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ABSTRACT |
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ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
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The immunomodulatory role of arachidonic acid metabolites in allergic sensitization is undefined. Prostaglandin E2 (PGE2), a product
of arachidonic acid metabolism through the cyclooxygenase pathway, has been reported to favor Type 2-like cytokine secretion profiles in murine and human CD4+ T cells by inhibiting the production of Type 1-associated cytokines. On the basis of these in vitro
data, we hypothesized that indomethacin, a nonselective cyclooxygenase inhibitor, would diminish allergen-induced production of
Type 2 cytokines in mice, and protect against airway hyperresponsiveness (AHR) to methacholine. We found that ovalbumin-sensitized mice that were treated with indomethacin (OVA-indomethacin mice) had significantly greater AHR (p < 0.05) and higher levels
of IL-5 (176 ± 52 versus 66 ± 4 pg/ml) and IL-13 (1,226 ± 279 versus 475 ± 65 pg/ml) in lung supernatants than mice sensitized with ovalbumin alone (OVA mice), while levels of IL-4 and serum IgE were not different. Lung mRNA expression of the C-C chemokine MCP-1 was increased in OVA-indomethacin mice, while there was no difference between the two groups in lung mRNA expression of eotaxin, MIP-1
, MIP-1
, or MIP-2. Histologic examination revealed greater pulmonary interstitial eosinophilia in OVA-indomethacin mice as well. Contrary to our expectations, we conclude that in the
BALB/c mouse, cyclooxygenase inhibition during allergen sensitization increases AHR, production of IL-5 and IL-13, and interstitial eosinophilia.
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INTRODUCTION |
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ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
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Cyclooxygenase is the critical enzyme necessary for the formation of prostaglandins and thromboxane (1). Prostaglandin E2
(PGE2), one of the products of the cyclooxygenase pathway, may favor a Type 2-like cytokine secretion profile in murine
and human CD4+ T cells by inhibiting production of the Type
1-associated cytokines interleukin 2 (IL-2) and interferon 
(IFN-) (2). In addition, in human monocytes PGE2 inhibits
production of IL-12, a potent inducer of T cell differentiation
toward Type 1 cytokine production (6). PGE2 in some reports
does not increase T lymphocyte IL-4 production in vitro (2, 3,
7, 8), while others have shown that IL-4 and IL-5 are upregulated by PGE2 in the presence of IL-2 (5). In summary, it is hypothesized that the upregulation of PGE2 resulting from decreased aspirin use enhances allergic sensitization and asthma
by amplifying the relative Type 2/Type 1 cytokine imbalance
in genetically predisposed individuals (9).
On the basis of these in vitro data, we hypothesized that mice in which cyclooxygenase activity was blocked by a nonsteroidal antiinflammatory drug (NSAID) during an allergen sensitization protocol would have a diminished production of the Type 2 cytokines IL-4, IL-5, and IL-13; decreased serum IgE production; and protection against allergen-induced airway hyperresponsiveness (AHR) to methacholine. To test this hypothesis, we used a well-characterized murine model of allergic sensitization and airway hyperresponsiveness, employing ovalbumin (OVA) as an antigen. To inhibit cyclooxygenase activity and prostaglandin synthesis, we treated the mice during the allergen sensitization protocol with indomethacin, a nonselective cyclooxygenase inhibitor. The dose of indomethacin used inhibits PGE2 production and does not cause illness in experimental animals (10). Our results suggest that cyclooxygenase inhibition during allergen sensitization in mice actually augments induction of the Type 2 cytokines IL-5 and IL-13 and enhances allergen-induced AHR.
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METHODS |
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ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
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Mice
Pathogen-free, 8-wk-old female BALB/c mice were purchased from Harlan (St. Louis, MO). They were shipped in filtered crates and housed in a high-efficiency particulate air (HEPA)-filtered Duo-flo laminar flow unit. Cages, bedding, food, and water were sterilized before use. Room temperature was maintained between 24 and 27° C and a 12-h-on, 12-h-off light cycle was provided. In caring for animals the investigators adhered to the Guide for the Care and Use of Laboratory Animals prepared by the Committee on Care and Use of Laboratory Animals of the Institute of Laboratory Animal Resources, National Research Council (revised 1996).
