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B/Rel Transcription Factors: c-Rel Promotes Airway Hyperresponsiveness and Allergic Pulmonary Inflammation1
*
Pulmonary and
Renal Divisions, Brigham and Women’s Hospital, Boston, MA 02115;
Cornell University Medical College, New York, NY 10021; and
§
Departments of Medicine and Pathology, Harvard Medical School, Boston, MA 02115
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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B/Rel family of transcription factors induces many genes
involved in immune and inflammatory responses. Mice with germline
deletions of individual NF-B/Rel subunits have different phenotypes,
suggesting that the NF-B/Rel transcription factors have different
functions. We tested whether c-Rel promotes allergic asthma using a
murine model of allergen-induced pulmonary inflammation and airway
hyperresponsiveness. Our investigation focused on c-Rel, which is
expressed in lymphoid cells and is important for lymphocyte activation.
In response to allergen sensitization and challenge, c-Rel-deficient
mice did not develop increases in pulmonary inflammation,
bronchoalveolar lavage fluid eosinophilia, or total serum IgE. c-Rel
deficiency also prevented the induction of airway hyperresponsiveness.
Allergen-treated wild-type mice had increased DNA binding to an NF-B
consensus site. Chemokine expression was altered in allergen-treated
c-Rel-deficient mice. Monocyte chemoattractant protein-1, which is
regulated by NF-B, was decreased in allergen-treated c-Rel-deficient
mice relative to wild-type controls. The increase in NF-B/Rel
transcription factors after allergen challenge in wild-type mice and
the decrease in allergen reactivity found in c-Rel-deficient mice
indicate that c-Rel promotes allergic inflammation. Alteration of
pulmonary chemokine expression in c-Rel-deficient mice may inhibit
allergen-induced pulmonary inflammation and airway
hyperresponsiveness. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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B/Rel family of transcription factors is of particular interest
because it induces many genes involved in immune and inflammatory
responses (7, 8, 9). Several studies have linked cytokines or
allergens with activation of NF-B (10, 11, 12, 13). Inhibition
of NF-B by glucocorticoids may be an important mechanism by which
steroids attenuate inflammatory diseases such as asthma (10, 14, 15).
NF-B has been implicated in the pathogenesis of asthma, but the
roles of specific NF-B/Rel transcription factors in the induction of
allergic pulmonary inflammation and AHR are not yet defined. There are
multiple NF-B/Rel family members: c-Rel, p65 (RelA); p50 and its
p105 precursor; p52 and its p100 precursor; RelB; v-Rel; and the
Drosophila proteins Dorsal and DIF (8, 16, 17).
The NF-B/Rel family members have distinct tissue distributions and
target gene specificities (18). The phenotypes of mice
lacking specific NF-B/Rel proteins are different, underscoring that
the NF-B/Rel transcription factors do not have redundant functions
(9, 19, 20, 21, 22, 23, 24, 25, 26). One study has found that p50-deficient
(p50-/-) mice do not develop allergen-induced
eosinophilic airway inflammation, concomitant with decreased IL-5 and
eotaxin production (27). Here we analyze c-Rel-deficient
(c-Rel-/-) mice, which have a phenotype
distinct from that of p50-/- mice, in a model
of allergen-induced pulmonary inflammation to investigate whether c-Rel
promotes allergic asthma (3). We focused on c-Rel because
it is expressed predominantly in lymphocytes and is necessary for
normal lymphocyte activation and proliferation.
Lymphocyte development and hemopoiesis are normal in c-Rel-/- mice, but mature T lymphocytes have proliferation defects in vitro related to decreased production of cytokines such as IL-2, IL-3, and GM-CSF (9, 19, 21). In the presence of exogenous IL-2, c-Rel-/- T cells proliferate normally and are able to differentiate into functional cytotoxic and helper T cells (21). c-Rel-/- mice have impaired humoral immunity as well: c-Rel is required for the synthesis of IgG1 and IgG2a, the transduction of survival and cell cycle progression signals in B cells, and for the generation of T cell-dependent humoral responses (9, 28). B cells deficient in the C-terminal trans-activation domain of c-Rel have defects in Ig class switching, including failure to switch to IgE (26). p50-/- mice have abnormal B cell activation, proliferation, Ab production, and Ig class switching (23, 24). The defects in isotype switching and germline CH RNA expression in p50-/- B cells, however, are different from those in B cells lacking the C-terminal trans-activation domain of c-Rel (20, 24). p50-/- T cells have proliferative defects that are less well characterized than the B cell defects (23).
Whether the immune response defects observed in c-Rel-/- lymphocytes in vitro would foster alterations in an in vivo model had not been previously studied. Specifically, we tested whether loss of c-Rel would lead to inhibition of allergic disease, including AHR.
Chemokines, known to regulate both NF-B and allergic asthma, were
altered in c-Rel-/- mice. For example, monocyte
chemoattractant protein-1 (MCP-1) was decreased in allergen-treated
c-Rel-/- mice relative to wild-type controls.
