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*
Department of Molecular Biology, DNAX Research Institute for Molecular and Cellular Biology, Palo Alto, CA 94304;
Department of Immunology, University of Cape Town, Cape Town, South Africa; and
Schering-Plough Research Institute, Lafayette, NJ 07848
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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, IP-10, MIG, CXCR3,
chemokine receptor-8, and IL-4. However, cotreatment with either IL-10
or anti-IL-12 mAb effectively blocked the development of colitis in
the presence of exogenous IL-4. These data indicate that IL-4 treatment
exacerbates a Th1-mediated disease rather than induces Th2-mediated
inflammation. As other cell types besides T cells express the receptor
for IL-4, the proinflammatory effects of IL-4 on host cells in
Rag2-/- recipients were assessed. IL-4 treatment was able
to moderately exacerbate colitis in Rag2-/- mice that
were reconstituted with IL-4R
-deficient (IL-4R-/-)
CD4+CD45RBhigh T cells, suggesting that the
IL-4 has proinflammatory effects on both non-T and T cells in this
model. IL-4 did not cause colitis in Rag2-/- mice in the
absence of T cells, but did induce an increase in MHC class II
expression in the lamina propria of the colon, which was blocked by
cotreatment with IL-10. Together these results indicate that IL-4 can
indirectly promote Th1-type inflammation in the
CD4+CD45RBhigh T cell transfer model of
colitis. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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(1, 2). When
uncontrolled, Th1 responses can result in chronic inflammatory
diseases, such as diabetes, arthritis, and multiple sclerosis. In
animal models, inhibition of IL-12 will prevent the development of
these autoimmune diseases by blocking the development of
IFN--producing CD4+ T cells (3, 4). Thus, the ability to control the development of Th1-type T
cells may be critical for the prevention of some chronic inflammatory
diseases. Th2-type responses are characterized by the development of CD4+ T cells, which secrete IL-4, IL-5, IL-10, and IL-13 (1, 2). Some Th2-type cytokines, especially IL-10 and IL-4, are known to suppress the development of Th1 T cells (3, 5, 6). IL-10 has been shown to suppress many Th1-type inflammatory responses in vivo, including LPS-induced endotoxic shock, contact hypersensitivity, experimental autoimmune encephalomyelitis, and collagen-induced arthritis (7, 8, 9, 10, 11). Like IL-10, IL-4 treatment can suppress or ameliorate the development of Th1-type inflammatory diseases. IL-4 treatment has been shown to suppress the development of proteoglycan-induced arthritis and to prevent cartilage destruction in a collagen-induced model of arthritis (12, 13). In addition, IL-4 inhibits the development of diabetes in both NOD and transgenic mouse models as well as the development of experimental autoimmune encephalomyelitis (5, 14, 15, 16, 17, 18). Furthermore, the combination of IL-4 and IL-10 is more effective than either cytokine alone in ameliorating glomerulonephritis and delayed-type hypersensitivity in mice primed with Leishmania major (19, 20). These data suggest that Th2-associated cytokines are potent inhibitors of Th1-type immune responses.
The ability of Th2-associated cytokines to inhibit the development of
deleterious Th1-mediated immune responses suggested that they may be
useful therapeutics for ablating certain autoimmune diseases (5, 6). However, there is increasing evidence that IL-4, in
particular, can aid in Th1-type inflammatory responses. IL-4 treatment
of rats significantly worsened the development of Th1-type experimental
autoimmune uveoretinitis and resulted in increased production of
IFN-, TNF-, and NO in recall responses in vitro to retinal Ag
(21). Furthermore, IL-4-deficient
(IL-4-/-) mice have severely impaired
anti-tumor responses that are characterized by an inability to
develop CD8+ anti-tumor CTLs and by decreased
production of splenic IFN- (22).
IL-4-/- mice are also unable to produce enough
IFN- and IL-12 to develop immunity to Candida albicans
infection (23). Recently, IL-4 has been shown to be
critical for the development of alloreactive CD4+
T cells, and blocking IL-4 prolongs the survival of allogeneic skin
grafts (24). These data indicate that IL-4 may be
important for the development of some Th1-type inflammatory
responses.
