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Is a Critical Mediator of Host Defense Against Invasive Pulmonary Aspergillosis in Neutropenic Hosts
Department of Medicine, Division of Pulmonary and Critical Care Medicine, University of Michigan Medical School, Ann Arbor, MI 48109
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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(MIP-1) is a CC chemokine with potent chemotactic
activity for various subsets of mononuclear leukocytes. We therefore
tested the hypothesis that the influx of mononuclear cells into the
lung in invasive pulmonary aspergillosis is in part mediated by
MIP-1, and the manipulation of this ligand alters the outcome of the
infection. We found that in both immunocompetent and neutropenic mice,
MIP-1 was induced in the lungs in response to intratracheal
administration of Aspergillus fumigatus conidia. In
neutrophil-depleted mice challenged with intratracheal conidia, there
was evidence of invasive fungal pneumonia associated with a
predominantly mononuclear leukocyte infiltrate. Ab-mediated depletion
of MIP-1 resulted in a 6-fold increase in mortality in neutropenic
mice, which was associated with a 12-fold increase in lung fungal
burden. Studies of single-cell suspensions of whole lungs revealed a
36% decrease in total lung leukocyte infiltration as a result of
MIP-1 neutralization. Flow cytometry on whole lung suspensions
showed a 41% reduction in lung monocyte/macrophages as a result of
MIP-1 neutralization, but no difference in other lung leukocyte
subsets. These studies indicate that MIP-1 is a critical mediator of
host defense against A. fumigatus in the setting of
neutropenia and may be an important target in devising future
therapeutic strategies against invasive
aspergillosis. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The commonest and best characterized risk factor for development of invasive aspergillosis is neutropenia (7, 8). The role of neutrophils in the innate host defense against Aspergillus species has been well documented in both in vitro (9, 10) and in vivo (11, 12) experiments. Other immune cells also play a role in host defense against Aspergillus species, however. In experimental animals, alveolar and peritoneal macrophages bind and engulf Aspergillus fumigatus conidia in vitro (10, 13, 14, 15) and produce IL-1 and TNF when exposed to either conidia or hyphae (16). In vitro studies of human peripheral monocytes (17, 18), monocyte-derived macrophages (19), and alveolar macrophages (20, 21) have also shown these cells to phagocytose conidia and kill both conidial and hyphal forms of A. fumigatus. One study has suggested a role for splenic macrophages in acquired immunity against i.v. administered A. fumigatus conidia in vivo (22). Lung histopathology specimens from immunocompromised mice (23) and neutropenic patients (24) with invasive aspergillosis also show prominent infiltration of mononuclear cells in association with fungal hyphae. Because the patients who develop invasive aspergillosis have a limited pool of available neutrophils to combat the pathogen, studies aimed at augmentation of the antifungal activity of other immune cells, such as the mononuclear phagocytes, may be relevant in devising new therapeutic strategies against this infection.
Macrophage inflammatory protein-
(MIP-1),3 a member
of the CC family of chemokines, exhibits potent chemotactic and
activating properties for several populations of leukocytes, including
monocytes, various lymphocyte subsets, mast cells, and eosinophils
(reviewed in Refs. 25 and 26). MIP-1 is
induced in patients with diverse inflammatory diseases, including
bacterial sepsis (27, 28), malaria (29), and
interstitial lung diseases (30), and in vivo
administration of MIP-1 to laboratory animals results in localized
infiltration of monocytes (31, 32, 33). In animal models of
infections, MIP-1 has been shown to play a critical role in the
cellular recruitment and inflammatory response to coxsackie and
influenza infections (34), endotoxemia (35),
bacterial pneumonia (36), and cryptococcosis (37, 38). Two previous studies have examined the expression of
MIP-1 in response to Aspergillus species; one showed that
rat alveolar macrophages express MIP-1 mRNA when exposed to A.
fumigatus conidia in vitro (39), and another
demonstrated that MIP-1 is induced in the lungs after intrapulmonary
challenge with conidia (40). We have previously shown that
neutralization of TNF in experimental pulmonary aspergillosis results
in increased mortality and decreased fungal clearance, which was
associated with decreased levels of lung MIP-1 (41).
