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 Previous Article | Table of Contents | Next Article 
Blood, Vol. 95 No. 10 (May 15), 2000:
pp. 3162-3167
IMMUNOBIOLOGY
Glucocorticoids transform CD40-triggering of dendritic cells into
an alternative activation pathway resulting in antigen-presenting cells
that secrete IL-10
Delphine Rea,
Cees van Kooten,
Krista E. van
Meijgaarden,
Tom H. M. Ottenhoff,
Cornelis J. M. Melief, and
Rienk Offringa
From the Department of Immunohematology and Blood Bank and the
Department of Nephrology, Leiden University Medical Center, Leiden, The
Netherlands.
 |
Abstract |
Dendritic cell (DC) activation through CD40-CD40 ligand interactions
is a key regulatory step for the development of protective T-cell
immunity and also plays an important role in the initiation of T-cell
responses involved in autoimmune diseases and allograft rejection. In
contrast to previous reports, we show that the immunosuppressive drug
dexamethasone (DEX) redirects rather than simply blocks this DC
activation process. We found that DCs triggered through CD40 in the
presence of DEX were unable to acquire high levels of costimulatory, adhesion, and major histocompatibility complex class I and II molecules
and failed to express the maturation marker CD83, whereas antigen
uptake was not affected. Moreover, DEX strikingly modified the
CD40-activated DC cytokine secretion profile by suppressing the
production of the proinflammatory cytokine interleukin (IL)-12 and
potentiating the secretion of the anti-inflammatory cytokine IL-10.
Accordingly, DEX-exposed CD40-triggered DCs displayed a decreased
T-cell allostimulatory potential and a dramatically impaired ability to
activate cloned CD4+ T helper 1 (Th1) cells. Moreover,
interaction between Th1 cells and these DCs rendered the T cells
hyporesponsive to further antigen-specific restimulation. Collectively,
our results demonstrate that DEX profoundly modulates CD40-dependent DC
activation and suggest that the resulting alternatively activated DCs
can be exploited for suppression of unwanted T-cell responses in vivo.
(Blood. 2000;95:3162-3167)
© 2000 by The American Society of Hematology.
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Introduction |
The remarkable immunostimulatory properties of
dendritic cells (DCs) reside in their ability to transport antigens
from peripheral tissues to lymphoid organs where they present these
antigens to T cells in an optimal costimulatory context.1
To achieve this complex sequence of events, DCs exist in different
functional stages. Immature DCs behave as sentinels in peripheral
tissues where they efficiently capture antigens. On pathogen invasion, induction of protective T-cell responses requires the activation of
immature DCs into mature immunostimulatory cells. DC activation is
triggered in inflamed tissues by cytokines, such as interleukin (IL)-1
and tumor necrosing factor  (TNF-), and by bacterial components,
such as lipopolysaccharide (LPS).2,3 Activated DCs migrate to T-cell areas in the lymph nodes while up-regulating their costimulatory capacities and optimizing their antigen-presenting functions. On interaction with antigen-specific T cells, DC
activation is further completed through engagement of the
receptor-ligand (L) pair CD40-CD40L, leading to the production of
IL-12,4,5,6 a key cytokine for T helper type 1 (Th1) and
cytotoxic T-lymphocyte priming.7
Antigen-presenting cell (APC) activation through
CD40-CD40L interactions represents a crucial immunoregulatory step for
the establishment of protective T-cell immunity against pathogens and
tumors.8,9,10 This process also plays a key role in the
onset of destructive T cell-mediated disorders, such as autoimmune diseases, allograft rejection, and graft versus host
disease.11,12,13 The current treatment of these disorders
largely relies on the administration of glucocorticoids (GCs), which
exert potent anti-inflammatory and immunosuppressive effects. Because
GCs negatively interfere with many aspects of T-cell activation, such
as IL-2-driven proliferation and inflammatory cytokine production
(reviewed in 14), activated T cells have long been considered as the
main targets for GC action. Several lines of evidence now indicate a
role for DCs in GC-induced immune suppression. Moser et
al15 found that GCs prevented the spontaneous activation of
murine DCs, thereby decreasing their T-cell stimulatory potential.