Crohn’s disease is a chronic inflammatory condition of the intestinal
tract that has been characterized as a Th1-mediated disease. T cells
isolated from the colons of patients with Crohn’s disease produce
large amounts of IFN- and TNF- and little IL-4 or IL-10
(25, 26, 27). Many mouse models of chronic intestinal
inflammation, or colitis, are also characterized by massive
infiltration of IFN--producing CD4+ T cells in
the colon. IL-10-/- mice,
IL-2-/- mice,
Gi2-/- mice,
Stat3-/- mice, Tg
26 mice reconstituted with
wild-type (WT)3 bone
marrow cells, immunodeficient recipients of
CD4+CD45RBhigh T cells, and
trinitrobenzenesulfonic acid (TNBS)-treated SJL/J mice all develop
Th1-type colitis (28, 29, 30, 31, 32). In many of these models disease
is either prevented, or at least ameliorated, by early treatment with
IL-10 or neutralizing Ab to IL-12 (26, 28, 30). The role
of IL-4 in Th1-type intestinal disease is less clear. In one study IL-4
treatment did not suppress the development of colitis in
scid mice transplanted with
CD4+CD45RBhigh T cells
(33). In contrast, infection of mice with adenovirus
containing the IL-4 gene reduced inflammation in rats with TNBS-induced
colitis (34).
We have evaluated the effect of IL-4 treatment in a modified version of the original T cell transfer model of colitis (29, 35). In this modified model the transfer of CD4+CD45RBhigh T cells from either WT mice or IL-10-/- mice with established Th1-type disease into 129 SvEv Rag2-/- mice results in colitis characterized by diffuse inflammatory cellular infiltrates, epithelial hyperplasia, and, when severe, ulcers and transmural inflammation. The resulting colitis can be significantly ameliorated by treatment with anti-IL-12 mAb or rIL-10 in vivo (36, 37). We transferred CD4+CD45RBhigh T cells from either WT or IL-10-/- mice into Rag2-/- immunodeficient mice and treated them daily with IL-4. Our data show that IL-4 treatment has the ability to exacerbate Th1-type colonic inflammation in this model.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Recombinase-activating gene 2-deficient
(Rag2-/-) mice on either 129 SvEv or BALB/c
background and WT 129 SvEv mice were obtained from Taconic Farms
(Germantown, NY) or from a colony maintained at the DNAX Animal
Care Facility. IL-10-/- 129 SvEv mice and IL-4R
-chain-deficient (IL-4R-/-) BALB/c mice
were from colonies maintained under specific pathogen-free conditions
at the DNAX Animal Care Facility (38, 39).
In vivo treatments
Purified recombinant murine IL-4 and IL-10 were made as previously described and contained <0.1 E.U/mg protein of endotoxin (37, 40). Mice were given 10–20 µg of IL-4 i.p. daily for 28 days. For IL-10 treatment, mice were given 20 µg i.p. daily. Saline (1x HBSS; BioWhittaker, Walkersville, MD) was given daily as a negative control. Purified anti-murine IL-12 mAb (C.17.8.20) or isotype control mAb (MP4–25D2, anti-human IL-4; no cross-reactivity with murine IL-4) was given to mice i.p. weekly, at 2 mg/dose for the duration of the experiment. All cytokine and Ab treatments were started on the day of T cell transfer (day 0).
Cell isolations and transfers
CD4+CD45RBhigh splenic T cells were obtained by cell sorting. Briefly, splenocytes were first enriched for CD4+ T cells by red cell lysis and magnetic bead depletion using lineage-specific rat mAbs supernatants (10%, v/v): B220 (B cells) and Ter119 (erythrocytes). mAb-stained cells were removed in a magnetic field using goat anti-rat IgG (Fc) and goat anti-rat IgG (H+L)-coated magnetic beads (PerSeptive Diagnostics, Cambridge, MA). The remaining cells were then stained with anti-CD4-FITC and anti-CD45RB-PE (both from PharMingen, San Diego, CA). Two-color cell sorting was performed using a FACStar Plus (Becton Dickinson, Mountain View, CA). The sorted CD4+CD45RBhigh T cells were >98% pure upon reanalysis. The purified CD4+CD45RBhigh cells (2 x 105) were injected i.p. into 129 SvEv or BALB/c Rag2-/- recipient mice, depending on the donor cells used. Four weeks after T cell transfer, mice were sacrificed and analyzed for the presence of enterocolitis.
