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Departments of
*
Experimental Internal Medicine and
Infectious Diseases, Tropical Medicine and AIDS, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands;
Department of Pathophysiology of Plasma Proteins, Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam, The Netherlands; and
§
Fujisaki Institute, Hayashibara Biochemical Laboratories, Okayama, Japan
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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production. To determine the effect of
IL-10 on IFN- production and related inflammatory responses in
humans, 16 healthy subjects received a bolus i.v. injection of LPS (4
ng/kg) in combination with either placebo or recombinant human IL-10
(25 µg/kg), administered just before or 1 h after LPS. IL-10
treatment, particularly when administered after LPS, enhanced
LPS-induced IFN- release, as well as the release of the
IFN--dependent chemokines IFN--inducible protein-10 and monokine
induced by IFN-, while inhibiting or not influencing the production
of IFN--inducing cytokines. In addition, IL-10 treatment enhanced
activation of CTLs and NK cells after LPS injection, as reflected by
increased levels of soluble granzymes. These data indicate that
high-dose IL-10 treatment in patients with inflammatory disorders can
be associated with undesired proinflammatory
effects. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Recently, it has been suggested that IL-10 has immunostimulatory
properties. In vitro, IL-10 stimulates proliferation and
differentiation of activated human B cells (11), and
preincubation of resting CD4+ lymphocytes with
IL-10 enhances their capacity to produce cytokines after activation
(12). Moreover, IL-10 enhances IL-2-driven proliferation
of preactivated human-purified CD8+ T cells
(13). Addition of IL-10 to mouse splenic NK cell cultures
stimulated with IL-12 and IL-18 results in enhanced IFN- production
(14). In mice, IL-10 injections accelerated graft-vs-host
disease and graft rejection in bone marrow recipients, probably
mediated by augmented IFN- production by T cells
(15).
These data indicate that, under certain conditions, IL-10 can have
stimulatory effects on CD4+,
CD8+ T cells, and/or NK cells, which may result
in increased IFN- production. Knowledge of potential proinflammatory
effects of IL-10 in humans in vivo is highly limited. Such knowledge is
important, because IL-10 has been advocated as a new treatment modality
for several diseases, including Crohn’s disease and rheumatoid
arthritis (16, 17). Therefore, we studied the effect of
IL-10 on IFN- production and related inflammatory responses during
human endotoxemia, a well accepted model of systemic inflammation in
humans.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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This study was performed simultaneously with investigations on the effects of recombinant human (rh)IL-10 on cytokine production, leukocytes, and the hemostatic mechanism, the results of which have been reported elsewhere (10, 18). A total of 16 healthy volunteers (mean 23, range 20–35 years) participated in a double-blind, cross-over, randomized, placebo-controlled study. Written informed consent was obtained from all study participants. The study was approved by the research and ethical committees of the Academic Medical Center. Medical history, physical and routine laboratory examination, chest x-ray, and electrocardiogram of all volunteers were normal. Each volunteer was studied on two occasions after endotoxin (LPS) injection, separated by a wash-out period of 6 wk; on one occasion in combination with placebo, on the other occasion in combination with rhIL-10. The participants were randomized into two groups of eight persons. Group 1 received placebo or rhIL-10 treatment 2 min before LPS, group 2 received placebo or rhIL-10 1 h after LPS administration.
rhIL-10 (Schering-Plough Research Institute, Kenilworth, NJ) was supplied as a sterile powder and reconstituted with sterile water. rhIL-10 was administered by i.v. injection at a dose of 25 µg/kg. The reconstituted placebo powder was identical in appearance and was administered in an identical manner. The LPS preparation, LPS reference standard lot G, Escherichia coli (United States Pharmacopeia Convention, Rockville, MD) was administered at a dose of 4 ng/kg over 1 min in an ante-cubital vein, contralateral to the site of rhIL-10.
Blood samples were drawn directly before LPS injection and at 0.5, 1, 1.5, 2, 3, 4, 5, 6, 8, and 12 h thereafter. Blood was collected in nonadditive vacutainer tubes (Becton Dickinson, Mountain View, CA); after clotting, samples were centrifuged at 2000 x g for 20 min at room temperature, and serum was stored at -70°C until assays were performed.
