Sunteți pe pagina 1din 8

Immunopharmacology 49 2000.

317324
www.elsevier.comrlocaterimmpharm

Modulation of collagen-induced arthritis by IL-4 and


dexamethasone: the synergistic effect of IL-4 and dexamethasone
on the resolution of CIA
Insoo Kang a,) , Won Woo Lee b, Yong-Soon Lee b
a

Department of Internal Medicine, College of Medicine, Hallym Uniersity, South Korea


b
College of Veterinary Medicine, Seoul National Uniersity, Seoul, South Korea
Received 20 March 2000; received in revised form 2 May 2000; accepted 3 May 2000

Abstract
We investigated effects of IL-4, dexamethasone DXM., and the combination of IL-4 and DXM, low- or high-dose, on
collagen-induced arthritis CIA. in DBAr1 mice and correlated severity of arthritis with changes in IL-10 and IFN-g.
Compared with control mice, mice treated with IL-4 had increased IL-10 with the same degree of arthritis, whereas mice
treated with high-dose DXM had decreased IL-10 and increased IFN-g production with less severe arthritis. Mice treated
with low-dose DXM showed the absence of IL-10 and increased IFN-g production with a trend toward the resolution of
arthritis. Mice treated with IL-4 and low-dose DXM had neither IL-10 nor IFN-g production but revealed less severe
arthritis, compared with mice treated with low-dose DXM alone. These results suggest that the beneficial effects of
high-dose DXM and the combination of IL-4 and DXM on CIA are independent of IL-10 and IFN-g. q 2000 Elsevier
Science B.V. All rights reserved.
Keywords: CIA; Steroid; IL-4

1. Introduction
Rheumatoid arthritis RA. is an autoimmune disease characterized by chronic inflammation in multiAbbreiations: RA, rheumatoid arthritis; CIA, collagen-induced arthritis; CFA, complete Freunds adjuvant; DXM, dexamethasone; IL, interleukin; IFN, interferon; TNF, tumor necrosis
factor; Th, T heler; SC, spleen cell; IFA, incomplete Freunds
adjuvant; PBS, phosphate-buffered saline; ELISA, enzyme-linked
immunoabsorbent assay
)
Corresponding author. 609 LCI Section of Rheumatology,
Department of Internal Medicine, Yale University School of
Medicine, 333 Cedar St., New Haven, CT 06520, USA. Tel.:
q1-7203-785-2454; fax: q1-7203-785-7053.
E-mail address: insoo.kang@yale.edu I. Kang..

ple joints with subsequent destruction of cartilage


and erosion of bone with extra-articular manifestations. Harris, 1996.. While this disease is not an
uncommon condition, with a prevalence of 1%
worldwide, its pathogenesis is not clear yet. Collagen-induced arthritis CIA. in DBAr1 mice has
been used as an experimental model of rheumatoid
arthritis, as it shares many of the histological and
immunological characteristics found in RA Holmdahl et al., 1990.. Intradermal injection of chicken or
bovine type II collagen emulsified in complete Freunds adjuvant CFA. into DBAr1 mice induces an
inflammatory polyarthritis in the articular joints,
characterized by hyperplasia of the synovium with
dense infiltration of neutrophils and mononuclear

0162-3109r00r$ - see front matter q 2000 Elsevier Science B.V. All rights reserved.
PII: S 0 1 6 2 - 3 1 0 9 0 0 . 0 0 2 4 8 - 4