Histologic analysis of colitis
Microscopic examination of mouse colons was performed in a blinded fashion by the same pathologist (M.W.L.) on formalin-fixed tissue sections stained with hematoxylin and eosin as previously described (36). Longitudinal sections of the entire length of the colon were evaluated, taking into account both the number of lesions and their severity. Five regions of the colon (cecum, ascending, transverse and descending colon, and rectum) were graded semiquantitatively as 0 (no change) to 5 (most severe change). The grading represents the incidence and severity of inflammatory lesions based on infiltrates, goblet cell loss, crypt abscesses, ulcerations, and fibrosis. The summation of the score for each of five segments of the colon provides a total disease score per mouse (from 0–25), where 0–1 indicates no change, 2–5 indicates mild disease, 6–10 indicates moderate disease, and 11–20 indicates severe disease. No mice in these studies have a score >20, because such severe disease results in death.
Quantitative mRNA (TaqMan) analysis in colon
Total RNA was isolated from whole colon samples using Qiagen
RNeasy columns (Qiagen, Valencia, CA), according to the manufacturer’s
instructions. Total RNA (5 µg) was reverse transcribed into cDNA
using random hexamers (Promega, Madison, WI). The expression of
IFN-, IL-4, MIG, IP-10, eotaxin, TARC, CCR3, CCR4, CCR8, CXCR3, and
TNF- was determined by a method for real-time quantitative PCR using
the ABI 7700 sequence detector system (Perkin-Elmer Applied Biosystems,
Foster City, CA). Briefly, 50 ng of total cDNA was in a reaction volume
of 25 µl that contained final concentrations of 1x PCR buffer; 200
µM dATP, dCTP, and dGTP; 400 µM dUTP; 4 mM
MgCl2; 1.25 U of AmpliTaq DNA polymerase; 0.5 U
of Amp-Erase uracil-N-glycosylase; 900 nM of each primer;
and 250 nM probe. The thermal cycling conditions included 50°C for 2
min and 95°C for 10 min, followed by 40 cycles of amplification at
95°C for 15 s and 55°C for 1 min for denaturing and
annealing-extension, respectively. Sense and antisense primers as well
as probes used for the detection of the genes of interest were
predeveloped TaqMan assay reagents (Perkin-Elmer Applied Biosystems).
Primers and probes were designed to ensure that no cross-reactivity
with other genes would occur. Each primer-probe pair was tested on a
panel of cDNA plasmids containing a variety of cytokines, chemokines,
and chemokine receptors. The probes for each message were labeled at
the 5' end with a reporter fluorescent dye, FAM, and at the 3' end with
a quench fluorescent dye, TAMRA. Fluorescence detection of FAM was
performed at the end of each cycle. The quantity of cDNA of the gene of
interest was directly related to the amount of FAM detected after 40
cycles. cDNA plasmids containing the gene of interest were used as a
standard curve, ranging from 100 to 0.01 pg. From this standard curve,
the amount of cDNA of the gene of interest was calculated in femtograms
per 50 ng of total cDNA. As an internal control, 18S ribosomal RNA
(rRNA) expression was measured in each sample in a multiplex assay; the
probe for rRNA was labeled at the 5' end with the reporter fluorescent
dye VIC. The amount of 18S rRNA was correlated to the cycle at which
VIC fluorescence was first detected (cycle threshold value). To correct
for any variation in the amount of RNA between individual samples, the
mean cycle threshold value for 18S rRNA was calculated for all samples
and subtracted from each individual cycle threshold value, then this
difference was raised to the second power and multiplied by the FAM
value (in femtograms per 50 ng of total cDNA) for each sample. Thus,
the amount of cDNA of the gene of interest in each sample could be
directly compared with the amounts detected in all other samples.
Immunohistochemical analysis
Briefly, colon pieces from saline- and IL-4-treated Rag2-/- mice were cleaned of feces and frozen in tissue-freezing medium (Triangle Biomedical Sciences, Durham, NC). Frozen blocks were cut on a cryostat, and 8-µm-thick sections were cut onto gelatinized glass slides, dried, and fixed for 10 min with acetone. Sections were preincubated with 1x PBS (GeneMate, Kaysville, UT) and 10% normal mouse serum (NMS; Jackson ImmunoResearch Laboratories, West Grove, PA) for 15 min. Rat anti-mouse I-Ab (PharMingen) was diluted in PBS/10% NMS and incubated on sections for 1 h at room temperature. Rat IgG2b (PharMingen) was used as an isotype control. Sections were rinsed in 1x PBS and stained with HRP-conjugated sheep anti-rat IgG (The Binding Site, Birmingham, U.K.) diluted in PBS/10% NMS for 30 min at room temperature. AEC (Vector, Burlingame, CA) was used as a substrate for HRP. Sections were counterstained with hematoxylin (Vector), and then sequentially rinsed in H20, 1% ammonium hydroxide, and PBS. Sections were mounted with coverslips and observed by light microscopy.
