* Department of Tumor Immunology, University Medical Center Nijmegen St. Radboud, Nijmegen, and
Department of Dermatology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
Correspondence: Gosse J. Adema, Department of Tumor Immunology, University Medical Center Nijmegen St. Radboud, Geert Grooteplein Zuid 28, 6525 GA Nijmegen, The Netherlands. E-mail: g.adema{at}dent.kun.nl
 TOP ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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, and thymus
and activation-regulated chemokine (TARC). Upon maturation, DCs
up-regulated the expression of DC-CK1 (60-fold), ELC (7-fold), and TARC
(10-fold). Activation of DCs by CD40 ligand further up-regulated the
expression of ELC (25-fold). We found that freshly isolated blood DCs
expressed only low levels of interleukin-8, lymphotactin, and MIP-1.
It is interesting that the chemokine profile expressed by activated
CD11c- lymphoid-like as well as CD11c+ myeloid
blood DCs mimics that of monocyte-derived DCs. Additionally, purified
Langerhans cells that had migrated out of the epidermis expressed a
similar chemokine pattern. These data indicate that different DC
subsets in vitro and in vivo can express the same chemokines to attract
leukocytes.
Key Words: human • real-time quantitative PCR • monocytes • blood DCs • Langerhans cells
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Following antigen uptake in the periphery, immature DCs change their chemokine receptor profile and migrate into secondary lymphoid organs, where they mature and must interact in an antigen-specific manner with B cells and T cells to initiate an immune response [4 5 ]. Chemokines are involved in the recruitment of B cells, T cells, and DCs into the secondary lymphoid organs and the attraction of these cells to each other. This process requires accurate regulation, which can be defined at three different levels: (1) regulation of chemokine production [6 ], (2) regulation of chemokine receptor expression and desensitization [5 7 8 9 ], and (3) chemokines acting as antagonists [10 ].
To unravel in detail the fine-tuning of chemokine regulation in microenvironments, we studied the regulation of chemokine production. Since DCs play a key role in the induction of an antigen-specific immune response, we analyzed chemokine expression by DCs. For this purpose, we used real-time quantitative polymerase chain reaction (PCR), a new technique used to precisely quantify mRNA levels. This approach utilizes a sequence-specific probe containing at its 5' end a reporter dye which is quenched by a second fluorescent dye. During specific extension of the PCR, Taq polymerase cleaves the probe, and the quencher dye is released. The subsequent increase of fluorescence of the reporter dye is proportional to the amount of amplified copies and can be monitored on-line [11 12 13 ]. We have determined the chemokine expression profile of in vitro-cultured immature and mature monocyte-derived DCs. Furthermore, we studied whether monocyte-derived DCs primed, at the cytokine level, to induce a T-helper 1 response exhibit a chemokine profile different from that of DCs primed to induce a T-helper 2 response. Chemokine expression by monocyte-derived DCs was compared with that of in vivo DC subsets, namely peripheral blood DCs and LCs. Strikingly, although we observed quantitative differences between chemokine expression levels, all tested DC subsets expressed similar chemokine expression profiles. This indicates that they are able to attract the same set of cells of the immune system.
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Generation of monocyte-derived DCs
Peripheral blood mononuclear cells (PBMC) of healthy individuals
were separated by Percoll density centrifugation and were allowed to
adhere for 1 h at 37°C in RPMI 1640 (Life Technologies, Inc.,
Grand Island, NY) enriched with 2% human serum in 75-cm2
tissue culture flasks (Costar, Badhoevedorp, The Netherlands). Adherent
monocytes were cultured in RPMI 1640 enriched with 10% fetal calf
serum (FCS) and a combination antibiotic-antimycotic (all from Life
Technologies, Inc.) in the presence of interleukin (IL)-4 (500 U/mL;
Schering-Plough, Amstelveen, The Netherlands) and
granulocyte-macrophage colony-stimulating factor (GM-CSF; 800 U/mL,
Schering-Plough) for 6 days. Fresh cytokine-containing culture medium
was added at day 3. At day 6, the immature DCs were harvested. To
generate mature DCs, immature DCs were resuspended in fresh
cytokine-containing culture medium and stimulated by addition of either
10 ng/mL of tumor necrosis factor (TNF-; Bender Wien, Vienna,
Austria), 2 µg/mL of lipopolysaccharide (LPS; Sigma Chemical Co., St.