Kitajima et al16 showed that GCs could hamper the T
cell-mediated activation of a murine DC line. Viera et al
17 reported that human DCs exposed to GCs were poor producers of IL-12 on LPS stimulation. These findings only concern loss of typical
DC features and, therefore, favor a simple inhibitory role of GCs on DC
activation. A more complex immunoregulatory action on the DC system has
not been considered.
In this study, we performed detailed analysis of the impact of GCs on
the CD40-mediated activation of monocyte-derived DCs. These DCs develop
after culture with granulocyte-macrophage colony-stimulating factor
(GM-CSF) and IL-42,18 or after transmigration through endothelial cells19 and are known to mature into the most
potent human Th1-type-inducing APC on CD40 ligation.5,20
Moreover, these APCs can easily be generated in large numbers and are
thereby the cells of choice for DC-based modulation of T-cell
immunity.21,22 In contrast to previous studies, our data
show that GCs do not merely prohibit DC activation. In the presence of
the GC hormone dexamethasone (DEX), CD40 ligation on human
monocyte-derived DCs is transformed into an alternative activation
pathway, equipping these cells with unique features that enable them to
downmodulate Th1-type responses in vitro.
| |
Materials and methods |
Generation of DCs
Immature DCs were generated from peripheral blood monocyte
precursors. Human peripheral blood mononuclear cells from healthy donors, isolated through Ficoll-Hypaque density centrifugation were
plated at 1.5 × 107 per well in 6-well plates
(Costar Corp, Cambridge, MA) in RPMI 1640 (Life Technologies, Paisley,
Scotland) supplemented with 2 mmol/L glutamine, 100 UI/mL penicillin,
and 10% fetal calf serum. After 2 hours at 37°C, the nonadherent
cells were removed and the adherent cells were cultured in medium
containing 500 U/mL IL-4 (Pepro Tech Inc, Rocky Hill, NJ) and 800 U/mL
GM-CSF (kindly provided by Dr S Osanto, LUMC, Leiden, The Netherlands)
for a total of 7 days.
Activation of immature DCs with a CD8-CD40L fusion protein
Activation of DCs though CD40 was performed with a fusion protein
made of the extracellular domain of human CD40L and of the murine
CD8 chain (CD8-CD40L). The CD8-CD40L complementary DNA described by
Garrone et al23 was transferred into an eukaryotic expression vector containing the hygromycin resistance gene and used
for the generation of stably transfected Chinese hamster ovary cells.
Culture supernatants containing the CD8-CD40L fusion protein were
concentrated with a pressurized stirred cell system (Amicon, Inc,
Beverly, MA), checked for binding to CD40, and tested for optimal
DC-activation conditions (not shown). DCs were incubated at
5 × 105/mL/well in a 24-well plate (Costar Corp,
Cambridge, MA) and activated in the presence of 1/10 CD8-CD40L
supernatant. Cells and supernatants were analyzed after 48 hours.
Importantly, the specificity of CD8-CD40L supernatants was checked by
comparison with control supernatants obtained from untransfected
Chinese hamster ovary cells. Unlike DCs activated with CD8-CD40L
supernatants, immature DCs cultured with control supernatants or with
medium alone failed to up-regulate the maturation marker CD83
(respective mean fluorescence intensities: 20, 6, and 5) and to secrete
IL-12 (respective IL-12 production per 2.5 × 105
DC: 30 510 pg/mL, 600 pg/mL, and 250 pg/mL). The action of CD8-CD40L supernatants is, therefore, specific for the CD8-CD40L fusion protein
and is not due to other factors present in the concentrated conditioned medium.
Dexamethasone and RU486 treatment of DCs
Seven-day immature DCs were treated with 10 6
mol/L DEX (Sigma, St Louis, MO) in the presence of GM-CSF and IL-4 or
GM-CSF alone. After 24 hours, DCs were analyzed or were further
stimulated via CD40 by adding the CD8-CD40L fusion protein to the
cultures as described above. In some experiments, the GC receptor
antagonist RU485 (Roussel-UCLAF, Romainville, France) was used at 10 µmol/L final concentration, alone or in combination with DEX.