Whole blood stimulation
In separate experiments, whole blood was collected aseptically
from six healthy donors using a sterile collecting system consisting of
a butterfly needle connected to a syringe (Becton Dickinson).
Anticoagulation was obtained using LPS-free heparin (Leo Pharmaceutical
Products, Weesp, The Netherlands; final concentration 10 U/ml blood).
Whole blood, diluted 1:1 in pyrogen-free RPMI 1640 (BioWhittaker,
Verviers, Belgium), was stimulated for 24 h at 37°C with LPS
(final concentration 10 ng/ml; from E. coli serotype
0111:B4; Sigma, St. Louis, MO) in the presence or absence of increasing
concentrations of rhIL-10 (0.01–1000 ng/ml; Schering-Plough). In
addition, the effect of IL-10 on LPS-induced IFN- production was
studied in the presence or absence of an anti-IL-12 and/or
anti-IL-18 mAb (both mouse IgG, R&D Systems, Abingdon, U.K.; final
concentration both 10 µg/ml). The concentrations of mAbs represent at
least a 1–2 log-unit excess neutralizing capacity over IL-12 and IL-18
concentrations detected after stimulation with LPS (information on the
neutralizing capacities of the mAbs used provided by the manufacturer).
Control mouse IgG (R&D Systems) was used in the appropriate
concentrations. In separate experiments, IL-10 (10 ng/ml) was added at
0.5, 1, 2, or 4 h after the addition of LPS to whole blood. The
effect of IL-10 on IFN- production was also studied during whole
blood stimulation with anti-CD3/anti-CD28 (1:1000; Central
Laboratory of the Netherlands Red Cross Blood Transfusion Service
(CLB), Amsterdam, The Netherlands), the superantigen SEB (1 µg/ml;
Sigma), or PHA (5 µg/ml; HA16; Murex Diagnostics, Dartford, U.K.).
After the incubation, supernatant was obtained after centrifugation and
stored at -20°C until assays were performed.
Assays
All measurements were done in duplicate using specific ELISAs.
Serum levels of IL-12p70 and IL-12p40 were measured as described
previously (19). In short, IL-12 p70 was measured using
anti-IL12 p70 mAb 20C2 as coating Ab, biotinylated anti-IL-12
p40 mAb C8.6 as detecting Ab, and rhIL-12 as standard (detection limit
6 pg/ml). IL-12p40 was measured identically to the IL-12p70 ELISA using
anti-IL-12p40 mAb C11.79 as coating Ab (detection limit 54 pg/ml).
The IL-12p40 ELISA recognizes the total amount of p40, i.e., complexed
as IL-12 heterodimer or as p40 mono- or homodimer. 20C2 was kindly
provided by Dr. M. K. Gately (Hoffmann-La Roche, Nutley, NJ); C8.6
and C11.79 were kindly donated by Dr. G. Trinchieri (The Wistar
Institute, Philadelphia, PA). IL-18 was measured as described
previously (Fujisaki Institute, Okayama, Japan; detection limit 10
pg/ml) (20). IFN- (CLB; detection limit 2.4 pg/ml),
IL-15, and IFN--inducible protein-10 (IP-10) (both R&D Systems; 8.2
and 20 pg/ml, respectively), and monokine induced by IFN- (Mig)
(PharMingen, San Diego, CA; 8.2 pg/ml) were measured according to the
instructions of the manufacturer. Levels of soluble granzyme A (GrA)
and GrB were measured by specific ELISAs exactly as described
previously (21).