The role and mechanism of action of MIP-1 in host defense against
A. fumigatus have not, however, been characterized.
In this study we hypothesized that the influx of mononuclear phagocytes
into the lung in invasive pulmonary aspergillosis is in part mediated
by MIP-1, and that the manipulation of this chemokine alters the
outcome of the infection. We tested this hypothesis by evaluating the
outcome and severity of the infection in neutropenic mice in the
setting of MIP-1 depletion and by determining the effect of MIP-1
neutralization on inflammatory cellular influx into the lungs.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Polyclonal anti-murine MIP-1 Abs used in in vivo
neutralization experiments and in ELISA were produced by immunization
of rabbits with carrier-free murine recombinant MIP-1 (R&D Systems,
Minneapolis, MN) in multiple intradermal sites with CFA, as previously
described (42, 43). Abs were purified over a protein A
column, and endotoxin contamination was excluded by Limulus
lysate assay (ICN Biomedical, Costa Mesa, CA). In neutralization
experiments, 0.5 ml of rabbit anti-murine MIP-1 serum or control
rabbit serum was administered i.p. 2 h before A.
fumigatus inoculation, and 0.25 ml was given on days 2 and 4 after
inoculation.
Animals
Specific pathogen-free C57BL/6 mice (6- to 8-wk-old females; The Jackson Laboratory, Bar Harbor, ME) were used in all experiments. All mice were housed in specific pathogen-free conditions within the animal care facility at the University of Michigan until the day of sacrifice.
Preparation and use of RB6-8C5 mAb for in vivo neutrophil depletion
RB6-8C5 is a rat anti-mouse monoclonal IgG2b directed against Ly-6G, previously known as Gr-1, an Ag on the surface of murine granulocytes. The Ag expression increases with cell maturity and is absent from precursor cells. The Ab originally used for flow cytometry is a complement-fixing isotype, well suited for in vivo cell depletion (44, 45, 46, 47, 48). TSD BioServices (Germantown, NY) produced the Ab by i.p. injection of RB6-8C5 hybridoma into nude mice and ascites collection. The resultant ascitic fluid was pooled, and the concentration of IgG was quantified by HPLC. One hundred microliters of a 1/200 dilution of the pooled ascites was administered i.p. 1 day before challenge with intratracheal (i.t.) A. fumigatus or vehicle. This resulted in peripheral blood neutropenia (absolute circulating neutrophil count, <50 cells/µl) by days 1 and 3 after Ab administration in both infected and control animals, with a return of peripheral counts to pretreatment levels by day 5.
Preparation and administration of A. fumigatus conidia
A. fumigatus strain 13073 (American Type Culture Collection, Manassas, VA) was used in all studies, as this strain has previously been shown to induce invasive aspergillosis in immunocompromised mice (49). The organism was grown on Sabouraud dextrose agar plates (Becton Dickinson, Cockeysville, MD) for 7–10 days at 37°C. The surface of each plate was then washed with 100 ml of sterile 0.1% Tween 80 (Sigma, St. Louis, MO) in normal saline. The resulting suspension of conidia was filtered through sterile gauze to remove clumps and hyphal debris, and then washed once and resuspended in 4 ml of 0.1% Tween 80. The concentration of Aspergillus conidia in the suspension was determined using a particle counter (Z2 particle analyzer, Coulter, Hialeah, FL). The suspension was then diluted to the desired concentration, and the concentration was again measured before administration. In preliminary experiments the number of particles determined by the particle counter was in close agreement with the number of viable CFU found by serial dilution and plating of the suspension. On the day of inoculation, each animal was anesthetized with 1.8–2 mg of pentobarbital i.p. Using standard aseptic technique, the trachea was exposed, and a 30-µl inoculum (A. fumigatus suspension or 0.1% Tween 80 vehicle) was administered via a sterile 26-gauge needle. The skin incision was closed with surgical staples. Animals were challenged with inocula ranging from 1–2 x 106 to 1–2 x 107 conidia in various experiments. In preliminary studies i.t. administration of the Tween vehicle to immunocompetent animals did not result in changes in histology or influx of neutrophils (data not shown).
