(Chest. 2001;120:1175-1183.)
© 2001
American College of Chest Physicians
Preoperative Steroid Therapy Inhibits Cytokine Production in the Lung Parenchyma in Asthmatic Patients*
Kazuko Mitsuta, MD;
Terufumi Shimoda, MD;
Chizu Fukushima, MD;
Yasushi Obase, MD;
Hiroyoshi Ayabe, MD;
Hiroto Matsuse, MD and
Shigeru Kohno, MD, FCCP
*
From The Second Department of Internal Medicine (Drs. Mitsuta, Shimoda, Fukushima, Obase, Matsuse, and Kohno), and The First Department of Surgery (Dr. Ayabe), Nagasaki University School of Medicine, Nagasaki, Japan.
Correspondence to: Terufumi Shimoda, MD, The Second Department of Internal Medicine, Nagasaki University School of Medicine, 1–7-1 Sakamoto, Nagasaki 852-8501, Japan; e-mail:inmed2nd{at}net.nagasaki-u.ac.jp
 |
Abstract
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Objectives: During or after surgery, asthma attacks due
to airway hyperresponsiveness (AHR) are likely to occur in patients
with bronchial asthma. Preoperative administration of corticosteroid
for prevention of perioperative asthma attacks is useful. We examined
the mechanism of prevention of perioperative asthma attacks by the
preoperative administration of corticosteroid in
vitro.
Design: Five patients with asthma were
treated with 20 mg of prednisolone orally for 2 preoperative days and
80 mg of methylprednisolone IV immediately before and after surgery. In
another five patients without asthma, no steroids were administered. A
noncarcinomatous part of the resected tissue from each patient with
lung cancer was passively sensitized with the serum of an atopic
patient. In the patients without asthma, the tissue was treated with or
without dexamethasone, and then mite antigen was added.
Measurements: The culture supernatant and lung tissue were
recovered, and the supernatant was assayed for histamine, leukotriene
E4 (LTE4), interleukin (IL)-5, and tumor
necrosis factor (TNF)-
. Degranulation of mast cells was measured by
tryptase staining of the lung tissue, and the expression of messenger
RNA (mRNA) of IL-5 and TNF- was determined by the reverse
transcriptase-polymerase chain reaction method.
Results: While preoperative administration of
corticosteroid did not suppress the release of histamine and
LTE4 from the lungs of asthmatic patients, it completely
suppressed IL-5 and TNF- production at the mRNA level. The same
results were obtained in lung tissues of nonasthmatic patients treated
in vitro with dexamethasone.
Conclusions: Our results suggest that corticosteroid
treatment reduces AHR and prevents perioperative attacks of asthma
primarily by suppressing the production of inflammatory
cytokines.
Key Words: airway hyperresponsiveness • corticosteroid • cytokines • interleukin-5 • perioperative asthma • tumor necrosis factor-
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Introduction
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Patients
with asthma are at increased risk of specific complications during and
after surgical operations, ie, acute bronchoconstriction
triggered by intubation, hypoxemia and possible hypercapnia, impaired
effectiveness of cough, atelectasis, and respiratory infection due to
airway hyperresponsiveness (AHR), airflow obstruction, and mucus
hypersecretion.1
Therefore, preoperative evaluation of the
state of asthma and systemic administration of corticosteroid to
maintain pulmonary function at its best are important precautions to
reduce the risk during the perioperative period.2
However,
corticosteroid administration increases the likelihood of respiratory
infections, wound infections, difficulties in wound healing,
complications of adrenal insufficiency; therefore, its dosage
preoperatively must be carefully evaluated. In our hospital, we have
administered corticosteroid preoperatively to > 100 asthmatic
patients and noted the effectiveness of such therapy in preventing
asthmatic attacks during the perioperative period (unpublished
observation; August 2000). The National Institutes of Health
also established the following guideline: "For patients who have
received systemic corticosteroids during the past 6 months, give 100 mg
hydrocortisone every 8 hours intravenously during the surgical period
and reduce the dose rapidly within 24 hours following
surgery."2
The mechanism of action of corticosteroid as an antiasthma agent is not
entirely clear, but it is considered to suppress eosinophilic
inflammation of the airway and AHR by controlling the production of
inflammatory cytokines such as interleukin (IL)-5 and tumor necrosis
factor (TNF)-.3
4
On the other hand, corticosteroid has
no inhibitory effect on the release of chemical mediators such as
histamine and leukotriene from mast cells, and it is considered to be
ineffective for immediate-type airway obstruction, in which chemical
mediators are involved.5
6
7
We hypothesized that corticosteroid administered preoperatively
prevents perioperative attacks of asthma by suppressing AHR through a
reduction of production of inflammatory cytokines. The present in
vitro study was conducted to verify this hypothesis by preparing a
passively sensitized lung model of lung tissues obtained from asthmatic
patients preoperatively treated with corticosteroid.