Statistics
All data were analyzed using a statistical program (InstatP,
GraphPad, San Diego, CA). Student’s t test or nonparametric
Mann-Whitney test was used to determine statistical significance
between groups, with p 
0.05 considered
significant.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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To test the effect of IL-4 on a Th1-type murine model of colitis,
WT CD4+CD45RBhigh T cells
were transferred into immunodeficient Rag2-/-
129 SvEv mice, and the recipient mice were treated daily with either
saline (vehicle control) or 20 µg IL-4. In this model untreated
recipient mice develop moderate to severe colitis 8–12 wk after T cell
transfer (29, 36, 37). However, IL-4 treatment exacerbated
the disease such that the majority of mice were moribund 4 wk after T
cell transfer. Therefore, the experiment was terminated, and the mice
were analyzed histologically for signs of colitis. Five of eight mice
had moderate to severe colitis by 4 wk, while saline-treated mice had
few histological signs of disease at that time point (Fig. 1
A). As WT
CD4+CD45RBhigh T cells are
uncommitted to either a Th1 or a Th2 phenotype, we considered the
possibility that IL-4 treatment was pushing naive T cells to an
inflammatory Th2 phenotype, rather than having a proinflammatory effect
on Th1-type T cells. Therefore, we repeated the experiment using
CD4+CD45RBhigh T cells from
4- to 6-mo-old IL-10-/- 129 SvEv mice that
already had signs of chronic Th1-type colitis (29, 40).
The IL-10-/-
CD4+CD45RBhigh T cells
caused mild colitis in recipient Rag2-/- mice
within 4 wk (Fig. 1B; mean colitis score, 3.3; Fig. 2A). However, IL-4 treatment
was able to clearly exacerbate the colitis seen in
Rag2-/- recipients of
IL-10-/-
CD4+CD45RBhigh T cells
(Figs. 1B and 2B; mean colitis score, 10.9).
Because, in our hands, IL-10-/-
CD4+CD45RBhigh T cells
cause more consistent and aggressive disease in
Rag2-/- recipients than T cells from WT mice,
we performed all subsequent transfer experiments with
CD4+CD45RBhigh T cells from
IL-10-/- mice. (M. Fort and D. Rennick,
unpublished observations).
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The ability of exogenous IL-4 to exacerbate colitis in this T cell
transfer model of colitis lead us to investigate whether IL-4 induced a
switch from a Th1-dependent to a Th2-dependent disease. Therefore, we
looked for differential gene expression in the colons of
Rag2-/- mice 28 days after reconstitution with
IL-10-/-
CD4+CD45RBhigh T cells and
treatment with either saline or IL-4. Using a very sensitive,
quantitative method of PCR analysis, colonic total RNA was tested for
the presence of mRNA of various Th1- and Th2-type-associated cytokines,
chemokines, and chemokine receptors. IL-4 treatment induced a
significant increase in the expression of TNF- and IFN- mRNA, as
well as in that of the IFN--induced chemokines MIG and IP-10 (Fig. 3 and data not shown). In addition, IL-4
treatment up-regulated the expression of mRNA for CXCR3, which is the
receptor for MIG and IP-10 and is expressed on Th1-type
CD4+ T cells (41). In all samples
tested the mRNA expressions of IL-12p35 and IL-12p40 were below the
detection limits of the system (data not shown). IL-4 treatment also
induced a significant increase in IL-4 mRNA expression in the colon,
but not in the mRNA expression of other Th2-associated molecules, such
as IL-5, IL-13, eotaxin, and TARC (Fig. 3 and data not shown). CCR4 and
CCR3, which are preferentially expressed on Th2-type T cells and are
the receptors for TARC and eotaxin, respectively, were not up-regulated
with IL-4 treatment (Fig. 3 and data not shown) (32).
Expression of CCR8, which is expressed on Th2-type T cells and
monocytes, was significantly increased (Fig. 3) (41, 42, 43).
