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Immunologic Research 2002;26/13:95105

Macrophage Signaling
and Respiratory Burst

Karen E. Iles
Henry Jay Forman


Key Words

Macrophages are key defenders of the lung and play an essential

role in mediating the inflammatory response. Critical to this is the
activation of the NADPH oxidase. Through receptor-mediated interactions, extracellular stimuli activate pathways that signal for the
phosphorylation and assembly of the NADPH oxidase. Once the
NADPH oxidase is activated, it produces superoxide and H2O2 in a
process known as the respiratory burst. The involvement of O 2 and
H2O2 in the antimicrobicidal function of macrophages has been
assumed for many years, but it is now clear that the H2O2 produced
by the respiratory burst functions as a second messenger and activates major signaling pathways in the alveolar macrophage. Both
the nuclear factor-B and activator protein-1 transcription factors
are activated by H2O2 produced by the respiratory burst, and, since
these control the inducible expression of genes whose products are
part of the inflammatory response, this may be a critical link between
the respiratory burst and other inflammatory responses. The c-Jun
N-terminal kinase (JNK) and extracellular-regulated kinase (ERK)
pathways, two members of the mitogen-activated protein kinase
family, are also activated by the respiratory burst. JNK is activated
by both exogenous and endogenously produced H2O2. Studies with
ERK have shown that specific agonists of the respiratory burst, but
not bolus H2O2, can activate this pathway. The ERK pathway also
modulates the expression of genes via phosphorylation of the transcription factor Elk-1 that controls the production of the c-Fos transcription factor. Although an understanding of the mechanism of
redox signaling is in its infancy, it is becoming clear that the reactive oxygen species produced by the respiratory burst have a profound effect on intracellular signaling pathways and ultimately in
modulating gene expression.

Signal transduction
NADPH oxidase
Nuclear factor-B
Mitogen-activated protein
Extracellular-regulated kinase
Activator protein-1

Dr. Henry Jay Forman

University of Alabama at Birmingham
Birmingham, AL 35294-0022.

Humana Press Inc.

Department of Environmental
Health Sciences, School of Public
Health, and Center for Free
Radical Biology, University
of Alabama at Birmingham


Alveolar macrophages are the major resident phagocytes in the lung, and as such are
the first line of defense against bacteria and
particles in the lung. Part of the killing mechanism employed by phagocytes depends on
the generation of the superoxide anion in a
metabolic process known as the respiratory
burst (1). This process depends on the assembly and activation of a multisubunit complex
known as the NADPH oxidase. Assembly of
the oxidase is dependent upon the phosphorylation of critical subunits and the translocation of cytosolic components to the plasma
membrane (2,3). The assembly and activation
of the NADPH oxidase have been shown to be
preceded by increases in intracel lular calcium,
activation of IP3-mediated signaling and protein kinase C (PKC) activation (4). Superoxide generation by the NADPH oxidase is
rapidly converted to H2O2 spontaneously and
enzymatically by superoxide dismutase. The
bactericidal properties of the reactive oxygen
species (ROS) are well established. However,
compared to neutrophils, the size of the respiratory burst (i.e., the amount of superoxide
and H 2 O 2 produced) is much reduced in
Information transfer within and between
cells is critical to defending the lung against
infection and other insults. The appropriate
response to stimuli requires the consistent, regulated flow of information from the cell surface, through intracellular signaling pathways,
to the nucleus where the original stimuli is
transduced into changes in gene expression
and the generation of new products (i.e.,
inflammatory cytokines) in response to the initial stimuli. This transfer of information is collectively known as signal transduction. This
process may entail complex pathways that
include second messengers, kinases, and phosphatases. H2O2 has clearly been established