Allergen Sensitization Protocol
Mice were injected intraperitoneally with 0.1 ml (10 µg) of ovalbumin (chicken OVA, grade V; Sigma, St. Louis, MO) complexed with 20 mg of Al(OH)3 on Day 0 (Figure 1). On Days 14 through 22, the mice were placed in an acrylic box and exposed to aerosols of 1% ovalbumin diluted in sterile phosphate-buffered saline (PBS), using an ultrasonic nebulizer (Ultraneb 99; DeVilbiss, Somerset, PA), for 40 min each day. Opposite the aerosol orifice was a small exhaust orifice vented into a chemical hood to ensure continuous air flow. Nonsensitized mice were injected intraperitoneally with Al(OH)3 on Day 0. Methacholine challenges were performed on Day 23.
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Indomethacin Administration
Indomethacin (30 µg/ml) was administered in the drinking water starting on Day 
1. An indomethacin stock was made by dissolving 150 mg
of indomethacin in 50 ml of ethanol. Three times per week throughout
the experimental protocol, 2 ml of the indomethacin stock solution was
added to 200 ml of water in the animals' water bottles. The water of the
control mice was also changed three times per week and 2 ml of ethanol
was added to 200 ml of water in the water bottles of those mice.
Measurement of Arachidonic Acid Metabolites
Arachidonic acid metabolites were measured in bronchoalveolar lavage (BAL) fluid harvested on Day 23. PGE2 was measured by a modified stable isotope dilution assay that used gas chromatography-negative ion chemical ionization-mass spectrometry as previously described (11). Leukotrienes C4/D4/E4 (LTC4/D4/E4) were quantified by enzyme immunoassay employing a peptido-leukotriene polyclonal antiserum (Cayman Chemical, Ann Arbor, MI) after prior purification on C18 columns (Altech, Los Altos, CA).
Cytokine and Chemokine Detection by RNase Protection Assay
Total lung RNA was isolated with guanidine thiocyanate. Probes for a
panel of cytokines (MCK-1; PharMingen, San Diego, CA) and chemokines (MCK-5; PharMingen) were used according to the manufacturer
instructions. Briefly, RNA was dissolved in 80% formamide, 0.4 M
NaCl, 1 mM EDTA, and 40 mM piperazine-N,N'-bis(2-ethanesulfonic acid), heated to 90° C for 5 min, and hybridized for 12-16 h with corresponding [-32P]UTP-labeled antisense probes at 56° C. The unhybridized RNA was digested with 100 µl of RNase T1+A (250 U/µl;
PharMingen) and 100 µl of RNase A (80 ng/ml; PharMingen) for 45 min at 30° C. After phenol-chloroform extraction and ammonium acetate-ethanol precipitation, the protected hybridized RNA was denatured and electrophoresed on a 5% polyacrylamide gel. The gel was
dried and exposed to film.
Quantitation of IL-4, IL-5, IL-13, and IL-6 in Lung Tissues
Levels of IL-4, IL-5, IL-13, and IL-6 in lung tissues of the four groups of mice were measured with commercially available enzyme-linked immunosorbent assay (ELISA) kits (IL-4, IL-5, and IL-6 [Endogen, Woburn, MA]; IL-13 [R&D Systems, Minneapolis, MN]) according to the manufacturer protocols. On Day 18, the lungs from four mice in each group were analyzed for cytokine levels. Briefly, one lung from each mouse was ground, using a mortar and pestle and ground glass. The solution of ground lung and ground glass was then centrifuged at 1,000 rpm for 10 min. The supernatant was then either frozen for later use or added to precoated wells, and incubated for 2 h. Dilutions of recombinant cytokine were included for generation of a standard curve. Peroxidase-labeled anti-cytokine antibody was added to detect bound cytokine, and the plates were developed by the addition of tetramethylbenzidene substrate. Concentrations of cytokines in the lung supernatants were calculated from the standard curve produced. The cytokine level from each lung was measured in duplicate.
Total IgE Concentrations
Before sacrifice on Day 23, sera were collected from sensitized mice; then analyzed by ELISA to determine levels of total IgE. To determine total IgE levels, 96-well Immunolon II plates (Nunc, Roskilde, Denmark) were coated with a 1:200 dilution of rat monoclonal anti-murine IgE clone LO-ME-3 (Serotec, Oxford, UK). Plates were washed with PBS-0.5% Tween and blocked with PBS-1% bovine serum albumin for 1 h. The plates were then washed before adding 100 µl of serum diluted 1:30 in PBS. Plates were incubated overnight and washed, and 100 µl of rat anti-mouse IgE clone LO-ME-2 (Serotec) diluted 1:2,000 was added to each well. After 1 h of incubation at 37° C, the plates were again washed and horseradish peroxidase (HRP) activity was determined with a tetramethylbenzidine (TMB; Sigma) developing solution (1% TMB in dimethyl sulfoxide [DMSO], 0.001 M sodium acetate, and 0.45% H2O2 final concentration). Substrate development was stopped with 2.5 M H2SO4 and optical density was measured at 450 nm (OD450). Concentration was extrapolated by use of an IgE standard (Maine Biotech, Portland, ME).