Statistical analysis
All data are expressed as mean ± SE. Changes in time were analyzed by one-way ANOVA (p value vs time). Differences between the placebo and the rhIL-10 treatment groups were analyzed by cross-over ANOVA for repeated measures (p value vs placebo). Data of the in vitro experiments were analyzed by Wilcoxon test. p < 0.05 was considered to represent a significant difference.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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and IFN--inducing
cytokines
Administration of LPS induced a transient elevation of IFN-
serum concentrations peaking after 4–5 h (4.8 ± 1.0 pg/ml in
group 1, 12.8 ± 7.8 pg/ml in group 2; both p <
0.05 vs time) (Fig. 1
). Remarkably, IL-10
administration enhanced the release of IFN-, which was more
pronounced for IL-10 posttreatment, resulting in peak levels of
14.4 ± 3.6 pg/ml (group 1) and 28.5 ± 7.4 pg/ml (group 2)
(both p < 0.001 vs placebo).
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is regulated by a coordinated action of a
number of monocyte/macrophage-derived cytokines (22).
IL-12 is a potent inducer of IFN- production, whereas IL-18
synergistically enhances IL-12 effects on IFN- synthesis
(23, 24, 25). In addition, IL-15 and TNF have been identified
as costimuli for optimal IFN- production (26, 27).
Having established that IL-10 enhances LPS-induced IFN- release in
vivo, we next determined the effect of IL-10 on IFN--inducing
cytokines.
IL-12p70 levels remained undetectable after LPS administration in all
but two volunteers. Neither IL-10 pretreatment nor IL-10 posttreatment
influenced IL-12p70 levels (data not shown). LPS injection resulted in
a transient increase in IL-12p40 concentrations peaking at 3 h
after LPS injection (group 1:1.53 ± 0.35 ng/ml, group 2:
1.47 ± 0.23 ng/ml; both p < 0.001 vs time) (Fig. 1
). IL-10 pretreatment completely prevented the LPS-induced increase of
IL-12p40 levels (p < 0.001 vs placebo),
whereas IL-10 posttreatment only partly reduced peak levels of IL-12p40
(0.83 ± 0.13 ng/ml; NS). Serum concentrations of IL-18 were
detectable before LPS injection (group 1: 290 ± 68 pg/ml, group2:
234 ± 45 pg/ml). Administration of LPS did not result in changes
in IL-18 levels in either group (data not shown). Also, IL-10 treatment
did not influence IL-18 concentrations. IL-15 levels were not
detectable before LPS administration and remained undetectable after
LPS injection with or without IL-10 treatment. As reported previously,
IL-10 pretreatment, but not posttreatment, inhibited LPS-induced TNF
release (10). Hence, the stimulatory effect of IL-10 on
IFN- release was not associated with a detectable increase in
IFN--inducing cytokines.
Effect of IL-10 on levels of the IFN--dependent chemokines IP-10
and Mig
To determine whether the IL-10-induced increase in IFN- levels
also resulted in increased IFN- activity, we measured serum
concentrations of IP-10 and Mig, CXC chemokines of which the production
is largely IFN- dependent (28). LPS injection caused a
transient increase in IP-10 peaking after 5 h (group 1: 3.47
± 0.88 ng/ml, group 2: 3.23 ± 0.70 ng/ml; both p
< 0.001 vs time) (Fig. 2). Although
IL-10 pretreatment did not affect IP-10 levels, IL-10 posttreatment
resulted in enhanced IP-10 release with peak levels of 4.77 ±
1.19 ng/ml (p < 0.001 vs placebo). LPS
administration caused increased levels of Mig, which remained elevated
until the end of the study period (12 h: group 1: 2.23 ± 0.41
ng/ml, group 2: 2.36 ± 0.58 ng/ml; both p <
0.001 vs time). IL-10 pretreatment reduced the early increase in Mig
levels between 4–6 h after LPS injection (p <
0.05 vs placebo), whereas levels at 12 h were not different. In
contrast, IL-10 posttreatment resulted in increased levels of Mig from
6 h until the end of the 12-h study period (4.45 ± 0.74
ng/ml; p < 0.05 vs placebo).
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We also studied whether IL-10 treatment was associated with
activation of CD8+ CTLs and/or NK cells, as has
been suggested in previous in vitro studies (13, 14).
Therefore, we measured serum levels of granzymes, proteins that are
released from granules of activated CTL and NK cells (29).