Louis, MO), and either 1 mg/mL of CD40 ligand (CD40L) trimer or 1 mg/mL
of CD40L in combination with 1,000 U/mL of 
interferon (IFN-;
Boehringer Ingelheim, Ingelheim am Rhein, Germany) in six-well culture
plates. Mature monocyte-derived DCs were used at day 8. DCs cultured in
autologous serum were prepared according to the same procedure, using
X-VIVO 15 medium (BioWhittaker, Walkersville, MD) enriched with 1%
autologous serum. Maturation was established by addition of the
combination of 10 ng/mL of TNF-, (Bender Wien), 10 µg/mL of
prostaglandin E2 (PGE2; Sigma), and 50% (v/v)
monocyte-conditioned medium [MCM; from immunoglobulin G
(IgG)-activated monocytes] [14
].
Isolation and stimulation of fresh blood DCs
DCs were isolated from PBMC by using a MACS Blood DC isolation
kit (CLB, Amsterdam, The Netherlands). Briefly, blood DCs were
pre-enriched from PBMC by immunomagnetic depletion of CD3+
T cells, CD11b+ monocytic cells, and CD16+
natural killer (NK) cells. During a second immunomagnetic separation,
CD4+ blood DCs were positively selected. To obtain
activated CD4+ CD11c+ blood DCs, freshly
isolated blood DCs were cultured for 3 days in RPMI 1640 (Life
Technologies, Inc.) enriched with 10% FCS and 50% (v/v) MCM
[15
]. Stimulated CD4+ CD11c-
blood DCs were obtained by depletion of CD11c+ cells from
the pre-enriched blood DC fraction, using anti-CD11c mAbs and magnetic
cell sorting (Dynal, Oslo, Norway) followed by a positive selection for
CD4. Isolated CD4+ CD11c- blood DCS were
activated in RPMI 1640 medium (Life Technologies, Inc.) enriched with
10% FCS and 100 U/mL of IL-3 for 4 days; 1 day before harvesting of
cells, 1 mg/mL of CD40L trimer was added [16
].
Isolation of LCs from the skin
Full-thickness human skin specimens were obtained from healthy
donors undergoing plastic surgery of the breast or abdomen. These
specimens were shaved into slices of 0.2-mm thickness by using a
dermatome. To enable separation of the epidermis from the dermis, the
slices were incubated with 0.2% dispase II (Boehringer Mannheim,
Mannheim, Germany) in phosphate-buffered saline at 37°C for 30 min.
Subsequently, epidermal sheets were rinsed in phosphate-buffered saline
and cultured dermal side down in Iscove’s modified Dulbecco’s medium
(IMDM; Life Technologies, Inc.) enriched with 10% FCS and 50 µg/mL
of gentamicin (Sigma). After 42 h, the cells that had migrated out
of the epidermal sheets were collected [17
]. LCs were
enriched using anti-HLA-DR mAbs and magnetic cell sorting (MACS; CLB)
and were >98% pure as analyzed by fluorescence-activated cell
sorting.
TaqmanTM primers and probes
TaqmanTM probes were synthesized by PE-Applied
Biosystems (Branchburg, NJ). The chemokine-specific probes were labeled
at the 5' end with a 6-carboxyfluorescein (FAM) fluorescent group and
at the 3' end with a 6-carboxytetramethylrhodamine (TAMRA) quencher
group. Probes specific for the glyceraldehyde-3-phosphate dehydrogenase
(GAPDH) and porphobilinogen deaminase (PBGD) housekeeping genes were
labeled at the 5' end with a VIC fluorescent group and at the 3' end
with 6-carboxytetramethylrhodamine. Sequences of all primers and probes
used in this study are given in Table 1
.