Methacholine Challenge
Mice were anesthetized with intraperitoneal injections of pentobarbital sodium (85 mg/kg) and a tracheostomy tube was placed. The internal jugular vein was cannulated and a microsyringe was attached to
intravenous tubing for methacholine administration. The mice were
then placed in a whole-body plethysmography chamber and mechanically ventilated (12). Transpulmonary pressure was measured as airway opening pressure referenced to pressure within the chamber. A
four-way connector was attached to the tracheostomy tube. One port
of the four-way connector was attached to the inspiratory side of a
ventilator (rodent ventilator, model 683; Harvard, South Natick, MA)
while another port was connected to the ventilator expiratory side
port. Mice were ventilated at a rate of 200 breaths/min with a tidal
volume of 5-6 ml/kg and a positive end-expiratory pressure of 2 cm
H2O. Lung volume changes were measured by detecting pressure
changes in the plethysmographic chamber (model MC 1-3-871; Validyne, Northridge, CA). Flow was measured by differentiation of the
volume signal. Pressure, flow, and volume changes were recorded.
Lung resistance was continuously computed (LabVIEW Graphical Programming for Instruments; National Instruments, Austin, TX) by
fitting flow, volume, and pressure to an equation of motion. Acetyl--methacholine (Sigma) was dissolved in normal saline and administered intravenously at a starting dose of 5 µg/kg. The average volume
per methacholine dose was approximately 35 µl. Threefold-increasing
concentrations of methacholine were administered at 2-min intervals,
and only after transpulmonary pressure and tidal volume returned to
within 10% of baseline. Pulmonary variables were recorded for at
least 10 breaths during the peak response, within 30 s after each intravenous methacholine dose. Methacholine dose-response curves were
obtained by calculating the mean ± standard error for individual animals at each methacholine dose.
Protocol for Examining Lung Sections
The mice were killed by cervical dislocation on Day 23 and the lung block was removed. The lung tissue was stored in 4% paraformaldehyde, paraffin embedded, cut in 6-µm sections, mounted, and stained with hematoxylin and eosin for routine histology, periodic acid-Schiff (PAS) to assess mucus, and Luna stain to specifically evaluate eosinophils (13). Slides were examined by one observer in a blinded fashion as previously described (14). The following compartments of the lung were assessed: alveolar spaces, airways at all levels, interstitium, and vessels (both arteries and veins). Inflammatory infiltrates were evaluated for location, severity, and composition (cell types: small mononuclear cells, transformed lymphocytes, histiocytes, neutrophils, and eosinophils). The degrees of inflammation were graded as follows: 0, no infiltrate; 1+, most vessels have an infiltrate up to four cells thick; 2+, most vessels have an infiltrate five to seven cells thick; 3+, most vessels have an infiltrate greater than seven cells thick. Interstitial alveolar cellularity was graded as follows: 0, no infiltrate; 1+, minimal increased cellularity without widening of septa; 2+, obvious increased cellularity with widening of septa; and 3+, markedly increased cellularity with thickened septa; this score also includes blood or edema fluid in the tissue space.
Statistical Analysis
Results are expressed as means ± standard error of the mean (SEM). Dose-response curves to methacholine were compared by repeated measures analysis of variance (ANOVA) with the Fisher least significant difference performed as a post hoc analysis. Measurements of PGE2, LTC4/D4/E4, cytokines by ELISA, chemokines by RNase protection assay (RPA), and total IgE were analyzed by ANOVA with the Fisher least significant difference performed as a post hoc analysis. Differences were considered to be significant if p < 0.05.
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RESULTS |
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ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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Indomethacin Administration during Allergen Sensitization Decreases PGE2 Levels and Increases Cysteinyl Leukotriene Levels in BAL Fluid
The PGE2 level measured in the BAL fluid on Day 18 was 241 ± 30 pg/ml in the OVA group and 102 ± 20 pg/ml in the OVA- indomethacin group (p < 0.05) as shown in Figure 2. In unsensitized mice not treated with indomethacin (NS), the PGE2 level in the BAL fluid was 94 ± 19 pg/ml. PGE2 was not measured in unsensitized mice treated with indomethacin (NS- indomethacin). In contrast, the level of LTC4/D4/E4 was 18 pg/ ml in the OVA-indomethacin group and 3 pg/ml in the OVA group (n = 4 in each group, results representative of three separate experiments). LTC4/D4/E4 was not measured in the BAL fluid of unsensitized mice. Thus, indomethacin treatment during allergen sensitization decreased the concentration of PGE2, yet increased the cysteinyl leukotrienes.
