 , and MCP-1 into Asthmatic
Airways Following Endobronchial Allergen Challenge
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
ABSTRACT |
|---|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
We have investigated the presence of regulated on activation, normal T-cell expressed and probably
secreted (RANTES), macrophage inflammatory peptide-1
(MIP-1), and macrophage chemotactic
peptide (MCP-1) in the bronchoalveolar lavage fluid (BALF) obtained from normal (n = 7) and stable
asthmatic subjects (n = 8), and studied their kinetic release into asthmatic airways following endobronchial allergen challenge (n = 18). Measurements of RANTES, MIP-1, and MCP-1 in 10 times
(10×) concentrated BALF showed that these three chemokines were present in both normal controls
and stable asthmatic patients, but no significant difference between the two groups was found in the
levels of the three chemokines. However, at 4 h after allergen challenge, BALF levels of RANTES, MIP-1, and MCP-1 were significantly increased in fluid obtained from the allergen-challenge site when
compared with the saline-challenge control site (median: 175 pg/ml versus 11.5 pg/ml, 258 pg/ml
versus 88 pg/ml, and 900 pg/ml versus 450 pg/ml, respectively). At 24 h, levels of the three chemokines returned to baseline values. To investigate whether cells in BALF obtained 4 h after allergen exposure release chemokines, they were cultured for 24 h. BALF cells from the allergen site released
more RANTES and MCP-1 than those from the saline site, but released similar amounts of MIP-1. These findings suggest that RANTES, MIP-1, and MCP-1 may regulate cell trafficking in asthma in response to allergen exposure.
| |
INTRODUCTION |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Leukocyte recruitment into the airways is a feature of bronchial asthma. Increased number of both activated T cells and eosinophils have been a consistent finding in bronchial biopsies (1), bronchoalveolar lavage fluid (BALF) (2, 3), and in peripheral blood (4) derived from asthmatic patients. Moreover, exposure of asthmatic patients to a relevant allergen further increases leukocyte recruitment into asthmatic airways through the upregulation of vascular adhesion molecules (5) and release of cytokines from mast cells (6) and activated Th-2-like lymphocytes (7). Eosinophils are considered to play a key role in leading to disruption of the bronchial epithelium, enhanced bronchial responsiveness, and airway obstruction (8, 9). Cytokines encoded in the interleukin-4 (IL-4) gene cluster on Chromosome 5, specifically IL-3, IL-5, and granulocyte-macrophage colony-stimulating factor (GM-CSF), play a key role in eosinophil recruitment, migration, and activation associated with allergen exposure (7, 10), although this does not preclude other cytokines and mediators from being involved.
Over the past decade, a new family of cytokine peptides,
known as the chemokines, has been identified. These chemokines are 8- to 10-kD, basic heprin-binding polypeptides that
have been subdivided into C-X-C, C-C, and C branches according to their amino-acid sequences (11, 12). The chemokines
regulated on activation, normal T-cell expressed and probably
secreted (RANTES), macrophage inflammatory peptide-1
(MIP-1), and macrophage chemotactic peptide (MCP-1) are
members of the C-C branch and are thought to play an important role in the allergic inflammatory process. RANTES
and MIP-1 are chemotactic for lymphocytes, monocytes, and
eosinophils (13, 14), whereas the activities of MCP-1 are restricted to monocytes, lymphocytes (15, 16), and basophils (17). In vivo evidence for the involvement of these chemokines in leukocyte recruitment has been obtained from experimental animal studies. Injection of RANTES into dog skin results in the recruitment of monocytes and eosinophils at the
injection site (18). Similarly, at 4 to 8 h after intratracheal administration of Schistosoma mansoni eggs into mouse airways,
there occurs recruitment of eosinophils in parallel with MIP-1
release (19). Increased levels of RANTES and MIP-1 have
been detected in the nasal secretions of atopic patients exposed to local allergen challenge (20), and MCP-1 has been reported to be increased in nasal washings of ragweed-sensitive subjects during the pollen allergen season (21). Recently, these three chemokines have been found to be present in increased
levels in the bronchoalveolar lavage fluid (BALF) of patients
with active asthma (22). Immunoreactivity for MCP-1 is also
increased in the bronchial epithelium of asthmatic patients
(23). More recently, we have reported that allergen challenge
leads to the release of RANTES and MIP-1 into asthmatic airways, at 4 h after challenge (24, 25), whereas Sur and colleagues
have reported RANTES release at 24 h after allergen challenge (26). In the present study we have investigated the presence of RANTES, MIP-1, and MCP-1 immunoreactivity in
BALF derived from normal subjects and patients with mild allergic asthma. We further studied the effect of endobronchial
(local) allergen challenge on the release of these chemokines,
and their relationship with leukocyte recruitment in BALF at
4 and 24 h after asthmatic subjects were exposed to allergen.
