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 Previous Article | Table of Contents | Next Article 
Blood, Vol. 91 No. 7 (April 1), 1998:
pp. 2223-2230
Regulation of Allergic Inflammation and Eosinophil Recruitment in Mice
Lacking the Transcription Factor NFAT1: Role of Interleukin-4 (IL-4)
and IL-5
By
João P.B. Viola,
Alexander Kiani,
Patricia T. Bozza, and
Anjana Rao
From the Center for Blood Research and the Department of Pathology,
Harvard Medical School, Boston, MA; and Departamento de Fisiologia e
Farmacodinâmica, Fundação Oswaldo Cruz, Rio de
Janeiro, RJ, Brazil.
 |
ABSTRACT |
Transcription factors of the NFAT (nuclear factor of activated T
cells) family regulate the expression of many genes encoding immunoregulatory cytokines and cell surface proteins during the immune
response. The NFAT protein NFAT1 (NFATp) is expressed and functional in
T cells, B cells, mast cells, and natural killer cells. Here we report
a detailed analysis of the enhanced eosinophil responses of
NFAT1-deficient mice, observed in an in vivo model of allergic
inflammation. In addition to the pleural eosinophilia described
previously, NFAT1 / mice that have been sensitized
with antigen display a significant increase, relative to wild-type
mice, in the numbers of eosinophils in bone marrow and peripheral
blood. After restimulation with antigen in vitro, antigen-responsive T
cells from the draining lymph nodes of NFAT1/ mice
show increased expression of mRNA encoding the Th2 cytokines interleukin-4 (IL-4), IL-5, and IL-13. Consistent with this finding, there is a pronounced increase in the levels of IL-5 and IL-13 in the
pleural cavities of sensitized NFAT1/ mice after
allergen challenge in vivo. Furthermore, development of eosinophilia
depends on overexpression of IL-4 and IL-5, because it is strongly
inhibited by administration of neutralizing antibodies to either of
these cytokines. These results indicate that NFAT1-deficient mice are
prone to develop a classically allergic phenotype characterized by
eosinophilia and increased production of Th2 cytokines. Thus, the
presence of NFAT1 might inhibit the allergic response, perhaps by
interfering with the development of Th2 immune responses, and the lack
or dysfunction of NFAT1 could potentially underlie certain cases of
atopic disease.
| |
INTRODUCTION |
ATOPY OR ALLERGIC disease is a complex
familial disorder with multiple manifestations, including allergic
asthma, rhinitis, conjunctivitis, and dermatitis.1-3 Both
eosinophils and mast cells have been implicated in the pathogenesis of
allergic disease. The hallmark of these diseases is a pronounced
increase in the level of serum IgE, reflecting the actions of the
cytokines interleukin-4 (IL-4) and IL-13 to promote B-cell isotype
switching to IgE.4 IgE produced by allergen-reactive B
cells binds to Fc receptors present on the surface of mast cells and
basophils; when challenged with allergen, these cells release
vasoactive mediators that directly damage the surrounding tissue, as
well as chemotactic factors and cytokines that promote leukocyte
infiltration and exacerbate the inflammatory response.2,4
Likewise, eosinophils accumulate at sites of allergic inflammation,
produce leukotrienes and other inflammatory mediators, and contribute
significantly to tissue damage at sites of allergic
inflammation.5 The magnitude of asthmatic responses is
related to the number of eosinophils present in the lung,6
and suppression of eosinophil accumulation at the site of inflammation
impairs the development of airway hyperreactivity.7,8
By promoting IgE production and eosinophil recruitment, respectively,
the cytokines IL-4 and IL-5 play a critical role in the development of
atopic disease. The genes encoding these cytokines are clustered on
human chromosome 5 and mouse chromosome 11, and the gene cluster is
closely linked to atopy in certain families.1 IL-4, IL-5,