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View this table: [in a new window] |
Table 1. Primers and Probes Used for Real-time Quantitative PCR
Analysis
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Chemokine sandwich ELISAs
The secretion of the chemokines DC chemokine 1 (DC-CK1),
macrophage-derived chemokine (MDC), and thymus and activation-regulated
chemokine (TARC) in culture supernatants was analyzed using specific
sandwich enzyme-linked immunosorbent assays (ELISAs). In the DC-CK1
ELISA, mouse anti-human DC-CK1 mAb (AZN-CK18B) (Lindhout, E., Torensma,
R., Guelen, L., Van Berkum, N., Trancikova, D., Looman, M., Ruers, T.,
Figdor, C.G., and Adema, G.A., unpublished data) was used as the
primary (catching) antibody (Ab) and goat anti-human PARC was used as
the secondary (detection) Ab (R&D Systems). The primary and secondary
antibodies for the MDC ELISA were mouse anti-human MDCs (clone 252Y; a
gift from R. Rodenburg) and rabbit anti-human MDCs (Preprotech Inc.,
Rocky Hill, NJ), respectively. In the TARC ELISA, rat anti-human TARC
mAb (clone 25D8; DNAX) and goat anti-human TARC (R&D Systems Europe,
Ltd., Abingdon, UK) were used as the primary and secondary antibodies,
respectively. Detection Abs were stained with horseradish peroxidase
(HRP)-conjugated donkey anti-goat IgG (Jackson Immunoresearch
Laboratory Inc., West Grove, PA) or HRP-conjugated goat anti-rabbit IgG
(Zymed Laboratories Inc., San Francisco, CA). HRP was visualized by
incubation with 3,3',5,5'-tetramethylbenzidine substrate. Absorption
was measured at 450 nm using a Titertek multiscan ELISA reader. The
detection limit for all ELISAs was 0.8 ng/mL.
IL-12p70 sandwich ELISA
The measurement of IL-12p70 (detection limit, 0.2 ng/mL) was
performed using a specific sandwich ELISA. Anti-human IL-12p70 mAb
(Endogen, Woburn, MA) was used as the primary Ab, and anti-human
IL-12p40/p70 biotin-labeled mAb (Endogen) was used as the detection Ab.
Streptavidin-HRP conjugate (CLB) was used to specifically stain the
biotin-labeled mAb and was visualized by incubation with
3,4,5-trimethoxybenzoic acid substrate. Absorption was measured at 450
nm.
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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We first analyzed chemokines expressed by monocyte-derived immature and
mature DCs. Immature DCs expressing major histocompatibility complex
(MHC) class I antigens, low levels of MHC class II antigens, and B7.2
(CD86) but neither B7.1 (CD80) nor CD83 were generated by culturing
monocytes in the presence of GM-CSF and IL-4. Immature DCs were
stimulated with LPS or TNF-, resulting in mature DCs that expressed
high levels of MHC class I and II antigens and were positive for B7.1,
B7.2, and CD83. Relative mRNA levels of chemokines produced by freshly
isolated monocytes, immature DCs, and mature DCs are depicted in
Figure 1
. Monocytes expressed high levels of IL-8 and smaller amounts of
macrophage-inflammatory protein (MIP)-1, MIP-3, and
lymphotactin (Fig. 1A)
. Expression of IL-8 in immature and mature DCs
was dramatically decreased (1,000-fold) compared to that in monocytes
(Fig. 1B)
. Interestingly, immature DCs expressed high levels of MDC and
TARC. TARC was up-regulated approximately 10-fold in mature DCs
compared to immature DCs, while MDC levels remained constant. DC-CK1
and EBI1-ligand chemokine (ELC) were expressed at intermediate levels
by immature DCs, but expression increased 60- and 7-fold, respectively,
upon maturation. The other chemokines tested were expressed at lower
levels, and no expression of stromal cell-derived factor (SDF) 1 or
SDF-1ß was detected in cultured DCs.
 View larger version (28K): [in a new window] |
Figure 1. Expression of chemokines by immature and mature monocyte-derived DCs.
Levels of chemokines expressed by freshly isolated monocytes (A) and by
immature monocyte-derived DCs (day 6) and DCs stimulated with LPS or
TNF- (B) were determined relative to those of PBGD. Each graph
represents data from one representative donor out of 3. (C) Sandwich
ELISA for DC-CK1, MDC, and TARC produced by immature DCs (for 48 h) and DCs stimulated with LPS or TNF- (for 48 h).