as a signaling molecule in many different cell

types and systems. There is growing evidence
that the products of the respiratory burst, particularly H2O2, are involved in activating signaling pathways in macrophages as well. It is
intriguing to postulate that the main role of the
respiratory burst in macrophages may be to
provide second messengers for signaling,
whereas in neutrophils the main role is defense.
A growing list of intracellular signaling pathways has been implicated in H2O2-mediated
signaling. Work in our laboratory has demonstrated that the nuclear factor-B (NF-B) (5)
and mitogen-activated protein kinase (MAPK)
signaling pathways are activated by the respiratory burst. The activity of c-Jun N-terminal
kinase (JNK) and extracellular-regulated kinase
(ERK), two members of the MAPK signaling
family, has been shown to increase with some,
although not all, agonists known to activate the
respiratory burst (6) (unpublished data). The
mechanism of activation of the respiratory burst
in alveolar macrophages and the changes in
intracellular signaling that this produces are the
focus of this review.
Activation of NADPH Oxidase
in Alveolar Macrophages
The term macrophage includes many different cell types of monocytic lineage that
acquire specific properties in response to their
environment. It is well known that alveolar
macrophages, which are located at the interface between the air and the lung tissue, are
directly exposed to high oxygen levels and
various pollutants. Consequently, they have
different characteristics than peritoneal macrophages, such as in their ability to release tumor
necrosis factor- (TNF-), interleukin-6
(IL-6), NO, and O 2 (7). All macrophages
express the multicomponent enzyme NADPH
oxidase and generate O 2 to varying degrees.
According to the classic model of NADPH

Iles and Forman

oxidase activation, receptor-mediated release

of diacylglycerol and inositol trisphosphate
(IP3) by phospholipase C, which causes release
of Ca2+ from the endoplasmic reticulum (ER),
causes the activation of PKC (4,8). The activation of PKC leads to the phosphorylation of
the cytosolic components of the NADPH oxidase, particularly p47phox (9). The cytosolic
complex, which is composed of p47phox/p67phox/
p40phox and the small guanosine 5-triphosphatase Rac1/Rac2, translocates to the plasma
membrane and forms a new complex with the
membrane-bound flavocytochrome subunits
gp91phox and p22phox. Homologs to the catalytic gp91phox have been found in other cell
types, now collectively known as Nox proteins (10). The assembled oxidase then uses
cytosolic NADPH to reduce extracellular O2
to O 2. The O 2 can then be dismutated into
H2O2 and O2 or react with other extracellular
Many studies regarding the effects of oxidants have been performed in primary cells
and cell lines exposed to high doses of exogenous H2O2 or other hydroperoxides. Because
the concentrations used often approached cytotoxicity, they are not necessarily good models
for physiologic activation of the respiratory
burst. Nonetheless, they may provide insight
into some of the initial changes that affect signaling for the priming and/or assembly of the
NADPH oxidase and the respiratory burst.
Such conditions cause substantial elevations
of free intracellular calcium ([Ca2+]i) (11,12).
Under physiologic conditions, adenosine
triphoposphatases (Ca2+ pumps) in the ER and
plasma membrane regulate the levels of [Ca2+]i
in unstimulated cells. Exposure to various concentrations of oxidant-generating systems, or
to bolus hydroperoxides or lipid peroxides,
has long been known to alter Ca2+ in many systems. This is relevant because many signaling
pathways are Ca2+ dependent, and modulation
of cellular function by oxidants may be owing

Macrophage Signaling
and Respiratory Burst

to activation of these pathways by oxidantmediated changes in [Ca2+]i.

In contrast to the high levels of peroxides
used in some studies, we have demonstrated
that in rat alveolar macrophages, relatively low
levels of ROOH can cause transient changes
in [Ca2+]i that resemble those occurring under
normal cell signaling (13). In general, the
mechanisms of ROOH induced increase
[Ca2+]i and the sources of the calcium are not
clearly defined. In studies with rat alveolar
macrophages, we have shown that Ca 2+
released in response to ROOH originated from
an intracellular source; the increase occurred
in the absence of extracellular calcium, and in
the presence of an extracellular calcium chelator (13). Pretreatment with thapsigarin, a compound that inhibits release of calcium from the
ER, had no effect on intracellular calcium,
ruling out the possibility that calcium is
being released from the ER in response to
ROOH (14). Confocal microscopy revealed
that the release of calcium mediated by tertiarybutylhydroperoxide (t-BOOH) appeared to
come from a Ca2+-binding protein on the inner
surface of the plasma membrane. Calcium
release from the interior of the plasma membrane correlated with the release of annexin
VI from the membrane (15). Annexin VI
is a member of a family of phospholipid and
Ca 2+-binding membrane proteins. Various
annexins, also known as lipocortins, are capable of inhibiting phospholipase A2 (PLA2),
which catalyzes the release of arachidonic acid,
which has been implicated in signaling for the
assembly of the NADPH oxidase in in vitro
studies and in some cell lines (14,1618).
Macrophages express a cytosolic form of
PLA2, which is active at physiologic concentrations of Ca2+, and is regulated by phosphorylation and dephosphorylation (19). Exposure
of macrophages to ROOH causes PLA2-mediated release of free arachidonic acid, which
may then in turn be converted into a whole