Lung harvest
At designated time points the mice were sacrificed by CO2 asphyxiation. The chest cavity was opened aseptically, and the pulmonary vasculature was perfused with PBS via the right ventricle. For histologic examination, lungs were perfused with 1 ml of 4% paraformaldehyde in PBS, inflated with 1 ml of 4% paraformaldehyde in PBS via the trachea, and then excised en bloc. Lungs for assays were perfused with 1 ml of PBS containing 5 mM EDTA, removed, frozen in liquid nitrogen, and stored at -20°C until the day of the assay. Lungs for various assays were homogenized in 1 ml of 2x complete protease inhibitor cocktail buffer (Roche, Mannheim, Germany) in PBS using a tissue homogenizer (Biospec Products, Bartlesville, OK). A 900-µl aliquot of PBS was added to 900 µl from each sample, sonicated for 10 s, and centrifuged at 500 x g for 10 min. Supernatants were passed through a 0.45-µ pore size filter (Gelman Sciences, Ann Arbor, MI) and stored at 4°C for ELISA.
Single-cell suspensions of the lungs were prepared as previously described (50). Briefly, freshly resected lungs were minced with scissors to a fine slurry and incubated at 37°C for 30 min in an RPMI solution (Sigma) containing 1 mg/ml type A collagenase (Roche) and 30 U/ml DNase (Sigma). The solutions were then drawn up and down 20 times in 10-ml syringes (Becton Dickinson, Franklin Lakes, NJ) to disperse the cells mechanically. The resulting cell suspensions were pelleted, resuspended, passed through Nitex mesh filters (Tetko, Kansas City, MO), and passed through a 20% Percoll gradient (Sigma) before cell counting under a hemocytometer. Cytospin slides of this suspension were then prepared and stained (Diff-Quik Stain Set, Dade Behring, Newark, DE), and differential cell counts were determined using a high power microscope. The absolute number of a leukocyte type was determined by multiplication of the percentage of that cell type by the total number of lung leukocytes in that sample.
Flow cytometric analyses
Total lung leukocytes were isolated from animals as described
above. For identification of total lymphocyte population, isolated
leukocytes were stained with FITC-labeled anti-ß-TCR,
anti-
-TCR, anti-CD19, and anti-NK1.1. For
identification of monocyte/macrophage populations, isolated leukocytes
were stained with FITC-labeled anti-CD11b (all reagents from
PharMingen, San Diego, CA). In addition, cells were stained with
anti-CD45-Tricolor (Caltag, South San Francisco, CA), allowing
discrimination of leukocytes from nonleukocytes and thus eliminating
any nonspecific binding of cell surface markers on nonleukocytes. The
samples were washed in staining buffer and fixed in 1%
paraformaldehyde (Sigma) in PBS. Stained samples were kept in the dark
at 4°C until analyzed on a FACScan cytometer (Becton Dickinson, San
Jose, CA) using CellQuest software (Becton Dickinson). Lymphocyte and
monocyte/macrophage populations were analyzed by first gating on
CD45-positive cells of the appropriate light scatter characteristics
and were then examined for FL1 and FL2 fluorescence expression. The
absolute number of a leukocyte type was determined by multiplication of
the percentage of that cell type by the total number of lung leukocytes
in that sample.
Lung chitin assay
Given that molds (including Aspergillus species) do not reliably form reproductive units in tissue, we employed a previously described assay for chitin to measure the burden of organisms in the lungs (41). Chitin, a component of the hyphal wall, is absent from conidia and mammalian tissue. The level of chitin, as detected by this assay, directly correlates with the weight of hyphae (51). Lungs were homogenized, centrifuged, and resuspended in sodium lauryl sulfate (3%, w/v), heated at 100°C for 15 min, washed and resuspended in KOH (120%, w/v), heated at 130°C for 60 min, cooled, incubated on ice after the addition of 8 ml of ice-cold 75% ethanol, pelleted after the addition of 0.3 ml of Celite suspension, washed, and resuspended in 0.5 ml of distilled water. NaNO2 (5%, w/v; 0.5 ml) and KHSO4 (5%, w/v; 0.5 ml) were added to each sample and to standards (consisting of distilled water and 10 µg/ml glucosamine). Aliquots (0.6 ml) of each were transferred to separate tubes, 0.2 ml of ammonium sulfamate and then 0.2 ml of 3-methyl-2-thiazolone hydrazone HCl monohydrate (50 mg in 10 ml of distilled water) were added, and samples were heated to 100°C for 3 min and cooled. FeCl3·6H2O (0.83%, w/v; 0.2 ml) was then added to each, and OD was measured at 650 nm after 25 min. Chitin content, in glucosamine equivalents, was then calculated as follows: chitin content = {[5 x (OD of organ - OD of control organ)]/(OD of glucosamine - OD of water)}.