Analysis of DC surface phenotype by flow cytometry
Cells were stained on ice with fluorescein
isothiocyanate (FITC) or
phosphatidylethanolamine (PE)-conjugated mouse monoclonal antibodies for 30 minutes in phosphate-buffered saline 1% fetal calf
serum and were analyzed on a FACScan® (Becton Dickinson, San Jose,
CA). The following monoclonal antibodies were used: FITC-anti-CD80
(BB1), PE-anti-CD86 (FUN-1), FITC-anti-CD40 (5C3), PE-anti-CD54 (HA
58), and PE- anti-CD58 (1C3) (Pharmingen, San Diego, CA);
PE-anti-CD14 (L243) and PE-anti-HLA-DR (M -P9) (Becton Dickinson);
PE-anti-CD83 (HB15A) (Immunotech, Marseille, France); and PE-anti-HLA
class I (Tu 149) (Caltag Laboratories, Burlingame, CA).
Antigen uptake experiments
DCs were resuspended in medium buffered with 25mmol/L Hepes.
FITC-beef serum albumin (BSA) and FITC-mannosylated BSA (both from
Sigma) were added at 1 mg/mL final concentration, and the cells were
incubated at 37°C or at 0°C to determine background uptake.
After 1 hour, DCs were washed extensively with ice-cold phosphate-buffered saline and analyzed by FACS® with the use of propidium iodide to eliminate dead cells.
Cytokine detection by enzyme-linked immunosorbent assay
Culture supernatants were analyzed in serial twofold dilutions in
duplicate. IL-12p70 was detected with the use of a solid phase sandwich
enzyme-linked immunosorbent assay (ELISA) kit (Diaclone Research,
Besancon, France) (sensitivity 3 pg/mL). For IL-12p40 and interferon
(IFN)- detection, capture monoclonal antibodies and polyclonal
biotinylated detection antibodies were obtained from Peter van de
Meijde (BPRC, Rijswijk, The Netherlands) (sensitivity 10 pg/mL). IL-10
was detected with the use of the Pelikine compact human IL-10 ELISA kit
(CLB, Amsterdam, The Netherlands) (sensitivity 3 pg/mL).
Allogeneic mixed lymphocyte reaction
Nonadherent allogeneic adult peripheral blood mononuclear cells from
an unrelated individual were cultured in 96-well flat-bottom plates
(Costar Corp, Cambridge, MA) at a density of
1.5 × 105/well with various numbers of
-irradiated (3000 rads) DCs, in triplicates. Proliferation was
assessed on day 5 by [3H]thymidine uptake (0.5 µCi/well, specific activity 5 Ci/mmol, Amersham Life
Science, Buckinghamshire, UK) during a 16-hour pulse.
Th1 stimulation assays
The Mycobacterium tuberculosis and M leprae
hsp65-specific, HLA-DR3-restricted CD4+ Th1 clone Rp15 1-1 used in this
study recognizes an hsp65 determinant corresponding to peptide residues 3 to 13 (p3-13).24 HLA-DR-matched DEX-treated immature DCs
and their DEX-untreated counterparts were pulsed with 10 µg/mL of p3-13 or with 10 µg/mL of hsp65 for 2 hours, washed extensively, and
stimulated through CD40 as described above. For Ag-pulsed DEX-treated
immature DCs, CD40 triggering was performed in the presence of DEX.
Hsp65-specific T cells (104) were cultured with different
numbers of -irradiated (3000 rads) DCs in 96-well flat-bottom plates
(Costar Corp) in triplicates for 3 days. [3H]thymidine
incorporation was measured on day 3 after a 16-hour pulse. Before the
addition of [3H]thymidine, 50 µL of supernatants was
collected from each well, and supernatants from triplicate wells were
pooled to measure IFN- production. To test hsp65-specific T-cell
responsiveness to a second antigenic challenge, 104 T cells
were first cultured for 5 days with 104 peptide-pulsed DCs
prepared as above and allowed to rest for 4 additional days in medium
containing low doses of IL-2 (5 U/mL). Subsequently, 104
viable T cells were restimulated with 104 peptide-pulsed
DCs generated from the same donor as used for the first culture and
tested for their ability to proliferate and to produce IFN- as
previously described.