range of new products classified as eicosanoids, including leukotrienes and prostaglandins. It is not known if Ca2+-mediated
activation of the PLA2-signaling pathway activates the respiratory burst in alveolar macrophages. However, since calcium is required
for the receptor-mediated stimulation of the
NADPH oxidase, this does provide a plausible pathway that connects changes in [Ca2+]i
to activation of the respiratory burst.
In rat alveolar macrophages, mild oxidative
stress has an inverse dose-dependent effect on
the activation of the respiratory burst. Low
concentrations of H2O2 or t-BOOH (<50 M )
act as a priming agent, enhancing the respiratory burst triggered by subsequent stimulation
(20). By contrast, higher concentrations of
H2O2 (>50 M ) inhibit the respiratory burst.
Although both high and low concentrations of
peroxide increased [Ca2+]i, the temporal and
concentration-dependent effects of ROOH on
[Ca2+]i correlated with subsequent enhancement or inhibition of O 2 production (20).
This dual response was also observed in
NR8383 cells, a rat alveolar macrophage cell
line, using an in vitro assay system in which
a single population of cells was exposed to a
gradient of H2O2 (21). On subsequent stimulation with known activators of the respiratory
burst such as adenosine 5-diphosphate (ADP),
phorbol myristate acetate (PMA), and
zymosan, inhibition of the respiratory burst
was observed in cells exposed to the highest
concentration of H2O2, whereas the strongest
activation of the burst was observed in cells
exposed to an intermediate level of H2O2 (21).
The relationship between increases in [Ca2+]i
and the modulation of the respiratory burst was
further investigated by buffering [Ca2+]i. Interestingly, both the enhancement and the inhibition of the respiratory burst by low and
high concentrations of t-BOOH were attenuated by buffering [Ca2+]i (22). The effect of
buffering [Ca 2+ ] i on the assembly of the


NADPH oxidase was also investigated. Multiple phosphorylation events and the translocation of p47phox from the cytoplasm to the
plasma membrane are essential to the activation of the respiratory burst in phagocytes (23).
t-BOOH alone or t-BOOH priming followed
by PMA stimulation did not affect the phosphorylation or translocation of p47phox. By contrast, PMA-induced translocation of p47phox
was increased by priming with 25 M t-BOOH
and decreased by pretreatment with 100 M
t-BOOH (24), consistent with the effect of these
concentrations of t-BOOH on activating or
inhibiting the respiratory burst. Buffering
[Ca2+]i abolished the ability of PMA, primed
with low-dose t-BOOH, to activate the translocation of p47phox (24) and to activate the respiratory burst (22). Thus, it appears that changes
in [Ca2+]i are fundamental to this process.
Activation of Signaling
by Respiratory Burst
The role of ROS produced by the respiratory burst on bacterial killing and tissue
damage is well established. Extensive literature also exists on the pathology inflicted by
high doses of exogenous agonists. In the alveolar macrophage, this may result in the production of lipid peroxides and other damaging
molecules that can alter cell function. For
example, ROS production by macrophages can
induce oxidation of low-density lipoproteins
(LDLs), which are taken up more readily than
normal LDLs. Oxidized LDL has recently been
shown to interfere with normal signaling in
macrophages (25).
By contrast, relatively few studies have been
conducted on the products of the NADPH oxidase per se, specifically on H2O2 as a second
messenger in the signaling pathways of alveolar macrophages. H2O2 is not a radical. It is
highly diffusible and relatively unreactive
unless it is in the presence of a reduced tran-