Thus, IL-4 treatment in vivo can up-regulate the colonic mRNA
expression of Th1-associated cytokines and chemokines as well as some
Th2-associated molecules.
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D and 4,
Rag2-/- recipients of
IL-10-/-
CD4+CD45RBhigh T cells
cotreated with IL-4 and anti-IL-12 mAb developed significantly less
disease than recipients cotreated with IL-4 and isotype control mAb
(mean colitis score, 2.4 and 10.9, respectively; p <
0.001). Quantitative PCR analysis of gene expression in the colons of
mice cotreated with anti-IL-12 mAb and saline or IL-4 showed
decreased mRNA expression of Th1-associated genes compared with isotype
control-cotreated animals (data not shown). The gene expressions of
MIG, IP-10, and CXCR3 were decreased with anti-IL-12 mAb
cotreatment by 35- to 125-fold, 8- to 32-fold, and 3-fold,
respectively. The expression of IFN- mRNA was completely suppressed
in the colons of recipients cotreated with saline and anti-IL-12
mAb, but was only partially suppressed (3-fold decrease) in colons of
recipients cotreated with IL-4 and anti-IL-12 mAb. This could
explain the mild disease seen in IL-4- and anti-IL-12 mAb-cotreated
mice compared with the almost complete absence of disease seen in
saline- and anti-IL-12 mAb-cotreated mice (Fig. 4). There was no
change in the gene expression of IL-4, CCR4, or CCR8 in the colons of
anti-IL-12 mAb-treated vs isotype control-treated mice (data not
shown).
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C and 4). These data
suggest that the colitis seen in IL-4-treated recipients was dependent
on the production of IL-12 and was inhibited by the presence of
exogenous IL-10, and thus fits the paradigm of a Th1-type inflammatory
disease. IL-4 need not act directly on T cells to augment a Th1-type inflammatory response
The receptor for IL-4 consists of the IL-4R -chain and IL-2R
-chain and is expressed on many cell types, including T cells, B
cells, monocytes, and nonhemopoietic cells, including intestinal
epithelial cells (44, 45, 46, 47). Therefore, IL-4 treatment
in vivo may accelerate colitis by acting on T cells, microenvironmental
cells, or both. To differentiate the effects of exogenous IL-4 on the
Rag2-/- host cells vs donor T cells, we made
use of T cells from mice that are deficient in the IL-4R -chain
(IL-4R-/-).
CD4+CD45RBhigh T cells from
IL-4R-/- BALB/c mice were transferred into
Rag2-/- BALB/c mice, and the mice were
treated for 28 days with either IL-4 or saline. As shown in Fig. 5, saline-treated recipients of
IL-4R-/- T cells developed mild-moderate
colitis (mean score, 4.8) within 4 wk. Of the IL-4-treated recipients,
9 of 13 mice had moderate to severe disease, even though the
transferred T cells could not respond to IL-4 (Fig. 5). However,
because of the large variability in disease score within each group,
the results were not statistically significant. Nevertheless, the trend
toward exacerbated disease following IL-4 treatment suggests a
proinflammatory effect on the microenvironmental cells that express
functional IL-4R.
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, 9 of 10 nonreconstituted
Rag2-/- mice treated with IL-4 remained disease
free, and only one mouse had very mild colitis. Histological analysis
failed to show obvious increases in mononuclear or polynuclear
infiltrates in the colons of IL-4-treated
Rag2-/- mice (Fig. 2, E and
F). To determine whether there were changes in macrophage or
dendritic cell populations, immunohistochemical analyses of frozen
sections from colon tissue were performed. No difference in the
expression of F4/80, CD11b, CD11c, or CD4 was found in colons from
IL-4-treated nonreconstituted Rag2-/- mice
compared with saline-treated controls (data not shown). However, there
was a consistent increase in MHC class II expression in the lamina
propria (LP) of IL-4-treated nonreconstituted
Rag2-/- mice (Fig. 6). Furthermore, cotreatment of
Rag2-/- with IL-10 suppressed this increase in
MHC class II expression in the LP by IL-4 (Fig. 6). Thus, IL-4
treatment may promote colitis in the presence of T cells by increasing
Ag presentation by resident macrophages and/or dendritic cells.
