactions. Blood 94:4132–42
170. Jozsef L, Zouki C, Petasis NA, Serhan CN, Filep JG. 2002. Lipoxin A4 and aspirin-triggered 15-epi-
lipoxin A4 inhibit peroxynitrite formation, NF-κB and AP-1 activation, and IL-8 gene expression in
human leukocytes. Proc. Natl. Acad. Sci. USA 99:13266–71
171. Godson C, Mitchell S, Harvey K, Petasis NA, Hogg N, Brady HR. 2000. Cutting edge: lipoxins rapidly
stimulate nonphlogistic phagocytosis of apoptotic neutrophils by monocyte-derived macrophages.
J. Immunol. 164:1663–67
172. Snelgrove RJ, Jackson PL, Hardison MT, Noerager BD, Kinloch A, et al. 2010. A critical role for LTA4H
in limiting chronic pulmonary neutrophilic inflammation. Science 330:90–94
173. Tanaka D, Kagari T, Doi H, Shimozato T. 2006. Essential role of neutrophils in anti-type II collagen
antibody and lipopolysaccharide-induced arthritis. Immunology 119:195–202
174. Chen M, Lam BK, Kanaoka Y, Nigrovic PA, Audoly LP, et al. 2006. Neutrophil-derived leukotriene B4
is required for inflammatory arthritis. J. Exp. Med. 203:837–42
175. Kim ND, Chou RC, Seung E, Tager AM, Luster AD. 2006. A unique requirement for the leukotriene
B4 receptor BLT1 for neutrophil recruitment in inflammatory arthritis. J. Exp. Med. 203:829–35
176. Chou RC, Kim ND, Sadik CD, Seung E, Lan Y, et al. 2010. Lipid-cytokine-chemokine cascade drives
neutrophil recruitment in a murine model of inflammatory arthritis. Immunity 33:266–78
177. Virchow R. 1862 /1863. Die krankhaften Geschwülste: 30 Vorlesungen gehalten während des Wintersemesters
1862/1863. Berlin: Hirschwald
178. Shacter E, Beecham EJ, Covey JM, Kohn KW, Potter M. 1988. Activated neutrophils induce prolonged
DNA damage in neighboring cells. Carcinogenesis 9:2297–304
179. Dizdaroglu M, Olinski R, Doroshow JH, Akman SA. 1993. Modification of DNA bases in chromatin of
intact target human cells by activated human polymorphonuclear leukocytes. Cancer Res. 53:1269–72
180. Gregory AD, McGarry Houghton A. 2011. Tumor-associated neutrophils: new targets for cancer ther-
apy. Cancer Res. 71:2411–16
181. Bellocq A, Antoine M, Flahault A, Philippe C, Crestani B, et al. 1998. Neutrophil alveolitis in bron-
chioloalveolar carcinoma: induction by tumor-derived interleukin-8 and relation to clinical outcome.
Am. J. Pathol. 152:83–92
488 Amulic et al.