Iles and Forman

sition metal; the hydroxyl radical can be produced via Fenton chemistry. Any OH produced is sufficiently reactive that it will oxidize
whatever is closest to it. As such, it cannot
function as a second messenger because it lacks
specificity. Conversely, because H2O2 is relatively unreactive, it can act with specificity,
and function effectively as a second messenger. H2O2 can also be elevated transiently and
regulated enzymatically, additional characteristics required of second messengers. Not
long ago, ROS were considered either as
mimics of signaling or more often as disruptive of signaling and other metabolic processes.
Today there is ample evidence that ROS function as signaling molecules (2630). Antioxidant enzymes such as superoxide dismutase
(SOD) and catalase, and the glutathione
(GSH/GSSG) and thioredoxin (Trx) systems
prevent the toxicity of O 2 and H2O2 and restore
the redox status of the cell. It is now known
that several key intracellular signaling pathways may be activated by the products of the
respiratory burst, in particular, H2O2. A new
concept is emerging that antioxidant enzymes,
by restoring the resting state concentrations
of ROS such as H2O2, control signal termination and, therefore, are essential to regulated
intracellular signaling.
Activation of NF-B Signaling Pathway
via Respiratory Burst
The NF-B signaling pathway was one of
the first major pathways shown to be activated
by H2O2 produced by the respiratory burst (5).
NF-B controls the inducible expression of a
variety of genes, particularly those involved
in inflammatory and other immune responses
(31,32). In resting cells, NF-B is predominantly found in the cytoplasm and is typically
a heterodimer composed of p50 and p65
(Rel A) subunits, where it is complexed with
IB, an inhibitory protein that prevents the
migration of NF-B into the nucleus (33).

Macrophage Signaling
and Respiratory Burst

When cells are stimulated, NF-B is translocated to the nucleus, where it binds to a B
transcription element and, along with the binding of other transcription factors and the RNA
polymerase II complex, initiates the transcription of NF-B-inducible genes. In the
classic pathway of NF-B activation, activation of the cell triggers the phosphorylation of
IB by IB-specific kinases (IKKs) (3436).
The IKKs are multicomponent complexes that
contain IKK, IKK, along with several other
proteins (36). Once IB is phosphorylated, it
is targeted for ubiquitination and degradation
(37). This facilitates the release of IB from
the NF-B complex, which is then translocated to the nucleus to initiate transcription of
B-dependent genes. Recently, NF-B activation independent of IB phosphorylation
has also been reported (38). In a given cell
type, IB-dependent and IB-independent
activation of NF-B may be inducible.
The NF-B/Rel family was the prototype
of the redox-sensitive transcription factor.
A model was proposed based on studies with
antioxidants that NF-B was redox sensitive,
and that regardless of the stimulus or agonist
it was activated owing to the production of
ROS (39). Later studies revealed that ROSmediated induction of NF-B is not a universal phenomenon (40). NF-B is activated by
many diverse compounds and stresses including proinflammatory cytokines such as TNF-
and IL-1, T and B cell mitogens, intact viruses,
viral proteins, double-stranded RNA, bacteria, lipopolysaccharide, PMA, ADP, and H2O2
(5,41 43). Several of these compounds have
also been shown to activate the respiratory
burst in alveolar macrophages and to increase
the production of ROS (5).
Early studies in nonmacrophage cell lines
first established that exogenous H2O2 can activate NF-B binding. This was first shown in
Jurkat T cells (39). As well, Schmidt et al. (44)
reported that TNF- and okadaic acidinduced


NF-B activation is diminished in cell lines

that overexpress catalase. However, it was soon
determined that H2O2 is not a universal activator of NF-B in all cell types. Suzuki et al.
(45) showed that overexpression of catalase
did not reduce PMA- and TNF- induced NFB activation in COS-1 cells, establishing that
in these cells at least, activation of NF-B is
not dependent on intracellular H2O2. Other studies that did demonstrate that H2O2 activated
NF-B binding used cytotoxic concentrations
of hydroperoxides (46), calling into question
any relevance of these studies to physiology
and normal cell signaling (46). However, in
some macrophage cell lines, relatively low
doses of exogenous H2O2 can activate NF-B.
In work done in our laboratory using
J774.A1 cells, a monocyte/macrophage cell
line, 25100 M H2O2 activated NF-B binding (5). The doses used were nontoxic to the
cells, as determined by adherence and staining
with vital dye. This was repeated using similar doses (50100 M ) of H2O2 with primary
rat alveolar macrophages (5). The mechanism
of increased NF-B binding in J774.A1 cells
was increased translocation of p65 to the
nucleus. Translocation of p65 to the nucleus
was preceded by activation of the respiratory
burst and increased with time following the
activation of the burst (47). The mechanism of
NF-B activation upstream of the assembly of
the NADPH oxidase is less clear. Specific PKC
inhibitors reduced basal but had little effect on
stimulated NF-B binding. Further, buffering
[Ca2+]i had no effect on NF-B binding.
Experiments using bolus H2O2 sometimes
serve as useful models of the respiratory burst,
but they are not necessarily mechanistically
synonymous with receptor- and agonistmediated activation of the respiratory burst.
Studies using the endogenous production of
H2O2 via activation of the respiratory burst
have confirmed that the H2O2 produced by
the respiratory burst is sufficient to activate