Loss of c-Rel function inhibited the allergic phenotype, including
allergen-induced physiological outcomes. Our findings suggest that the
NF-B/Rel transcription factor c-Rel promotes allergic pulmonary
inflammation, may regulate the expression of chemokines that are
relevant to the pathogenesis of asthma, and plays a critical role in
the induction of allergic AHR.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Mice deficient in c-Rel (c-Rel-/-) were generated as previously described (9, 21, 28). Male wild-type BALB/c and C57BL/6 mice (5 wk old) were purchased from The Jackson Laboratory (Bar Harbor, ME). All mice were maintained according to the guidelines of the Committee on Animals at Harvard Medical School and the Committee on Care and Use of Laboratory animals of the Institute of Laboratory Animal Resources, National Research Council.
Ag sensitization and challenge
As previously described, mice were immunized with i.p. injections of 10 µg OVA (Sigma, St. Louis, MO) and 1 mg of Al(OH)3 (alum, J. T. Baker Chemical, Phillipsburg, NJ) on days 0 and 7 and underwent aerosol treatments with 6% OVA for 20 min/day on days 14 through 20 (2, 3). Control mice received i.p. injections and aerosol treatments with PBS.
Determination of lung resistance and airway reactivity
One day after the final aerosol challenge, lung resistance and dynamic compliance were determined as previously described (2, 3, 29, 30). In brief, mice were anesthetized with i.p. injections of pentobarbital sodium (Anthony Products, Arcadia, CA). The trachea was cannulated. An internal jugular vein was cannulated with a catheter attached to a microsyringe (Hamilton, Reno, NV) and used to administer methacholine (acetyl-ß-methylcholine chloride, Sigma). As previously reported, methacholine exerted no significant systemic side effects (29). Dose-response curves to methacholine were obtained by administering increasing doses of methacholine (33 to 1000 µg/kg). Each new dose of methacholine was delivered only after the pulmonary resistance had returned to baseline. In addition, the effective methacholine dose that would have resulted in a doubling of pulmonary resistance, the log ED200, was calculated by log-linear interpolation and is used as an index of AHR.
Analysis of pathology
After determination of lung resistance and airway reactivity, mice were exsanguinated by cardiac puncture. Lungs were inflated with OCT compound (Miles, Elkhart, IN), fixed, stained with hematoxylin and eosin, and examined by light microscopy.
Determination of bronchoalveolar lavage fluid (BAL) cell differential counts
Mice underwent BAL after plethysmography. PBS (1 ml) with 0.6 M EDTA was instilled into the lungs and withdrawn three times via the tracheal cannula. The total number of live cells was determined by trypan blue exclusion with a hemocytometer. Cytospins were prepared (Shandon Scientific, Cheshire, U.K.), fixed, and stained with Diff-Quick (Dade Diagnostics of PR, Aguada, PR). An investigator who was unaware of the treatment groups performed duplicate counts of 100 cells to determine the percentages of each cell type.
Determination of serum total IgE
Blood was obtained by cardiac puncture. Serum levels of total
IgE were measured by an ELISA assay. Microtiter plates (Marsh
Biomedical Products, Rochester, NY) were coated with anti-IgE Ab (2
µg/ml in 0.1 M NaHCO3) (PharMingen, San Diego,
CA), incubated overnight at 4°C, washed three times with PBS/0.05%
Tween, blocked with 3% BSA (Sigma) in PBS, and washed again. Serum,
diluted 1/20 in 1% BSA/PBS, and mouse IgE isotype standards were
added to the wells, and the samples were incubated overnight at 4°C.
Plates were washed and the secondary Ab (biotin anti-mouse
light chain, PharMingen) diluted to 2 µg/ml was added. After
incubation at room temperature for 1 h, samples were washed and
treated with avidin-peroxidase (Sigma). The plates were incubated at
room temperature for 1 h, washed, treated with
o-phenylenediamine dihydrochloride, and read at 450 nm
(Model 2550, Bio-Rad, Richmond, VA). Murine serum IgE concentrations
were determined by the standard curve generated by analysis of the
commercial standard.
Electrophoretic mobility shift assays (EMSA) and supershift assays
Nuclear extracts were prepared as previously described from
thoracic lymphocytes from OVA-sensitized and challenged or PBS-treated
mice (31, 32). EMSA (6) were performed as
previously described (32) with the NF-B consensus
sequence (5'-AGTTGAGGGGACTTTCCCAGGC-3') (Promega,
Madison, WI), which was end-labeled with
[
-32P]ATP. Nuclear extracts were normalized
for protein (Bio-Rad protein assay), and equal amounts were added
within experiments. DNA-protein complexes were resolved in a 5%
polyacrylamide gel in 45 mM Tris, 45 mM boric acid, 1 mM EDTA (0.5x
TBE) and run at 11 V/cm with cooling. The gels were dried and underwent
autoradiography with intensifying screens at -70°C. For competition
analysis, nuclear extracts were preincubated with water or competitor
DNA for 5 min at room temperature; all other components were then
added, and the samples were processed as for EMSA. The competitors were
homologous unlabeled NF-B probe or a non-NF-B oligonucleotide
(CTLA4) of the same size. mAbs (0.5–1.0 µg) specific to NF-B/Rel
family members (p50, p52, p65, Rel-B, and c-Rel) or to other
transcription factors (Fos, Jun, JunB, JunD) (Santa Cruz Biotechnology,
Santa Cruz, CA) were used for supershift analysis.