and IL-13 are coordinately produced by Th2 cells, which are recognized
as key regulators of allergy and asthma.1-3,9 Th2 cells are
the predominant CD4+ T cells in the inflamed tissues of
patients with atopic dermatitis and in the bronchoalveolar lavage
fluids of patients with allergic asthma.10,11 In a murine
model of allergic airway inflammation, severe combined immunodeficiency
disease (SCID) mice lacking T and B cells showed no antigen-induced
airway hyperreactivity,12 and similarly depletion of
CD4+ T cells prevented antigen-induced airway
hyperreactivity and pulmonary eosinophilia.13 In contrast,
neither IgE-deficient nor mast-cell-deficient mice showed significant
diminution of eosinophilic inflammation and airway
hyperresponsiveness,14-16 confirming the importance of Th2
cells in these processes. The differentiation of uncommitted T-cell
precursors into Th2 cells is largely driven by IL-4.17,18
In murine models of airway hyperreactivity, BALB/c mice that had been
treated with anti-IL-4 during antigen sensitization showed a
significant decrease of airway hyperreactivity in response to antigen
challenge,12 whereas IL-4-deficient C57BL/6 mice showed
almost no eosinophilia and much less peribronchial
inflammation.14
Proteins belonging to the NFAT (nuclear factor of activated T cells)
family of transcription factors control the transcription of many genes
involved in allergy and eosinophil function.19 NFAT
proteins are expressed in T cells, B cells, mast cells, and natural
killer (NK) cells, where they are activated by stimulation of
calcium-mobilizing antigen and Fc receptors and regulate the expression
of appropriate inducible genes.20-24 Binding sites for NFAT
proteins have been described in the promoter/enhancer regions of
diverse inducible genes encoding cytokines and cell-surface receptors,
including interferon  (IFN-), IL-2, IL-3, IL-4, IL-5, IL-13,
tumor necrosis factor- (TNF-), granulocyte-macrophage colony-stimulating factor (GM-CSF), CD40L, FasL, and
CTLA-4.19 Given these observations, it was surprising that
mice lacking the family member NFAT1 (NFATp) showed increased pleural
accumulation of eosinophils and increased serum IgE levels in an in
vivo model of allergic inflammation25 and displayed an
unexpected pattern of IL-4 gene expression.26 Here we show
that the pleural eosinophilia and inflammation observed in
NFAT1-deficient mice depends on overexpression of both IL-4 and IL-5
and is characterized by the development of a Th2 cytokine profile.
Thus, NFAT1 deficiency in mice leads to the development of a
classically allergic phenotype, unlike that seen in nonallergic or
intrinsic asthma where IL-5 is important but there is no involvement of
IgE or IL-4.11 Given the known genetic predisposition of
allergic disease in humans, it is possible that certain forms of human
atopy could involve a deficiency in NFAT1.
| |
MATERIALS AND METHODS |
Animals.
NFAT1-deficient and control wild-type+/+ mice were
generated on a mixed genetic 129/Sv × C57BL/6
background.25 Eight- to 12-week-old wild-type and
NFAT1/ mice, bred and maintained independently, were
used in all experiments. C57BL/6 and 129/Sv mice were purchased from
Jackson Laboratory (Bar Harbor, ME).
Sensitization and allergic pleurisy.
Animal sensitization and allergic pleurisy were assessed as described
with minor modifications.27 Mice were immunized in the hind
footpad by subcutaneous injection of 0.1 mL of ovalbumin (OVA; 200 µg) emulsified in complete Freund's adjuvant. Allergic pleurisy was
induced in ovalbumin-sensitized mice by intrathoracic injection of 0.1 mL OVA (12 µg) in phosphate-buffered saline (PBS) 14 days after
sensitization. Control animals were injected intrathoracically with PBS
alone. Animals were killed 24 hours later, and the thoracic cavity was
rinsed with 0.3 mL PBS containing heparin (10 U/mL). The pleural washes
were collected, their volume was measured, and the cells were evaluated
as described below.