318

I. Kang et al.r Immunopharmacology 49 (2000) 317324

cells. In severe cases, destruction of cartilage and


bony erosions are observed. The development of
CIA in DBAr1 mice requires both B and T cells
Seki et al., 1988; Taylor et al., 1995.. Several
studies showed the importance of cytokines in the
pathogenesis of CIA by depleting or administering a
specific cytokine in mice with CIA Bessis et al.,
1996; Boissier et al., 1995; Kasama et al., 1995;
Yoshino, 1998.. Blockades of IL-1 or TNF-a by
administrating of cytokine-specific monoclonal antibodies or cytokine receptor antagonists to mice with
CIA alleviated arthritis, whereas injection of these
cytokines in mice with CIA aggravated arthritis Hom
et al., 1988; Piguet et al., 1992; Wooley et al., 1993..
Based on cytokine secretion patterns, T-helper
Th. cells can be divided into three subsets. Th1 cells
predominantly secrete the pro-inflammatory cytokines IL-2 and IFN-g, whereas Th2 cells secrete
anti-inflammatory cytokines such as IL-4, IL-5, IL-10
and IL-13, and Th0 cells secrete both IFN-g and
IL-4 Mosmann and Sad, 1996.. In CIA, it was
reported that mice developed a Th0rTh1 dominant
response 15 days after immunization, while T cells
produced predominantly IL-4, a Th2 cytokine, 30
days after the first immunization, a time point just
before the onset of arthritis Doncarli et al., 1997..
This suggests the potential role of Th1rTh2 cytokines in the pathogenesis of CIA. In fact, administration of IL-4 to mice with CIA accelerated the
resolution of arthritis when it was given during the
late stage of arthritis only or in combination with
IL-10 Joosten et al., 1997a., whereas its administration did not affect arthritis when it was injected
during the early active stage of arthritis Marcelletti
et al., 1991.. These findings indicate that IL-4 alone
is not sufficient to alter the course of CIA during the
active stage of the disease and that it may need an
additional factor for the resolution of arthritis. Glucocorticoids, potent anti-inflammatory drugs commonly used for the treatment of RA, could be drugs
that are able to enhance the anti-inflammatory effect
of IL-4. Anti-inflammatory actions of glucocorticoids are mediated by several different mechanisms
including the modulation of Th1rTh2 cytokine balance Wilckens and De Rijk, 1997.. The effect of
glucocorticoids on Th1rTh2 immune responses does
not appear to consist simply in inducing a Th1 or
Th2 immune response but is more complicated

Wilckens and De Rijk, 1997.. In fact, glucocorticoids can suppress or enhance IL-4 and IL-10 productions depending on the stage of T-cell development Brinkmann and Kristofic, 1995..
In this study, we investigate the synergistic effect
of glucocorticoids and IL-4 on the resolution of CIA
by measuring the macroscopic and microscopic
severity of arthritis. Changes in the Th1rTh2 cytokine balance were also studied by measuring IL-10
and IFN-g productions in spleen cells SCs. of these
mice, and correlating them with the severity of
arthritis. We report that the systemic administration
of dexamethasone DXM. and IL-4 synergistically
accelerates the resolution of CIA, which may be due
to blocking of DXM-induced IFN-g production by
IL-4.

2. Materials and methods


2.1. Mice
Inbred 7- to 8-week-old male DBAr1 mice were
purchased from Charles River Japan and housed at
the Seoul National University College of Veterinary
Medicine animal facility. Mice were fed standard
mouse food and water ad libitum.
2.2. Induction and gross assessment of CIA
Immunization and arthritis evaluation were performed as described in detail with some modifications Doncarli et al., 1997; Joosten et al., 1997b..
DBAr1 mice were injected intradermally at the base
of the tail with 100 mg type II bovine collagen
Chondrex, Seattle, USA. emulsified in CFA and
boosted with 100 mg type II bovine collagen in
incomplete Freunds adjuvant IFA. on day 21 after
the first immunization. To accelerate the development of arthritis, 40 mg of LPS Sigma, USA. in
sterile PBS was intraperitoneally injected in mice
that had not developed arthritis until day 28. Mice
were observed daily for the presence of arthritis and
the clinical severity of arthritis was scored for each
paw on a scale of 0 to 4: 0 s normal; 1 s mild
swelling and erythema; 2 s moderate swelling and
erythema; 3 s severe swelling and erythema of entire paw including digits; 4 s ankylosis. Because of a