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Materials and Methods
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Subjects
The subjects were five asthmatic and five nonasthmatic patients.
All 10 patients had lung cancer. Table 1
shows the characteristics of each patient. The diagnosis of bronchial
asthma and the grading of its severity were based on the guideline set
by Global Initiative for Asthma.8
Patients 2, 4, and 9 had
pulmonary emphysema. None of the patients had received preoperative
anticancer chemotherapy or radiotherapy. Corticosteroid was
administered preoperatively to the asthmatic patients using the
following regimen: 20 mg of oral prednisolone for 2 days before the
operation and 80 mg of IV methylprednisolone immediately before and
after the operation. No steroid was administered to the control
nonasthmatic patients. Induction (fentanyl) and maintenance (nitrous
oxide sevoflurane) of anesthesia were achieved using the same
drugs in both groups of subjects, and no significant difference was
observed between the two groups with regard to age, male/female ratio,
preoperative pulmonary function, and mean operation time. Only mild
wheezing appeared in one asthmatic patient after extubation, but no
asthma attacks occurred in the remaining four patients. The absence of
side effects of corticosteroid therapy either during or after surgery
was confirmed by a review of the surgical records and interviews of the
patients by the attending physicians. The mean operation time of both
groups of patients was 3 h and 47 min, and the mean period from
the beginning of surgery to resection of the specimens was 2 h.
The study was carried out after the approval by the Ethical Committee
of Nagasaki University School of Medicine, and informed consent was
obtained from each patient prior to participation in the study.
Culture of Lung Tissues
The noncarcinomatous parts of surgically resected lung tissues
were cut and passively sensitized. For this purpose, the resected lung
tissue was minced into fragments of 300 mg and washed twice with
RPMI1640 (Gibco; Grand Island, NY) to prevent contamination by blood
cells. For stimulation via IgE receptors, the lung tissue was passively
sensitized with serum of an atopic patient who showed a mite
radioallergosorbent test score of 
5 over 2 h, and
cultured in RPMI1640 alone or RPMI1640 containing 1.5 µg/mL of mite
antigen for 48 h. Dexamethasone (Sigma Chemical; St. Louis, MO)
was added at 10-6 mol/L for in
vitro treatment of lung tissues of nonasthmatic patients. The lung
tissues of asthmatic patients, who had already been treated with
corticosteroid before surgery, were not treated in vitro
with dexamethasone. To examine whether the reaction was induced via
IgE, the lung tissues of nonasthmatic patients without passive
sensitization and treatment in vitro with dexamethasone were
directly stimulated with mite antigen. The culture supernatant and lung
tissues were recovered after a predetermined period following
stimulation; histamine, leukotriene E4
(LTE4), IL-5, and TNF- concentrations in the
supernatant were determined. The lung tissues were stained for
tryptase, mast cells were counted, total RNA was extracted from part of
the tissues, and the expression of messenger RNA (mRNA) of IL-5 and
TNF- was measured by the reverse transcriptase-polymerase chain
reaction (RT-PCR) method. Cultures were prepared invariably at 37°C
in 5% CO2 with the addition of 100 U/mL of
penicillin (Meiji Seika; Tokyo, Japan) and 100 µg/mL of streptomycin
sulfate to RPMI1640. The endotoxin level in the culture medium measured
using Toxicolor test (SRL; Tokyo, Japan) was 
50 pg/mL.