182. Jensen HK, Donskov F, Marcussen N, Nordsmark M, Lundbeck F, von der Maase H. 2009. Presence of
intratumoral neutrophils is an independent prognostic factor in localized renal cell carcinoma. J. Clin.
Oncol. 27:4709–17
183. Ji H, Houghton AM, Mariani TJ, Perera S, Kim CB, et al. 2006. K-ras activation generates an inflam-
matory response in lung tumors. Oncogene 25:2105–12
184. Pekarek LA, Starr BA, Toledano AY, Schreiber H. 1995. Inhibition of tumor growth by elimination of
granulocytes. J. Exp. Med. 181:435–40
185. Shojaei F, Singh M, Thompson JD, Ferrara N. 2008. Role of Bv8 in neutrophil-dependent angiogenesis
in a transgenic model of cancer progression. Proc. Natl. Acad. Sci. USA 105:2640–45
186. Huh SJ, Liang S, Sharma A, Dong C, Robertson GP. 2010. Transiently entrapped circulating tumor
cells interact with neutrophils to facilitate lung metastasis development. Cancer Res. 70:6071–82
187. Fridlender ZG, Sun J, Kim S, Kapoor V, Cheng G, et al. 2009. Polarization of tumor-associated neu-
trophil phenotype by TGF-β: “N1” versus “N2” TAN. Cancer Cell 16:183–94
188. De Santo C, Arscott R, Booth S, Karydis I, Jones M, et al. 2010. Invariant NKT cells modulate the
suppressive activity of IL-10-secreting neutrophils differentiated with serum amyloid A. Nat. Immunol.
11:1039–46
189. Gabrilovich DI, Nagaraj S. 2009. Myeloid-derived suppressor cells as regulators of the immune system.
Nat. Rev. Immunol. 9:162–74
190. Zea AH, Rodriguez PC, Atkins MB, Hernandez C, Signoretti S, et al. 2005. Arginase-producing myeloid
suppressor cells in renal cell carcinoma patients: a mechanism of tumor evasion. Cancer Res. 65:3044–48
191. Rodriguez PC, Ernstoff MS, Hernandez C, Atkins M, Zabaleta J, et al. 2009. Arginase I-producing
myeloid-derived suppressor cells in renal cell carcinoma are a subpopulation of activated granulocytes.
Cancer Res. 69:1553–60
192. Kusmartsev S, Nefedova Y, Yoder D, Gabrilovich DI. 2004. Antigen-specific inhibition of CD8+
T cell response by immature myeloid cells in cancer is mediated by reactive oxygen species. J. Immunol.
172:989–99
193. Raza K, Scheel-Toellner D, Lee CY, Pilling D, Curnow SJ, et al. 2006. Synovial fluid leukocyte apoptosis
is inhibited in patients with very early rheumatoid arthritis. Arthritis Res. Ther. 8:R120
194. Harper L, Cockwell P, Adu D, Savage CO. 2001. Neutrophil priming and apoptosis in anti-neutrophil
cytoplasmic autoantibody-associated vasculitis. Kidney Int. 59:1729–38
195. Ren Y, Tang J, Mok MY, Chan AW, Wu A, Lau CS. 2003. Increased apoptotic neutrophils and
macrophages and impaired macrophage phagocytic clearance of apoptotic neutrophils in systemic lupus
erythematosus. Arthritis Rheum. 48:2888–97
196. Brouwer E, Huitema MG, Mulder AH, Heeringa P, van Goor H, et al. 1994. Neutrophil activation in
vitro and in vivo in Wegener’s granulomatosis. Kidney Int. 45:1120–31
197. Hacbarth E, Kajdacsy-Balla A. 1986. Low density neutrophils in patients with systemic lupus erythe-
matosus, rheumatoid arthritis, and acute rheumatic fever. Arthritis Rheum. 29:1334–42
198. Bennett L, Palucka AK, Arce E, Cantrell V, Borvak J, et al. 2003. Interferon and granulopoiesis signatures
in systemic lupus erythematosus blood. J. Exp. Med. 197:711–23
199. Yasutomo K, Horiuchi T, Kagami S, Tsukamoto H, Hashimura C, et al. 2001. Mutation of DNASE1
in people with systemic lupus erythematosus. Nat. Gen. 28:313–14
200. Chen G, Zhuchenko O, Kuspa A. 2007. Immune-like phagocyte activity in the social amoeba. Science
317:678–81
201. Ribeiro C, Brehelin M. 2006. Insect haemocytes: What type of cell is that? J. Insect Physiol. 52:417–29
202. Martin JS, Renshaw SA. 2009. Using in vivo zebrafish models to understand the biochemical basis of
neutrophilic respiratory disease. Biochem. Soc. Trans. 37:830–37
203. Robert J, Ohta Y. 2009. Comparative and developmental study of the immune system in Xenopus. Dev.
Dyn. 238:1249–70
204. Hawkey CM. 1985. Analysis of hematologic findings in healthy and sick adult chimpanzees
(Pan troglodytes). J. Med. Primatol. 14:327–43