We also report the production of RANTES, MIP-1, and MCP-1 by BALF cells.
| |
METHODS |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Subjects
Seven normal subjects (six male and one female; median age: 24 yr;
age range: 20 to 40 yr) and 25 patients with mild asthma (18 male and
seven female; median age: 31; age range: 20 to 55 yr) were recruited for
the study; their clinical characteristics are summarized in Tables 1 and
2. At the time of enrollment, all of the asthmatic subjects had stable
pulmonary function, with a baseline FEV1 
70% of that predicted
for their age and height. None of the asthmatic subjects had been
treated with inhaled or oral corticosteroids, sodium cromoglycate, or
theophylline for at least 6 wk prior to participation in the study. All of
asthmatic subjects were atopic as defined by a > 3 mm skin-wheal response to one or more of five common allergens (Dermatophagoides
pteronyssinus, mixed grass pollen, dog, feathers, and cat dander (Hollister Stier, Elkhart, IN). All of the asthmatic subjects were hyperresponsive to inhaled methacholine, with their geometric mean concentration of methacholine needed to evoke a 20% decline in FEV1
(PC20) value ranging from 0.1 to 2.9 mg/ml (Tables 1 and 2). The study
was approved by the Southampton Hospitals and University Ethical
Committee, and patients gave their written informed consent.
|
|
Study Design
The asthmatic patients were divided into four groups; five individuals formed part of more than one data group (27). Group A (n = 8) had a single bronchoscopy for bronchoalveolar lavage (BAL). Group B (n = 9) had allergen instilled into the middle lobe (ML) and saline into the right upper lobe (RUL), followed 4 h later by a second bronchoscopy for BAL of the ML and RUL. Group C (n = 8) underwent an identical procedure to that for Group B, but with the second bronchoscopy undertaken 24 h after allergen challenge and saline instillation. In order to determine the effects of bronchoscopy and BAL independently of the effects of allergen, Group D (n = 5) was treated exactly as Group B, with the exception that both the ML and RUL were exposed to saline, with BAL being performed in the ML 4 h later.
The allergen extract (mixed grass pollen or D. pteronyssinus)
(Hollister Stier) used for local bronchial challenge was that which produced the largest wheal response on skin-prick testing. In each subject, a skin-wheal dose-response series was undertaken, using 10-fold
dilutions of allergen, and the allergen concentration chosen for the
segmental bronchial challenge was 10 times lower than the concentration that produced a 3-mm wheal response. The median concentration
of allergen inducing a 3-mm wheal response was 10
4 (range: 103 to
106) (Table 2).
Bronchoscopy and Local Challenge
Fiberoptic bronchoscopy was undertaken as previously described (28). All subjects received 2.5 mg salbutamol by nebulization 15 min prior to the procedure, and intravenous atropine 0.6 mg and midazolam 3 to 8 mg. Oxygen (100%) was administered via nasal prongs throughout the procedure, and oxygen saturation (SaO2) was monitored with a digital oximeter (Minolta, Middlesex, UK). Fiberoptic bronchoscopy was done with an Olympus IT-20 bronchoscope (Olympus Optical Co., Tokyo, Japan). The sham challenge was undertaken in the RUL with 20 ml of prewarmed saline, and the allergen challenge was performed in the medial segment with 20 ml of allergen solution. Either 4 or 24 h later, the second bronchoscopy was performed and BAL was undertaken with six 20-ml aliquots of pre-warmed 0.9% saline solution. On completion of the procedure, subjects were observed for 3 h, and additional nebulized salbutamol was given as necessary.
Processing of BAL Fluid
The BAL fluid (BALF) obtained from bronchoscopies was immediately filtered through a 0.9 mm-gauge sieve and centrifuged at 400 × g
for 10 min at 4° C to separate cells from fluid. BALF thus processed
was then aliquoted into plastic tubes and frozen at 
70° C. The cell
pellet was resuspended in 5 ml of RPMI-1640 (containing 5% human
AB serum with 100 U/ml streptomycin), and the cells were counted in
a hemocytometer. A 100 µl aliquot of cells (1 × 106/ml) was subjected
to cytocentrifugation (Cytospin, Shandon Southern, Runcorn, UK)
and air dried to obtain differential cell counts after staining with May-
Grunwald-Giemsa stain. Six hundred cells were counted per cytospin.
BAL Cell Culture
BAL cells obtained from asthmatic subjects undergoing endobronchial allergen challenge were washed in RPMI-1640, resuspended in
culture medium (106 cells/ml), and cultured at 37° C in an incubator
containing 5% CO2. The culture medium consisted of RPMI-1640
with 5% human AB serum, 1 mM 1-glutamine, 2 mM sodium pyruvate, 100 U/ml streptomycin, and 0.5 µg/ml fungizone (Gibco, Paisley,
UK). After 24 h incubation, culture supernatant of BAL cells was collected and kept at 70° C for measurements of chemokines.
Measurements of Chemokines
Measurement of the chemokines RANTES, MIP-1, and MCP-1 in
both BALF (10 × concentrated, using 3-kDA-cutoff Centricon filters; Amicon Inc., Beverly, MA) and BAL cell culture supernatant (diluted up to 5-fold to get the sample concentration onto the linear part of the
enzyme-linked immunoassay [ELISA] standard curves) was done
with ELISA kits according to the manufacturer's protocol (R&D Systems, Abingdon, UK). Study samples and standard dilutions of specific cytokine were assayed in duplicate. Each assay had demonstrated
no measurable cross-reactivity to other cytokines as determined by
ourselves and as stated by the manufacturer. The limits of detection
for RANTES, MIP-1, and MCP-1 were 5 pg/ml, 10 pg/ml, and 15 pg/
ml, respectively. Concentrations of chemokines in BAL cell culture
supernatant were corrected for their initial dilution, whereas concentrations of chemokines in BALF were given as measured in 10×-concentrated BALF.