Measurement of Chemical Mediators and Cytokines in Supernatant of
Lung Fragments
Based on the method described previously,9
radioimmunoassay was used to measure histamine concentration 30 min
after stimulation with mite antigen and LTE4
concentration after 60 min, when their productions reach peak levels in
this model. IL-5 concentration 24 h after stimulation with mite
antigen and TNF- concentration 4 h after stimulation were
determined using a commercial enzyme-linked immunosorbent assay kit
(Quantikine; R&D Systems; Minneapolis, MN) when their
productions reach peak levels in this model. All results were expressed
as production per 300 mg of lung tissue.
RT-PCR
To analyze the expression of cytokine mRNA, we first extracted
the total cellular RNA by the guanidinium thiocyanate method, and then
amplified complementary DNA (cDNA) by reverse transcription of total
cellular RNA, as previously described.9
After stimulation
with RPMI or mite antigen for 30 min, 1 h, or 24 h, the lung
tissues were cut with scissors into small pieces on a Petri dish placed
on ice, then placed in a glass homogenizer with TRIzol reagent (1
mL/100 mg lung tissue; Gibco, BRL) and homogenized. The homogenate was
mixed with 0.2 mL of TRIzol/mL of chloroform, and then centrifuged at
12,000g for 15 min. In the next step, the liquid phase was
collected and mixed with isopropanol to extract total RNA. RNA pellets
were washed in 75% ethanol and dried, and then dissolved in diethyl
pyrocarbonate-treated water and stored at - 80°C.
Total cellular RNA was reverse transcribed with oligo deoxythymidine,
10x reverse transcriptase reaction buffer,
deoxynucleotide triphosphates (deoxyadenosine triphosphate,
deoxycitadine triphosphate, deoxyguanosine triphosphate, deoxythymidine
triphosphate), SuperScript II RNAase H-reverse
transcriptase (Gibco), 0.1 mol/L dithiothreitol, and 25 mM
MgCl2 to prepare cDNA. RNA was reacted at
42°C for 50 min, and then at 70°C for 15 min. Polymerase chain
reaction (PCR) buffer containing MgCl2, 25 mM
deoxynucleotide triphosphate mix, Taq polymerase, and diethyl
pyrocarbonate-treated water comprised a volume of 42 µL. To this
solution, we added 3 µL of cDNA, 2.5 µL of cytokine 3-primer, and
2.5 µL of cytokine 5-primer (total volume, 50 µL). Specific
cytokine oligonucleotide PCR primers were designed based on published
cDNA sequences (Genbank; Los Alamos National Laboratory; Los Alamos,
NM). For all primers, an amplifier set (Clontech Laboratories; Palo
Alto, CA) was used. Thirty cycles of reactions were induced using a
thermocycler under the following conditions: IL-5, denaturation at
94°C for 30 s, annealing at 49°C for 30 s, and extension
at 72°C for 60 s; TNF-, denaturation at 94°C for 30 s,
annealing at 47°C for 30 s, and extension at 72°C for 60
s. The last extension was performed at 72.5°C, and the final products
were stored at 4°C until electrophoresis. For electrophoresis, 10
µL of PCR-amplified product was placed on a 2% agarose gel, and then
visualized by ultraviolet illumination.
Immunohistochemistry
Lung tissues were stained for tryptase within 30 min after
stimulation with RPMI or mite antigen. The recovered lung fragments
were fixed with 20% formalin and embedded in paraffin. Tissue sections
were placed on glass slides, deparaffinized, and treated with trypsin.