LPS injection induced a transient increase in serum levels of GrA,
peaking after 1.5–3 h (group 1: 30.1 ± 4.2 pg/ml, group 2:
26.2 ± 5.3 pg/ml; both p < 0.05 vs time) (Fig. 3). IL-10 pretreatment did not influence
the early rise in GrA levels, but resulted in increased levels of GrA
from 6 h after LPS until the end of the 12-h study period
(p < 0.001 vs placebo). IL-10 posttreatment
slightly delayed and increased peak levels of GrA (28.2 ± 2.5
pg/ml), and levels remained elevated until the end of the study period
(p < 0.01 vs placebo).
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Effect of IL-10 on LPS-induced production of IFN- and
IFN--inducing cytokines in vitro
In an attempt to study the mechanisms involved in IL-10-induced
enhancement of IFN- production, we determined the effect of IL-10
during whole blood stimulations with LPS, an in vitro system that is
considered to most closely resemble the in vivo situation, leaving
interactions between different cell populations and serum factors
intact (30). Incubation of whole blood for 24 h
without stimulus did not result in detectable levels of IFN-,
IL-12p70, or IL-15, whereas low levels of IL-18 (35.2 ± 5.0
pg/ml) were measured. Incubation with IL-10 alone did not change these
levels (data not shown). LPS stimulated the production of IFN-
(14.9 ± 4.2 ng/ml), IL-12p70 (38.1 ± 13.2 pg/ml), and IL-18
(80.8 ± 7.7 pg/ml) (all p < 0.05 vs incubation
without LPS), whereas IL-15 remained undetectable. Addition of IL-10
resulted in a strong dose-dependent inhibition of LPS-induced IFN-
release (IL-10 1 ng/ml: 2.8 ± 0.4% of IFN- levels measured
after incubation without IL-10; p < 0.05) (Fig. 4A). In addition, IL-10
inhibited the production of IL-12p70 and IL-18 with similar potency
(both p < 0.05) (Fig. 4A). In the in vivo
experiments, the enhancement of IFN- release occurred in the absence
of detectable changes in IL-12p70 and IL-18 levels. In whole blood in
vitro, the IL-10-induced reduction in IL-12p70 and IL-18
concentrations, which is expected to result in a strong inhibition of
IFN- release, could have masked a more direct stimulating effect of
IL-10 on IFN- production. Therefore, we were interested in
determining the effect of IL-10 on IFN- production in whole blood in
the absence of IL-12 and/or IL-18 activity. For this purpose, we
performed whole blood stimulations with LPS and IL-10 in the presence
or absence of an anti-IL-12 mAb and/or an anti-IL-18 mAb.
Although addition of a control Ab did not have any effect, addition of
anti-IL-12 or anti-IL-18 resulted in a strong inhibition of
IFN- release, which was most pronounced for anti-IL-12 (Fig. 4B; both p < 0.05). The combination of
anti-IL-12 and anti-IL-18 resulted in an additional inhibitory
effect (p < 0.05 vs anti-IL-12 only). In
the presence of anti-IL-12 or anti-IL-18, IL-10 still inhibited
LPS-induced IFN- production, although this inhibition was
significantly less in the presence of anti-IL-12 (Fig. 4C). The inhibitory effect of IL-10 was further reduced when
both anti-IL-12 and anti-IL-18 were added
(p < 0.05).
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release, we also studied the effect of
IL-10 added at different time points after the start of incubation of
whole blood with LPS. Addition of IL-10 at 0.5, 1, 2, or 4 h after
LPS still potently inhibited IFN- production, although this effect
was less strong when IL-10 was added 4 h after LPS (data not
shown). The inhibitory effect of IL-10 in vitro was not specific for
LPS-induced IFN- production because IL-10 also inhibited
anti-CD3/anti-CD28, SEB-, or PHA-stimulated IFN- release
(data not shown). |
Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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release, which was
associated with elevated levels of the IFN--dependent chemokines
IP-10 and Mig. The stimulatory effect of IL-10 on IFN- release was
not associated with a detectable increase in IFN--inducing
cytokines. In addition, IL-10 treatment enhanced activation of CTL and
NK cells after LPS injection, as reflected by elevated levels of
soluble GrA and GrB. The stimulatory effect of IL-10 on LPS-induced
IFN- release could not be reproduced in whole blood in vitro, in
which IL-10 dose-dependently reduced IFN- production at least in
part by inhibiting the synthesis of IL-12 and IL-18. These data
indicate that high-dose IL-10 therapy in patients with inflammatory
disorders can be associated with undesired proinflammatory effects
in vivo.