Lung myeloperoxidase (MPO) activity
Lung MPO activity was measured as a marker of neutrophil sequestration, as described previously (52). Briefly, a 100-µl aliquot of each lung homogenate was added to 100 µl of a buffer containing 50 mM potassium phosphate (pH 6.0), 5% hexadecyltrimthylammonium bromide, and 5 mM EDTA. Samples were sonicated for 10 s and centrifuged at 3000 x g for 15 min. The supernatant was mixed 1:15 with a buffer containing 1 M monobasic potassium phosphate, 1 M dibasic potassium phosphate, 3% H2O2, and o-dianisidine hydrochloride, and read at 490 nm. MPO units were calculated as the change in absorbency over time.
MIP-1 ELISA
Murine MIP-1 level was quantified using a modification of a
double-ligand method as described previously (36).
Briefly, flat-bottom 96-well microtiter plates (Immuno-Plate I 96-F,
Nunc, Copenhagen, Denmark) were coated with 50 µl/well of rabbit
anti-MIP-1 Ab (1 µg/ml in 0.6 M NaCl, 0.26 M
H3BO4, and 0.08 N NaOH, pH
9.6) for 16 h at 4°C and then washed with PBS, pH 7.5, and
0.05% Tween 20 (wash buffer). Microtiter plate nonspecific binding
sites were blocked with 2% BSA in PBS and incubated for 90 min at
37°C. Plates were rinsed four times with wash buffer, and diluted
(undiluted and 1/10) cell-free supernatants (50 µl) in duplicate were
added, followed by incubation for 1 h at 37°C. Plates were
washed four times, followed by the addition of 50 µl/well
biotinylated rabbit anti-MIP-1 Abs (3.5 µg/ml in PBS (pH 7.5),
0.05% Tween 20, and 2% FCS), and plates were incubated for 30 min at
37°C. Plates were washed four times, streptavidin-peroxidase
conjugate (Bio-Rad, Richmond, CA) was added, and the plates were
incubated for 30 min at 37°C. Plates were washed again four times and
chromogen substrate (Bio-Rad) was added. The plates were incubated at
room temperature to the desired extinction, and the reaction was
terminated with 50 µl/well of 3 M
H2SO4 solution. Plates were
read at 490 nm in an ELISA reader. Standards were half-log dilutions of
recombinant murine MIP-1 from 1 pg/ml to 100 ng/ml. This ELISA
method consistently detected MIP-1 at concentrations >25 pg/ml. The
ELISAs did not cross-react with IL-1, IL-2, IL-4, IL-6, IL-10, or
IFN-. In addition, the ELISA did not cross-react with other members
of the murine CXC or CC chemokine families.
Statistical analysis
Data were analyzed by a Power Macintosh 8600/300 computer using the InStat version 2.01 statistical package (GraphPad Software, San Diego, CA). Survival data were compared using Fisher’s exact test. All other data were expressed as the mean ± SEM and compared using an unpaired two-tailed Mann-Whitney (nonparametric) test. Values of p < 0.05 were considered statistically significant.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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We first examined the influx of inflammatory cells into the lungs
in response to intratracheal challenge with A. fumigatus
conidia in both normal and neutrophil-depleted C57BL/6 mice. Lungs were
harvested at various time points after challenge for histology. In
normal animals, histology showed evidence of a patchy peribronchial
infiltration of inflammatory cells within both the interstitial and
alveolar compartments on day 2, which consisted predominantly of
neutrophils. By day 4 after challenge, the cellular infiltrate had
decreased and consisted of both neutrophils and mononuclear cells
(Figs. 1
, A and B).
As previously described, fungal hyphae did not form in tissues of
immunocompetent animals, and the inflammatory cellular infiltrates were
associated with areas of conidia deposition (41, 53).
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, C and inset). These
inflammatory cells occurred in association with fungal hyphae as
previously described (41, 53). By day 4 after challenge,
the patchy cellular infiltrate was extremely dense and consisted of
both neutrophils and mononuclear cells associated with fungal
hyphae (Fig. 1D).
Expression of MIP-1 in the lungs in response to A.
fumigatus
To address the mechanism of influx of mononuclear cells into the
lungs in response to challenge with A. fumigatus in normal
and neutropenic animals, we next measured concentrations of MIP-1 in
lung homogenates at various times after i.t. administration of conidia.
MIP-1 was induced in the lungs of both normal and neutropenic
animals after inoculation with conidia compared with animals challenged
with vehicle. MIP-1 levels reached a higher peak in neutropenic
animals compared with normal animals (Fig. 2), but were undetectable in plasma in
both groups, indicating a compartmentalized response.