Statistical analysis
Covariance analysis was used to compare T-cell proliferation and
IFN- production as a function of DC number, between DEX-treated CD40-triggered DCs, immature DCs, and CD40-triggered DCs (Figure 5).
| |
Results |
Impairment of CD40-CD40L-mediated phenotypic changes by DEX
We explored the impact of DEX on the phenotypic changes induced by
CD40 ligation on immature monocyte-derived DCs. In the absence of DEX,
the fusion protein CD8-CD40L, which triggers DCs specifically through
their CD40 receptor, induced a strong up-regulation of the
costimulatory molecules CD80, CD86, and CD40; of the major histocompatibility complex (MHC) class I and II molecules; of the
adhesion markers CD54 and CD58; and of the DC maturation marker CD83
(Figure 1). In the presence of DEX, these
CD8-CD40L-induced phenotypic changes were dramatically impaired: the
up-regulation of CD80, CD86, CD40, CD54, CD58, and of the MHC class I
and II molecules was largely inhibited and CD83 was not expressed
(Figure 1). Importantly, DEX-treated DCs did not revert to a
monocyte/macrophage stage as shown by the lack of expression of CD14
(Figure 1). Titration of DEX showed a complete inhibition of
CD40-mediated phenotypic changes at 106 mol/L and
107 mol/L, a partial blockade at
108 mol/L, and no effect at 109
mol/L and 1010 mol/L (data not shown). In addition,
DEX action depended on binding to the GC receptor, since it was
abolished by simultaneous addition of the GC-receptor antagonist RU486
(data not shown). In experiments performed with LPS or TNF- as
activation agents, similar results were obtained. However, the
combination of DEX and TNF- induced a massive cell death (viable
cell recovery 5% to 10% of control cultures), a phenomenon that was
not observed when DEX-treated DCs were stimulated with LPS or through
CD40 (viable cell recovery 60%-100% of control cultures) (not shown).
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| Fig 1.
Pretreatment with DEX inhibits the phenotypic changes
induced by CD40 ligation.
Seven-day immature DCs were cultured for 24 hours in the absence or the
presence of 106 mol/L DEX and activated via CD40
with the CD8-CD40L fusion protein for 48 hours. The comparison with
immature DCs maintained in medium alone is shown. Empty histograms show
the background staining with isotype controls monoclonal antibody, and
solid histograms represent specific staining of the indicated
cell-surface markers. Specific mean fluorescence intensities are
indicated. Mean fluorescence intensities of isotype controls were
between 3 and 4. Data are representative of 4 independent experiments.
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We next analyzed whether activated DCs could still be affected by DEX.
DCs incubated with CD8-CD40L for 48 hours and further exposed to DEX
maintained a stable activated phenotype (Figure 2).
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| Fig 2.
DCs triggered through CD40 maintain an activated
phenotype on a subsequent DEX exposure.
Immature DCs were activated with the CD8-CD40L fusion protein. DEX
(106 mol/L) or medium control was added 48 hours
later, and cells were analyzed after 2 additional days of culture. The
comparison with immature DCs maintained in medium alone is shown. Empty
histograms show the background staining with isotype controls
monoclonal antibody, and solid histograms represent specific staining
of the indicated cell-surface markers. Specific mean fluorescence
intensities are indicated. Mean fluorescence intensities of isotype
controls were between 3 and 5. Data are representative of 2 independent
experiments.
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We conclude that DEX prevents the phenotypic changes induced by CD40
signals on immature DCs and that already activated DCs are resistant to
DEX action.