Albumin Measurements in BALF
Measurements of albumin in BALF samples (nonconcentrated BALF) were undertaken with rocket immunoelectrophoresis (29). Sensitivity of the assay was 1 µg/ml, and the coefficient of variation (cv) of repeat measurements was < 5%.
Statistical Analyses
Statistical analyses of RANTES, MIP-1, and MCP-1 in BALF and
BAL cell supernatant was performed with Wilcoxon's paired rank test. Spearman's rank correlation coefficient was used to investigate any relationship between cell counts and RANTES, MIP-1, and
MCP-1 concentrations. A value of p < 0.05 was regarded as statistically significant.
| |
RESULTS |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Steady-state Asthma
BALF and cell population. BALF was obtained from all subjects taking part in the study. There was no significant difference in the volume of lavage fluid recovered from the normal and the asthmatic subjects (59.3 ± 4.3 ml versus 60.8 ± 5.1 ml, p > 0.05). Differential cell counts showed that BALF from the asthmatic subjects contained more inflammatory cells; however, only eosinophil numbers reached statistical significance (p < 0.05) (Table 3). Baseline measurement of albumin in BALF failed to reveal any significant difference between the normal controls and the subjects with mild asthma (30.5 µg/ml, range: 15 to 70 µg/ml; versus 47.2 µg/ml, range: 15 to 95 µg/ml, respectively).
|
Chemokine concentrations in BALF. Measurement of the
chemokines RANTES, MIP-1, and MCP-1 in BALF with
ELISA showed that these chemokines were present in both
normal controls and stable asthmatic patients. No difference
between the two groups was found in the baseline levels of the
three chemokines (Figure 1). Because the recovery of BALF
can vary from one subject to another, levels of RANTES,
MIP-1, and MCP-1 were also analyzed in relation to concentration of albumin (data not shown). The comparative results obtained did not differ from those that were expressed per
milliliter of BALF. There was no correlation between the
level of any of the chemokines and the total number of leukocytes or leukocyte subsets present in BALF (data not shown).
|
Allergen-induced Asthma
BALF and cell population. There was no significant difference in the volume of fluid recovered from saline- or allergen-exposed bronchial segments (54.3 ± 3.3 ml versus 45.5 ± 4 ml, respectively). Eosinophil numbers were significantly increased in BALF from the allergen-challenged as compared with the saline-challenged segments. At 4 h after allergen challenge (Group B), there was a 5-fold increase in eosinophils (p < 0.05), whereas at 24 h (Group C) the eosinophil count was 19-fold greater than at the saline-exposed site (p < 0.02). No significant difference was observed in the numbers of macrophages, lymphocytes, or neutrophils at the two sites (Table 4). As compared with baseline lavage (Group D), there was a significant increase in the number of neutrophils at the saline-challenge site, p < 0.005 (Table 4). Levels of albumin in BALF from the saline and allergen lavages were not significantly different at either 4 h (72.5 µg/ml, range: 31 to 124 µg/ml; versus 79.0 µg/ml, range: 33 to 225 µg/ml, respectively) or 24 h (59 µg/ ml, range: 21 to 136 µg/ml; versus 84 µg/ml, range: 25 to 177 µg/ ml, respectively).
|
Chemokine concentrations in BALF. As compared with
those at the saline site, the levels of RANTES, MIP-1, and
MCP-1 were greatly increased at the allergen-challenge site 4 h
after challenge (median: 11.5 pg/ml versus 175 pg/ml, p = 0.02;
88 pg/ml versus 258 pg/ml, p < 0.05; and 450 pg/ml versus 900 pg/ml, p < 0.05, respectively (Figure 2). At the 24-h time
point, the three chemokines were still detectable at both the
allergen- and saline-exposed sites, although the levels were
substantially lower than those measured at 4 h, there being no
statistically significant differences between the concentrations
of the three chemokines at the two sites at this later time
point. Concentrations of the three chemokines in BALF obtained 4 h after challenge were significantly increased over
those observed at 24 h, p 
0.02 (Figure 2). The double saline challenge also induced the release of MIP-1 and MCP-1, but
not of RANTES, into the BALF (Figure 3). However, levels
of these chemokines induced by saline were substantially
lower than those observed in the allergen-challenged segments. As compared with the double saline challenge, allergen
challenge induced 2- to 3-fold greater levels of MIP-1 and
MCP-1 in BALF (Figures 2 and 3).
|
|
There was a significant correlation between the concentrations of RANTES and the numbers of eosinophils in BALF at
4 h after allergen exposure (r = 0.68, p = 0.04), but not after
saline exposure (r = 0.21, p = 0.60). In contrast, at 24 h after
challenge, no correlations were found between RANTES levels and eosinophil numbers at the saline- (r = 0.07, p = 0.86)
or allergen-challenged (r = 0.08, p = 0.91) sites. There were
no correlations between MIP-1 and eosinophil numbers at
the saline- or allergen-challenge sites at either 4 h (r = 0.06,
p = 0.06, versus r = 0.03, p = 0.93) or 24 h (r = 0.07, p = 0.86, versus r = 0.81, p = 0.01). Similarly, BALF concentrations of RANTES, MIP-1, and MCP-1 did not correlate with
either the number of lymphocytes or of macrophages obtained from the saline- and allergen-challenged sites at the two time points (data not shown).