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neutralization on the course of infection in
neutropenic mice challenged with A. fumigatus
To assess the contribution of MIP-1 to host defense against
A. fumigatus in neutropenic animals, we next assessed the
survival of animals pretreated with rabbit anti-MIP-1 serum. In
normal (nonneutropenic) animals, administration of anti-MIP-1
serum followed by i.t. challenge with 1–2 x
107 conidia did not result in any deaths.
However, administration of anti-MIP-1 serum to neutropenic
animals 2 h before challenge with 1–2 x
106 conidia resulted in a >6-fold increase in
mortality compared with neutropenic animals receiving control serum
(Fig. 3).
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-treated neutropenic animals, mice were sacrificed on
day 2 after challenge with conidia, and lungs were harvested for
histology and chitin content. Compared with those in animals treated
with control serum, lungs of anti-MIP-1-treated animals
contained a substantially greater number of hyphal forms, as determined
by histologic examination (Fig. 4). To
quantify the burden of hyphae present in the two groups, lung chitin
levels were measured 2 days after challenge with conidia in neutropenic
animals receiving anti-MIP-1 or control serum. A 12-fold
increase in lung chitin content was noted in animals treated with
anti-MIP-1 compared with control animals (Fig. 5). Chitin levels were undetectable in
nonneutropenic animals treated with anti-MIP-1 or control serum
(data not shown).
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depletion on influx of immune cells into the
lungs in neutropenic mice challenged with A. fumigatus
To determine whether impaired fungal clearance in
MIP-1-depleted animals occurred as a result of changes in lung
neutrophil influx, lungs from neutropenic animals treated with
anti-MIP-1 or control serum were harvested for estimation of
myeloperoxidase activity, a surrogate measure of neutrophil presence.
On the basis of studies showing time of maximum expression of MPO (not
shown), MPO levels were evaluated 2 days after inoculation in
RB6-8C5-treated animals. There was no significant difference in the
lung MPO content of neutropenic animals treated with anti-MIP-1
or control serum 2 days after challenged with 1–2 x
106 conidia, indicating that differences in
neutrophil influx did not explain the worse outcome with MIP-1
neutralization.
We next assessed whether the poor outcome of infection in
MIP-1-depleted neutropenic animals was attributable to differences
in the influx of other cell populations into the lungs, by examining
whole lung single cell suspensions. Normal animals, neutropenic animals
pretreated with control serum, and neutropenic animals pretreated with
anti-MIP-1 serum were sacrificed 2 days after i.t. challenge
with 1–2 x 106 A. fumigatus
conidia, and lung cell suspensions were prepared and compared with
those from normal uninfected animals. The total number of cells in the
lungs of neutropenic animals pretreated with anti-MIP-1 was 36%
less than the number in neutropenic mice treated with control serum
(Fig. 6A). The number of lung
neutrophils, however, did not differ significantly between the two
groups (Fig. 6B), thus confirming the findings of earlier
experiments that employed MPO as a surrogate for neutrophil presence.
Flow cytometry of lung homogenates showed that
anti-MIP-1-treated neutropenic animals had 41% fewer lung
monocyte/macrophages compared with neutropenic animals treated with
control serum (Fig. 6C), while the total numbers of lung
lymphocytes were not significantly different in the two groups (mean,
1.16 x 107 and 7.49 x
106 cells, respectively; p =
0.39).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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We have previously shown that depletion of TNF in murine invasive
pulmonary aspergillosis resulted in increased mortality and lung fungal
burden, which was associated with decreased lung levels of the CXC
chemokine, MIP-2, and the CC chemokines, MIP-1 and JE
(41). We subsequently demonstrated that neutralization of
the common murine receptor for the glutamic
acid-leucine-arginine-positive CXC chemokines, CXCR-2, resulted in a
markedly increased susceptibility to this infection in nonneutropenic
animals, which occured in association with a decreased influx of
neutrophils into the lungs. In addition, the intrapulmonary transgenic
expression of the glutamic acid-leucine-arginine-positive CXC
chemokine, KC, reduced the susceptibility to this infection
(53). In the present study we demonstrated that in the
setting of pulmonary aspergillosis in transiently neutropenic mice,
MIP-1 plays a critical role in host defense. Specifically,
neutralization of MIP-1 resulted in a reduction of lung mononuclear
cell influx, identified as monocyte/macrophages by flow cytometry.