IL-10 is a potent inhibitor of the production of proinflammatory
cytokines including TNF, IL-1
, IL-1ß, IL-6, and IL-8 (1, 4, 9, 10). We demonstrate here that IL-10 stimulates the production
of the proinflammatory cytokine IFN- during human endotoxemia.
IFN- is mainly produced by CD4+ Th1 cells,
CD8+ T cells, and NK cells (22).
IL-12 and IL-18 positively regulate IFN- production (23, 24). IL-12 is the most potent inducer of IFN- synthesis,
whereas IL-18 synergistically enhances the IL-12 effect (25, 31). IL-12 is a heterodimeric cytokine consisting of a p35 and a
p40 subunit (23). Upon stimulation, IL-12p40 is found in
large excess over the biologically active IL-12p70 heterodimer. In our
study, IL-12p70 levels remained undetectable after LPS injection,
whereas IL-12p40 concentrations transiently increased. Conceivably, LPS
induces a rise in circulating IL-12p70 levels only when higher doses
are administered. Indeed, during severe Gram-negative bacteremia in
baboons, the plasma concentrations of both IL-12p70 and IL-12p40
increase, albeit the former to a lesser extent (32). In
mice, IL-12p40 immunoreactivity detected after LPS injection comprises
both p40 monomers and p40 homodimers, the latter compromising 20–40%
of the total p40 produced (33). To our knowledge, it has
not been demonstrated that human IL-12p40 homodimers can be detected
during human endotoxemia. On human T cell lines, IL-12p40 homodimers
have been found to inhibit binding of IL-12 without mediating
biological activity (34). It has been found that murine
IL-12p40 homodimers can function as IL-12 receptor antagonists
(33, 35), but may have immunostimulatory effects on
CD8+ cells, resulting in IFN- production
(36). Because IL-10 strongly reduced LPS-induced IL-12p40
release, it is not likely that the positive effect of IL-10 on IFN-
secretion was mediated by an effect on IL-12p40 production. Similarly,
because IL-18 concentrations remained unchanged in all subjects, IL-10
effects on IL-18 production likely did not play a role. Other cytokines
implicated in IFN- production either remained undetectable (IL-15)
or were inhibited (TNF) by IL-10 (the present study and Ref.
10). Together, these data suggest that the stimulatory
effect of IL-10 on IFN- production in vivo is mediated by other
pathways or a direct effect of IL-10 on IFN- producing cells.
Results from our study are in contrast with data found in a previous
study in which human volunteers were injected with increasing doses (1,
10, or 25 µg/kg) of rhIL-10 (37, 38). PBMCs, isolated at
3 h and 6 h after IL-10 administration at a dose of 10 or 25
µg/kg, produced less IFN- during in vitro stimulation with PHA and
PMA compared with PBMCs isolated from volunteers receiving placebo
(38). However, the inhibitory effect was stronger when
IL-10 was administered at a dose of 10 µg/kg, indicating that
high-dose IL-10 may not only be associated with antiinflammatory
effects. In this study, PHA and PMA were used as stimuli, which leads
to T cell stimulation, whereas we used LPS as a stimulus, which targets
monocytes/macrophages. Because IFN- is produced by T and NK cells,
and LPS has no direct effect on these cells, it could be expected that
other stimuli could result in a similar stimulating effect. However, it
seems likely that priming of T and NK cells, either through direct or
indirect stimulation, is needed for an immunostimulatory effect of
IL-10 administration in vivo.