Pulmonary chemokine mRNA analysis
Total RNA was isolated from the lungs of OVA-treated wild-type and c-Rel-/- mice and their PBS-treated controls using RNAzol. Chemokine RNA analysis was performed via the RiboQuant MultiProbe RNase protection assay (RPA) (7) System (PharMingen). RNA was hybridized with 32P-labeled mck-5 probe according to the supplier’s directions. After RNase treatment and purification, protected probes were run on a denaturing 5% polyacrylamide gel. RNA (20 µg/sample) was run in each lane. The gels were developed in a PhosphorImager (Molecular Dynamics, Sunnyvale, CA). The identity of each protected fragment was established by analyzing its migration distance against a standard curve of the migration distance vs the log nucleotide length for each undigested probe. Samples were normalized to the housekeeping genes, L32 and GAPDH. Densitometry analysis was performed using ImageQuant software (Molecular Dynamics).
Statistical analysis
All data are reported as means ± SE. Data were analyzed with the JMP 3.0 statistical package (SAS Institute, Cary, NC). Parametric data were analyzed with the Tukey-Kramer test; nonparametric data were analyzed using the Wilcoxon/Kruskal-Wallis rank sum test. A p value of <0.05 was considered significant.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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To determine whether c-Rel plays a role in allergic pulmonary
inflammation, pulmonary histology from c-Rel-/-
and wild-type mice was analyzed. Histological sections of lungs from
allergen-treated c-Rel-/- mice (Fig. 1
c) had minimal inflammation,
appearing similar to PBS control wild-type mice (Fig. 1A)
and to PBS control c-Rel-/- mice (not shown).
OVA-treated c-Rel-/- mice did not have
peribronchial or perivascular eosinophilia. Lungs from wild-type mice
developed inflammatory changes typical of our model in response to
allergen treatment: peribronchial and perivascular infiltrates composed
of eosinophils, neutrophils, and lymphocytes (Fig. 1B)
(2).
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). Eosinophils were not detected
in either PBS control group.
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Allergen-treated c-Rel-/- mice had very
low levels of serum IgE, similar to levels in
c-Rel-/- PBS controls (5.9 ± 2.2 and
4.65 ± 2.1 ng/ml, respectively) (Fig. 3). In contrast, allergen-treated
wild-type mice demonstrated a significant increase in total serum IgE
compared with their PBS controls (56.4 ± 14.4 and 11.9 ±
3.7 ng/ml, respectively) (*, p = 0.002). Serum IgE
levels in allergen-treated c-Rel-/- mice were
significantly lower than in allergen-treated wild-type mice (**,
p = 0.001).
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To determine whether loss of c-Rel influences allergen-induced
physiological outcomes, we measured AHR which we have previously shown
to be significantly increased after allergen treatment (2, 3). Previous analyses of specific NF-B/Rel subunits after
allergen challenge have not reported measurements of AHR.
Allergen-treated c-Rel-/- mice did not develop
AHR. Importantly, their pulmonary resistance in response to
methacholine was not significantly different from PBS control
c-Rel-/- mice (Fig. 4). In contrast, allergen-treated
wild-type mice developed significant increases in pulmonary resistance
at all doses of methacholine compared with PBS control wild-type mice.
The pulmonary resistance of allergen-treated wild-type mice was
significantly higher than similarly treated
c-Rel-/- mice at each dose of methacholine, as
expressed by the difference between the log ED200
for each group (p < 0.0001).
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B/Rel is induced after allergen sensitization and challenge
in wild-type mice
To test the hypothesis that NF-B/Rel transcription factors are
induced in allergen-treated mice without a germline deletion of c-Rel,
EMSA analysis was performed. There was an increase in binding to an
NF-B consensus sequence by nuclear extracts of thoracic lymphocytes
from allergen-treated wild-type mice compared with PBS controls (Fig. 5A, lanes 1 and 2).
To analyze binding specificity, competition assays with homologous
unlabeled NF-B oligonucleotide showed a decrease in binding (Fig. 5A, lanes 3 and 4), which was not paralleled by
competition with a non-NF-B oligonucleotide (Fig. 5A, lanes
5 and 6). When nuclear extracts of thoracic lymphocytes
from allergen-treated mice were incubated with mAbs to the different
NF-B/Rel family members, supershifted bands were detected with the
use of Abs to c-Rel, p50, and p65 but not to p52, Rel-B, or
non-NF-B/Rel transcription factors (Fig. 5B). These
results confirm that NF-B/Rel transcription factors, including
c-Rel, are induced in our OVA model of allergic pulmonary inflammation
and AHR.