Designated groups of mice were injected intraperitoneally with 1 mg
neutralizing anti-IL-4 monoclonal antibody (MoAb; 11B11; gift from
Biological Response Modifiers Program, National Cancer Institute,
Frederick, MD) weekly during the sensitization protocol as well as 30 minutes before the intrathoracic injection. Neutralizing anti-IL-5
MoAb (TRFK-5; PharMingen, San Diego, CA) was administered intraperitoneally 30 minutes before the intrathoracic injection at two
different concentrations (1 mg/kg and 5 mg/kg). There was no difference
in OVA-induced eosinophil accumulation in sensitized animals when
nontreated animals were compared with animals treated with an isotype
control antibody (rat IgG1; PharMingen). Values for wild-type mice
(mean ± SEM eosinophils × 105/cavity) were 5.2 ± 1.4 in nontreated mice, 5.8 ± 1.6 in mice treated with 1 mg/kg
control IgG1, and 7.9 ± 2.9 in mice treated with 5 mg/kg IgG1,
whereas the corresponding values for NFAT1/ mice were
17.8 ± 5.0 for nontreated mice, 18.1 ± 6.5 for mice treated with 1 mg/kg control IgG1, and 17.4 ± 5.8 for mice treated with 5 mg/kg
control IgG1.
Collection of bone marrow and peripheral blood cells.
Bone marrow cells were isolated from the left femur. The femoral head
and condyles were removed, and the displaceable cells were recovered by
flushing the marrow cavity of the femur shaft with 3 mL PBS containing
heparin (10 U/mL). Blood samples were collected from the mice tail
vein. Total and differential nucleated bone marrow and peripheral blood
cells were evaluated as described below.
Cell counts.
Total cell counts were obtained after dilution of the pleural fluid,
bone marrow cell suspension, and peripheral blood in Turk solution (2%
acetic acid). Differential analysis of leukocytes was done under oil
immersion objective in cytocentrifuged smears stained with Diff-Quik
(Baxter, FL). Eosinophils, macrophages/monocytes, lymphocytes, and
neutrophils were enumerated based on morphology and staining
characteristics.
Cytokine analysis.
Pleural fluid obtained as described above was centrifuged (1,000g, 10 minutes), and the cell-free supernatant was assessed for cytokine
content. IL-4 (Genzyme, Cambridge, MA), IL-5 (Endogen, Woburn, MA), and
IL-13 (R&D Systems, Minneapolis, MN) protein levels were determined by
enzyme-linked immunosorbent assay (ELISA).
For cytokine mRNA analysis, wild-type and NFAT1/ mice
were sensitized with OVA as described above. Ten days later, draining lymph nodes were removed, and cells were incubated without or with 1 mg/mL OVA for 6 hours. Cytokine mRNA expression in total cellular RNA
from each sample was analyzed with a multiple cytokine RNase protection
assay (Ribo-Quant; PharMingen).
IgE measurement.
Blood serum for IgE analysis was obtained by cardiac puncture 15 days
after sensitization with ovalbumin as described above. Serum IgE was
measured by sandwich ELISA using anti-mouse IgE MoAb R35-72 as capture
antibody and biotinylated anti-mouse IgE R32-92 as detection antibody
(PharMingen).
Statistical analysis.
Statistical analysis of values from wild-type and
NFAT1/ and between control and treated groups was
determined using an unpaired Student's t-test for single
comparison or an analysis of variance followed by the
Student-Newman-Keuls for multiple comparisons. P < .05 was
considered to be statistically significant as indicated in the figure
legends.
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RESULTS |
Allergic response to OVA sensitization and challenge in NFAT1-deficient
mice.
Eosinophil tissue infiltration at the site of inflammation is one of
the major events in the allergic response. We previously showed that
NFAT1-deficient mice display increased eosinophil accumulation in a
model of allergic pleurisy.25 To investigate if this
phenotype was related to the mixed genetic background of these mice, we
compared them with mice of the 129/Sv and C57BL/6 parental strains. As
shown in Fig 1, eosinophil accumulation in the pleural cavity in response to antigen challenge was
indistinguishable in the parent strains and wild-type mice, whereas
eosinophil accumulation in the pleural cavity of
NFAT1/ mice was significantly greater under the same
conditions. These results suggested that the increased eosinophil
infiltration in NFAT1/ mice is not related to their
mixed genetic background, but rather to their lack of expression of
NFAT1 protein.
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| Fig 1.