Previous in vitro studies have demonstrated that IL-10 can have a
direct stimulatory effect on activated CD8+ T
cells and NK cells. IL-10 enhances IL-2-stimulated proliferation of
purified human CD8+ cells (13).
Addition of IL-10 to mouse NK cells enhances IFN- production when
they are stimulated with IL-12 and IL-18 (14). A recent
study showed that IL-10 enhanced the capacity of IL-18 to stimulate
IFN- production, cytotoxicity, and proliferation of murine NK cells
(39). Also, in transgenic mice expressing human IL-10,
tumor cell growth was importantly inhibited compared with tumor growth
in nontransgenic controls, an effect that was mediated in part by
IL-10-activated CD8+ T cells (40).
To determine whether IL-10 treatment during endotoxemia involves
increased activation of CD8+ T cells and NK cells
in vivo, we measured concentrations of GrA and GrB. Granzymes are a
family of serine proteinases present in cytoplasmic granules of CTL and
NK cells (29, 41) that are released upon activation and
can trigger pathways of apoptosis in the target cells (29, 42). Measurements of levels of soluble granzymes is considered
to reflect the involvement of CTL and NK cells in various diseases
states, and increased plasma levels of soluble granzymes have been
found in patients with rheumatoid arthritis, EBV, or HIV-1 infection,
and during primary cytomegalovirus infection (21, 43).
Levels of GrA and GrB were modestly elevated after LPS administration,
but IL-10 treatment induced a significant increase in levels of both
GrA and GrB. These data suggest that IL-10 treatment results in
enhanced activation of cytotoxic lymphocytes leading to increased
IFN- production.
The increase in IFN- levels after IL-10 posttreatment was
accompanied by enhanced release of IP-10 and Mig, both members of the
CXC chemokine family. In vitro, IP-10 and Mig are produced by a variety
of cells in response to IFN- (28). IP-10 and Mig are
closely related and share a common receptor, CXCR3, which is
preferentially expressed on activated Th1 cells. IP-10 and Mig are
potent chemoattractants for activated T cells and hereby play an
important role in inflammatory processes. We here report that levels of
IP-10 and Mig increase during human endotoxemia, indicating that these
chemokines may play a role during systemic infection. IP-10
concentrations peaked early, whereas levels of Mig remained elevated
until the end of the study period, indicating that these proteins are
differentially regulated. Although IL-10 pretreatment had no effect on
IP-10 levels and inhibited the early increase of Mig, IL-10
posttreatment enhanced both IP-10 and Mig. Because only IL-10
posttreatment increased IP-10 and Mig levels, whereas IFN- was also
increased with IL-10 pretreatment, these data suggest that in vivo,
besides IFN-, other factors are involved in IP-10 and Mig
production. In vitro, TNF has been shown to be a costimulus for
IFN--induced IP-10 and Mig production (44). Previously,
we have reported that only pretreatment with IL-10 inhibited TNF
production during human endotoxemia (10), suggesting that
IL-10 posttreatment was more effective in stimulating IP-10 and Mig
production due to enhancement of IFN- release in the absence of a
concurrent reduction in TNF concentrations.