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IL-5 and IL-13 Are Increased in Indomethacin-treated OVA-sensitized Mice, while IL-4 and IL-6 Are Unchanged
Cytokines were measured in the ground lung supernatants on
Day 18, the fifth day of 8 d of ovalbumin aerosol treatment. We have previously found that cytokine levels peak on this day
even though aerosol treatments continued for three more days
(data not shown). Levels of IL-4, IL-5, or IL-13 were not detected in the lung supernatants of the NS and NS-indomethacin
groups. Indomethacin treatment had no significant effect on
IL-4 protein levels in lung supernatants of the allergically sensitized mice (Figure 3). However, indomethacin treatment of
ovalbumin-sensitized BALB/c mice significantly increased IL-5
(176 ± 52 versus 66 ± 4 pg/ml) and IL-13 (1,226 ± 279 versus
474 ± 65 pg/ml) protein levels in lung supernatants (p < 0.05;
n = 4 in each group, results representative of three separate
experiments). Thus, prostaglandin inhibition increased, rather
than decreased, cytokines that are considered to be central in
allergen-induced AHR. IL-6 was also measured in the ground
lung supernatants on Day 18. There was a trend for the OVA-
indomethacin group to have a decrease in IL-6 (146 ± 5 versus
208 ± 33 pg/ml) protein in lung supernatants (p = 0.1; n = 4).
Levels of IFN- were undetectable in the ground lung supernatants from any of the groups. RNase protection assays
(RPAs) were performed on the lungs from mice harvested on
Day 18 of the protocol (Figures 4A and 4B). There were no
differences in the levels of cytokine mRNA in the lung tissue
for IL-4 and IL-5 between the OVA-indomethacin and OVA
groups; however, the level of IL-13 mRNA was significantly increased in the OVA-indomethacin group. There was a trend
for the IL-6 mRNA to be increased in the OVA group compared with the OVA-indomethacin group (p = 0.08). There
was no detectable mRNA for IFN- in either the OVA or
OVA-indomethacin group.
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Indomethacin Treatment Increased Lung MCP-1 mRNA in OVA-sensitized Mice
RNase protection assays (RPAs) were performed on the lungs
from mice harvested on Day 18 of the protocol. There was no
difference in mRNA levels for eotaxin, MIP-1, MIP-1, or
MIP-2 between the OVA-indomethacin and OVA groups (Figures 5A and 5B), although the lung mRNA for these chemokines was significantly greater in the OVA-indomethacin and
OVA groups compared with the two groups of mice that were
not OVA sensitized. However, the lung mRNA for MCP-1 was
significantly greater in the OVA-indomethacin group compared
with the OVA group (p < 0.05). In the nonsensitized mice there
was no difference in MCP-1 lung mRNA expression between the mice treated with indomethacin and those that were not.
Thus indomethacin treatment increased MCP-1 mRNA expression only in those mice that were OVA sensitized.
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Indomethacin Did Not Increase Total IgE Levels in the Sera of OVA-sensitized Mice
On Day 23, the day after the last OVA aerosol treatment, blood was drawn after the methacholine challenge for measurement of total IgE levels. The average total IgE level was 9.34 ± 1.43 µg/ml in the sera of the OVA-indomethacin mice and 4.95 ± 1.00 µg/ml in the OVA group (n = 4 in each group, results representative of two separate experiments). This represents a trend for an increase in total IgE in the OVA-indomethacin mice compared with the OVA group (p = 0.08) (Figure 6). Mice that are not sensitized have no detectable IgE in their sera (data not shown).
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Indomethacin Increased Allergen-induced Lung Pathology in the Lung Interstitium
The OVA-indomethacin group had increased interstitial inflammation compared with the OVA group (Figure 7). This inflammation was composed of a large number of eosinophils (2+), macrophages (2+), and small lymphocytes (2+), while there were only a few macrophages (1+) in the interstitium of the OVA group. There was no difference in the inflammation in the bronchovascular or perivenous spaces between the two groups. Both the OVA and OVA-indomethacin groups had a significant inflammation in the bronchovascular and perivenous compartments with extensive infiltration of eosinophils, small lymphocytes, and plasma cells. There was also a large amount of mucus in the airways of both the OVA and OVA-indomethacin groups. There was essentially no inflammation in the bronchovascular, perivenous, and interstitial compartments of the two groups of nonsensitized mice, nor was there any mucus in the airways of these two groups.