Release of Chemokines by BAL Cells
To investigate whether BAL cells spontaneously release
RANTES, MIP-1, and MCP-1, concentrations of these
chemokines were measured in the 24 h culture supernatant of
BAL cells obtained 4 h after challenge. BAL cells derived
from the allergen-challenge site released more RANTES and
MCP-1 than did BAL cells from the saline-challenge site (median: 660.0 pg/ml versus 105.0 pg/ml, and 5.7 ng/ml versus 2.6 ng/ml; respectively, p < 0.05 (Figure 4). BAL cells derived
from the allergen-challenge site also released greater levels of
MIP- than did cells from the saline-challenge site (5.5 ng/ml
versus 4.4 ng/ml), but this difference failed to reach statistical
significance.
|
| |
DISCUSSION |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Bronchial asthma is a chronic inflammatory disease that is
characterized by infiltration of eosinophils and lymphocytes
into the airway tissue. The present study has demonstrated that
the chemokines RANTES, MIP-1, and MCP-1 are present in
the epithelial lining fluid of both normal and mildly asthmatic
subjects, with no difference in the concentrations of the three
chemokines in the two populations. Of importance is that endobronchial allergen challenge of sensitized asthmatic airways
induced the release of these chemokines in BALF at concentrations that were greatly increased at 4 h. However, by 24 h,
levels of the chemokines had almost returned to normal values. In demonstrating ongoing release of RANTES and MCP-1
by BAL cells from the allergen-exposed site after 24 h of culture, we suggest that these chemokines may recruit leukocytes
into asthmatic airways.
To study leukocytes infiltrating asthmatic airways, we performed endobronchial allergen challenge in patients with mild asthma. This allowed us to demonstrate that eosinophil recruitment is already evident at 4 h after allergen exposure, and that the numbers of eosinophils increase further at 24 h. In concurrence with our previous study, we found a nonspecific recruitment of neutrophils, which was predominantly related to the bronchoscopic procedure (27). Allergen challenge did not induce a significant increase in the number of lymphocytes or macrophages.
RANTES, MIP-1, and MCP-1 are potent leukocyte chemoattractants in vitro, and they are thought to be involved in the
allergic inflammatory process. In the present study we have
shown that these three cytokines are constitutively present in
the airways of both normal subjects and patients with steady-state asthma, suggesting that they may regulate cell trafficking
under physiologic conditions. Our findings fail to support Alam
and colleagues' report that these chemokines are increased in
BALF from patients with stable asthma (22). Since our patients were to undergo segmental allergen challenge, they
were deliberately chosen to have milder disease as compared
with those taking part in Alam and colleagues' study. We have
previously demonstrated that endobronchial allergen challenge leads to the release of RANTES and MIP-1 at 4 h after
challenge, and have shown that these two chemokines have biologic activity on eosinophils and lymphocytes, respectively (24, 25). The present study extends these observations by demonstrating that allergen challenge also induces the release of MCP-1 into asthmatic airways. Of particular interest in this study was the finding that levels of these chemokines were
high at the 4-h time point and had returned to baseline values
at 24 h after allergen exposure. Our findings do not support
those in a report from Sur and associates indicating that allergen challenge induces RANTES release at 24 h after challenge (26).
In a previous study we demonstrated that endobronchial
challenge with both allergen and saline causes release of the
C-X-C chemokine IL-8 into asthmatic airways (27). However,
in this previous study, we were unable to show any difference
in IL-8 levels in BALF with the two exposures, suggesting that
instrumentation and saline washing of the airways alone is sufficient to generate IL-8, probably from the epithelium. The
present study extends this observation by showing that saline
challenge alone induced the release of small amounts of MIP-1
and MCP-1, but not of RANTES, at 4 h after challenge; however, in contrast to the case with IL-8, allergen exposure results in a further increase in lavage levels of the C-C chemokines.
To investigate whether RANTES, MIP-1 and MCP-1 are
involved in leukocyte recruitment, we sought correlations between these chemokines and cells infiltrating asthmatic airways.
A significant correlation was found at the allergen-challenge
site between the concentration of RANTES in BALF and the
number of eosinophils recovered from the exposed segment at
4 h but not at 24 h. We have recently purified RANTES from
BALF of six asthmatic subjects exposed to segmental allergen
challenge, and have demonstrated its biologic activity on eosinophils (24). In this last study, levels of RANTES were also
found to correlate with eosinophil numbers in BALF. These
observations are consistent with the release of RANTES being involved in eosinophil recruitment during the early phase
of the allergen-induced late asthmatic response (4 h after challenge). Chemoattractants other than RANTES may dominate eosinophil recruitment at 24 h after challenge. In a recent study it was suggested that IL-5 is the major eosinophil attractant associated with eosinophil recruitment at this time point (26). However, the possibility cannot be excluded that RANTES
could have bound proteoglycans on the extracellular matrix
(30), in which case this cytokine could recruit eosinophils by a
hapotactic mechanism.