Treatment of nonreconstituted Rag2-/- mice with
IL-4 did cause a 3-fold increase in spleen size (Table I) due to
extramedullary hemopoiesis, as has been previously described by others
(48). The splenomegaly was not significantly altered by
cotreatment with IL-10. As IL-10 effectively blocked the exacerbation
of colitis and the increased MHC class II expression in the LP, the
development of extramedullary hemopoiesis and splenomegaly appeared to
be a distinct effect of IL-4 treatment. These data show that IL-4 can
have effects on hemopoietic cells in the absence of T or B cells, but
the development of colitis is strictly T cell dependent in this
model.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The conclusion that IL-4 treatment promoted a Th1-type inflammatory
response is further supported by the greatly enhanced gene expression
of IFN-, MIG, IP-10, and CXCR3 in the colons of IL-4-treated mice.
Furthermore, anti-IL-12 mAb cotreatment reduced the expression of
Th1-associated genes as well as the severity of disease. MIG and IP-10
are both produced by monocytes and lymphocytes in response to IFN-.
MIG has chemotactic activity for activated T cells, while IP-10 is a
chemoattractant for activated CD4+ T cells,
monocytes, and NK cells (52). CXCR3 is the receptor for
both MIG and IP-10 and has been shown to be expressed on Th1-type
CD4+ T cells, but not Th2-type cells
(41). In addition, of the strictly Th2-associated
molecules examined, only the expression of IL-4 was increased. The
increase in expression of CCR8 could be due to an influx of Th2-type T
cells or monocytes into the colon (42, 43). Because the
expressions of CCR4 and CCR3, which are specific to Th2 cells, were not
also up-regulated, we favor the possibility that the increased
expression of CCR8 was by monocytes. Together, these data indicate that
the exacerbation of colitis by IL-4 treatment is due to an influx of
Th1-type T cells and, potentially, monocytes.
The ability of IL-4 to exacerbate a T cell transfer model of colitis is
contrary to the observations by others that IL-4 gene transfer
significantly ameliorated inflammation in a rat model of TNBS-induced
colitis (34). Possible explanations for the disparity in
the results with IL-4 treatment include differences in the animals used
(rats vs mice), induction of disease (chemical vs T cell), and method
of administration of IL-4 (adenoviral vector vs daily injections of
protein). Another critical difference may be the dose of IL-4 received.
For example, we observed that saline-treated recipients of
IL-4R-/- T cells developed more severe
colitis than saline-treated recipients of either WT or
IL-10-/- T cells (compare Figs. 1 and 5). This
observation could be explained by the hypothesis that low amounts of
endogenous IL-4 are inhibitory to the development of Th1-type
inflammatory responses, but that higher amounts of IL-4 augment those
same responses. An alternative explanation is the strain differences
between the IL-4R-/- donor mice (BALB/c) and
the WT and IL-10-/- donor mice (129 SvEv).
Therefore, IL-4 may have either suppressive or proinflammatory effects,
depending on the dose, genetic background, and cell types present at
the site of inflammation.
The proinflammatory effects of IL-4 on Th1 responses in vivo appear to
contradict considerable evidence that IL-4 inhibits such responses in
vitro. IL-4 is known to enhance T cell proliferation, to drive the
differentiation of naive CD4+ T cells toward a
Th2 phenotype, and to suppress the production of IFN- by
CD4+ T cells (46). IL-4 also has
been shown to inhibit IFN--induced responses by macrophages and
their production of proinflammatory molecules, such as IL-1, IL-5,
IL-12, GM-CSF, and TNF- (45, 46). However, under
certain in vitro conditions, IL-4 has been shown to possess
proinflammatory activities on APC. Monocytes that have been pretreated
with IL-4 for an extended period actually have enhanced, rather than
suppressed, IL-12 production in response to LPS or Staphylococcus
aureus (53). IL-4 also has been shown to act
synergistically with IL-12 in vitro to stimulate IFN- secretion by
splenic dendritic cells (54). Furthermore, IL-4 inhibits
IL-10 production by monocytes and up-regulates MHC class II, B7.1, and
B7.2 expression on APC (45, 46).