NF-B binding (5). Production of TNF-,

which is NF-B dependent, can be induced in
primary alveolar macrophages and in the RAW
264.7 cell line and is inhibited by the addition
of SOD and catalase (48). This finding suggests that superoxide is involved in NF-B
activation in these cells. In mouse peritoneal
macrophages, silica-stimulated activation of
the respiratory burst activated NF-B (49) that
was accompanied by the classic tyrosine phosphorylation of IB (50). Both SOD and catalase inhibited these changes, indicating a direct
role of superoxide in NF-B activation. In primary rat alveolar macrophages and in J774.A1
cells, PMA and ADP activated the respiratory
burst and increased NF-B binding. However,
while this was abolished by the addition of
exogenous catalase, SOD had no effect (5).
Thus, in these cells, H2O2 is responsible for
the activation of NF-B binding. Together,
these data suggest that the type of the stimulus (i.e., particulate vs soluble) may induce
localized or diffuse production of ROS and
may dictate which product of the respiratory
burst is responsible for NF-B binding.
Several novel mechanisms have been proposed for H2O2-induced activation of NF-B.
In some cell systems, tyrosine phosphorylation of IB is correlated with increased
NF-B binding (51,52), and in stimulated
macrophages, H2O2 has been shown to increase
tyrosine phosphorylation. Recently, it has been
demonstrated that IKK is directly activated by
H2O2 (53). This would facilitate increased
phosphorylation of IB, and increased translocation of NF-B to the nucleus. However, IBindependent pathways of NF-B activation
have also been described. H2O2 may induce
the phosphorylation of p105, the precursor
protein to p50, and facilitates cleavage into the
active form, and p65 phosphorylation may be
stimulated by H2O2 (38,54). Mutational studies of phosphorylation sites in p65 revealed
that abolishing phosphorylation greatly

Iles and Forman

impaired nuclear translocation (54). Overall,

results so far indicate cell- and stimulusspecific phosphorylation of NF-B subunits
via H2O2 and other agonists of the burst.
Activation of MAPK Signaling Pathways
via Respiratory Burst
The MAPKs are a family of signaling pathways, including the ERKs, JNKs, p38MAPKs,
and big MAPK, and are among the most extensively studied signal transduction pathways.
Oxidant-mediated activation of the MAPK signaling pathways is well established. Many
diverse cellular functions and the expression
of many genes are controlled by the MAPKs.
MAPKs are activated by phosphorylation cascades and, conversely, are inactivated by
dephosphorylation. Although many stimuli are
known to activate more than one pathway, generalizations can be made about the type of
stimuli that will typically activate a given
MAPK pathway.
Activation of ERK Pathway
and Respiratory Burst
The ERK pathway is predominantly activated by growth factors and endocrine hormones. However, activation of the ERKs by
exogenous H2O2 has been reported in many
different cell types (5558). In alveolar
macrophages and in the macrophage cell line
NR8383, increased production of ROS either
by transfection of constitutively active Ras
or by addition of the redox cycling quinone,
menadione, correlated with increased activation of the ERKs (59). Other studies have also
shown that stimuli that activate the respiratory
burst increase phosphorylation of the ERKs.
ERK activation may have a role in the assembly of the NADPH oxidase, be increased as a
result of the activation of the respiratory burst,
or both. In rat alveolar macrophages, both
ERK1 and ERK2 were activated by zymosanactivated serum, a source of C5a, a known acti-