I. Kang et al.r Immunopharmacology 49 (2000) 317324

variation in the total number of arthritic joint in each


mouse, we followed the change of arthritis arthritic
score. in each limb like in another study Lubberts et
al., 1999..
2.3. Treatment regimens
At day 32 after the first immunization, mice
developing arthritis in at least one joint were selected
and divided into four groups. Mice injected with LPS
were equally assigned to each group. All injections
in mice were done intraperitoneally for 5 days. Mice
in group A received PBS as a negative control and
group B received 200 ng of IL-4 Pharmingen, San
Diego, USA.. Mice in group C received DXM
Sigma, USA. 0.25 mgrkg high-dose experiment.
or 0.025 mgrkg low-dose experiment.. Mice in
group D had both IL-4 and DXM with the same
dosage as for groups B and C. The experiment was
done separately for high-dose DXM treatment and
low-dose DXM treatment with a different batch of
mice. All mice were sacrificed and bled to obtain
sera by cardiac puncture on the day after the last
dose of treatment. Spleens were harvested and pooled
for cytokine analysis like a previous study for cytokine analysis in lymph nodes Kang et al., 1997.,
and front paws and hind limbs were submitted to
histopathological examinations.
2.4. Cytokine analysis
Spleens were processed into single-cell suspensions in Clicks medium supplemented with 5% fetal
bovine serum, 0.04 mM 2-mercaptoethanol, 200 mM
L-glutamine, 100 U of penicillinrml, and 100 mg of
streptomycinrml 5% Clicks medium.. SCs were
washed with 5% Clicks medium, resuspended in
Clicks medium with 10% fetal calf serum and supplements 10% Clicks medium. at 5 = 10 6 cellsrml,
and aliquoted into a 24-well total volume of 1
mlrwell. tissue culture plate Nalge Nunc International, Rochester, New York.. SCs were stimulated
with T-cell proliferation assay-grade bovine type II
collagen Chondrex, Seattle, USA. at 30 mgrml,
with an equivalent volume of PBS as a negative
control, or with concavalin A at 10 mgrml as a
positive control. SCs were incubated at 378C, 5%
CO 2 for 72 h and supernatants were harvested. IL-10

319

and IFN-g in supernatants were measured by the


sandwich enzyme-linked immunosorbent assay
ELISA. as specified by the manufacturer Serotec,
UK.. Concentrations of IL-4 and IFN-g were calculated based on standard curves obtained from serial
dilutions of recombinant IL-4 and IFN-g Serotec,
UK..
2.5. Histopathological testing
The joints of front paws and hind limbs were
immersion fixed in neutral-buffered formalin pH
7.2., demineralized, and then processed and stained
with hematoxylineosin by routine histological technique Kang et al., 1997.. The joints were scored for
arthritis severity on a scale of 0 normal. to 3
severe. based on the presence of pannus, and the
degree of leukocytic infiltration, cartilage destruction
and bony erosion, by an observer blinded to the
source of slides.

3. Results
3.1. Deelopment of CIA
In our experiments, about 20% of mice had developed CIA before the injection of LPS at day 28 after
the first immunization with bovine type II collagen.
The subsequent LPS injection to mice without CIA
at day 28 increased the incidence of CIA to 95% in
mice.
3.2. Effects of high-dose DXM and IL-4 on CIA
On macroscopic examination, mice treated with
IL-4 had the same degree of arthritic index as mice
treated with PBS throughout the period of treatment
p ) 0.05, Fig. 1a., whereas mice treated with highdose DXM showed a significant reduction in arthritic
index mean " S.D.. at days 2 0.563 " 0.727., 3
0.500 " 0.730. and 4 0.375 " 0.500. of treatment,
compared with mice treated with PBS at the same
time points p - 0.05, Fig. 1a.. Also, mice treated
with both IL-4 and high-dose DXM had less severe
arthritis at days 3 0.438 " 0.629. and 4 0.375 "
0.619. than did mice treated with PBS and mice
treated with IL-4 alone p - 0.05, Fig. 1a.. No

320

I. Kang et al.r Immunopharmacology 49 (2000) 317324

mean " S.D. in mice treated with IL-4 1.159 "