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Chemokines are a superfamily of structurally related cytokines
that promote the adhesion, chemotaxis, and activation of multiple cell
types in inflammatory states (33). Because NF-B/Rel
transcription factors induce proinflammatory cytokines, we investigated
whether loss of c-Rel alters the chemokine mRNA profile in the lungs of
allergen-exposed mice. RNA was isolated from the lungs of
allergen-treated wild-type and c-Rel-/- mice as
well as their PBS controls. The lungs were harvested 24 h after
the final aerosol challenge. RPA analysis demonstrated that allergen
treatment led to up-regulation of the chemokines
macrophage-inflammatory protein-2 (1.9-fold), IFN--inducible
protein-10 (3.3 fold), and monocyte chemoattractant protein-1 (MCP-1)
(2.5-fold) in wild-type mice (Fig. 6,
lanes 1 and 2). c-Rel-/-
mice had a different pattern of chemokine mRNA expression. Although
macrophage-inflammatory protein-2 was up-regulated in allergen-treated
c-Rel-/- mice (1.3-fold), IFN--inducible
protein-10 was expressed constitutively (Fig. 6, lanes 3 and
4). Interestingly, MCP-1 was not up-regulated in
allergen-treated c-Rel-/- mice (Fig. 6, lanes 3 and 4). MCP-1 has previously been
implicated in asthma; Abs that neutralize MCP-1 decrease
allergen-induced AHR and pulmonary inflammation in a murine model
(34). Taken together, these data suggest that
c-Rel-/- mice do not have global defects in
pulmonary chemokine mRNA expression and that their blunted response to
allergen may be due in part to chemokine modulation.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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B/Rel family of transcription factors regulates the
expression of multiple genes that have been implicated in immune and
inflammatory responses, including asthma (8, 16).
Analyzing the effects of germline deletions or mutations of specific
NF-B/Rel subunits will elucidate their roles in various inflammatory
diseases, including asthma. The present study demonstrates that absence
of c-Rel is associated not only with significant attenuation of
allergen-induced pulmonary and BAL eosinophilia but also with
inhibition of total IgE and the physiological correlate of allergic
airway inflammation, AHR. p50-/- mice do not
develop eosinophilic airway inflammation in response to allergen
sensitization and challenge, concomitant with a lack of IL-5 and
eotaxin production; total serum IgE and AHR were not examined
(27). The phenotype and immune system defects of
c-Rel-/- mice are distinct from those of
p50-/- mice (23, 24).
One explanation for the blunted response of
c-Rel-/- mice to allergen sensitization and
aerosol challenge is their defects in lymphocyte activation and
proliferation. T lymphocyte activation is an important mediator of
allergic pulmonary inflammation and AHR (2, 35, 36).
Interruption of T lymphocyte costimulatory signals inhibits AHR,
pulmonary inflammation, total serum IgE, and Th2
cytokine production (3, 37, 38). c-Rel is not necessary
for normal hemopoiesis, lymphocyte development, or T cell effector
function (9); however, it controls early activation events
mediated by Ag receptor signals in T lymphocytes (21).
c-Rel also regulates the expression of various cytokine genes, either
directly (IL-2, IL-3, GM-CSF) or indirectly (IL-5, TNF-
, IFN-),
through its effect on T cell proliferation (9, 19, 21).
Current data support the predominance of Th2 cytokines (IL-4, IL-5,
IL-10, IL-13) in allergic asthma, but some studies suggest that Th1
cytokines such as IL-2, IFN-, and TNF- may also promote allergic
airway inflammation (39, 40). IL-3 and GM-CSF prolong
eosinophil survival and are elevated in the airways of asthmatics
(41, 42, 43). c-Rel thus appears to be essential for T cell
activation and the production of multiple cytokines involved in the
pathogenesis of allergic asthma.
We found a significant reduction in the allergen-induced rise in total
serum IgE in c-Rel-/- mice compared with
wild-type mice. To our knowledge, this is the first reported
measurement of IgE in c-Rel-deficient mice. Previous investigators have
measured the Ab response to antigenic challenge in wild-type and
c-Rel-/- mice and found a significant reduction
in the IgG1 response to a T cell-dependent Ag in
c-Rel-/- mice, but IgE levels were not reported
(9). Normal induction of CD23, the low affinity receptor
for IgE in c-Rel-/- mice, has been reported
(44); however, no characterization of the high affinity
IgE receptor in c-Rel-/- mice has been
performed. B cells deficient in the C-terminal activation domain of
c-Rel have marked defects in switching to IgE despite normal levels of
germline CH
RNA, suggesting an important role
for c-Rel in the regulation of Ig class switching
(20).
Given the significant reduction in total serum IgE in c-Rel-/- mice, we did not measure OVA-specific IgE as this would likely also have been reduced. Low levels of OVA-specific IgE in c-Rel-/- mice may have led to inadequate sensitization of mast cells and basophils in the lungs and therefore a weak response to allergen challenge; however, the role of Ag-specific Abs in murine models of allergic inflammation is unresolved. In one study, passive sensitization of mice by iv injections of anti-OVA IgE or IgG1 followed by airway challenge with OVA resulted in pulmonary eosinophilia and AHR (45). In contrast, allergen-induced AHR and pulmonary eosinophilia still develop in B cell-deficient (46), IgE-deficient (47), and mast cell-deficient (48) mice. It has been proposed that AHR and pulmonary eosinophilia may develop in murine models by mast cell-dependent or CD4+ T cell-dependent pathways (46). C57BL/6 mice, the background of the wild-type and c-Rel-/- mice used in the present study, are genetically deficient in mast cell mediators and may therefore be more dependent on T cells than on mast cells (46, 49). One possibility is that c-Rel is required for a normal IgG1/IgE response to allergen sensitization and challenge but that allergen-specific IgE is not essential for the development of AHR and pulmonary eosinophilia in our T cell-dependent model (3, 4).