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Indomethacin Increased Allergen-induced AHR
Methacholine challenge was performed on Day 23, 1 d after the last OVA aerosol treatment. The AHR was significantly greater in the OVA-indomethacin group compared with the OVA group (Figure 8). At the highest dose of methacholine, the OVA-indomethacin mice had a lung resistance of 27.5 ± 3.6 cm H2O/ml/s, while the lung resistance of the OVA mice was 17.6 ± 2.8 cm H2O/ml/s. Indomethacin treatment did not increase AHR in either the NS or NS-indomethacin group.
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DISCUSSION |
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ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
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Prostaglandins influence many important inflammatory processes (15). These mediators are produced by the phospholipase A2 conversion of membrane phospholipids to arachidonic acid, followed by the cyclooxygenase conversion of
arachidonic acid to prostaglandin H2 (PGH2). PGH2 may then
be converted by specific synthases or isomerases to either
prostaglandin D2 (PGD2), prostaglandins F2
(PGF2), prostaglandin E2 (PGE2), prostaglandin I2 (PGI2), or thromboxane
(15). The prostaglandins and thromboxane have diverse effects on the inflammatory cascade and are presumed to have a
pathogenic role in asthma (16, 17). PGD2 is increased in the
BAL fluid of subjects with asthma (18) while both PGD2 and
PGF2 contract human airway smooth muscle in vitro and are
potent bronchoconstrictors in vivo (19). Thromboxane also causes human airway smooth muscle contraction in vitro (20). On the other hand, PGI2 causes relaxation of isolated precontracted human bronchi (19), but has minimal effects on airway
function in vivo (21). PGE2 has a bronchoprotective effect in
humans (22). Many groups have reported that PGE2 in vitro
inhibits lymphocyte production of the Type 1 cytokines IL-2
and interferon , thus promoting T cell differentiation toward
a Type 2 cytokine profile (2).
In our study, we found that cyclooxygenase inhibition by indomethacin treatment during allergen sensitization decreased PGE2 and increased LTC4/D4/E4 in BAL fluid, and increased the lung supernatant protein levels of the Type 2 cytokines IL-5 and IL-13. To our knowledge, this is the first time that prostaglandin synthesis inhibition has been reported to selectively increase specific Type 2 cytokines in the setting of allergic sensitization. We found that indomethacin treatment during allergen sensitization caused a trend for an increase in IL-4 protein levels in lung supernatants and on serum total IgE production, although these were not significant changes. These data corroborate the in vitro and in vivo demonstrations that IL-4 is required for the production of IgE and is the principal cytokine that stimulates switching of B cells to the IgE heavy chain (23). Others have reported that indomethacin treatment for 60 d decreased IL-6 production in a model of mineral oil-induced chronic intraperitoneal inflammation (10). In our model of allergic inflammation, we found that indomethacin treatment caused a trend toward decreased IL-6 production (p = 0.1) in mice that were exposed to aerosolized ovalbumin for 8 d. It is possible that the duration of the inflammatory stimulus in our model was too short for indomethacin to have exerted a significant change in IL-6 concentrations.
Indomethacin treatment during ovalbumin sensitization decreased PGE2 while increasing the cysteinyl leukotrienes. The increase in the cysteinyl leukotrienes could result from "shunting" of arachidonate to the 5-lipoxygenase pathway (11). Alternatively, inhibition of PGE2 by indomethacin is known to augment leukotriene-C4 synthase activity, thereby increasing the production of the cysteinyl leukotrienes (24). Similarly, the increase in the cysteinyl leukotriene products of the 5-lipoxygenase pathway could result from an IL-5-mediated activity (25). IL-5 increases the expression of the 5-lipoxygenase-activating protein and translocates 5-lipoxygenase to the nucleus in normal blood eosinophils in vitro. This is associated with an increased capacity for cysteinyl leukotriene expression and is similar to the increase in 5-lipoxygenase expression seen in the eosinophils from subjects with allergic asthma (25).
This increase in the cysteinyl leukotrienes may be important in the increased lung interstitial eosinophilic inflammation seen in the OVA-indomethacin-treated mice, as these
mediators are described as being involved in eosinophil recruitment (26). In addition, indomethacin caused an increase
in IL-5, a cytokine that increases eosinophil accumulation (27,
28) and is an important mediator causing eosinophil growth,
recruitment, and survival (23). However, none of the chemokines thought to be responsible for eosinophil chemotaxis were
elevated in the OVA-indomethacin group compared with the
OVA mice. The lung mRNA expression of eotaxin, MIP-1,
and MIP1 were not different between the OVA-indomethacin and OVA groups. MCP-1 is a member of the C-C chemokine
family and binds to the CCR2 receptor (29). CCR2 receptors
are present on basophils, monocytes, activated T cells, dendritic
cells, and natural killer cells, thus allowing for MCP-1-mediated
biologic effects (29). Although the upregulation of MCP-1 as a
result of indomethacin treatment may have recruited an increased number of activated T cells expressing IL-5 to the lung
in the OVA-indomethacin group, this possibility remains unexplored as we did not perform immunohistochemistry to delineate activated from nonactivated T cells.