1.167. was not significantly lower than that in mice
treated with PBS 1.500 " 1.269, p ) 0.05, Fig. 1b..
The histopathological index significantly decreased
in mice treated with high-dose DXM 0.136 " 0.351.,
and mice treated with the combination of IL-4 and
high-dose DXM 0.222 " 0.428., compared with
mice treated with PBS 1.500 " 1.269, p - 0.05,
Fig. 1b.. The histopathological index of mice treated
with the combination of IL-4 and high-dose DXM
was not lower than that of mice treated with highdose DXM alone Fig. 1b.. Together, the results
show that administration of high-dose DXM but not
of IL-4 can accelerate the resolution of CIA, and that
adding IL-4 to high-dose DXM has no synergistic
effect on the resolution of CIA.

3.3. Effects of low-dose DXM and IL-4 on CIA


Fig. 1. Effects of IL-4, high-dose DXM 0.25 mgrkg., and the
combination of IL-4 and high-dose DXM on the severity of
arthritis measured by the arthritic index a. and the histopathological index b.. Statistical analysis was done with the MannWhitney test. a. In mice treated with high-dose DXM, the arthritic
index was significantly lower than in mice treated with PBS at
days 2, 3, and 4 of treatment as indicated by single asterisk
p- 0.05 for each time point.. b. The histopathological index
was significantly lower in mice treated with high-dose DXM than
in mice treated with PBS at day 5 of treatment p- 0.05.. The
letter n indicates the number of limbs a. or joints b. observed in
each group. PBS group had four mice and other groups had five
mice per group. Results are expressed as the mean"SD a. or the
meanqSD b..

difference in the arthritic index was noticed between


mice treated with DXM alone and mice treated with
IL-4 and high-dose DXM. At day 5 of treatment,
mice treated with high-dose DXM and mice treated
with both IL-4 and high-dose DXM had lower
arthritic indices 0.3750 " 0.7190 and 0.3130 "
0.4790, respectively. than mice treated with IL-4
alone 1.603 " 1.1240. or PBS 1.200 " 1.2290., but
this difference was not statistically significant p s
0.056 for PBS vs. DXM, p s 0.053 for PBS vs.
combination, Fig. 1a.. The histopathological severity
in the joints of four groups of mice correlated with
macroscopic findings. The histopathological index

In separate experiments, where mice treated with


low-dose DXM alone had a trend to show a decrease
in the arthritic index, this was not significantly lower
than that in mice treated with PBS p ) 0.05, Fig.
2a.. Like the high-dose DXM and IL-4 experiment,
IL-4 alone was not more effective than PBS in
reducing the severity of arthritis p ) 0.05, Fig. 2a..
However, at days 3 1.200 " 0.862., 4 0.866 "
0.516. and 5 0.733 " 0.594. of treatment, mice
treated with the combination of IL-4 and low-dose
DXM had lower arthritic indices than did mice treated
with PBS. At day 5 of treatment, a statistically
significant difference in arthritic indices was noticed
between mice treated with low-dose DXM alone
1.154 " 0.594., and mice treated with the combination of IL-4 and low-dose DXM 0.733 " 0.594,
p s 0.0038, Fig. 2a., showing a synergistic effect on
the resolution of CIA in DBAr1 mice. The results of
the histopathological examination on arthritic joints
correlated with the macroscopic findings. Both
groups of mice treated with only IL-4 or DXM did
have the same degree of arthritis compared with
mice treated with PBS, whereas mice treated with
the combination of IL-4 and low-dose DXM had less
severe arthritis 1.188 " 0.795. than mice treated
with PBS 2.050 " 0.887. or low-dose DXM alone
1.800 " 0.696, p - 0.05, Fig. 2b.. These results

I. Kang et al.r Immunopharmacology 49 (2000) 317324

321

by high levels of IL-10 and low levels of IFN-g Fig.