We have not been able to show a correlation between the
high level of MIP-1 after allergen challenge and eosinophil
numbers. MIP-1 has been reported to be an eosinophil
chemoattractant in vitro, but as compared with RANTES it is
relatively weak (31). RANTES, MIP-1, and MCP-1 exert
chemotactic effects on monocytes when studied in vitro; however, on allergen challenge, there was no increase in the number of these cells in BALF at either 4 h or 24 hours after challenge. Similarly, the three chemokines have also been shown
to exert chemotactic effects on basophils and on both CD4+
and CD8+ T lymphocytes. Although this matter was not specifically examined in the present study, increased numbers of
activated T cells and basophils are known to occur from 4 to
24 h after allergen challenge of asthmatic airways (5, 32).
In the present study we did not investigate the cellular
source of the chemokines detected in BALF. However, a
number of cells, including alveolar macrophages, neutrophils,
endothelial cells, and bronchial epithelial cells are known
sources of these cytokines (12), and as such could contribute
to airway chemokine release. Interestingly, free cells lavaged
from the airways at 4 h after allergen as compared with saline
exposure generated appreciably more RANTES and MCP-1
but not MIP-1 after ex vivo culture for 24 h. The finding that
BAL cells from the allergen- and saline-challenge sites release
similar amounts of MIP-1 suggests that the bronchoscopic procedure itself recruits and activates cells into asthmatic airways, and that these cells subsequently release this cytokine
into the BAL cell culture supernatant.
Other chemokines that may play an important role in allergic inflammation include eotaxin, MCP-3, and MCP-4. However, there are no reagents commercially available for measuring these cytokines in biologic fluids. The development of sensitive assays for detecting these novel chemokines may allow their role in bronchial asthma to be defined.
In summary, the present study has demonstrated that
RANTES, MIP-1, and MCP-1 are constitutively present in
the BALF of both normal and asthmatic subjects. Endobronchial allergen challenge of asthmatic airways leads to release
of additional amounts of these chemokines. Moreover, BAL
cells derived from the allergen-challenge site spontaneously
release RANTES and MCP-1. These findings suggest that the
C-C chemokines contribute in the regulation of cell trafficking into asthmatic airways, especially after exposure to allergen, such as occurs during the late asthmatic response.
| |
Footnotes |
|---|
Supported by a project grant from the National Asthma Campaign and a program grant from the British Medical Research Council.
Correspondence and requests for reprints should be addressed to Professor S. T. Holgate, University Medicine, Level D, Centre Block, Southampton General Hospital, Southampton S016 6YD, UK.
(Received in original form October 21, 1996 and in revised form May 1, 1997).
| |
References |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
1. Azzawi, M., B. Bradley, P. K. Jeffery, A. J. Frew, A. J. Wardlaw, G. Knowles, B. Assoufi, J. V. Collins, S. Durham, and A. B. Kay. 1990. Identification of activated T lymphocytes and eosinophils in bronchial biopsies in stable atopic asthma. Am. Rev. Respir. Dis 142: 1407-1413 [Medline].
2.
Wilson, J. W.,
R. Djukanovi
,
P. H. Howarth, and
S. T. Holgate.
1992.
Lymphocyte activation in bronchoalveolar lavage and peripheral
blood in atopic asthma.
Am. Rev. Respir. Dis
145:
958-960
[Medline].
3. Bousquet, J., P. Chanez, J. Y. Lacoste, G. Barneon, N. Ghavanian, I. Enander, P. Venge, A. Ahlstedt, J. Simony-Lafontaine, P. Godard, and F. B. Michael. 1990. Eosinophilic inflammation in asthma. N. Engl. J. Med 323: 1033-1039 [Abstract].
4. Corrigan, C. J., and A. B. Kay. 1990. CD4 T-lymphocyte activation in acute severe asthma: relationship to disease severity and atopic status. Am. Rev. Respir. Dis 141: 970-977 [Medline].
5. Montefort, S., C. Gratziou, D. Goulding, R. Polosa, D. O. Haskard, P. H. Howarth, and S. T. Holgate. 1994. Bronchial biopsy evidence for leukocyte infiltration of leukocyte-endothelial cell adhesion molecules 6 h after local allergen challenge of sensitized asthmatic airways. J. Clin. Invest 93: 1411-1421 .
6.
Bradding, P.,
J. A. Roberts,
K. M. Britten,
S. Montefort,
R. Djukanovi,
C. Heusser,
P. H. Howarth, and
S. T. Holgate.
1994.
Interleukin-4, -5, -6 and TNF in normal and asthmatic airways: evidence for the human
mast cell as an important source of these cytokines.
Am. J. Respir. Cell
Mol. Biol
10:
471-480
[Abstract].