Our results are in agreement with those of Gavett and colleagues, who reported their findings in mice lacking either prostaglandin H synthase 1 (PGHS-1) or PGHS-2 (30). In comparison with wild-type mice, they found a heightened degree of eosinophilic inflammation, higher levels of IgE, decreased PGE2 and increased leukotriene B4 in BAL fluid, and increased airway responsiveness in the mice lacking PGHS-1. Our findings with indomethacin are consistent with an inhibition of a prostanoid, e.g., PGE2, which may restrain allergic inflammation. Our results extend theirs to show that this effect is not the result of alterations in immune responses in transgenic mice that have never been exposed to PGHS-1 products, and by our finding that the Type 2 cytokines IL-5 and IL-13 are elevated as well, and may play a role in the increased inflammation and AHR we found in indomethacin-treated mice.
IL-5 and IL-13 are reported to be critical for the development of allergic airway inflammation and hyperresponsiveness (31). The mechanism by which prostaglandin inhibition leads to the increase in IL-5 and IL-13 is unknown, but several possibilities exist. Perhaps in an in vivo system, PGE2 does not increase a Type 2 cytokine profile as suggested by in vitro studies, and instead may downregulate Type 2 cytokine production. Another possibility is that the cysteinyl leukotrienes may have an effect on T cell function and could perhaps regulate IL-5 and IL-13 production. The cysteinyl leukotriene 1 receptor has been cloned and the highest expression of the mRNA for this receptor is in spleen and peripheral blood leukocytes (38). To our knowledge, there have been no reports on the role of the cysteinyl leukotrienes in T cell development or on the distribution of the cysteinyl leukotriene 1 receptor on T cell subsets. We have previously shown that mice lacking the 5-lipoxygenase gene produce less IgE in response to OVA than do wild-type mice, suggesting that the leukotrienes could be involved in allergic inflammation (12).
Our report is the first to describe that cyclooxygenase inhibition during allergic sensitization increases IL-5 and IL-13 production and that this increase in these Type 2 cytokines is associated with an increase in allergen-induced AHR. Our findings suggest that the products of arachidonic acid metabolism have important immunomodulatory effects on allergic sensitization in vivo.
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Footnotes |
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Correspondence and requests for reprints should be addressed to James R. Sheller, M.D., Center for Lung Research, T-1217 MCN, Vanderbilt University Medical Center, Nashville, TN 37215. E-mail: James.Sheller{at}mcmail.vanderbilt.edu
(Received in original form November 15, 1999 and in revised form February 14, 2000).
Acknowledgments: Supported by K08-HL-03730, GM 15431, and R01-AI-45512.
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References |
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ABSTRACT
INTRODUCTION
METHODS
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REFERENCES
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1. Vane, J. R.. 1976. The mode of action of aspirin and similar compounds. J. Allergy Clin. Immunol. 58: 691-712 [Medline].
2. Betz, M., and B. S. Fox. 1991. Prostaglandin E2 inhibits production of Th1 lymphokines but not of Th2 lymphokines. J. Immunol. 146: 108-113 [Abstract].
3. Snijdewint, F. G., P. Kalinski, E. A. Wierenga, J. D. Bos, and M. L. Kapsenberg. 1993. Prostaglandin E2 differentially modulates cytokine secretion profiles of human T helper lymphocytes. J. Immunol. 150: 5321-5329 [Abstract].
4. Katamura, K., N. Shintaku, Y. Yamauchi, T. Fukui, Y. Ohshima, M. Mayumi, and K. Furusho. 1995. Prostaglandin E2 at priming of naive CD4+ T cells inhibits acquisition of ability to produce IFN-gamma and IL-2, but not IL-4 and IL-5. J. Immunol. 155: 4604-4612 [Abstract].
5. Hilkens, C. M., H. Vermeulen, R. J. van Neerven, F. G. Snijdewint, E. A. Wierenga, and M. L. Kapsenberg. 1995. Differential modulation of T helper type 1 (Th1) and T helper type 2 (Th2) cytokine secretion by prostaglandin E2 critically depends on interleukin-2. Eur. J. Immunol. 25: 59-63 [Medline].