3a.. In spite of the absence of resolution of arthritis
in mice treated with IL-4, administration of IL-4
increased the level of IL-10 and decreased the level
of IFN-g in high-dose DXM experiment Fig. 3a and

Fig. 2. Effects of IL-4, low-dose DXM 0.025 mgrkg., and the


combination of IL-4 and low-dose DXM on the severity of
arthritis measured by the arthritic index a. and the histopathological index b.. The statistical analysis was done with the Mann
Whitney test. a. The arthritic index was significantly lower in
mice treated with the combination of IL-4 and low-dose DXM
than in mice treated with DXM at day 5 of treatment as indicated
by double asterisks p- 0.05.. b. In mice treated with the
combination of IL-4 and DXM at day 5 of treatment p- 0.05..
The letter n indicates the number of limbs a. or joints b.
observed in each group. Each group had four mice. Results are
expressed as the mean"SD a..

show that the combined treatment with IL-4 and


low-dose DXM synergistically accelerates the resolution of CIA.
3.4. Correlation of arthritis seerity with cytokine
profiles
To correlate the severity of arthritis with cytokine
profiles, the IL-10 and IFN-g productions from SCs
stimulated for 72 h in the presence of type II bovine
collagen were measured. No IL-10 or IFN-g was
detected in SCs stimulated with PBS, and IL-10
andror IFN-g was detected in SCs stimulated with
concavalin A data not shown.. In the high-dose
DXM experiment, mice treated with PBS showed
predominantly a Th2 immune response characterized

Fig. 3. IL-10 and IFN-g productions by spleen cells in mice


treated with IL-4, high- a. or low-dose b. DXM, and the
combination of IL-4 and high a. or low-dose b. DXM. Experiments for graph A high-dose DXM. and graph B low-dose
DXM. were done separately with a different batch on mice at a
different time. DBAr1 mice with CIA had Th2 predominant
immune response characterized by high levels of IL-10 and low
levels a. or absence b. of IFN-g production. In mice treated with
the combination of IL-4 and high-dose a and b.. a. Treatment of
mice with high-dose DXM decreased IL-10 production but increased IFN-g production. In mice treated with the combination
of IL-4 and high-dose DXM, the level of IL-10 decreased and
IFN-g production was below the level of detection by our assay.
a. Treatment of mice with low-dose DXM reduced IL-10 production below the level of detection but increased IFN-g production.
No IL-10 of IFN-g was detected in mice treated with the combination of IL-4 and low-dose DXM by our assay. Spleens were
harvested on the day after day 5 of treatment, pooled and restimulated for 72 h in the presence of bovine type II collage. Five mice
per group except PBS group with four mice for experiment A and
four mice per group for experiment B. The bars represent the
mean results from duplicate wells, as determined by ELISA,
qstandard error of the mean. The levels of detection were 156
pgrml for IL-10 and 56 pgrml for IFN-g.

322

I. Kang et al.r Immunopharmacology 49 (2000) 317324

b., whereas mice treated with high-dose DXM, which


showed significant resolution of arthritis, had lower
levels of IL-10 and higher levels of IFN-g than did
mice treated with PBS Fig. 3a.. In mice treated with
the combination of IL-4 and high-dose DXM, the
level of IL-10 was lower than in mice treated with
IL-4 but higher than in mice treated with DXM,
while no IFN-g production was detected Fig. 3a..
Despite differences in cytokine profiles between mice
treated with high-dose DXM and mice treated with
both IL-4 and DXM, both groups of mice had the
same severity of arthritis Fig. 1a and b.. Similar to
the high-dose DXM experiment, mice treated with
PBS in the low-dose DXM experiment, separately
done with a different batch of mice at a different
time, had a predominant Th2 immune response with
the presence of IL-10 and the absence of IFN-g Fig.
3b.. IFN-g but not IL-10 was detected in mice
treated with low-dose DXM, which showed a trend
of decrease in the severity of arthritis Fig. 2a and b..
Despite accelerated resolution of arthritis in mice
treated with both IL-4 and low-dose DXM, neither
IL-4 nor IFN-g was detected Fig. 3b..