The IL-10-induced enhancement of IFN- release in vivo could not be
reproduced during human whole blood stimulation with LPS in vitro. We
used whole blood stimulation rather than stimulation of isolated cells
because the former system is considered to mimic in vivo conditions
best, containing hormones, cytokines, and other soluble factors that
may influence cytokine production (30). This method has
been adopted by many different groups, and has been used extensively to
study the regulation of cytokine production (19, 37, 38, 45, 46, 47, 48, 49). In the whole blood experiments, IL-10 induced a strong
dose-dependent inhibition of LPS-induced IFN- production, which was
associated with a concurrent reduction in IL-12 and IL-18
concentrations. These findings confirm and extent a previous study in
which IL-10 was reported to attenuate IFN- and IL-12 production by
LPS-stimulated PBMCs (50). Because we considered it
possible that IFN- release after low dose LPS administration to
humans occurs at least in part independently from IL-12 and IL-18
(i.e., circulating concentrations of IL-12p70 and IL-18 remained
undetectable and unchanged respectively), we also evaluated the effect
of IL-10 on LPS-induced IFN- release in whole blood in vitro in the
absence of IL-12 and IL-18 activity. However, in the presence of
anti-IL-12 and/or anti-IL-18, IL-10 still inhibited LPS-induced
IFN- production, although to a lesser extent, indicating that IL-10
inhibits IFN- release in vitro in part through inhibition of
IFN--inducing cytokines. Hence, the effects found of IL-10 on
IFN- production in vivo likely involve cell populations or
mediators, which are not present or present in low quantities in
peripheral blood. Discrepancies between effects of experimental
interventions observed in vivo or in vitro have been described
previously. Addition of anti-TNF Abs has been reported to have no
effect on LPS-induced IL-6 and IL-1ß release during human whole blood
stimulation in vitro (45). In contrast, during
experimental endotoxemia in chimpanzees, injection of anti-TNF
strongly inhibited the production of IL-6 (51). Also, Abs
to TNF importantly reduced the release of IL-6 and IL-1ß during
experimental bacteremia in baboons (52). Furthermore, we
previously reported that incubation with rhIL-10 did not significantly
influence LPS-stimulated monocyte chemoattractant protein (MCP)-1
production in whole blood in vitro, whereas administration of rhIL-10
during experimental human endotoxemia strongly attenuated MCP-1 release
(53).
IL-10 is evaluated as a new adjuvant therapy for several inflammatory
diseases (16, 17). Treatment of Crohn’s disease with
IL-10 has been found to be associated with a bell-shaped dose-response
curve (54). At a dose of 5–10 µg/kg s.c., IL-10 has
been reported to induce clinical remissions, but at higher-dose IL-10
(20 µg/kg) the beneficial effects are lost. In the present study, we
demonstrate that IL-10 used at a dose of 25 µg/kg, particularly when
administered shortly after an inflammatory stimulus, has
proinflammatory effects in humans in vivo. Therefore, these data
indicate that the potential IFN--enhancing effect of IL-10 may
especially warrant caution for the use of high-dose IL-10 therapy for
Th1-mediated illnesses like Crohn’s disease and rheumatoid
arthritis.
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Fanny N. Lauw, Department of Experimental Internal Medicine, Room G2–105, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands.
3 Abbreviations used in this paper: SEB, staphylcoccal enterotoxin B; rh, recombinant human; IP-10, IFN--inducible protein-10; Mig, monokine induced by IFN-; GrA, granzyme A; GrB, granzyme B.
Received for publication November 2, 1999. Accepted for publication June 20, 2000.
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 E. Giacomini, A. Sotolongo, E. Iona, M. Severa, M. E. Remoli, V. Gafa, R. Lande, L. Fattorini, I. Smith, R. Manganelli, et al. Infection of Human Dendritic Cells with a Mycobacterium tuberculosis sigE Mutant Stimulates Production of High Levels of Interleukin-10 but Low Levels of CXCL10: Impact on the T-Cell Response. Infect. Immun., June 1, 2006; 74(6): 3296 - 3304. [Abstract] [Full Text] [PDF]