IY30-Frontmatter ARI 17 February 2012 11:21
Annual Review of
Immunology
Contents Volume 30, 2012
Decisions About Dendritic Cells: Past, Present, and Future

Ralph M. Steinman p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 1
The Basel Institute for Immunology
Fritz Melchers p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p23

Regulation of Immune Responses by mTOR
Jonathan D. Powell, Kristen N. Pollizzi, Emily B. Heikamp,
and Maureen R. Horton p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p39
Sphingosine-1-Phosphate and Lymphocyte Egress from Lymphoid Organs
Jason G. Cyster and Susan R. Schwab p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p69
Selection of Self-Reactive T Cells in the Thymus
Gretta L. Stritesky, Stephen C. Jameson, and Kristin A. Hogquist p p p p p p p p p p p p p p p p p p p p p p p95
Adaptive Immunity to Fungi
Marcel Wüthrich, George S. Deepe, Jr., and Bruce Klein p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 115
Microbial Translocation Across the GI Tract
Jason M. Brenchley and Daniel C. Douek p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 149
The Response to and Repair of RAG-Mediated DNA Double-Strand Breaks
Beth A. Helmink and Barry P. Sleckman p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 175
VLR-Based Adaptive Immunity
Thomas Boehm, Nathanael McCurley, Yoichi Sutoh, Michael Schorpp,
Masanori Kasahara, and Max D. Cooper p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 203
Immune Regulatory Function of B Cells
Claudia Mauri and Anneleen Bosma p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 221
Lung Dendritic Cells in Respiratory Viral Infection and Asthma: From
Protection to Immunopathology
Bart N. Lambrecht and Hamida Hammad p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 243
Tolerance of Infections
Janelle S. Ayres and David S. Schneider p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 271
microRNA Regulation of Inflammatory Responses
Ryan M. O’Connell, Dinesh S. Rao, and David Baltimore p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 295
ix
IY30-Frontmatter ARI 17 February 2012 11:21
Reflex Principles of Immunological Homeostasis

Ulf Andersson and Kevin J. Tracey p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 313
Chromatin Topology and the Regulation of Antigen Receptor Assembly
Claudia Bossen, Robert Mansson, and Cornelis Murre p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 337
Siglecs and Immune Regulation
Shiv Pillai, Ilka Arun Netravali, Annaiah Cariappa, and Hamid Mattoo p p p p p p p p p p p p p 357
Monogenic Autoimmunity
Mickie H. Cheng and Mark S. Anderson p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 393
Germinal Centers
Gabriel D. Victora and Michel C. Nussenzweig p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 429
Neutrophil Function: From Mechanisms to Disease

Borko Amulic, Christel Cazalet, Garret L. Hayes, Kathleen D. Metzler,
and Arturo Zychlinsky p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 459
Signaling by Myeloid C-Type Lectin Receptors in Immunity and
Homeostasis
David Sancho and Caetano Reis e Sousa p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 491
Regulatory T Cells: Mechanisms of Differentiation and Function
Steven Z. Josefowicz, Li-Fan Lu, and Alexander Y. Rudensky p p p p p p p p p p p p p p p p p p p p p p p p p p 531
Pathogenesis of Human B Cell Lymphomas
Arthur L. Shaffer III, Ryan M. Young, and Louis M. Staudt p p p p p p p p p p p p p p p p p p p p p p p p p p p 565
Autophagy and the Immune System
Petric Kuballa, Whitney M. Nolte, Adam B. Castoreno, and Ramnik J. Xavier p p p p p p p 611
Innate Lymphoid Cells: Emerging Insights in Development, Lineage
Relationships, and Function
Hergen Spits and Tom Cupedo p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 647
Cancer and Inflammation: An Old Intuition with Rapidly Evolving New
Concepts
Giorgio Trinchieri p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 677
Transcriptional and Epigenetic Control of T Helper Cell Specification:
Molecular Mechanisms Underlying Commitment and Plasticity
Yuka Kanno, Golnaz Vahedi, Kiyoshi Hirahara, Kentner Singleton,
and John J. O’Shea p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 707
Induced CD4+ Foxp3+ Regulatory T Cells in Immune Tolerance
Angelina M. Bilate and Juan J. Lafaille p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 733
The Microbiome in Infectious Disease and Inflammation
Kenya Honda and Dan R. Littman p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 759
x Contents