The response of c-Rel-/- mice to allergen
sensitization and aerosol challenge may be blunted due to differences
in chemokine expression in the lungs as well. For example, MCP-1 mRNA
is up-regulated in the allergen-treated wild-type mice but not in the
allergen-treated c-Rel-/- mice (Fig. 6
).
Several studies have found elevated expression of MCP-1 in the airways
or BAL fluid of allergic asthmatics (50, 51, 52, 53, 54). MCP-1
induction concomitant with NF-B activation have been found in
several models of inflammatory diseases (55, 56, 57, 58). The
promoters of both the human MCP-1 gene and its murine homologue have
NF-B sites (59). In a model of allergic airway
inflammation, inhibition of MCP-1 significantly decreased bronchial
hyperresponsiveness, pulmonary inflammation, and the recruitment of
eosinophils and T lymphocytes to the airways (34). Absence
of c-Rel may prevent maximal transcription of the MCP-1 gene after
allergen challenge, representing a potential mechanism for loss of the
allergic phenotype in c-Rel-/- mice.
Our examination of pulmonary eotaxin mRNA showed that at 24 h after the final allergen challenge, there was no significant difference in pulmonary eotaxin mRNA expression between PBS and allergen-treated wild-type mice. Our results in wild-type mice differ from those of other investigators who have found that eotaxin mRNA expression peaks 3–6 h after allergen challenge and remains elevated for 24 h (27, 34, 60). Potential reasons for the different results include different allergen sensitization and challenge protocols, time points examined, and murine strains. At different time points, c-Rel-/- mice might be deficient in eotaxin production relative to wild-type mice. Nonetheless, eotaxin mRNA expression in c-Rel-/- mice is at least detectable after allergen challenge, in contrast to p50-/- mice (27).
Factors other than eotaxin mRNA expression might be responsible for the observed attenuation in eosinophilia. Decreased translation of eotaxin mRNA or defective eotaxin receptor-response coupling are two possible mechanisms that have not yet been investigated. Other chemokines and cytokines that attract eosinophils and prolong their survival in allergic inflammation may be very relevant to our model, including MCP-1, IL-3, IL-5, and GM-CSF (41, 42, 43). As already noted, MCP-1 mRNA is up-regulated in OVA-treated wild-type mice but not in OVA-treated c-Rel-/- mice. Inhibition of MCP-1 prevented pulmonary and BAL eosinophilia in response to OVA but not the expression of eotaxin mRNA in a murine model; hence, MCP-1 may regulate eosinophil recruitment to the lungs via mediators other than chemokines (34). c-Rel is required by T cells for the production of IL-3 and GM-CSF (9, 21). We postulate that the most likely mechanisms for attenuated pulmonary eosinophilia in c-Rel-deficient mice are failure to up-regulate MCP-1 expression and decreased production of IL-3 and GM-CSF.
Our EMSA analysis of thoracic lymphocytes from allergen-treated
wild-type mice supports a role for NF-B/Rel transcription factors in
the response to allergen sensitization and challenge. In a previous
study, increased NF-B DNA binding was detected in the lungs of
allergen-treated rats, but specific NF-B/Rel family members were not
analyzed (12). The EMSA in this report is unique in its
analysis of nuclear extracts from murine thoracic lymphocytes,
previously shown to have allergen-specific responsiveness after
aerosolized allergen challenge (3). NF-B/Rel is induced
after allergen exposure in wild-type mice (Fig. 5A).
Supershift analysis demonstrated that c-Rel, p50, and p65 were present
in the NF-B/Rel complexes induced by allergen treatment (Fig. 5B). In vitro stimulation of T cells from wild-type mice via
the TCR and CD28 costimulatory pathway induced significant increases in
NF-B/Rel complexes over those of unstimulated controls
(21). Consistent with our results, the complexes were
supershifted or inhibited by Abs against p50, p65, and c-Rel. As
expected, in T cells from c-Rel-/- mice, the
complexes were inhibited only by Abs against p50 and p65. The in vitro
experiments are relevant to our in vivo disease model in which T cell
activation and CD28 costimulation are essential (3, 4).
The increase in NF-B/Rel transcription factors after allergen
treatment in normal mice and the decrease in allergen reactivity
associated with loss of c-Rel function suggest that c-Rel is a mediator
in the inflammatory cascade leading to allergic asthma. Absence of the
c-Rel transcription factor prevents development of the allergic
phenotype in our studies, supporting the notion of a critical role for
NF-B/Rel regulation of lymphocyte activation, proliferation, and
cytokine production in the response to allergen sensitization and
aerosolized challenge. Modulation of the expression of chemokines such
as MCP-1 by c-Rel may play an essential role in the pathogenesis of
allergic pulmonary inflammation. We show for the first time that an
NF-B/Rel transcription factor, c-Rel, is required for
allergen-induced AHR.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Patricia W. Finn, Pulmonary and Critical Care Division, Brigham and Women’s Hospital, 75 Francis Street, Boston, MA 02115. E-mail address:
3 Abbreviations used in this paper: AHR, airway hyperresponsiveness; MCP-1, monocyte chemoattractant protein-1; BAL, bronchoalveolar lavage; EMSA, electrophoretic mobility shift assay; RPA, RNase protection assay.