Endogenous peroxidase activity was blocked by incubating slides in
methanol containing 3% hydrogen peroxidase for 30 min, followed by
washing in phosphate-buffered saline solution. After blocking
with 10% normal rabbit serum for 20 min, sections were incubated
overnight at 4°C with a mouse monoclonal antihuman mast cell tryptase
antibody (AA1; Dako; Tokyo, Japan) at a dilution of 1 x 500 of a 105
µg/mL solution. Following application of the primary antibody, we
applied a biotinylated rabbit antimouse antibody (Nichirei; Tokyo,
Japan), followed by the addition of peroxidase-conjugated avidin for 20
min at room temperature. Chromogen-fast diaminobenzidine (DAB; Sigma
Chemical) was used for 5 min, and the slides were counterstained in
hematoxylin and mounted.
Statistical Analysis
Data are presented as mean ± SEM. Differences between the
asthmatic lungs and nonasthmatic lungs were compared by two-group
unpaired t test, and differences in the same patient between
measurements performed before and after stimulation or treatment were
compared by two-group paired t test. A p value < 0.05 was
considered significant.
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Results
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Lung Histology
Figure 1
shows hematoxylin-eosin–stained lung sections. No inflammatory cell
infiltration including eosinophils or detachment of the airway
epithelium was noted in the asthmatic lungs probably because of
adequate preoperative control of asthma and preoperative administration
of corticosteroid.
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Figure 1.. Sections of the resected lung.
Top, a: without asthma; this patient was
not administered corticosteroid before surgery. Middle,
b: mild persistent asthma; this patient was treated with
oral administration of pranlukast and inhalation of sodium cromoglycate
plus salbutamol via nebulizer. Bottom, c:
moderate persistent asthma; this patient was treated with inhalation of
fluticasone propionate, 400 µg/d, and inhalation of sodium
cromoglycate plus salbutamol via nebulizer. The patients in
b and c were administered corticosteroid
preoperatively. No eosinophil infiltration or detachment of the airway
epithelium was observed in the asthmatic lungs, indicating satisfactory
control of asthma (hematoxylin-eosin, original x 800).
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Release of Chemical Mediators Induced by Dermatophagoides
farinae Allergen Stimulation
The release of chemical mediators from the passively sensitized
lung induced by stimulation with D farinae
allergen via IgE was examined. Significant release of histamine was
observed in the lungs of passively sensitized asthmatic subjects
preoperatively treated with corticosteroid and the lungs of the
passively sensitized nonasthmatic patients not treated with
corticosteroid, and the amount of histamine release was
comparable between the two groups. Furthermore, in vitro
stimulation of the lungs of passively sensitized nonasthmatic patients
with mite antigen after treatment with dexamethasone failed to suppress
the release of histamine (Fig
2
, left, A). The release of
LTE4 was similar to that of histamine (Fig 2
,
right, B). To examine whether the
reaction was induced via IgE, the lung tissues of nonasthmatic patients
without passive sensitization and treatment in vitro with
dexamethasone were directly stimulated with mite antigen. Both
histamine and LTE4 levels were below the
detectable ranges, and the release of chemical mediators by stimulation
with the mite antigen in the passive sensitization model was considered
a reaction via IgE.
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Figure 2.. Left, A: Histamine
concentration in the lungs of passively sensitized asthmatic and
nonasthmatic patients 30 min after stimulation with the medium and
Dermatophagoides antigen. Significant histamine release was induced by
antigen stimulation in the lungs of the passively sensitized asthmatic
patients who received preoperative steroid therapy, those of passively
sensitized nonasthmatic patients, and those of passively sensitized
nonasthmatic patients that received in vitro steroid
treatment. The amount of histamine release was comparable among the
three groups. Bars represent the mean ± SEM values of five subjects
in each group. *p < 0.01, not significant. Right,
B: LTE4 concentrations in the lungs of
passively sensitized asthmatic and nonasthmatic patients measured
1 h after medium stimulation and Dermatophagoides antigen
stimulation. Significant LTE4 production was induced by
antigen stimulation in the lungs of the passively sensitized asthmatic
patients who received preoperative steroid therapy, those of the
passively sensitized nonasthmatic patients, and those of the passively
sensitized nonasthmatic patients that received in vitro
steroid therapy. The amount of LTE4 production was
comparable among the three groups. Bars represent the mean ± SEM
values of five subjects in each group. Bars without SD mean the values
under limit of measurement. *p < 0.01, not significant.