Allergic pleurisy in wild-type, NFAT1/,
and parental strain mice. Shown are the total number of eosinophils in
the pleural cavity of C57BL/6, 129/Sv, wild-type (+/+), and
NFAT1/ mice sensitized with OVA and challenged in the
pleural cavity with PBS or OVA. Data are expressed as mean ± SEM of
values from 5 mice. (*) Indicates significantly greater response
relative to wild-type mice (P < .05).
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Several studies have indicated an association of atopic disease with
bone-marrow response to allergen. Myeloid progenitor cells
(granulocyte-macrophage colony-forming units [GM-CFU])
are increased in response to Ascaris suum antigen in
canines,28 and in atopic human blood there are higher
numbers of eosinophil-basophil colony-forming units
(Eo/B-CFU).29,30 Indeed, it has been suggested that there
is a mobilizable pool of bone marrow eosinophils that can be rapidly
recruited to blood and migrate into tissues in response to
chemoattractant.31 To evaluate eosinophil differentiation in response to allergen, wild-type and NFAT1/ mice
were sensitized with OVA, and eosinophil counts in bone marrow and
peripheral blood were assessed. As shown in Fig
2, NFAT1/ mice that had
been sensitized with ovalbumin displayed increased eosinophil numbers
in bone marrow and peripheral blood when compared with wild-type mice.
Moreover, the number of bone marrow and blood eosinophils in sensitized
mice showed a positive correlation with eosinophil numbers in the
pleural cavity in response to allergen challenge (data not shown).
These data indicated that the enhanced eosinophil accumulation at the
site of allergic inflammation was due not merely to a local reaction,
but rather reflected a general upregulation of the allergic response.
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| Fig 2.
Analysis of bone marrow response to OVA sensitization.
Shown are the total number of eosinophils in bone marrow and peripheral blood of wild-type (+/+) and NFAT1/ mice that
were either not sensitized (none) or were sensitized with OVA. Data are
expressed as mean ± SEM of values from 5 to 6 mice. (*) Indicates
significantly greater response relative to wild-type mice
(P < .05).
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OVA-sensitized cells in lymph nodes of NFAT1/ mice
display a Th2 cytokine profile.
Eosinophil migration into inflamed tissue is coordinated by an
interacting network of cytokines, chemokines, adhesion molecules, and
inflammatory mediators, and chemoattractant production at the site of
inflammation is essential for eosinophil recruitment.5 The
cytokine IL-5, which is produced by T cells and mast cells as well as
by eosinophils themselves, is recognized as a primary regulator of
eosinophil growth, differentiation, chemotaxis, and activation.32-34 Furthermore, several studies have shown
higher expression of different cytokines, such as IL-4, IL-5, and IL-13 at sites of allergic inflammation.35-38 To determine
whether the eosinophilia seen in NFAT1/ mice was
related to differential cytokine expression, we used an RNase
protection assay where multiple cytokines can be assessed in a sample
of very few cells. Wild-type and NFAT1/ mice were
immunized in the footpads with OVA, using the same protocol as for
antigen sensitization for later intrapleural challenge, cells from the
draining lymph node were restimulated in vitro with OVA, and the
expression of cytokines was assessed. As shown in Fig
3, no cytokines were detected
in unstimulated cells (lanes 1 and 2). However, although only a small
fraction of lymph node cells were expected to be OVA-responsive,
NFAT1/ mice showed a striking increase in production
of the Th2 cytokines IL-4, IL-5, and IL-13, relative to wild-type mice,
after restimulation with OVA (Fig 3, compare lanes 3 and 4).
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| Fig 3.
Analysis of cytokine expression by
OVA-sensitized wild-type (+/+) and NFAT1/ cells.
Mice were sensitized in vivo with OVA, the draining lymph node cells
were left unstimulated ( ) or restimulated in vitro with 1 mg/mL of
OVA for 6 hours and the total cellular RNA was analyzed by RNase
protection assay for cytokine mRNA levels. The detected cytokines
(IL-4, IL-5, IL-10, IL-13, IL-2, and IFN-) and the housekeeping
genes (L32 and GAPDH) are indicated. These results are representative
of experiments using three different pairs of wild-type and
NFAT1-deficient mice.