Given the short duration of the response (2 days), this increase in
lung monocyte/macrophages almost certainly represents active
recruitment rather than proliferation. In addition to changes in the
numbers of lung mononuclear phagocytes after MIP-1 neutralization,
the activation state of these cells is also likely to have changed.
Although we did not observe qualitative changes in total lung
lymphocytes, a similar change in the activation state cannot be
excluded. With regard to neutrophils, we found that neutralization of
MIP-1 in nonneutropenic mice did not result in mortality or invasive
disease. Furthermore, in transiently neutrophil-depleted animals, we
found no difference in the numbers of neutrophils in the lungs of
MIP-1-depleted and control animals either by direct cell count or
lung MPO activity. Collectively, these observations suggest that the
effects of MIP-1 in this model are not mediated through alterations
in neutrophil recruitment.
Like many chemokines, MIP-1 exhibits receptor promiscuity, binding
to both CCR-1 and CCR-5. A study of mice with targeted deletion of
CCR-1 has shown increased susceptibility of these animals to i.v.
injected A. fumigatus, a finding that was interpreted as
evidence of abnormal neutrophil-mediated host defense
(54). Importantly, we did not observe any increase in
susceptibility to intratracheal A. fumigatus in CCR-1
knockout animals compared with wild-type controls (data not shown). Our
data therefore indicate that CCR-1 is not essential for protective
innate immunity against A. fumigatus in the lung. The
discrepancy between our findings and those of Gao et al. may be
explained by several considerations, which are not mutually exclusive.
First, the i.v. administration of Aspergillus conidia, as
employed by Gao et al. (55), has previously been shown to
result in foci of infection predominantly in the kidney and brain
rather than the lung. Second, in the setting of disseminated and
overwhelming infection in a normal host, the innate immune response is
likely to differ from the response to intrapulmonary challenge in a
neutropenic host and may absolutely require CCR-1 expression by
neutrophils and possibly CCR-1 ligands other than MIP-1.
A caveat to all studies of chemokines in murine models is the potential
for dissimilarities in chemokine biology between human and murine
systems; such differences have been well characterized in the CXC
chemokines (56). This study has identified MIP-1 as a
critical mediator of host defense against invasive aspergillosis in the
setting of neutropenia. Because the beneficial effect of MIP-1 in
invasive pulmonary aspergillosis is most likely to be observed with
compartmentalized overexpression, studies aimed to define the
therapeutic potential of lung-specific overexpression of MIP-1 are
in progress.
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Borna Mehrad, University of Michigan Medical Center, 6301 MSRB III, 1150 West Medical Center Drive, Ann Arbor, MI 48109-0360.
3 Abbreviations used in this paper: MIP-1, macrophage inflammatory protein-1; CCR, CC chemokine receptor; CXCR, CXC chemokine receptor; i.t., intratracheal; MCP, monocyte chemoattractant protein; MPO, myeloperoxidase.
Received for publication December 29, 1999. Accepted for publication April 25, 2000.