DerfAg = D farinae allergen;
DEX = dexamethasone; NS = not significant.
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Immunostaining for Tryptase
Mast cells in the lung tissues were counted by immunostaining
using antihuman tryptase antibody. The number of mast cells per 200
epithelial cells of the bronchial epithelium and mucosa was not
significantly different between asthmatic and nonasthmatic lungs (Fig 3
). No significant difference was noted in the degree of degranulation of
mast cells after antigen stimulation between asthmatic and nonasthmatic
lungs. Tryptase-positive cells decreased due to degranulation after
antigen stimulation in the passively sensitized asthmatic lungs
preoperatively treated with corticosteroid and the passively sensitized
nonasthmatic lungs treated in vitro with dexamethasone (Fig 4
). These results indicate that preoperative or in vitro
corticosteroid administration did not suppress degranulation of mast
cells in the lungs of passively sensitized asthmatic or nonasthmatic
patients.
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Figure 3.. Effect of treatment on the number of
mast cells. Mast cells among 200 epithelial cells of the bronchial
epithelium and mucosa were counted in tryptase-stained sections of the
lungs of passively sensitized asthmatic and nonasthmatic patients and
those of passively sensitized nonasthmatic patients who received
in vitro steroid therapy 30 min after medium stimulation
and Dermatophagoides antigen stimulation. The number of mast cells was
not significantly different among the three groups. After antigen
stimulation, mast cells decreased due to degranulation, and no
difference was observed in the degree of degranulation among the three
groups. Bars represent the mean ± SEM values of five subjects in
each group. *p < 0.01, not significant. See Figure 2
legend for
expansion of abbreviations.
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Figure 4.. Sections of lung tissues from passively sensitized
asthmatic patients and passively sensitized nonasthmatic patients who
received in vitro dexamethasone treatment 30 min after
medium stimulation and Dermatophagoides antigen stimulation. Top
left, a: Sensitization plus RPMI (asthma).
Top right, b: Sensitization plus D
farinae (asthma). Bottom left, c:
Sensitization plus RPMI plus dexamethasone, 10-6
mol/L (nonasthma). Bottom right,
d: sensitization plus D farinae plus
dexamethasone, 10-6 mol/L (nonasthma). After antigen
stimulation, tryptase-positive cells decreased in number due to
degranulation in the lungs of both groups (tryptase,
original x 1,000).
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Cytokine Production by D farinae Allergen
Stimulation
No IL-5 production was noted when the lungs of the passively
sensitized asthmatic patients preoperatively treated with
corticosteroid were stimulated with D farinae allergen. In
contrast, significant IL-5 production was induced by D
farinae allergen stimulation in the lungs of passively sensitized
nonasthmatic patients who did not receive preoperative corticosteroid
administration, and this IL-5 production was suppressed by in
vitro pretreatment with dexamethasone (Fig 5
, left, A). Similar changes were observed with
regard to TNF- production (Fig 5
, right, B).
To examine whether the reaction was induced via IgE, the lung tissues
of nonasthmatic patients without passive sensitization and treatment
in vitro with dexamethasone were directly stimulated with
mite antigen. Both IL-5 and TNF- levels were below the detectable
ranges, and the production of cytokines by stimulation with the mite
antigen in the passive sensitization model was considered a reaction
via IgE. Mite extracts often contain a large amount of endotoxin, but
the endotoxin level in the culture medium containing D
farinae allergen measured using Toxicolor test (SRL; Tokyo, Japan)
was 50 pg/mL. The result suggested that the production of cytokines
was not affected by endotoxins in mite extracts.