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Analysis of cytokine production in the in vivo allergic response.
Because lymph node cells from NFAT1/ mice
overexpressed IL-4, IL-5, and IL-13 after OVA stimulation in vitro, we
investigated the production of these cytokines in vivo in the model of
allergic pleurisy. Pleural washes from OVA-sensitized wild-type and
NFAT1/ mice were assessed by ELISA for IL-4, IL-5,
and IL-13 protein content. The levels of IL-5 and IL-13 in the
wild-type mice challenged with OVA increased 6- and 15-fold,
respectively, when compared with mice challenged with PBS. However, in
our experimental conditions only the increase in IL-13
(P = .005), but not in IL-5 (P = .065), reached
significant values (P < 0.05). In NFAT1-deficient animals, the levels of both cytokines IL-5 (P = .018) and IL-13
(P = .018) were significantly different when OVA-challenged
and PBS-injected mice were compared (Fig
4). Interestingly, NFAT1/
mice presented increased levels of IL-5 and IL-13 in the pleural cavity
after OVA challenge when compared with wild-type mice (Fig 4), whereas
IL-4 was not detected in either mouse group (data not shown). The lack
of detection of IL-4 in the pleural washes, although this cytokine is
overexpressed in vitro (Fig 3), could be due to IL-4 consumption in the
in vivo response or an insufficient sensitivity of the ELISA for IL-4.
The finding of IL-5 overproduction in the pleural cavity of
NFAT1-deficient animals is consistent with the enhanced eosinophil
infiltration observed in these mice. IL-5 is a selective
chemoattractant for eosinophils33 and has the ability to
prime and activate these cells.32-34 In addition, IL-5-deficient mice do not show eosinophilia39 and fail to
develop airway hyperresponsiveness and eosinophil infiltration in an
experimental model of asthma.8
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| Fig 4.
Analysis of in vivo cytokine production during allergic
pleurisy. Measurement of IL-5 and IL-13 production in pleural cavity of
wild-type (+/+) and NFAT1/ mice sensitized with
OVA and challenged in the pleural cavity with PBS or OVA. Pleural
washes were obtained 24 hours after OVA challenge and assessed for
cytokine levels by ELISA. Data are expressed as mean ± SEM of values
from 5 mice. (*) Indicates significantly greater values relative to
wild-type mice (P < .05).
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Role of IL-5 and IL-4 in the enhanced in vivo allergic response.
Both IL-4 and IL-5 genes are known targets for NFAT
proteins.19 To investigate the involvement of these
cytokines in mediating eosinophil recruitment in NFAT1-deficient mice,
we used an in vivo treatment with neutralizing anti-IL-4 and
anti-IL-5 MoAbs in the model of allergic pleurisy. Wild-type and
NFAT1/ mice were sensitized with OVA and treated with
anti-IL-5 MoAb at two different concentrations before allergen
challenge. As shown in Fig 5, a low
concentration of anti-IL-5 (1 mg/kg) completely abrogated OVA-induced
eosinophil recruitment in the wild-type mice but only slightly
inhibited the eosinophil accumulation observed in
NFAT1/ mice under the same conditions. Interestingly,
treatment with a fivefold higher dose of anti-IL-5 (5 mg/kg)
completely inhibited eosinophil accumulation in the
NFAT1/ mice (Fig 5), consistent with the observed
IL-5 overproduction in the pleural cavity of these mice (Fig 4). Thus,
IL-5 is likely to be the major mediator contributing to the high
numbers of eosinophils present in NFAT1-deficient mice in the in vivo
allergic response (Fig 1). Although various chemokines and lipid
mediators including eotaxin, RANTES, platelet-activating
factor, and leukotriene B4 have been implicated in eosinophil
recruitment, our results indicate that these other chemoattractants do
not act independently of IL-5 in NFAT1/ or wild-type
mice. However, they could synergize promoting high levels of eosinophil
recruitment. In fact, it has been shown that IL-5 and eotaxin synergize
as chemoattractants for eosinophils in an in vivo model of eosinophil
migration.31,40
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| Fig 5.