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 R. W.-Y. Chan, F. M.-M. Lai, E. K.-M. Li, L.-S. Tam, K.-M. Chow, P. K.-T. Li, and C.-C. Szeto The effect of immunosuppressive therapy on the messenger RNA expression of target genes in the urinary sediment of patients with active lupus nephritis Nephrol. Dial. Transplant., June 1, 2006; 21(6): 1534 - 1540. [Abstract] [Full Text] [PDF]
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 S. Mocellin, F. M. Marincola, and H. A. Young Interleukin-10 and the immune response against cancer: a counterpoint J. Leukoc. Biol., November 1, 2005; 78(5): 1043 - 1051. [Abstract] [Full Text] [PDF]
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R. W. DePaolo, R. Lathan, B. J. Rollins, and W. J. Karpus The Chemokine CCL2 Is Required for Control of Murine Gastric Salmonella enterica Infection Infect. Immun., October 1, 2005; 73(10): 6514 - 6522. [Abstract] [Full Text] [PDF]
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 M. N. Sharif, I. Tassiulas, Y. Hu, I. Mecklenbrauker, A. Tarakhovsky, and L. B. Ivashkiv IFN-{alpha} Priming Results in a Gain of Proinflammatory Function by IL-10: Implications for Systemic Lupus Erythematosus Pathogenesis J. Immunol., May 15, 2004; 172(10): 6476 - 6481. [Abstract] [Full Text] [PDF]
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 V. Deleuze, J. Lefort, M. F. Bureau, D. Scherman, and B. B. Vargaftig LPS-induced bronchial hyperreactivity: interference by mIL-10 differs according to site of delivery Am J Physiol Lung Cell Mol Physiol, January 1, 2004; 286(1): L98 - L105. [Abstract] [Full Text] [PDF]
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C. Herrero, X. Hu, W. P. Li, S. Samuels, M. N. Sharif, S. Kotenko, and L. B. Ivashkiv Reprogramming of IL-10 Activity and Signaling by IFN-{gamma} J. Immunol., November 15, 2003; 171(10): 5034 - 5041. [Abstract] [Full Text] [PDF]
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 K. Asadullah, W. Sterry, and H. D. Volk Interleukin-10 Therapy--Review of a New Approach Pharmacol. Rev., June 1, 2003; 55(2): 241 - 269. [Abstract] [Full Text] [PDF]
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 M. de Fost, R. A. Hartskeerl, M. R. Groenendijk, and T. van der Poll Interleukin 12 in Part Regulates Gamma Interferon Release in Human Whole Blood Stimulated with Leptospira interrogans Clin. Vaccine Immunol., March 1, 2003; 10(2): 332 - 335. [Abstract] [Full Text] [PDF]
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A.-F. Petit-Bertron, C. Fitting, J.-M. Cavaillon, and M. Adib-Conquy Adherence influences monocyte responsiveness to interleukin-10 J. Leukoc. Biol., January 1, 2003; 73(1): 145 - 154. [Abstract] [Full Text] [PDF]
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R. Lang, D. Patel, J. J. Morris, R. L. Rutschman, and P. J. Murray Shaping Gene Expression in Activated and Resting Primary Macrophages by IL-10 J. Immunol., September 1, 2002; 169(5): 2253 - 2263. [Abstract] [Full Text] [PDF]
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G. S. Deepe, Jr., and R. S. Gibbons Cellular and Molecular Regulation of Vaccination with Heat Shock Protein 60 from Histoplasma capsulatum Infect. Immun., July 1, 2002; 70(7): 3759 - 3767. [Abstract] [Full Text] [PDF]
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 H Herfarth and J Scholmerich IL-10 therapy in Crohn's disease: at the crossroads Gut, February 1, 2002; 50(2): 146 - 147. [Full Text] [PDF]
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H Tilg, C van Montfrans, A van den Ende, A Kaser, S J H van Deventer, S Schreiber, M Gregor, O Ludwiczek, P Rutgeerts, C Gasche, et al. Treatment of Crohn's disease with recombinant human interleukin 10 induces the proinflammatory cytokine interferon {gamma} Gut, February 1, 2002; 50(2): 191 - 195. [Abstract] [Full Text] [PDF]
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D. G. Mordue, F. Monroy, M. La Regina, C. A. Dinarello, and L. D. Sibley Acute Toxoplasmosis Leads to Lethal Overproduction of Th1 Cytokines J. Immunol., October 15, 2001; 167(8): 4574 - 4584. [Abstract] [Full Text] [PDF]
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K. Emmanuilidis, H. Weighardt, S. Maier, K. Gerauer, T. Fleischmann, X. X. Zheng, W. W. Hancock, B. Holzmann, and C.-D. Heidecke Critical Role of Kupffer Cell-Derived IL-10 for Host Defense in Septic Peritonitis J. Immunol., October 1, 2001; 167(7): 3919 - 3927. [Abstract] [Full Text] [PDF]
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