4. Discussion
In this study, we investigated effects of IL-4,
DXM, and the combination of IL-4 and DXM on
CIA in DBAr1 mice and correlated the macroscopic
and microscopic severity of arthritis with changes in
the Th1 and Th2 immune responses by measuring
IL-10 and IFN-g productions from SCs. During the
active phase of CIA, mice treated with PBS had a
Th2 predominant immune response characterized by
high level of IL-10 and low level or absence of
IFN-g in spite of active inflammation in joints. This
is consistent with the finding by Doncarli et al.
1997., who showed that CD4 q T cells in type II
collagen-immunized DBAr1 mice converted from a
dominant Th0rTh1 immune response to a Th2 immune response during the development of CIA. The
significance of this Th2 immune response in the
pathogenesis of CIA is not completely understood,
but some reports claimed that injections of IL-4
accelerated the resolution of arthritis and that a
neutralization of IL-4 or IL-10 with monoclonal

antibodies aggravated arthritis Kasama et al., 1995;


Marcelletti et al., 1991;Yoshino, 1998.. However,
not all studies with IL-4 injections had positive
results in CIA. The timing of IL-4 administration
appeared a critical factor in determining the effect of
IL-4 on CIA since mice treated with IL-4 during the
late stage of CIA, but not during the early stage,
accelerated resolution of CIA compared with control
mice Joosten et al., 1997a; Marcelletti et al., 1991..
This is consistent with the findings of our study
where mice treated with IL-4 during the first 10 days
after the onset of CIA did not show an improvement
in CIA compared with control mice, indicating that
administration of IL-4, with subsequent increase in
IL-10, does not have an immediate effect on CIA
during the active phase of disease. In our study, as
expected, mice treated with high-dose DXM showed
accelerated resolution of CIA on macroscopic and
microscopic exams, compared with mice treated with
PBS. However, the cytokine profile in these mice
characterized by a higher level of IFN-g and a lower
level of IL-10 than in mice treated with PBS was
somewhat surprising since IFN-g, a prototype Th1
cytokine, has a strong pro-inflammatory property.
Effects of glucocorticoids on Th1rTh2 immune responses are variable and depend on the status of
T-cell development and disease models Brinkmann
and Kristofic, 1995; Wilckens and De Rijk, 1997..
The benefit of glucocorticoids to both Th1-driven
diseases such as experimental allergic encephalitis,
and Th2-driven diseases such as asthma suggests that
the effect of glucocorticoids on Th1rTh2 does not
follow a simple unidirectional mechanism but is
more complicated. In fact, in effector T cells derived
from the naive subset, glucocorticoids induced the
production of IL-4 and IL-10, but blocked the production of IL-5 and IFN-g, whereas in effector T
cells from the memory subset, glucocorticoids
blocked the production of IL-4, IL-5, and IL-10 but
inhibited the production of IFN-g by only 50%
Brinkmann and Kristofic, 1995.. This could be an
explanation for the findings of our study, showing
suppressed IL-10 and increased IFN-g productions
from bovine type II collagen-restimulated SCs in
mice treated with high or low-dose DXM, which
might be due to the suppression of Th2 cells by
DXM and subsequent activation of Th1 cells. Despite an increase in IFN-g, a pro-inflammatory cy-