DEX does not interfere with the regulation of DC-antigen uptake
machinery
Unlike activated DCs, immature DCs efficiently internalize antigens
through macropinocytosis and mannose receptor-mediated endocytosis.2,3,25,26 We analyzed whether DEX could affect the DC-antigen capture machinery and its down-regulation following CD40
cross-linking. As shown in Figure 3,
incorporation of FITC-BSA and FITC-mannosylated BSA by immature DCs and
by DEX-treated immature DCs was comparable. On CD40 triggering, a
similar decrease of FITC-BSA and FITC-mannosylated BSA uptake by both
DEX-treated and untreated DCs was observed (Figure 3). These results
were the first to indicate to us that DEX does not block all aspects of
DC activation, since it does not interfere with the
down-regulation of the DC-antigen capture machinery.
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| Fig 3.
Pretreatment with DEX does not affect the regulation of
DC-antigen uptake machinery.
Immature DCs were incubated in the absence or the presence of
106 mol/L DEX for 24 hours and further activated or
not via CD40 with the CD8-CD40L fusion protein for 48 hours. Cells were
pulsed for 1 hour with medium containing either 1 mg/mL FITC-BSA or 1 mg/mL FITC-mannosylated BSA. Empty histograms show the background
autofluorescence, gray-filled histograms show the background uptake at
0°C, and black-filled histograms show the specific uptake at
37°C. Data are representative of 3 independent experiments.
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Altered cytokine secretion profile of DEX-treated CD40-triggered DCs
A key feature of CD40-triggered DCs for initiating T-cell immunity
resides in their ability to produce the pro-inflammatory cytokine
IL-12.5,6,27 We investigated whether DEX affected IL-12
production by DCs stimulated through CD40, and we explored the
possibility that DEX could promote the secretion of the
anti-inflammatory cytokine IL-10. As shown in Figure
4, CD40 triggering of DCs strongly induced
IL-12p40 and IL-12p70 secretion (up to 120 ng/mL and 170 pg/mL,
respectively) but only poorly stimulated the production of IL-10 (up to
68 pg/mL). In contrast, CD40 triggering of DEX-treated DCs resulted in
a dramatically reduced IL-12p40 production (up to 100-fold) and in the
complete suppression of IL-12p70 secretion, whereas IL-10 production
was strongly enhanced (up to 50-fold) (Figure 4). Immature DCs and
their DEX-treated counterparts failed to secrete detectable amounts of
IL-12 and IL-10 (Figure 4). Therefore, CD40 ligation of DCs in the
presence of DEX triggers the secretion of high levels of the
anti-inflammatory cytokine IL-10 instead of IL-12.
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| Fig 4.
Pretreatment with DEX alters the cytokine secretion
profile of CD40-triggered DCs.
DEX-exposed or control immature DCs were left in culture without
further treatment or stimulated with the CD8-CD40L fusion protein.
Culture supernatants were harvested 48 hours later, and IL-10,
IL-12p40, and IL-12p70 secretion were analyzed by specific
enzyme-linked immunosorbent assay. Data are representative from 6 independent experiments.
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Modulation of DC T-cell stimulatory capacities by DEX
The strikingly modified response of DCs to CD40 ligation in the
presence of DEX prompted us to compare the T-cell stimulatory potential
of these cells with that of their DEX-untreated counterparts. In an
allogeneic mixed lymphocyte reaction, CD40-triggered DCs induced a
strong proliferative T-cell response, whereas the addition of DEX prior
to CD40 triggering reduced their T-cell stimulatory capacity to that of
immature DCs (Figure 5). When tested for
their ability to stimulate an hsp65-specific CD4 ± Th1 clone,
CD40-triggered DCs pulsed with the hsp65 protein or with the specific
peptide epitope p3-13 were found to be highly potent inducers of both T-cell proliferation and T-cell dependent IFN- production (Figure 5). In the presence of Ag-pulsed DEX-treated CD40-triggered DCs, T-cell
proliferation and IFN- production were significantly lower (P  .001), and the T-cell stimulatory capacity of these DCs
was even lower than that of immature DCs (P .01). We next
investigated whether DEX-treated CD40-triggered DCs were simply poor
stimulators of Th1 cells or whether they could exert suppressive
effects on these T cells. We, therefore, tested hsp65-specific T cells
stimulated with p3-13-pulsed DEX-treated CD40-triggered DCs for their
capacity to respond to a second potent antigenic challenge. Figure
6 shows that preculturing T cells with
CD40-triggered DCs or with immature DCs led to a strong T-cell
proliferation and IFN- production on second antigen-specific
restimulation. In contrast, preculture with DEX-treated CD40-triggered
DCs resulted in a dramatically reduced proliferative and IFN-
production capacity of Th1 cells. Thus, CD40 triggering of DCs in the
presence of DEX results in APC that are not merely poor inducers of
T-cell responses but that also induce a state of hyporesponsiveness in
Th1 cells.