Received for publication April 27, 1999. Accepted for publication October 5, 1999.
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 C. Pantano, J. L. Ather, J. F. Alcorn, M. E. Poynter, A. L. Brown, A. S. Guala, S. L. Beuschel, G. B. Allen, L. A. Whittaker, M. Bevelander, et al. Nuclear Factor-{kappa}B Activation in Airway Epithelium Induces Inflammation and Hyperresponsiveness Am. J. Respir. Crit. Care Med., May 1, 2008; 177(9): 959 - 969. [Abstract] [Full Text] [PDF]
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 M. Elser, L. Borsig, P. O. Hassa, S. Erener, S. Messner, T. Valovka, S. Keller, M. Gassmann, and M. O. Hottiger Poly(ADP-Ribose) Polymerase 1 Promotes Tumor Cell Survival by Coactivating Hypoxia-Inducible Factor-1-Dependent Gene Expression Mol. Cancer Res., February 1, 2008; 6(2): 282 - 290. [Abstract] [Full Text] [PDF]
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K. Y. Jen, M. Campo, H. He, S. S. Makani, G. Velasco, D. M. Rothstein, D. L. Perkins, and P. W. Finn CD45RB Ligation Inhibits Allergic Pulmonary Inflammation by Inducing CTLA4 Transcription J. Immunol., September 15, 2007; 179(6): 4212 - 4218. [Abstract] [Full Text] [PDF]
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 H. Hayakawa, M. Hayakawa, A. Kume, and S.-i. Tominaga Soluble ST2 Blocks Interleukin-33 Signaling in Allergic Airway Inflammation J. Biol. Chem., September 7, 2007; 282(36): 26369 - 26380. [Abstract] [Full Text] [PDF]
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Z. Bao, S. Lim, W. Liao, Y. Lin, C. Thiemermann, B. P. Leung, and W. S. F. Wong Glycogen Synthase Kinase-3beta Inhibition Attenuates Asthma in Mice Am. J. Respir. Crit. Care Med., September 1, 2007; 176(5): 431 - 438. [Abstract] [Full Text] [PDF]
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 A. P Rogerio, C. R Cardoso, C. Fontanari, M. A Souza, S. R Afonso-Cardoso, E. V. Silva, N. S Koyama, F. L Basei, E. G Soares, J. B Calixto, et al. Anti-asthmatic potential of a D-galactose-binding lectin from Synadenium carinatum latex Glycobiology, August 1, 2007; 17(8): 795 - 804. [Abstract] [Full Text] [PDF]
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H.-Y. Cho, D. L. Morgan, A. K. Bauer, and S. R. Kleeberger Signal Transduction Pathways of Tumor Necrosis Factor-mediated Lung Injury Induced by Ozone in Mice Am. J. Respir. Crit. Care Med., April 15, 2007; 175(8): 829 - 839. [Abstract] [Full Text] [PDF]
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W. M. Abraham, A. Ahmed, I. Serebriakov, I. T. Lauredo, J. Bassuk, J. A. Adams, and M. A. Sackner Whole-Body Periodic Acceleration Modifies Experimental Asthma in Sheep Am. J. Respir. Crit. Care Med., October 1, 2006; 174(7): 743 - 752. [Abstract] [Full Text] [PDF]
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 N. Wang, M. Campo, L. Ting, C. Fleming, C. Terhorst, and P. W. Finn The Costimulatory Molecule SLAM Is Critical for Pulmonary Allergic Responses Am. J. Respir. Cell Mol. Biol., August 1, 2006; 35(2): 206 - 210. [Abstract] [Full Text] [PDF]
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B. D. Medoff, B. Seed, R. Jackobek, J. Zora, Y. Yang, A. D. Luster, and R. Xavier CARMA1 Is Critical for the Development of Allergic Airway Inflammation in a Murine Model of Asthma. J. Immunol., June 15, 2006; 176(12): 7272 - 7277. [Abstract] [Full Text] [PDF]
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 D. H. Broide, T. Lawrence, T. Doherty, J. Y. Cho, M. Miller, K. McElwain, S. McElwain, and M. Karin Allergen-induced peribronchial fibrosis and mucus production mediated by I{kappa}B kinase {beta}-dependent genes in airway epithelium PNAS, December 6, 2005; 102(49): 17723 - 17728. [Abstract] [Full Text] [PDF]
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 T. M Lindstrom and P. R Bennett The role of nuclear factor kappa B in human labour Reproduction, November 1, 2005; 130(5): 569 - 581. [Abstract] [Full Text] [PDF]
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J. M. Lora, D. M. Zhang, S. M. Liao, T. Burwell, A. M. King, P. A. Barker, L. Singh, M. Keaveney, J. Morgenstern, J. C. Gutierrez-Ramos, et al. Tumor Necrosis Factor-{alpha} Triggers Mucus Production in Airway Epithelium through an I{kappa}B Kinase {beta}-dependent Mechanism J. Biol. Chem., October 28, 2005; 280(43): 36510 - 36517. [Abstract] [Full Text] [PDF]