Neutrophil Function - From Mechanisms To Disease

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Neutrophil Function - From Mechanisms To Disease

Uploaded by

Copyright:

Available Formats

IY30CH19-Zychlinsky ARI 17 February 2012 13:38

• Other articles in this volume

• Our comprehensive search

and Arturo Zychlinsky∗

Annu. Rev. Immunol. 2012. 30:459–89 Keywords

The Annual Review of Immunology is online at Abstract

INTRODUCTION early and enthusiastic evolutionary biologist in-

of the neutrophil also inspired Metchnikoff to

phonuclear leukocytes (or PMNs), a title that

460 Amulic et al.

Unfortunately, in vivo studies of neutrophil infection. Indeed, the number of neutrophils

a comprehensive understanding of the neu- to reducing leakage of their dangerous cargo

www.annualreviews.org • Neutrophil Functions 461

abundant; these compounds stimulate the selectins, chemoattractants, cytokines, and

462 Amulic et al.

subsequent to degranulation; however, conclu- point of high chemoattractant concentration,

neutrophil activation. In the interstitial space, physiology at different concentrations, a

www.annualreviews.org • Neutrophil Functions 463

In fact, one could be (erroneously) led to

a Capture b Rolling c Firm adhesion

464 Amulic et al.

neutrophil maturation. They are named for

Primary Secondary Tertiary Secretory

Stage of Myeloblast Promyelocyte Myelocyte Metamyelocyte Band cell

Characteristic Lysozyme Complement receptor 1

Other Cathepsin G, PR3, Gp91phox/p22phox, Gp91phox/p22phox, Gp91phox/p22phox,

www.annualreviews.org • Neutrophil Functions 465

466 Amulic et al.

www.annualreviews.org • Neutrophil Functions 467

Table 1 Mechanism of action of neutrophil antimicrobial proteins

Proteinase 3 (PR3) Mechanism independent of a proteolytic activity by binding to the

468 Amulic et al.

scaffold for DNA and as antimicrobials, their Reactive Oxygen Species

tain several serine proteases (including PR3, autoinﬂammation

www.annualreviews.org • Neutrophil Functions 469

470 Amulic et al.

maturation process. Our understanding of importance of phagocytosis in the innate

www.annualreviews.org • Neutrophil Functions 471

472 Amulic et al.

www.annualreviews.org • Neutrophil Functions 473

474 Amulic et al.

such interactions in vivo is still missing.

T cell responses by cross-presenting exogenous

www.annualreviews.org • Neutrophil Functions 475

476 Amulic et al.

lipoxins also impact neutrophil activation by Disorders Associated with

www.annualreviews.org • Neutrophil Functions 477

Neutrophils are prominent players in the innate

conferring the enzyme with anti-inﬂammatory

478 Amulic et al.

proinﬂammatory state, causing a reduction in in the tumor microenvironment could repre-

neutrophils underwent a complete reversal in

www.annualreviews.org • Neutrophil Functions 479

bodies may trigger a switch from apoptosis to

480 Amulic et al.

der Leukocyten. Z. Klin. Med. 1:553–60

www.annualreviews.org • Neutrophil Functions 481

migration, activation, and apoptosis. Clin. Infect. Dis. 41(Suppl. 7):S421–26

the biology of the human neutrophil. J. Leukoc. Biol. 77:886–92

482 Amulic et al.

www.annualreviews.org • Neutrophil Functions 483

for methionine at the reactive center. J. Biol. Chem. 254:4022–26

for regulation of serine proteinases by myeloperoxidase. J. Biol. Chem. 280:29311–21