I. Kang et al.r Immunopharmacology 49 (2000) 317324

tokine, and a decrease in IL-10, an anti-inflammatory


cytokine, in mice treated with high-dose DXM, the
severity of arthritis decreased. Similar to mice treated
with high-dose DXM, mice treated with low-dose
DXM had a Th1 predominant immune response
characterized by the absence of IL-10 and the presence of IFN-g production from SCs, showing a trend
of improvement in the arthritic index without reaching a statistically significant level. It is not clear
whether DXM-induced IFN-g production is detrimental to CIA, since mice treated with high and
low-dose DXM have showed a statistically significant resolution of arthritis and a trend toward the
resolution of arthritis respectively, compared with
mice treated with PBS. In fact, IFN-g receptor
knock-out DBAr1 mice developed more severe CIA
compared with wild type mice, suggesting no direct
effect of IFN-g on the pathogenesis of CIA Vermeire
et al., 1997.. In our experiment, the improvement of
CIA in mice treated with high or low-dose DXM
was most likely due to other anti-inflammatory effects of glucocorticoids such as suppression of the
phospholipase A2, cyclooxygenase 2, and nitric oxide synthase 2 genes, resulting in a decrease in the
production of prostanoids, platelet activating factor
and nitric oxide, which might be stronger than the
pro-inflammatory effect of IFN-g Boumpas et al.,
1993.. Findings of a predominantly Th1 immune
response in mice treated with low- or high-dose
DXM leading to an improvement of CIA suggest
that the resolution of arthritis by DXM may not
depend on Th1rTh2 immune responses, but on other
anti-inflammatory mechanisms.
The synergistic effect of IL-4 and DXM was
observed in mice treated with a combination of
low-dose DXM and IL-4 but not in mice treated with
high-dose DXM and IL-4, which could be due to the
stronger anti-inflammatory effect of high-dose DXM
than low-dose DXM. From SCs of mice treated with
both IL-4 and low-dose DXM, neither IL-10 nor
IFN-g production was detected by our assay. This
might be due to suppression of IFN-g by the exogenous IL-4, whereas absence of IL-10 could be due to
suppression of IL-10 production by low-dose DXM,
as seen in mice treated with only low-dose DXM,
suggesting the mutual suppression of IL-10 and IFNg by DXM and IL-4 respectively. Similarly, in a
separate experiment, a mutual suppression of IL-10

323

and IFN-g was noticed in mice treated with high-dose


DXM and IL-4.
In this study, we have shown that DBAr1 mice
with CIA have a Th2 predominant immune response
during the active stage of the disease and that exogenous IL-4 injections in these mice do not accelerate
the resolution of CIA despite increase in IL-10, a
Th2 cytokine. On the other hand, high-dose DXM
injections in DBAr1 mice with CIA accelerate the
resolution of CIA, despite an increase in IFN-g and a
decrease in IL-10, suggesting that the modulation of
these cytokines by DXM is not the principal mechanism for the resolution of CIA. Finally, a combination therapy of IL-4 and low-dose DXM in DBAr1
mice with CIA shows the synergistic effect on the
resolution of CIA, and this requires further investigation to find the mechanism.

Acknowledgements
This work was supported by the Hallym University Medical Center Research Grant.

References
Bessis, N., Boissier, M.C., Ferrara, P., Blankenstein, T., Fradelizi,
D., Fournier, C., 1996. Attenuation of collagen-induced arthritis in mice by treatment with vector cells engineered to secrete
interleukin-13. Eur. J. Immunol. 26, 23992403.
Boissier, M.C., Chiocchia, G., Bessis, N., Hajnal, J., Garotta, G.,
Nicoletti, F., Fournier, C., 1995. Biphasic effect of interferongamma in murine collagen-induced arthritis. Eur. J. Immunol.
25, 11841190.
Boumpas, D.T., Chrousos, G.P., Wilder, R.L., Cupps, T.R., Balow,
J.E., 1993. Glucocorticoid therapy for immune-mediated diseases: basic and clinical correlates. Ann. Intern. Med. 119,
11981208.
Brinkmann, V., Kristofic, C., 1995. Regulation by corticosteroids
of Th1 and Th2 cytokine production in human CD4q effector
T cells generated from CD45ROy and CD45ROq subsets. J.
Immunol. 155, 33223328.
Doncarli, A., Stasiuk, L.M., Fournier, C., Abehsira-Amar, O.,
1997. Conversion in vivo from an early dominant Th0rTh1
response to a Th2 phenotype during the development of
collagen-induced arthritis. Eur. J. Immunol. 27, 14511458.
Harris, E.D. Jr., 1996. In: Kelly, W.H., Harris, E.D. Jr., Ruddy,
S., Sledge, C.B. Eds.., Text Book of Rheumatology. Saunders, Philadelphia, pp. 898932.