6.
van der Pouw Kraan, T. C., L. C. Boeije, R. J. Smeenk, J. Wijdenes, and
L. A. Aarden.
1995.
Prostaglandin-E2 is a potent inhibitor of human
interleukin 12 production.
J. Exp. Med.
181:
775-779
7. Sottile, A., I. Venza, M. Venza, A. Valenti, and D. Teti. 1996. PGE2- induced immunoregulation mediated by cytokine production from cultures of human peripheral T lymphocytes. Immunopharm. Immunotoxicol. 18: 27-36 [Medline].
8. Khan, M. M.. 1995. Regulation of IL-4 and IL-5 secretion by histamine and PGE2. Adv. Exp. Med. Biol. 383: 35-42 [Medline].
9. Varner, A. E.. 1999. The cyclooxygenase-2 theory of atopy and asthma. Pediatr. Asthma Allergy Immunol. 13: 43-50 .
10.
Shacter, E.,
G. K. Arzadon, and
J. Williams.
1992.
Elevation of interleukin-6 in response to a chronic inflammatory stimulus in mice: inhibition by indomethacin.
Blood
80:
194-202
11. Dworski, R., J. R. Sheller, N. E. Wickersham, J. A. Oates, K. Brigham, L. J. Roberts II, and G. A. Fitzgerald. 1989. Allergen-stimulated release of mediators into sheep bronchoalveolar lavage fluid: effect of cyclooxygenase inhibition. Am. Rev. Respir. Dis. 139: 46-51 [Medline].
12. Chen, X.-S., J. R. Sheller, E. N. Johnson, and C. D. Funk. 1994. Role of leukotrienes revealed by targeted disruption of the 5-lipoxygenase gene. Nature (London) 372: 179-182 [Medline].
13. Luna, L. G. 1979. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. McGraw-Hill, New York. 111.
14. Peebles, R. S. Jr., J. R. Sheller, J. E. Johnson, D. B. Mitchell, and B. S. Graham. 1999. Respiratory syncytial virus infection prolongs methacholine-induced airway hyperresponsiveness in ovalbumin sensitized mice. J. Med. Virol. 57: 186-192 [Medline].
15. Smith, W. L., and D. L. Dewitt. 1996. Prostaglandin endoperoxide H synthases-1 and -2. Adv. Immunol. 62: 16-215 .
16. Holtzman, M. J.. 1991. Arachidonic acid metabolism: implications of biological chemistry for lung function and disease. Am. Rev. Respir. Dis. 143: 188-203 [Medline].
17. Wenzel, S. E.. 1997. Arachidonic acid metabolites: mediators of inflammation in asthma. Pharmacotherapy 17: 3S-12S [Medline].
18. Murray, J. J., A. B. Tonnel, A. R. Brash, L. J. Roberts II, P. Gosset, R. Workman, A. Capron, and J. A. Oates. 1986. Release of prostaglandin D2 into human airways during acute antigen challenge. N. Engl. J. Med. 315: 800-804 [Abstract].
19. Gardiner, P. J., and H. O. Collier. 1980. Specific receptors for prostaglandins in airways. Prostaglandins 19: 819-841 [Medline].
20. Armour, C. L., P. R. Johnson, M. L. Alfredson, and J. L. Black. 1989. Characterization of contractile prostanoid on human airway smooth muscle. Eur. J. Pharmacol. 165: 215-222 [Medline].
21. Hardy, C., C. Robinson, R. A. Lewis, A. E. Tattersfield, and S. Holgate. 1985. Airway and cardiovascular responses to inhaled prostacyclin in normal and asthmatic subjects. Am. Rev. Respir. Dis. 131: 18-21 [Medline].
22.
Gauvreau, G. M.,
R. M. Watson, and
P. M. O'Byrne.
1999.
Protective effects of inhaled PGE2 on allergen-induced airway responses and airway inflammation.
Am. J. Respir. Crit. Care Med.
159:
31-36
23. Abbas, A. K., A. H. Lichtman, and J. S. Pober. 1997. Cytokines. In A. K. Abbas, A. H. Lichtman, and J. S. Pober, editors. Cellular and Molecular Immunology. W.B. Saunders, Philadelphia, PA. 249-277.
24. Docherty, J. C., and T. W. Wilson. 1987. Indomethacin increases the formation of lipoxygenase products in calcium ionophore stimulated human neutrophils. Biochem. Biophys. Res. Commun. 148: 534-538 [Medline].
25.
Cowburn, A. S.,
S. T. Holgate, and
A. P. Sampson.
1999.