The results of our studies in vivo support observations that IL-4 can
have proinflammatory effects on accessory cells. For example, IL-4
treatment was able to moderately exacerbate colitis when the T cells
could not respond directly to IL-4. In parallel studies, we observed an
increase in MHC class II expression on cells in the colonic LP of
IL-4-treated nonreconstituted Rag2-/- mice,
which is in agreement with the ability of IL-4 to up-regulate MHC class
II expression on monocytes in vitro (46). In addition,
IL-10 was able to block the IL-4-induced increased MHC class II
expression in the LP and, more importantly, abrogated the ability of
IL-4 to exacerbate colitis upon T cell transfer. It is also possible
that IL-10 may be able to block enhanced IL-12 production by APC
(45). Nevertheless, our studies with
IL-4R-/- T cells show that IL-4 may have
some direct proinflammatory effect on Th1 cells. It is also likely that
IL-4 has proinflammatory effects on nonhemopoietic cells. Colonic
epithelial cells express IL-4R and can increase ICAM-1 expression in
response to IL-4, and thus may contribute to the exacerbation of
disease in this model (47, 55).
While there is much evidence to support the role of IL-4 in allergic
Th2-type responses, the role of IL-4 in the development of an
IL-12-dependent, IFN--producing immune response in vivo needs to be
more fully elucidated. In particular, which microenvironmental
conditions cause IL-4 to favor a Th2 vs Th1 inflammatory response
remains to be determined. By developing a better understanding of the
interaction of Th1-associated and Th2-associated cytokines, more
effective therapeutics for chronic inflammatory diseases can be
designed.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Madeline M. Fort, DNAX Research Institute, 901 California Avenue, Palo Alto, CA 94304.
3 Abbreviations used in this paper: WT, wild type; IL-10-/-, IL-10-deficient, IL-4R-/-, IL-4R -chain-deficient; Rag2-/-, recombinase-activating gene-2-deficient; TNBS, trinitrobenzenesulfonic acid; IP-10; rRNA, ribosomal RNA; NMS, normal mouse serum; LP, lamina propria.
Received for publication July 14, 2000. Accepted for publication December 8, 2000.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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  J. Barbi, F. Brombacher, and A. R. Satoskar T Cells from Leishmania major-Susceptible BALB/c Mice Have a Defect in Efficiently Up-Regulating CXCR3 upon Activation J. Immunol., October 1, 2008; 181(7): 4613 - 4620. [Abstract] [Full Text] [PDF]
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 G. Yao, W. Chen, H. Luo, Q. Jiang, Z. Xia, L. Zang, J. Zuo, X. Wei, Z. Chen, X. Shen, et al. Identification of core functional region of murine IL-4 using peptide phage display and molecular modeling Int. Immunol., January 1, 2006; 18(1): 19 - 29. [Abstract] [Full Text] [PDF]
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 G. Bamias, M. R. Nyce, S. A. De La Rue, and F. Cominelli New Concepts in the Pathophysiology of Inflammatory Bowel Disease Ann Intern Med, December 20, 2005; 143(12): 895 - 904. [Full Text] [PDF]
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 B. Wei, P. Velazquez, O. Turovskaya, K. Spricher, R. Aranda, M. Kronenberg, L. Birnbaumer, and J. Braun Mesenteric B cells centrally inhibit CD4+ T cell colitis through interaction with regulatory T cell subsets PNAS, February 8, 2005; 102(6): 2010 - 2015. [Abstract] [Full Text] [PDF]
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 C. Van Kampen, J. Gauldie, and S. M. Collins Proinflammatory properties of IL-4 in the intestinal microenvironment Am J Physiol Gastrointest Liver Physiol, January 1, 2005; 288(1): G111 - G117. [Abstract] [Full Text] [PDF]
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 K. Kitamura, Y. Nakamoto, S. Kaneko, and N. Mukaida Pivotal roles of interleukin-6 in transmural inflammation in murine T cell transfer colitis J. Leukoc. Biol., December 1, 2004; 76(6): 1111 - 1117. [Abstract] [Full Text] [PDF]
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Y. P. de Jong, S. T. Rietdijk, W. A. Faubion, A. C. Abadia-Molina, K. Clarke, E. Mizoguchi, J. Tian, T. Delaney, S. Manning, J.-C. Gutierrez-Ramos, et al. Blocking inducible co-stimulator in the absence of CD28 impairs Th1 and CD25+ regulatory T cells in murine colitis Int. Immunol., February 1, 2004; 16(2): 205 - 213. [Abstract] [Full Text] [PDF]