Expression of Cytokine mRNAs
We examined the expression of IL-5 and TNF- mRNAs in the lungs
of passively sensitized asthmatic and nonasthmatic patients after
stimulation with mite antigen by using the RT-PCR method. Figure 6
shows typical gels. No expression of mRNA of either IL-5 or TNF- was
noted 30 min after antigen stimulation. One hour after antigen
stimulation, no expression of mRNA of either IL-5 or TNF- was noted
in the lungs of the passively sensitized asthmatic patients
preoperatively treated with corticosteroid while mRNA of both IL-5 and
TNF- was clearly expressed in the lungs of passively sensitized
nonasthmatic patients who did not receive preoperative corticosteroid
administration and was still expressed even after 24 h. The latter
expression was suppressed by in vitro dexamethasone
treatment. These results suggest that preoperative and
in vitro corticosteroid treatments suppress the production
of cytokine proteins and expression of mRNA induced by D
farinae allergen stimulation in the lungs of passively sensitized
asthmatic and nonasthmatic patients.
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Discussion
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The major findings of our study were as follows: (1) production of
histamine and LTE4 from human mast cells was not
reduced by corticosteroid treatment after stimulation with specific
allergen; (2) production of IL-5 and TNF- was, however,
significantly suppressed in lung tissues of passively sensitized
asthmatic patients preoperatively treated with corticosteroid, compared
with those in lung tissues of nonasthmatic patients without
corticosteroid treatment; and (3) production of IL-5 and TNF- was
suppressed in the lung tissues of passively sensitized nonasthmatic
patients treated in vitro with corticosteroid similar to the
lung tissues of the passively sensitized asthmatic patients
preoperatively treated with corticosteroid.
Bronchial asthma is a chronic eosinophilic inflammatory disorder of the
airway, and inhalation of specific allergens causes biphasic airway
obstruction. Chemical mediators such as histamine and leukotriene
produced by mast cells via IgE antibody are involved in immediate
allergic response that occurs shortly after inhalation of allergen. On
the other hand, the degree of late allergic response (LAR) observed
about 6 h after inhalation of allergen is related to the degree of
eosinophil infiltration, and the involvement of inflammatory cytokines
in this process has been suggested. In LAR, granular proteins are
thought to be continuously released from eosinophils accumulating in
the airway, and the consequent impairment of the airway epithelium is
speculated to induce AHR. Although corticosteroid is the most effective
antiasthmatic treatment available today, its precise mechanism of the
action has not yet been clarified. Treatment of asthmatics with
corticosteroid results in suppression of LAR but not immediate
asthmatic response.10
It has also been reported that
in vitro corticosteroid treatment does not suppress the
IgE-dependent release of histamine and cysteinyl-leukotrienes from mast
cells.5
6
Our results showed that the release of chemical
mediators induced by stimulation with specific allergen was not
suppressed either by preoperative treatment of asthmatics with
corticosteroid or in vitro corticosteroid treatment of lungs
of nonasthmatic patients. The results of tryptase staining also
suggested that degranulation of mast cells after stimulation with
specific allergen could not be prevented by corticosteroid. Thus, our
results indicate that corticosteroid does not act as antiasthmatic
agent by inhibiting the release of chemical mediators.
In contrast to its effect on chemical mediators, preoperative
corticosteroid therapy resulted in almost complete suppression of
production of IL-5 and TNF- proteins and inhibition of their mRNA
expressions in response to stimulation with specific allergen. Similar
results were obtained also in the lungs of nonasthmatic patients after
in vitro corticosteroid treatment. IL-5 and TNF- are
inflammatory cytokines that increase locally on allergen administration
through the airway of asthmatic patients and play important roles in
eosinophilic airway inflammation through activation of various
eosinophils and expression of adhesion molecules,
respectively.9
11
12
As mentioned above,
corticosteroid has a suppressive effect on LAR and
alleviates AHR by a short-term high-dose therapy.3
These
findings support our hypothesis that preoperative corticosteroid
therapy prevents perioperative asthmatic attacks primarily by reducing
the production of inflammatory cytokines such as IL-5 and TNF- and,
thus, suppressing AHR.