IL-5 increases
expression of 5-lipoxygenase-activating protein and translocates 5- lipoxygenase to the nucleus in human blood eosinophils.
J. Immunol.
163:
456-465
26.
Hisada, T.,
M. Salmon,
Y. Nasuhara, and
K. F. Chung.
1999.
Cysteinyl-leukotrienes partly mediate eotaxin-induced bronchial hyperresponsiveness and eosinophilia in IL-5 transgenic mice.
Am. J. Respir. Crit.
Care Med.
160:
571-575
27. van Oosterhout, A. J., D. Fattah, I. van Ark, G. Hofman, T. L. Buckley, and F. P. Nijkamp. 1995. Eosinophil infiltration precedes development of airway hyperreactivity and mucosal exudation after intranasal administration of interleukin-5 to mice. J. Allergy Clin. Immunol. 96: 104-112 [Medline].
28. Terada, N., A. Konno, H. Tada, K. Shirotori, K. Ishikawa, and K. Togawa. 1992. The effect of recombinant human interleukin-5 on eosinophil accumulation and degranulation in human nasal mucosa. J. Allergy Clin. Immunol. 90: 160-168 [Medline].
29.
Luster, A. D..
1998.
Chemokines
chemotactic cytokines that mediate
inflammation.
N. Engl. J. Med.
338:
436-445
30. Gavett, S. H., S. L. Madison, P. C. Chulada, P. E. Scarborough, W. Qu, J. E. Boyle, H. F. Tiano, C. A. Lee, R. Langenbach, V. L. Roggli, and D. C. Zeldin. 1999. Allergic lung responses are increased in prostaglandin H synthase-deficient mice. J. Clin. Invest. 104: 721-732 [Medline].
31. Coyle, A. J., G. Le Gros, C. Bertrand, S. Tsuyuki, C. H. Heusser, M. Kopf, and G. P. Anderson. 1995. Interleukin-4 is required for the induction of lung Th2 mucosal immunity. Am. J. Respir. Cell Mol. Biol. 13: 54-59 [Abstract].
32.
Foster, P. S.,
S. P. Hogan,
A. J. Ramsay,
K. I. Matthaei, and
I. G. Young.
1996.
.Interleukin 5 deficiency abolishes eosinophilia, airways hyperreactivity, and lung damage in a mouse asthma.
J. Exp. Med.
183:
195-201
33.
Corry, D. B.,
H. G. Folkesson,
M. L. Warnock,
D. J. Erle,
M. A. Matthay,
J. P. Wiener-Kronish, and
R. M. Locksley.
1996.
Interleukin 4, but not interleukin 5 or eosinophils, is required in a murine model of
acute airway hyperreactivity.
J. Exp. Med.
183:
109-117
34. Brusselle, G. G., J. C. Kips, J. H. Tavernier, J. G. van der Heyden, C. A. Cuvelier, R. A. Pauwels, and H. Bluethmann. 1994. Attenuation of allergic airway inflammation in IL-4 deficient mice. Clin. Exp. Allergy 24: 73-80 [Medline].
35. Kips, J. C., G. G. Brusselle, G. F. Joos, R. A. Peleman, R. R. Devos, J. H. Tavernier, and R. A. Pauwels. 1995. Importance of interleukin-4 and interleukin-12 in allergen-induced airway changes in mice. Int. Arch. Allergy Immunol. 107: 115-118 [Medline].
36.
Grunig, G.,
M. Warnock,
A. E. Wakil,
R. Venkayya,
F. Brombacher,
D. M. Rennick,
D. Sheppard,
M. Mohrs,
D. D. Donaldson,
R. M. Locksley, and
D. B. Corry.
1998.
Requirement for IL-13 independently of IL-4 in experimental asthma.
Science
282:
2261-2263
37.
Wills-Karp, M.,
J. Luyimbazi,
X. Xu,
B. Schofield,
T. Y. Neben,
C. L. Karp, and
D. D. Donaldson.
1998.
Interleukin-13: central mediator of
allergic asthma.
Science
282:
2258-2261
38. Lynch, K. R., G. P. O'Neill, Q. Liu, D. S. Im, N. Sawyer, K. Metters, N. Coulombe, M. Abramovitz, D. J. Figueroa, Z. Zeng, B. M. Connolly, C. Bai, C. P. Austin, A. Chateauneuf, R. Stocco, G. M. Greig, S. Kargman, S. B. Hooks, E. Hosfield, D. L. Williams Jr., A. W. Ford-Hutchinson, C. T. Caskey, and J. F. Evans. 1999. Characterization of the human cysteinyl leukotriene CysLT1 receptor. Nature (London) 399: 789-793 [Medline].
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