Analysis of allergic eosinophil accumulation into the
pleural cavity after treatment with anti-IL-5 and anti-IL-4
neutralizing MoAb. Total numbers of eosinophils in the pleural cavity
of wild-type (+/+) and NFAT1/ mice were assessed
in animals sensitized with OVA and challenged in the pleural cavity
with PBS or OVA. Mice were left untreated (none) or treated with
anti-IL-5 or anti-IL-4 neutralizing MoAb as described in Materials
and Methods. Different concentrations of anti-IL-5 (1 mg/kg and 5 mg/kg) were used for treatment as indicated. Data are expressed as mean ± SEM of values from 5 to 12 mice. (*) Indicates significantly
greater values relative to the PBS-challenged controls (P < .05),
and (+) indicates significantly smaller values than the groups
challenged with OVA but not treated with antibody
(P < .05).
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The cytokine IL-4 has been implicated as a major cytokine
in Th2 differentiation leading to IL-5 production, and it also promotes IgE class switching in B lymphocytes.17,18 To investigate
the importance of IL-4 in eosinophil tissue infiltration, wild-type and
NFAT1/ mice were treated with neutralizing anti-IL-4
MoAb during the sensitization period, and the allergic pleurisy was
evaluated. As shown in Fig 5, anti-IL-4 treatment significantly
inhibited eosinophil accumulation in both wild-type and
NFAT1/ mice, although the inhibition was not
complete. Furthermore, anti-IL-4 treatment also inhibited the
antigen-induced increase of serum IgE levels in the wild-type and
NFAT1/ mice (data not shown). It is important to note
that anti-IL-5 treatment specifically inhibited eosinophil recruitment
in the allergic pleurisy, whereas anti-IL-4 treatment also had an
effect on migration of other leukocytes (data not shown). These results suggest that the overexpression of IL-4 plays a key role during the
immunization process, which drives the immune response of NFAT1-deficient mice toward Th2 differentiation and an increased allergic response.
| |
DISCUSSION |
Our results indicate that the pleural eosinophilia observed in
NFAT1/ mice correlates with a classically Th2 pattern
of cytokine expression, involving the cytokines IL-4, IL-5, and IL-13.
The eosinophilia is abrogated or considerably inhibited by treatment of
the mice with anti-IL-5 and anti-IL-4 neutralizing MoAbs,
respectively (Fig 5). Moreover, antigen-responsive cells from the
pleural cavity and from draining lymph nodes of
NFAT1/ mice show a striking upregulation of these
three cytokines relative to wild-type mice (Figs 3 and 4). Furthermore,
NFAT1/ mice show increased numbers of eosinophils in
bone marrow and peripheral blood after allergen sensitization (Fig 2).
The differentiation, maturation, and proliferation of bone marrow
eosinophils is promoted by GM-CSF, IL-3, and IL-5.41-45 The
role of these cytokines in increased bone marrow eosinophil levels in
NFAT1/ mice remains to be determined. Although IL-3
and GM-CSF might play a role in eosinophilia following allergen
challenge, our data on IL-5 mRNA (Fig 3) and protein (Fig 4)
overexpression, and more importantly the ability of anti-IL-5 MoAb to
completely inhibit the enhanced eosinophil accumulation in
NFAT1/ mice (Fig 5), indicate that IL-5 is a major
effector molecule for eosinophil accumulation in NFAT1-deficient mice.
However, we cannot rule out a parallel or synergistic effect for IL-3, GM-CSF, or other effector molecules.
Earlier controversy about the relative importance of IL-4 and IL-5 in
mediating allergic responses8,12 was resolved by proposing
at least two different cellular mechanisms for the allergic response,
one dependent on IgE and IL-4 and mediated by mast cells, and another
dependent on IL-5 and mediated by eosinophils.2 These
different mechanisms were linked with the genetic background of the
experimental animal strain, where BALB/c mice are high IgE responders
and C57BL/6 are naturally hyporesponsive.2 Our results
indicate that NFAT1/ mice, genetically of mixed
129/Sv × C57BL/6 background, combine both the proposed mechanisms
that they show a characteristic allergic phenotype with high
eosinophilia that is associated with increased serum IgE
levels25 and is dependent on overexpression of both Th2
cytokines IL-4 and IL-5. In effect, the loss of NFAT1 protein leads a
hyporesponsive mouse strain to develop a high IgE- and IL-4-dependent
allergic response.