Macrophage Signaling
and Respiratory Burst

vator of the respiratory burst (6). The addition

of catalase, but not SOD, almost completely
blocked the phosphorylation and activation of
the ERKs. However, stimulation of the burst
with ADP, which causes a similar amount of
ROS production as zymosan-activated serum,
did not activate ERK1 and ERK2 in primary
rat alveolar macrophages (6). This suggests
that activation of the ERKs, in rat alveolar
macrophages, is not necessary for the activation of the respiratory burst, but, rather, that
activation of the ERKs is a downstream consequence of the burst. The mechanism by which
ERKs are activated by H2O2 is still largely
unknown, but it is thought to be indirect, either
through inhibition of a tyrosine phosphatase
or through activation of an upstream activator
that intersects with a pathway leading to ERK
activation that can be stimulated by zymosanactivated serum but not ADP.
Activation of the ERK pathway controls the
expression of many genes through the phosphorylation of Elk-1. Elk-1 is part of a complex that binds to the serum response element,
thereby functioning as a key regulator of gene
expression (60). One particular gene of interest that is regulated by Elk-1 via activation
of the ERK pathway is c-fos. The c-Fos protein is in itself of considerable importance to
oxidant-regulated gene expression. It often
comprises one half of the regulatory transcription factor complex activator protein-1
(AP-1), which is also activated by the respiratory burst, as described later.
Activation of JNK Pathway by ROS
In contrast to the ERK pathway, the JNK
pathway typically is not activated by endocrine
hormones and growth factors and has traditionally been linked to stress signals. In fact,
another name for the JNKs is the stressactivated protein kinases, or SAPKs. In numerous studies, JNK has been shown to be activated by ROS, in particular H2O2. The precise


mechanism of JNK activation by ROS is not

known, but very elegant experiments by Saitoh
et al. (61) suggested a role for the upstream
modulator apoptosis signalregulating kinase
(ASK-1). In quiescent cells, Trx is bound to
ASK-1, maintaining ASK-1 in an inactive
form. Oxidation of Trx by ROS causes it to
dissociate from ASK-1 and facilitates the activation of ASK-1 (61). Another group has
shown that the production of ROS by TNF-
induces dimerization and activation of ASK-1
(62). Since TNF- had also been shown to
increase JNK phosphorylation, this is consistent with ASK-1 being the upstream activator
in this case. JNK has also been shown to be
kept inactive in nonstressed cells by the
monomeric form of glutathione S-transferase
Pi (GSTp). Production of ROS removes this
inhibitory effect, possibly through the
oligomerization of GSTp, and JNK is activated
(27). Studies have also been conducted that
specifically consider the effects of H2O2. It is
well documented that H2O2 can produce the
lipid peroxidation product 4-hydroxynonenal
(4HNE) (63,64). Exposure to exogenous
4HNE also activates JNK, possibly through
the formation of a 4HNE-JNK adduct (65);
however, another report suggests that exposure to 4HNE increases JNK phosphorylation
by increasing H2O2 production (66). Clearly,
the interplay between 4HNE and H2O2 is complex and warrants further study.
Activation of JNK Pathway
by Respiratory Burst
A principal target of activated JNK, and the
one for which it is named, is the N-terminal
portion of the transcription factor, c-Jun.
c-Jun and other Jun family proteins are an
essential component of the transcription factor
complex AP-1. In NR8383 cells, bolus H2O2
(50200 M ), activated the JNK pathway, as
determined by c-Jun phosphorylation (unpublished data). These concentrations were pre-


viously shown to be nontoxic to the cell (21).

Using electrophoretic mobility shift assays, it
was determined that these doses of H2O2 also
increased AP-1 binding in NR8383 cells. Concentrations of ADP previously shown to activate the respiratory burst in NR8383 cells also
produced increased c-Jun phosphorylation
and increased AP-1 binding. The increase in
AP-1 binding was blocked by the addition of
catalase, which suggests that the H2O2 produced by the respiratory burst is sufficient to
activate the JNK pathway. It is also suggestive that it is sufficient to activate transcription and to modulate gene expression.
This review summarizes studies showing
the effects of ROS, particularly H2O2, on physiologic responses in alveolar macrophages.
H2O2 is a product of the respiratory burst and,
as such, acts as a second messenger in alveolar macrophages activating the NF-B and
AP-1 transcription factors and some members
of the MAPK signaling pathways. Low concentrations of exogenous H 2O 2 can prime
assembly of the NADPH oxidase and, consequently, the respiratory burst. Thus, it is postulated that the primary function of the
respiratory burst in alveolar macrophages is
not as a killing mechanism but, rather, to generate signaling molecules that coordinate the
production of inflammatory mediators, as well
as other appropriate defensive responses.
We wish to thank Dr. Martine Torres and
Dr. Julio Giron-Calle for their contributions
to this work, Dr. Dale Dickinson for critical
review of the manuscript, and many others
whose contributions could not be cited in this
short review. This work was supported by grant
HL05511 from the National Institutes of

Iles and Forman

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