7. Robinson, D. S., Q. Hamid, A. M. Bentley, S. Ying, A. B. Kay, and S. R. Durham. 1993. CD4+ T cell activation, eosinophil recruitment and interleukin-4 (IL-4), IL-5 and GM-CSF messenger RNA expression in bronchoalveolar lavage after allergen inhalation challenge in atopic asthmatics. J. Allergy Clin. Immunol 92: 313-324 [Medline].
8. Flavahan, N. A., R. Slifman, and G. J. Gleich. 1988. Human eosinophil basic protein causes hyperreactivity of respiratory smooth muscle: role of the epithelium. Am. Rev. Respir. Dis 138: 685-688 [Medline].
9. Holgate, S. T.. 1997. Mechanisms of allergic airway disease. J. Jpn. Bronchoesophagol. Soc. 48: 51-66 .
10.
Van Leeuwen, B. H.,
M. E. Martinson,
G. C. Webb, and
I. G. Young.
1989.
Molecular organisation of the cytokine gene cluster, involving
the human IL-3, IL-4, IL-5 and GM-CSF genes on human chromosome 5.
Blood
73:
1142-1148
11. Schall, T. J., and K. B. Bacon. 1995. Chemokines, leukocyte trafficking and inflammation. Curr. Opin. Immunol 6: 865-873 .
12. Teran, L. M., and D. D. E. Davies. 1996. The chemokines: their potential role in allergic inflammation. Clin. Exp. Allergy 126: 1005-1019 .
13. Schall, T. J., K. Bacon, K. J. Toy, and D. V. Goeddel. 1990. Selective attraction of monocytes and T lymphocytes of memory phenotype by cytokine RANTES. Nature 347: 669-671 [Medline].
14.
Kameyoshi, Y.,
A. Dorschner,
A. I. Mallet,
E. Christophers, and
J. Schroder.
1992.
Cytokine RANTES released by thrombin-stimulated
platelets is a potent attractant for human eosinophils.
J. Exp. Med
176:
587-592
15.
Matsushima, K.,
C. G. D. Larsen, and
J. J. Oppenheim.
1989.
Purification and characterization of a novel monocyte chemotactic and activating factor produced by a human myelomonocyte cell line.
J. Exp.
Med
169:
1485-1490
16.
Carr, M. W.,
S. J. Roth,
E. Luther,
S. S. Rose, and
T. A. Springer.
1995.
Monocyte chemoattractant protein-1 acts as a T-lymphocyte.
Proc. Natl.
Acad. Sci. U.S.A.
91:
3652-3656
17. Kuna, P., S. R. Reddigari, T. J. Schall, D. Rucinski, M. Sadick, and A. P. Kaplan. 1993. Characterization of the human basophil response to cytokines, growth factors, and histamine releasing factors of the intercrine/chemokine family. J. Immunol 150: 1932-1943 [Abstract].
18.
Meurer, R.,
G. Van Riper,
W. Feeney,
P. Cunningham,
D. Hora Jr.,
M. S. Springer,
D. E. MacIntyre, and
H. Rosen.
1993.
Formation of
eosinophilic and monocytic intradermal inflammatory sites in the dog
by injection of human RANTES but not human monocyte chemoattractant protein 1, human macrophage inflammatory protein 1 alpha,
or human interleukin 8.
J. Exp. Med
178:
1913-1921
19.
Lukacs, N. W.,
R. M. Strieter,
C. L. Shaklee,
S. W. Chensue, and
S. L. Kunkel.
1995.
Macrophage inflammatory protein-1 influences recruitment in antigen-specific airway inflammation.
Eur. J. Immunol
25:
245-251
[Medline].
20. Sim, T. C., L. M. Reece, K. A. Hilsmeier, J. A. Grant, and R. Alam. 1995. Secretion of chemokines and other cytokines in allergen-induced nasal responses: inhibition by topical steroid treatment. Am. J. Respir. Crit. Care Med 152: 927-933 [Abstract].
21. Kuna, P., M. Lazarovich, and A. P. Kaplan. 1996. .Chemokines in seasonal allergic rhinitis. J. Allergy Clin. Immunol 97: 104-112 [Medline].
22.
Alam, R.,
J. York,
M. Boyars,
S. Stafford,
J. A. Grant,
J. Lee,
P. Forsythe,
T. Sim, and
N. Ida.
1996.
Increased MCP-1, RANTES and MIP-1 in bronchoalveolar lavage fluid of allergic asthmatic patients.
Am.
J. Respir. Crit. Care Med
153:
1398-1404
[Abstract].
23. Sousa, A. R., S. Lane, J. A. Nakhostee, T. Yoshimura, T. H. Lee, and R. N. Poston. 1994. Increased expression of the monocyte chemoattractant protein-1 in bronchial tissue from asthmatic subjects. Am. J. Respir. Cell Mol. Biol 10: 142-147 [Abstract].
24. Teran, L. M., N. Noso, M. P. Carroll, D. E. Davies, S. T. Holgate, and J.-M. Schröder. 1996. Eosinophil recruitment following endobronchial allergen challenge is associated with the release of RANTES into asthmatic airways. J. Immunol 157: 1806-1812 [Abstract].
25.