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H. Liu, B. Hu, D. Xu, and F. Y. Liew CD4+CD25+ Regulatory T Cells Cure Murine Colitis: The Role of IL-10, TGF-{beta}, and CTLA4 J. Immunol., November 15, 2003; 171(10): 5012 - 5017. [Abstract] [Full Text] [PDF]
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U. P. Singh, S. Singh, D. D. Taub, and J. W. Lillard Jr. Inhibition of IFN-{gamma}-Inducible Protein-10 Abrogates Colitis in IL-10-/- Mice J. Immunol., August 1, 2003; 171(3): 1401 - 1406. [Abstract] [Full Text] [PDF]
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 M. H. Myles, R. S. Livingston, B. A. Livingston, J. M. Criley, and C. L. Franklin Analysis of Gene Expression in Ceca of Helicobacter hepaticus-Infected A/JCr Mice before and after Development of Typhlitis Infect. Immun., July 1, 2003; 71(7): 3885 - 3893. [Abstract] [Full Text] [PDF]
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 J. H. Bream, R. E. Curiel, C.-R. Yu, C. E. Egwuagu, M. J. Grusby, T. M. Aune, and H. A. Young IL-4 synergistically enhances both IL-2- and IL-12-induced IFN-{gamma} expression in murine NK cells Blood, July 1, 2003; 102(1): 207 - 214. [Abstract] [Full Text] [PDF]
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C. S. Bonder and P. Kubes The Future of GI and Liver Research: Editorial Perspectives: II. Modulating leukocyte recruitment to splanchnic organs to reduce inflammation Am J Physiol Gastrointest Liver Physiol, May 1, 2003; 284(5): G729 - G733. [Abstract] [Full Text] [PDF]
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G. Das, M. M. Augustine, J. Das, K. Bottomly, P. Ray, and A. Ray An important regulatory role for CD4+CD8alpha alpha T cells in the intestinal epithelial layer in the prevention of inflammatory bowel disease PNAS, April 29, 2003; 100(9): 5324 - 5329. [Abstract] [Full Text] [PDF]
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A. Finnegan, M. J. Grusby, C. D. Kaplan, S. K. O'Neill, H. Eibel, T. Koreny, M. Czipri, K. Mikecz, and J. Zhang IL-4 and IL-12 Regulate Proteoglycan-Induced Arthritis Through Stat-Dependent Mechanisms J. Immunol., September 15, 2002; 169(6): 3345 - 3352. [Abstract] [Full Text] [PDF]
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H. P. Jones, L. Tabor, X. Sun, M. D. Woolard, and J. W. Simecka Depletion of CD8+ T Cells Exacerbates CD4+ Th Cell-Associated Inflammatory Lesions During Murine Mycoplasma Respiratory Disease J. Immunol., April 1, 2002; 168(7): 3493 - 3501. [Abstract] [Full Text] [PDF]
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F. J. D. Mennechet, L. H. Kasper, N. Rachinel, W. Li, A. Vandewalle, and D. Buzoni-Gatel Lamina Propria CD4+ T Lymphocytes Synergize with Murine Intestinal Epithelial Cells to Enhance Proinflammatory Response Against an Intracellular Pathogen J. Immunol., March 15, 2002; 168(6): 2988 - 2996. [Abstract] [Full Text] [PDF]
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J. Major, J. E. Fletcher, and T. A. Hamilton IL-4 Pretreatment Selectively Enhances Cytokine and Chemokine Production in Lipopolysaccharide-Stimulated Mouse Peritoneal Macrophages J. Immunol., March 1, 2002; 168(5): 2456 - 2463. [Abstract] [Full Text] [PDF]
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 N. Iqbal, J. R. Oliver, F. H. Wagner, A. S. Lazenby, C. O. Elson, and C. T. Weaver T Helper 1 and T Helper 2 Cells Are Pathogenic in an Antigen-specific Model of Colitis J. Exp. Med., January 7, 2002; 195(1): 71 - 84. [Abstract] [Full Text] [PDF]
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B. Cautain, J. Damoiseaux, I. Bernard, E. Xystrakis, E. Fournie, P. van Breda Vriesman, P. Druet, and A. Saoudi The CD8 T Cell Compartment Plays a Dominant Role in the Deficiency of Brown-Norway Rats to Mount a Proper Type 1 Immune Response J. Immunol., January 1, 2002; 168(1): 162 - 170. [Abstract] [Full Text] [PDF]
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, saline-treated animals; 
,
IL-4-treated animals. Data are representative of two separate
experiments, with each bar representing the average of four or five
mice. *, Significantly different from saline-treated mice
(p < 0.05).