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K. J. Carpenter and C. M. Hogaboam Immunosuppressive Effects of CCL17 on Pulmonary Antifungal Responses during Pulmonary Invasive Aspergillosis Infect. Immun., November 1, 2005; 73(11): 7198 - 7207. [Abstract] [Full Text] [PDF]
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X. Zeng, T. A. Moore, M. W. Newstead, R. Hernandez-Alcoceba, W. C. Tsai, and T. J. Standiford Intrapulmonary Expression of Macrophage Inflammatory Protein 1{alpha} (CCL3) Induces Neutrophil and NK Cell Accumulation and Stimulates Innate Immunity in Murine Bacterial Pneumonia Infect. Immun., March 1, 2003; 71(3): 1306 - 1315. [Abstract] [Full Text] [PDF]
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B. Mehrad, M. Wiekowski, B. E. Morrison, S.-C. Chen, E. C. Coronel, D. J. Manfra, and S. A. Lira Transient Lung-Specific Expression of the Chemokine KC Improves Outcome in Invasive Aspergillosis Am. J. Respir. Crit. Care Med., November 1, 2002; 166(9): 1263 - 1268. [Abstract] [Full Text] [PDF]
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M. Kobayashi, H. Takahashi, A. P. Sanford, D. N. Herndon, R. B. Pollard, and F. Suzuki An Increase in the Susceptibility of Burned Patients to Infectious Complications Due to Impaired Production of Macrophage Inflammatory Protein 1{alpha} J. Immunol., October 15, 2002; 169(8): 4460 - 4466. [Abstract] [Full Text] [PDF]
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U. Maus, K. von Grote, W. A. Kuziel, M. Mack, E. J. Miller, J. Cihak, M. Stangassinger, R. Maus, D. Schlondorff, W. Seeger, et al. The Role of CC Chemokine Receptor 2 in Alveolar Monocyte and Neutrophil Immigration in Intact Mice Am. J. Respir. Crit. Care Med., August 1, 2002; 166(3): 268 - 273. [Abstract] [Full Text] [PDF]
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M. Santosuosso, M. Divangahi, A. Zganiacz, and Z. Xing Reduced tissue macrophage population in the lung by anticancer agent cyclophosphamide: restoration by local granulocyte macrophage-colony-stimulating factor gene transfer Blood, February 15, 2002; 99(4): 1246 - 1252. [Abstract] [Full Text] [PDF]
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J. M. Schuh, K. Blease, and C. M. Hogaboam CXCR2 Is Necessary for the Development and Persistence of Chronic Fungal Asthma in Mice J. Immunol., February 1, 2002; 168(3): 1447 - 1456. [Abstract] [Full Text] [PDF]
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M. ELLIS, R. WATSON, A. MCNABB, M.L. LUKIC, and M. NORK Massive intracerebral aspergillosis responding to combination high dose liposomal amphotericin B and cytokine therapy without surgery J. Med. Microbiol., January 1, 2002; 51(1): 70 - 75. [Abstract] [Full Text] [PDF]
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Z. Wang, L. Zheng, H. Liu, Q. Wang, M. Hauser, S. Kauffman, J. M. Becker, and P. J. Szaniszlo WdChs2p, a Class I Chitin Synthase, Together with WdChs3p (Class III) Contributes to Virulence in Wangiella (Exophiala) dermatitidis Infect. Immun., December 1, 2001; 69(12): 7517 - 7526. [Abstract] [Full Text] [PDF]
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D. M. Lindell, T. J. Standiford, P. Mancuso, Z. J. Leshen, and G. B. Huffnagle Macrophage Inflammatory Protein 1{alpha}/CCL3 Is Required for Clearance of an Acute Klebsiella pneumoniae Pulmonary Infection Infect. Immun., October 1, 2001; 69(10): 6364 - 6369. [Abstract] [Full Text] [PDF]
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S. K. Bliss, L. C. Gavrilescu, A. Alcaraz, and E. Y. Denkers Neutrophil Depletion during Toxoplasma gondii Infection Leads to Impaired Immunity and Lethal Systemic Pathology Infect. Immun., August 1, 2001; 69(8): 4898 - 4905. [Abstract] [Full Text] [PDF]
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K. Blease, B. Mehrad, N. W. Lukacs, S. L. Kunkel, T. J. Standiford, and C. M. Hogaboam Antifungal and Airway Remodeling Roles for Murine Monocyte Chemoattractant Protein-1/CCL2 During Pulmonary Exposure to Asperigillus fumigatus Conidia J. Immunol., February 1, 2001; 166(3): 1832 - 1842. [Abstract] [Full Text] [PDF]
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 F. Reichenberger, J.M. Habicht, A. Gratwohl, and M. Tamm Diagnosis and treatment of invasive pulmonary aspergillosis in neutropenic patients Eur. Respir. J., January 1, 2001; 19(4): 743 - 755. [Abstract] [Full Text] [PDF]
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 U. A. Maus, M. A. Koay, T. Delbeck, M. Mack, M. Ermert, L. Ermert, T. S. Blackwell, J. W. Christman, D. Schlondorff, W. Seeger, et al. Role of resident alveolar macrophages in leukocyte traffic into the alveolar air space of intact mice Am J Physiol Lung Cell Mol Physiol, June 1, 2002; 282(6): L1245 - L1252. [Abstract] [Full Text] [PDF]
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) were not significantly different from those in uninfected
animals. Af, A. fumigatus; RB6-8C5, rat anti-mouse
Ly-6G IgG2b mAb. Data shown represent the mean ± SEM of six
animals for each group at each time point. *, p
< 0.05 compared with animals receiving i.t. vehicle. The data shown
are representative of three separate experiments.