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| Fig 5.
Pretreatment with DEX impairs the T-cell stimulatory
capacities of DCs activated via CD40.
Allogeneic mixed lymphocyte reaction: nonadherent allogeneic peripheral
blood mononuclear cells were cultured with different numbers of
CD40-triggered DCs, DEX-treated CD40-triggered DCs, or immature DCs.
The proliferative response was measured on day 5. Th1 stimulation
assays: Hsp65-specific T cells were cultured with different numbers of
HLA-DR matched CD40-triggered DCs or with DEX-treated CD40-triggered
DCs, or with immature DCs, pulsed with the hsp65 protein or with the
specific p3-13 peptide epitope. The proliferative response and the
T-cell dependent IFN- production were analyzed on day 3. Data are
representative of 3 independent experiments.
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| Fig 6.
DEX-treated DCs triggered through CD40 induce a state of
hyporesponsiveness in Th1 cells.
Hsp65-specific T cells precultured with CD40-triggered DCs or with
immature DCs or with DEX-treated CD40-triggered DCs pulsed with the
p3-13 peptide epitope were harvested after 5 days, allowed to rest in
the presence of 5 U/mL IL2 for 4 days, and restimulated with
p3-13-pulsed DCs or with unpulsed DCs (medium control). The
proliferative response and IFN- production were measured on day 3. Similar results were obtained in 2 independent experiments.
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| |
Discussion |
In this study, we demonstrate that DEX profoundly affects
the CD40-dependent maturation of human monocyte-derived DCs, not only by preventing the up-regulation of costimulatory, adhesion, and MHC surface molecules but also by causing these cells to secrete the anti-inflammatory mediator IL-10 instead of the Th1 stimulatory cytokine IL-12. In agreement with these phenotypic and functional changes, DCs triggered through CD40 in the presence of DEX are poor
stimulators of Th1-type responses and are able to induce a state of
hyporesponsiveness in Th1 cells, suggesting that such DCs might
contribute to active suppression of Th1-type immunity.
The impact of GCs on DCs has been the subject of several previous
studies by others. However, in contrast with our data, these studies
only highlighted inhibitory effects of GCs on the DC system. DEX was
found to block the up-regulation of CD80, CD86, and MHC class II
molecules upon activation of murine spleen DCs,15,16 whereas very recently DEX was demonstrated to also prevent the differentiation of DCs from monocyte precursors.28 In these studies, the inability of DCs to acquire high expression of
costimulatory and MHC molecules was accompanied with a decrease in
their T-cell stimulatory potential, but the effect of GCs on
IL-12 production was not investigated. Conversely, Viera et
al17 found that the effect of GCs on LPS-induced DC
activation consisted in a 4-fold reduction of IL-12p70 synthesis. This
partial effect on IL-12 secretion contrasts with the complete
suppression of IL-12p70 production described in our work and can be
explained by the fact that their GC-treated immature DCs were
extensively washed prior to LPS stimulation. We indeed found that on
removal of GCs, the effects of these drugs on immature DCs were rapidly
reversible. The continuous presence of GCs during CD40 triggering of
DCs was absolutely required to stably and completely modulate DC
activation (data not shown). Taken together, previous findings
indicated that the impact of GCs on the DC system should be merely
interpreted as an inhibitory event.