Extending this line of thinking to asthma and atopy, two different
types of human asthma have been described, one allergic and another
unrelated form known as intrinsic or nonallergic.9 Whereas
allergic asthma usually manifests at a young age and shows a strong
familial occurrence and correlation with IgE levels, intrinsic asthma
usually begins in adulthood, is characteristically not familial, and is
not related to high levels of serum IgE.9,11 Nevertheless,
patients with intrinsic asthma resemble patients with allergic asthma
in that they show increased numbers of eosinophils in the peripheral
blood and bronchoalveolar fluid and similar bronchial
histopathologies.11 Allergic asthmatics display a characteristic Th2 cytokine profile, with increased levels of both IL-4
and IL-5 in peripheral blood and bronchoalveolar fluid, whereas the
corresponding fluids from patients with intrinsic asthma show increases
of IL-2 and IL-5 but no increase in IL-4.11 Our results
suggest that NFAT1-deficient mice display a phenotype resembling that
of allergic rather than that of nonallergic asthma in being
characterized by a Th2 cytokine profile. The genetic basis for atopic
disease and the propensity to develop a type 2 cytokine response is
currently under intense investigation in several
laboratories,1,46,47 and it is possible that certain forms
of human atopy could also involve a selective deficiency in the
transcription factor NFAT1.
The immune phenotype of NFAT1-deficient mice illustrates three
important points. First, these mice are immunocompetent rather than
immunodeficient and do not show any gross impairment in the production
of NFAT-dependent cytokines, indicating that the lack of NFAT1 is
compensated for by the presence of other NFAT proteins. Second, the
increased secondary immune responses and increased cell proliferation
observed in NFAT1-deficient mice suggests that NFAT1 may actually have
an overall negative effect on immune responsiveness in normal mice.
This behavior is not unprecedented. For example, in signal transduction
pathways, kinases that are activated early during a response often
activate feedback processes that contribute to the late downregulation
of the same response. Finally, the unusual hypereosinophilia of
NFAT1-deficient mice in a model of allergy, and their tendency toward
the late production of Th2-type cytokines suggests that NFAT1
critically influences Th differentiation during the normal immune
response. NFAT1 could act to promote the transcription of genes
encoding cytokines or effector proteins that skew T-cell
differentiation toward the Th1 pathway or cytokines that suppress
differentiation toward the Th2 pathway. Alternatively, NFAT1 could
inhibit the production of effector proteins having the opposite effect.
These possibilities are not mutually exclusive. Given the importance of
Th1-Th2 cytokine production in asthma, allergy, and other clinical
situations, it is of considerable interest to understand the mechanisms
by which NFAT1 exerts its profound effects on T-cell differentiation
and function. Moreover, NFAT1-deficient mice represent a good model in
which to study the effect of background genes and NFAT1 itself on the
process of T-cell differentiation toward Th1 and Th2 phenotypes.
| |
FOOTNOTES |
Submitted October 31, 1997;
accepted December 23, 1997.
Supported by National Institutes of Health Grant Nos. RO1 CA42471 and
PO1 AI35297 to A.R.; A.K. was supported by a fellowship of the Deutsche
Forschungsgemeinschaft (Germany); P.T.B. is a Research Fellow of the
Pew Foundation (USA) and CNPq (Brazil); and A.R. is Scholar of the
Leukemia Society of America (USA).
Address reprint requests to Anjana Rao, PhD, The Center for Blood
Research, Harvard Medical School, 200 Longwood Ave, Boston, MA, 02115.
The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" is accordance with 18 U.S.C. section 1734 solely to indicate this fact.
| |
ACKNOWLEDGMENT |
The authors thank Drs A. Lichtman and A. Abbas for helpful discussions.
| |
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Abstract
Introduction
Abstract