There are three problems with this study. Firstly, the dosage of
steroid used in the present study might be significantly greater than
that given in the National Institutes of Health guideline. Based on our
clinical experiences, we had determined the dosage of steroid. Any
steroid-related severe adverse effect had never occurred. Secondly, our
population samples did not include asthmatic patients who did not
receive preoperative corticosteroid administration. To evaluate
preoperative corticosteroid administration to asthmatic patients, it is
necessary to examine a control group of asthmatic patients who had not
received corticosteroid preoperatively. However, we encountered one
patient with mild intermittent asthma and two patients with mild
persistent asthma who underwent surgery under general anesthesia
without preoperative administration of corticosteroid. Surgery was
discontinued in the first patient because of bronchospasm triggered by
intubation, the second patient had an asthma attack with confusion
after surgery, and the third patient had bronchospasm triggered by
removal of the tube. These findings suggest that surgery under
general anesthesia is a risk for asthmatic patients with airway
hyperreactivity even when symptoms are stable. Therefore, we could not
ethically study the tissues of asthmatic patients who had not been
administered corticosteroids preoperatively. Previous
studies13
14
15
16
17
18
have shown that asthmatic patients exhibit a
high number of mast cells in their airway, enhanced sensitivity to
immunologic stimulation, and increased spontaneous release of
inflammatory mediators, compared with healthy individuals. In this
study, however, there were no differences in the number of mast cells
present in the airway epithelial and mucosal layer, the degree of mast
cell degranulation induced by stimulation with specific antigen, and
concentrations of chemical mediators in the supernatant between
asthmatics and nonasthmatic patients. Long-term administration of
corticosteroid has been reported to reduce the number of mast cells in
the airway and, consequently, to reduce the response to allergen
challenge.4
19
In this study, some patients were
receiving long-term corticosteroid inhalation therapy before surgery,
and corticosteroid was also administered immediately before operation.
Such a treatment could possibly explain the lack of significant
difference between the two groups. The third problem is that
cytokine-producing cells were not identified. All cells with IgE
receptors in lung tissues are potential cytokine-producing cells, but
it has been reported that mast cells are most important among
them.20
Previously, we confirmed that cytokines were not
produced by stimulation with CD3 (1 µg/mL) in the same
experimental model.21
Therefore, T cells were considered
noncytokine-productive cells. Eosinophils also have low-affinity IgE
receptor, but no eosinophils were detected in the
hematoxylin-eosin–stained lung tissues. Alveolar macrophages also have
low-affinity IgE receptors, but the number of IgE receptors is very low
or undetectable in healthy subjects without atopy.22
These
findings suggest that mast cells are the main source of the cytokines
measured in this study. Identification of cytokine-producing cells by
either in situ hybridization or in situ PCR is
necessary in future studies.
In conclusion, we have demonstrated in the present study that
preoperative corticosteroid therapy did not suppress histamine or
LTE4 production induced by specific allergen in
lung tissues of asthmatic patients, but it significantly suppressed
both IL-5 and TNF- production. Our results suggest that
corticosteroid prevents perioperative asthmatic attacks by suppressing
the inflammatory cytokine production and, thus, alleviating AHR.
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Footnotes
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Abbreviations:
AHR = airway hyperresponsiveness; cDNA = complementary DNA;
IL = interleukin; LAR = late asthmatic response;
LTE4 = leukotriene E4; mRNA = messenger
RNA; PCR = polymerase chain reaction; RT-PCR = reverse
transcriptase-polymerase chain reaction; TNF = tumor necrosis factor
Received for publication January 2, 2001.
Accepted for publication April 16, 2001.
|
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Abstract
Introduction
x 174/Hae;
bp = base pair