324

I. Kang et al.r Immunopharmacology 49 (2000) 317324

Holmdahl, R., Andersson, M., Goldschmidt, T.J., Gustafsson, K.,


Jansson, L., Mo, J.A., 1990. Type II collagen autoimmunity in
animals and provocations leading to arthritis. Immunol. Rev.
118, 193232.
Hom, J.T., Bendele, A.M., Carlson, D.G., 1988. In vivo administration with IL-1 accelerates the development of collagen-induced arthritis in mice. J. Immunol. 141, 834841.
Joosten, L.A., Lubberts, E., Durez, P., Helsen, M.M., Jacobs,
M.J., Goldman, M., van den Berg, W.B., 1997a. Role of
interleukin-4 and interleukin-10 in murine collagen-induced
arthritis. Protective effect of interleukin-4 and interleukin-10
treatment on cartilage destruction. Arthritis Rheum. 40, 249
260.
Joosten, L.A., Lubberts, E., Helsen, M.M., van den Berg, W.B.,
1997b. Dual role of IL-12 in early and late stages of murine
collagen type II arthritis. J. Immunol. 159, 40944102.
Kang, I., Barthold, S.W., Persing, D.H., Bockenstedt, L.K., 1997.
T-helper-cell cytokines in the early evolution of murine Lyme
arthritis. Infect. Immun. 65, 31073111.
Kasama, T., Strieter, R.M., Lukacs, N.W., Lincoln, P.M., Burdick, M.D., Kunkel, S.L., 1995. Interleukin-10 expression and
chemokine regulation during the evolution of murine type II
collagen-induced arthritis. J. Clin. Invest. 95, 28682876.
Lubberts, E., Joosten, L.A., van Den Bersselaar, L., Helsen,
M.M., Bakker, A.C., van Meurs, J.B., Graham, F.L., Richards,
C.D., van Den Berg, W.B., 1999. Adenoviral vector-mediated
overexpression of IL-4 in the knee joint of mice with
collagen-induced arthritis prevents cartilage destruction. J. Immunol. 163, 45464556.
Marcelletti, J.F., Ohara, J., Katz, D.H., 1991. Collagen-induced
arthritis in mice. Relationship of collagen-specific and total
IgE synthesis to disease. J. Immunol. 147, 41854191.
Mosmann, T.R., Sad, S., 1996. The expanding universe of T-cell

subsets: Th1, Th2 and more. Immunol. Today 17, 138144,


wsee commentsx.
Piguet, P.F., Grau, G.E., Vesin, C., Loetscher, H., Gentz, R.,
Lesslauer, W., 1992. Evolution of collagen arthritis in mice is
arrested by treatment with anti-tumour necrosis factor TNF.
antibody or a recombinant soluble TNF receptor. Immunology
77, 510514.
Seki, N., Sudo, Y., Yoshioka, T., Sugihara, S., Fujitsu, T., Sakuma,
S., Ogawa, T., Hamaoka, T., Senoh, H., Fujiwara, H., 1988.
Type II collagen-induced murine arthritis I. Induction and
perpetuation of arthritis require synergy between humoral and
cell-mediated immunity. J. Immunol. 140, 14771484.
Taylor, P.C., Plater-Zyberk, C., Maini, R.N., 1995. The role of the
B cells in the adoptive transfer of collagen-induced arthritis
from DBAr1 H-2q. to SCID H-2d. mice. Eur. J. Immunol.
25, 763769.
Vermeire, K., Heremans, H., Vandeputte, M., Huang, S., Billiau,
A., Matthys, P., 1997. Accelerated collagen-induced arthritis
in IFN-gamma receptor-deficient mice. J. Immunol. 158,
55075513.
Wilckens, T., De Rijk, R., 1997. Glucocorticoids and immune
function: unknown dimensions and new frontiers. Immunol.
Today 18, 418424.
Wooley, P.H., Whalen, J.D., Chapman, D.L., Berger, A.E.,
Richard, K.A., Aspar, D.G., Staite, N.D., 1993. The effect of
an interleukin-1 receptor antagonist protein on type II collagen-induced arthritis and antigen-induced arthritis in mice.
Arthritis Rheum. 36, 13051314.
Yoshino, S., 1998. Treatment with an anti-IL-4 monoclonal antibody blocks suppression of collagen-induced arthritis in mice
by oral administration of type II collagen. J. Immunol. 160,
30673071.

S-ar putea să vă placă și