Cruikshank, W. W.,
A. Long,
R. E. Tarpy,
H. Kornfeld,
M. P. Carroll,
L. M. Teran,
S. T. Holgate, and
D. M. Center.
1995.
Early identification of interleukin-16 (lymphocyte chemoattractant factor) and macrophage inflammatory protein-1 (MIP-1) in bronchoalveolar lavage fluid of antigen-challenged asthmatics.
Am. J. Respir. Cell Mol.
Biol
13:
738-747
[Abstract].
26. Sur, S., K. Hirohito, G. J. Gleich, T. C. Chenier, and L. W. Hunt. 1990. Eosinophil recruitment is associated with IL-5, but not with RANTES, twenty-four hours after allergen challenge. J. Allergy Clin. Immunol 97: 1272-1278 .
27. Teran, L. M., M. P. Carroll, A. J. Frew, A. E. Redington, D. E. Davies, I. Lindley, P. H. Howarth, M. K. Church, and S. T. Holgate. 1996. Leukocyte recruitment following local endobronchial allergen challenge in asthma: its relationship to procedure and to airway interleukin-8 release. Am. J. Respir. Crit. Care Med 154: 469-476 [Abstract].
28. Krug, N., L. M. Teran, A. E. Redington, C. Gratziou, S. Montefort, R. Polosa, H. Brewster, P. H. Howarth, S. T. Holgate, A. J. Frew, and M. P. Carroll. 1996. Safety aspects of local endobronchial allergen challenge in asthmatic patients. Am. J. Respir. Crit. Care Med 153: 1391-1397 [Abstract].
29. Weeke, B.. 1973. Rocket immuno-electrophoresis. Scand. J. Immunol 2: 37-46 [Medline].
30. Gilat, D., R. Hershkoviz, Y. A. Mekori, I. Vlodavsky, and O. Lider. 1994. Regulation of adhesion of CD4+ T lymphocytes to intact or heparinase-treated subendothelial extracellular matrix by diffusible or anchored RANTES and MIP-1 beta. J. Immunol 153: 4899-4906 [Abstract].
31.
Rot, A.,
M. Krieger,
T. Brunner,
S. C. Bischoff,
T. J. Schall, and
C. A. Dahinden.
1992.
RANTES and macrophage inflammatory protein 1 alpha induce the migration and activation of normal human eosinophil granulocytes.
J. Exp. Med
176:
1489-1495
32. Liu, M. C., W. C. Hubbard, D. Proud, B. A. Stealey, S. J. Galli, A. Kagey-Sobotka, E. R. Bleecker, and L. M. Lichtenstein. 1991. Immediate and late inflammatory responses to ragweed antigen challenge of the peripheral airways in allergic asthmatics: cellular mediator, and permeability changes. Am. Rev. Respir. Dis 144: 51-58 [Medline].
This article has been cited by other articles:
 |
|
 |
|
  P. Joubert, S. Lajoie-Kadoch, M. Welman, S. Dragon, S. Letuvee, B. Tolloczko, A. J. Halayko, A. S. Gounni, K. Maghni, and Q. Hamid Expression and Regulation of CCR1 by Airway Smooth Muscle Cells in Asthma J. Immunol., January 15, 2008; 180(2): 1268 - 1275. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Y. Thomas, A. Banerji, B. D. Medoff, C. M. Lilly, and A. D. Luster Multiple Chemokine Receptors, Including CCR6 and CXCR3, Regulate Antigen-Induced T Cell Homing to the Human Asthmatic Airway J. Immunol., August 1, 2007; 179(3): 1901 - 1912. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Montes-Vizuet, A. Vega-Miranda, E. Valencia-Maqueda, M. C. Negrete-Garcia, J. R. Velasquez, and L. M. Teran CC chemokine ligand 1 is released into the airways of atopic asthmatics Eur. Respir. J., July 1, 2006; 28(1): 59 - 67. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Woszczek, R. Pawliczak, H.-Y. Qi, S. Nagineni, S. Alsaaty, C. Logun, and J. H. Shelhamer Functional Characterization of Human Cysteinyl Leukotriene 1 Receptor Gene Structure J. Immunol., October 15, 2005; 175(8): 5152 - 5159. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. M. Lilly, H. Tateno, T. Oguma, E. Israel, and L. A. Sonna Effects of Allergen Challenge on Airway Epithelial Cell Gene Expression Am. J. Respir. Crit. Care Med., March 15, 2005; 171(6): 579 - 586. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. K. Lundy, S. A. Lira, J. J. Smit, D. N. Cook, A. A. Berlin, and N. W. Lukacs Attenuation of Allergen-Induced Responses in CCR6-/- Mice Is Dependent upon Altered Pulmonary T Lymphocyte Activation J. Immunol., February 15, 2005; 174(4): 2054 - 2060. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. E. John, M. S. Thomas, A. A. Berlin, and N. W. Lukacs Temporal Production of CCL28 Corresponds to Eosinophil Accumulation and Airway Hyperreactivity in Allergic Airway Inflammation Am. J. Pathol., February 1, 2005; 166(2): 345 - 353. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Liu, N. N. Jarjour, W. W. Busse, and E. A. B. Kelly Enhanced Generation of Helper T Type 1 and 2 Chemokines in Allergen-induced Asthma Am. J. Respir. Crit. Care Med., May 15, 2004; 169(10): 1118 - 1124. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Dakhama, M. Kraft, R. J. Martin, and E. W. Gelfand Induction of Regulated upon Activation, Normal T Cells Expressed and Secreted (RANTES) and Transforming Growth Factor-{beta}1 in Airway Epithelial Cells by Mycoplasma pneumoniae Am. J. Respir. Cell Mol. Biol., September 1, 2003; 29(3): 344 - 351. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. M. Phillips, V. E. L. Stubbs, M. R. Henson, T. J. Williams, J. E. Pease, and I. Sabroe Variations in Eosinophil Chemokine Responses: An Investigation of CCR1 and CCR3 Function, Expression in Atopy, and Identification of a Functional CCR1 Promoter J. Immunol., June 15, 2003; 170(12): 6190 - 6201. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. B. Vargaftig and M. Singer Leukotrienes, IL-13, and chemokines cooperate to induce BHR and mucus in allergic mouse lungs Am J Physiol Lung Cell Mol Physiol, February 1, 2003; 284(2): L260 - L269. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Cheng, M. Arima, K. Honda, H. Hirata, F. Eda, N. Yoshida, F. Fukushima, Y. Ishii, and T. Fukuda Anti-Interleukin-9 Antibody Treatment Inhibits Airway Inflammation and Hyperreactivity in Mouse Asthma Model Am. J. Respir. Crit. Care Med., August 1, 2002; 166(3): 409 - 416. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P.J. Barnes Cytokine modulators as novel therapies for airway disease Eur. Respir. J., July 2, 2001; 18(34_suppl): 67S - 77s. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Tateno, H. Nakamura, N. Minematsu, K. Amakawa, T. Terashima, S. Fujishima, A.D. Luster, C.M. Lilly, and K. Yamaguchi Eotaxin and monocyte chemoattractant protein-1 in chronic eosinophilic pneumonia Eur. Respir. J., May 1, 2001; 17(5): 962 - 968. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. W. Busse and R. F. Lemanske Asthma N. Engl. J. Med., February 1, 2001; 344(5): 350 - 362. [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
 |
|
 A. M A. El-Asrar, S. Struyf, S. A Al-Kharashi, L. Missotten, J. Van Damme, and K. Geboes Chemokines in the limbal form of vernal keratoconjunctivitis Br. J. Ophthalmol., December 1, 2000; 84(12): 1360 - 1366. [Abstract] [Full Text]
|
|
|
|
|
|
Z.-Z. Pan, L. Parkyn, A. Ray, and P. Ray Inducible lung-specific expression of RANTES: preferential recruitment of neutrophils Am J Physiol Lung Cell Mol Physiol, October 1, 2000; 279(4): L658 - L666. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Ochi, N. H. De Jesus, F. H. Hsieh, K. F. Austen, and J. A. Boyce IL-4 and -5 prime human mast cells for different profiles of IgE-dependent cytokine production PNAS, September 5, 2000; (2000) 180318697. [Abstract] [Full Text]
|
|
|
|
|
|
I. TILLIE-LEBLOND, H. HAMMAD, S. DESURMONT, J. PUGIN, B. WALLAERT, A.-B. TONNEL, and P. GOSSET CC Chemokines and Interleukin-5 in Bronchial Lavage Fluid from Patients with Status Asthmaticus . Potential Implication in Eosinophil Recruitment Am. J. Respir. Crit. Care Med., August 1, 2000; 162(2): 586 - 592. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Blease, B. Mehrad, T. J. Standiford, N. W. Lukacs, S. L. Kunkel, S. W. Chensue, B. Lu, C. J. Gerard, and C. M. Hogaboam Airway Remodeling Is Absent in CCR1-/- Mice During Chronic Fungal Allergic Airway Disease J. Immunol., August 1, 2000; 165(3): 1564 - 1572. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. BOUSQUET, P. K. JEFFERY, W. W. BUSSE, M. JOHNSON, and A. M. VIGNOLA Asthma . From Bronchoconstriction to Airways Inflammation and Remodeling Am. J. Respir. Crit. Care Med., May 1, 2000; 161(5): 1720 - 1745. [Full Text]
|
|
|
|
|
|
S. C. Lee, M. E. Brummet, S. Shahabuddin, T. G. Woodworth, S. N. Georas, K. M. Leiferman, S. C. Gilman, C. Stellato, R. P. Gladue, R. P. Schleimer, et al. Cutaneous Injection of Human Subjects with Macrophage Inflammatory Protein-1{alpha} Induces Significant Recruitment of Neutrophils and Monocytes J. Immunol., March 15, 2000; 164(6): 3392 - 3401. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. E. Donovan, D. A. Mark, H. Z. He, H.-C. Liou, L. Kobzik, Y. Wang, G. T. De Sanctis, D. L. Perkins, and P. W. Finn NF-{kappa}B/Rel Transcription Factors: c-Rel Promotes Airway Hyperresponsiveness and Allergic Pulmonary Inflammation J. Immunol., December 15, 1999; 163(12): 6827 |