Importantly, our results clearly demonstrate that GCs do not simply
suppress DC activation but rather redirect this process toward a
distinct functional program. Even though DCs triggered through CD40 in
the presence of DEX maintain an immature-like surface phenotype, an
important aspect of CD40-mediated DC activation, the shut-down of
DC-antigen uptake machinery, still takes place. Furthermore, DEX does
not only abrogate the production of bioactive IL-12 induced by CD40
cross-linking, but it also exerts a strong synergistic effect with CD40
signals on IL-10 secretion. A change in the IL-12/IL-10 balance in
favor of IL-10 induced by GCs, although less striking than in our
study, has also been observed in human peripheral blood mononuclear
cells and purified monocytes upon LPS stimulation.29,30
The varying effects of DEX on the multiple aspects of DC activation
suggest that DEX differentially affects the intracellular signals that
are elicited when DCs are triggered through their CD40 receptor. The
analysis of CD40 signaling events, although so far restricted to B
cells, has shown that binding of CD40L to CD40 triggers multiple
signaling pathways leading to the activation of transcription factors,
such as NF- B, AP-1, and NF-AT (reviewed in 31). These transcription
factors control the expression of many genes involved in immune
responses and are also major targets for GC-induced transcriptional
repression. For instance, GCs inhibit NF-B activation through the
induction of IB synthesis.32,33 The NF-B binding site
present in the IL-12p40 gene promoter region plays an important role
for CD40-dependent induction of IL-12.34,35 It is,
therefore, conceivable that DEX blocks IL-12 secretion by preventing
NF-B activation induced by CD40 ligation. Importantly, our data
clearly show that DEX does not block all CD40-dependent aspects of DC
activation since the CD40-mediated down-regulation of DC-antigen
capture machinery is unaffected. In addition, GCs may even enhance some
CD40 signals, as shown by the synergistic effect of DEX on
CD40-mediated IL-10 secretion.
DC activation through engagement of CD40-CD40L is a key stimulatory
event for the generation of effective Th1 and CD4-dependent cytotoxic
T-lymphocyte responses in vivo.10,36-38 This pathway, however, is also involved in the development of unwanted T-cell responses leading to autoimmune disease or organ-transplant
rejection.11-13 Until now, treatment of patients with such
disorders largely relies on the systemic administration of GC hormones.
This treatment does not only suppress pathogenic T-cell responses but
also induces a general state of immunosuppression and metabolic and
endocrine side effects. Activation of human monocyte-derived DCs
through CD40 in the presence of DEX results in the generation of
IL-10-producing APCs with low costimulatory capacities. Unlike immature
DCs, which mature upon interaction with Th1 cells,5 and
CD40-triggered DCs, these DCs are poor stimulators for Th1-type
responses and even confer hyporesponsiveness to Th1 cells. Although
experiments addressing the mechanisms underlying the unique phenotypic
and functional features of such DCs are under way in our laboratory, it
can already be envisioned that administration of such DCs loaded with
appropriate antigens may be exploited as a novel approach for
specifically down-regulating unwanted T-cell responses in vivo. The
feasibility of this approach is currently being tested in murine
autoimmune and transplantation models.
| |
Acknowledgments |
We thank Francine Briere and Pierre Garrone (Schering Plough, Dardilly,
France) for kindly giving us the opportunity to use the CD8-CD40L
fusion protein; Andrea Woltman (Department of Nephrology, LUMC) for
expert help with the CD8-CD40L protein production; and A.H. Zwindermann
for statistical analysis.
| |
Footnotes |
Submitted June 29, 1999; accepted January 10, 2000.
Supported by grants from the European Union, TMR contract
FMRX-CT96-0053, and from the Netherlands Leprosy Foundation (NSL).
Reprints: Delphine Rea, Department of Immunohematology and
Blood Bank, Leiden University Medical Center, Albinusdreef 2, Postbus 9600, 2300 RC Leiden, The Netherlands; e-mail:
d.g.rea{at}immunohematology.medfac.leidenuniv.nl.
The publication costs of this
article were defrayed in part by
page charge payment. Therefore,
and solely to indicate this fact,
this article is hereby marked
"advertisement"
in accordance with 18 U.S.C.
section 1734.
| |
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Abstract
Introduction