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M. A. Birrell, E. Hardaker, S. Wong, K. McCluskie, M. Catley, J. De Alba, R. Newton, S. Haj-Yahia, K. T. Pun, C. J. Watts, et al. I{kappa}-B Kinase-2 Inhibitor Blocks Inflammation in Human Airway Smooth Muscle and a Rat Model of Asthma Am. J. Respir. Crit. Care Med., October 15, 2005; 172(8): 962 - 971. [Abstract] [Full Text] [PDF]
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C. Desmet, P. Gosset, E. Henry, V. Garze, P. Faisca, N. Vos, F. Jaspar, D. Melotte, B. Lambrecht, D. Desmecht, et al. Treatment of Experimental Asthma by Decoy-mediated Local Inhibition of Activator Protein-1 Am. J. Respir. Crit. Care Med., September 15, 2005; 172(6): 671 - 678. [Abstract] [Full Text] [PDF]
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K. El Bakkouri, A. Wullaert, M. Haegman, K. Heyninck, and R. Beyaert Adenoviral Gene Transfer of the NF-{kappa}B Inhibitory Protein ABIN-1 Decreases Allergic Airway Inflammation in a Murine Asthma Model J. Biol. Chem., May 6, 2005; 280(18): 17938 - 17944. [Abstract] [Full Text] [PDF]
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G. Velasco, M. Campo, O. J. Manrique, A. Bellou, H. He, R. S. S. Arestides, B. Schaub, D. L. Perkins, and P. W. Finn Toll-Like Receptor 4 or 2 Agonists Decrease Allergic Inflammation Am. J. Respir. Cell Mol. Biol., March 1, 2005; 32(3): 218 - 224. [Abstract] [Full Text] [PDF]
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 M. Y. Kimura, H. Hosokawa, M. Yamashita, A. Hasegawa, C. Iwamura, H. Watarai, M. Taniguchi, T. Takagi, S. Ishii, and T. Nakayama Regulation of T helper type 2 cell differentiation by murine Schnurri-2 J. Exp. Med., February 7, 2005; 201(3): 397 - 408. [Abstract] [Full Text] [PDF]
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M. E. Poynter, R. Cloots, T. van Woerkom, K. J. Butnor, P. Vacek, D. J. Taatjes, C. G. Irvin, and Y. M. W. Janssen-Heininger NF-{kappa}B Activation in Airways Modulates Allergic Inflammation but Not Hyperresponsiveness J. Immunol., December 1, 2004; 173(11): 7003 - 7009. [Abstract] [Full Text] [PDF]
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I. M. Adcock, K. Ito, and P. J. Barnes Glucocorticoids: Effects on Gene Transcription Proceedings of the ATS, November 1, 2004; 1(3): 247 - 254. [Abstract] [Full Text] [PDF]
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C. Desmet, P. Gosset, B. Pajak, D. Cataldo, M. Bentires-Alj, P. Lekeux, and F. Bureau Selective Blockade of NF-{kappa}B Activity in Airway Immune Cells Inhibits the Effector Phase of Experimental Asthma J. Immunol., November 1, 2004; 173(9): 5766 - 5775. [Abstract] [Full Text] [PDF]
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 I M Verma Nuclear factor (NF)-{kappa}B proteins: therapeutic targets Ann Rheum Dis, November 1, 2004; 63(suppl_2): ii57 - ii61. [Full Text] [PDF]
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M. Inami, M. Yamashita, Y. Tenda, A. Hasegawa, M. Kimura, K. Hashimoto, N. Seki, M. Taniguchi, and T. Nakayama CD28 Costimulation Controls Histone Hyperacetylation of the Interleukin 5 Gene Locus in Developing Th2 Cells J. Biol. Chem., May 28, 2004; 279(22): 23123 - 23133. [Abstract] [Full Text] [PDF]
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 H. A. Haeberle, A. Casola, Z. Gatalica, S. Petronella, H.-J. Dieterich, P. B. Ernst, A. R. Brasier, and R. P. Garofalo I{kappa}B Kinase Is a Critical Regulator of Chemokine Expression and Lung Inflammation in Respiratory Syncytial Virus Infection J. Virol., March 1, 2004; 78(5): 2232 - 2241. [Abstract] [Full Text] [PDF]
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 A. KUMAR, S. LNU, R. MALYA, D. BARRON, J. MOORE, D. B. CORRY, and A. M. BORIEK Mechanical stretch activates nuclear factor-kappaB, activator protein-1, and mitogen-activated protein kinases in lung parenchyma: implications in asthma FASEB J, October 1, 2003; 17(13): 1800 - 1811. [Abstract] [Full Text] [PDF]
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 P. J. Barnes and I. M. Adcock How Do Corticosteroids Work in Asthma? Ann Intern Med, September 2, 2003; 139(5_Part_1): 359 - 370. [Full Text] [PDF]
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M. E. Poynter, C. G. Irvin, and Y. M. W. Janssen-Heininger A Prominent Role for Airway Epithelial NF-{kappa}B Activation in Lipopolysaccharide-Induced Airway Inflammation J. Immunol., June 15, 2003; 170(12): 6257 - 6265. [Abstract] [Full Text] [PDF]
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A. H. Boulares, A. J. Zoltoski, Z. A. Sherif, P. Jolly, D. Massaro, and M. E. Smulson Gene Knockout or Pharmacological Inhibition of Poly(ADP-Ribose) Polymerase-1 Prevents Lung Inflammation in a Murine Model of Asthma Am. J. Respir. Cell Mol. Biol., March 1, 2003; 28(3): 322 - 329. [Abstract] [Full Text] [PDF]
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 L. Da Costa, G. Narla, T.-N. Willig, L. L. Peters, M. Parra, J. Fixler, G. Tchernia, and N. Mohandas Ribosomal protein S19 expression during erythroid differentiation Blood, January 1, 2003; 101(1): 318 - 324. [Abstract] [Full Text] [PDF]
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W. R. Henderson Jr., E. Y. Chi, J.-L. Teo, C. Nguyen, and M. Kahn A Small Molecule Inhibitor of Redox-Regulated NF-{kappa}B and Activator Protein-1 Transcription Blocks Allergic Airway Inflammation in a Mouse Asthma Model J. Immunol., November 1, 2002; 169(9): 5294 - 5299. [Abstract] [Full Text] [PDF]
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P. S. Kabouridis, M. Hasan, J. Newson, D. W. Gilroy, and T. Lawrence Inhibition of NF-{kappa}B Activity by a Membrane-Transducing Mutant of I{kappa}B{alpha} J. Immunol., September 1, 2002; 169(5): 2587 - 2593. [Abstract] [Full Text] [PDF]
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 J. Caamano and C. A. Hunter NF-{kappa}B Family of Transcription Factors: Central Regulators of Innate and Adaptive Immune Functions Clin. Microbiol. Rev., July 1, 2002; 15(3): 414 - 429. [Abstract] [Full Text] [PDF]
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L. Vicentini, P. Mazzi, E. Caveggion, S. Continolo, L. Fumagalli, J. A. Lapinet-Vera, C. A. Lowell, and G. Berton Fgr Deficiency Results in Defective Eosinophil Recruitment to the Lung During Allergic Airway Inflammation J. Immunol., June 15, 2002; 168(12): 6446 - 6454. [Abstract] [Full Text] [PDF]
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H. A. Haeberle, F. Nesti, H.-J. Dieterich, Z. Gatalica, and R. P. Garofalo Perflubron Reduces Lung Inflammation in Respiratory Syncytial Virus Infection by Inhibiting Chemokine Expression and Nuclear Factor-{kappa}B Activation Am. J. Respir. Crit. Care Med., May 15, 2002; 165(10): 1433 - 1438. [Abstract] [Full Text] [PDF]
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 M. E. Poynter, C. G. Irvin, and Y. M. W. Janssen-Heininger Rapid Activation of Nuclear Factor-{kappa}B in Airway Epithelium in a Murine Model of Allergic Airway Inflammation Am. J. Pathol., April 1, 2002; 160(4): 1325 - 1334. [Abstract] [Full Text] [PDF]
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 K. J. Haley, A. Ciota, J. P. Contreras, M. R. Boothby, D. L. Perkins, and P. W. Finn Alterations in lung collectins in an adaptive allergic immune response Am J Physiol Lung Cell Mol Physiol, March 1, 2002; 282(3): L573 - L584. [Abstract] [Full Text] [PDF]
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P. W. Finn, J. R. Stone, M. R. Boothby, and D. L. Perkins Inhibition of NF-{kappa}B-Dependent T Cell Activation Abrogates Acute Allograft Rejection J. Immunol., November 15, 2001; 167(10): 5994 - 6001. [Abstract] [Full Text] [PDF]
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C. M. Fleming, H. He, A. Ciota, D. Perkins, and P. W. Finn Administration of Pentoxifylline During Allergen Sensitization Dissociates Pulmonary Allergic Inflammation from Airway Hyperresponsiveness J. Immunol., August 1, 2001; 167(3): 1703 - 1711. [Abstract] [Full Text] [PDF]
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M. Stassen, C. Muller, M. Arnold, L. Hultner, S. Klein-Hessling, C. Neudorfl, T. Reineke, E. Serfling, and E. Schmitt IL-9 and IL-13 Production by Activated Mast Cells Is Strongly Enhanced in the Presence of Lipopolysaccharide: NF-{{kappa}}B Is Decisively Involved in the Expression of IL-9 J. Immunol., April 1, 2001; 166(7): 4391 - 4398. [Abstract] [Full Text] [PDF]
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F. Bureau, S. Delhalle, G. Bonizzi, L. Fievez, S. Dogne, N. Kirschvink, A. Vanderplasschen, M.-P. Merville, V. Bours, and P. Lekeux Mechanisms of Persistent NF-{kappa}B Activity in the Bronchi of an Animal Model of Asthma J. Immunol., November 15, 2000; 165(10): 5822 - 5830. [Abstract] [Full Text] [PDF]
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