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Chemokine expression by cultured DCs is independent of FCS
The above data demonstrate that DCs cultured in medium
containing FCS expressed high levels of chemokines. To exclude the
involvement of FCS in the induction of chemokine expression, DCs were
cultured in autologous serum and subsequently analyzed for chemokine
expression. Figure 2
shows that immature DCs cultured in autologous serum (MHC class
I+ MHC class IIlow B7.2low
B7.1- CD83-) generally expressed levels of
chemokines comparable to those expressed by immature DCs cultured in
FCS. However, IL-8 was expressed to a higher level and TARC was
expressed to a lower level than in DCs cultured in FCS. In DCs
maturated with TNF-, PGE2, and MCM (MHC class
Ihigh MHC class IIhigh B7.2+
B7.1+ CD83+), expression of chemokines was
up-regulated; especially DC-CK-1, IL-8, MDC, and TARC were highly
expressed. These results are consistent with the chemokine levels
measured in mature DCs cultured in FCS and demonstrate that chemokine
expression by cultured DCs is independent of FCS.
 View larger version (31K): [in a new window] |
Figure 2. Relative mRNA expression levels of monocyte-derived DCs cultured in
autologous serum. Immature DCs were generated by culturing monocytes
with IL-4 and GM-CSF in the presence of autologous serum for 6 days.
Immature DCs were maturated using the combination of MCM, TNF-, and
PGE2. One representative experiment out of 4 is shown.
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differed
(Fig. 3
and Fig. 1B
). Activation with CD40L increased the expression of
ELC to high levels (up to 25-fold higher than those of immature DCs).
Furthermore, we found that, compared with immature DCs, CD40L-activated
DCs did not up-regulate or only slightly up-regulated DC-CK1. These
findings demonstrate that different stimuli up-regulate specific
chemokines in DCs.
 View larger version (30K): [in a new window] |
Figure 3. ELC but not DC-CK1 is up-regulated in CD40L-activated DCs. Relative
mRNA levels of chemokines expressed by immature DCs (day 6) and DCs
activated with CD40L were determined. One representative experiment out
of 4 is shown.
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produce large
amounts of IL-12 [22
, 23
]. We investigated
whether DCs expressing high levels of the T-helper-1-inducing cytokine
IL-12 down-regulate the expression of MDC and TARC. Immature DCs were
stimulated with CD40L and with a combination of CD40L and IFN-. Both
immature and CD40L-stimulated DCs expressed little IL-12; however, DCs
stimulated with a combination of IFN- and CD40L produced large
amounts of IL-12 (Fig. 4A
). Interestingly, we found that addition of IFN- to
CD40L-stimulated DCs had no effect on the expression of MDC and TARC
(Fig. 4B)
or on any chemokine in our panel. These data demonstrate that
both DC types produce high levels of MDC and TARC despite the
difference in their IL-12 production levels.
 View larger version (42K): [in a new window] |
Figure 4. MDC and TARC are highly expressed in DCs that produce high levels of
IL-12. (A) IL-12p70 production by immature DCs (day 6) and by DCs
stimulated with CD40L or a combination of CD40L and IFN-, measured
by ELISA. (B) Relative mRNA levels of DC-CK1, ELC, MDC, and TARC
expressed by these DCs. One representative experiment out of 2 is
shown.
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while the other tested chemokines were
expressed not at all or at minor levels (Fig. 5B)
. To investigate
whether chemokine expression is up-regulated in activated blood DCs,
CD11c- DCs were stimulated with IL-3 and CD40L
[16
]. CD11c+ DCs were stimulated with MCM,
which preferentially supports the maturation of CD11c+
blood DCs [15
]. Stimulated CD11c- DCs
clearly up-regulated the expression of ELC (100-fold) and MDC
(4,500-fold) and induced a relatively high level of expression of TARC
(Fig. 5B)
. In addition, these DCs expressed low levels of IL-8,
lymphotactin, and MIP-1. Interestingly, CD11c- DCs did
not express DC-CK1. Mature CD11c+ DCs expressed ELC, IL-8,
MDC, MIP-1, and TARC and, to a lesser extent, DC-CK1, lymphotactin,
and MIP-3 (Fig. 5B)
. ELISAs confirmed the secretion of MDC and TARC
by blood DCs (Fig. 5C)
, while the production of DC-CK1 did not exceed
background levels (data not shown). Altogether, these data indicate
that the chemokine profiles of activated blood DCs are surprisingly
similar to the chemokine profile of monocyte-derived DCs, although the
expression levels are significantly lower.
 View larger version (36K): [in a new window] |
Figure 5. Relative mRNA levels of chemokines expressed by freshly isolated blood
DCs. (A) Freshly isolated blood DCs were stained with PE-conjugated Abs
to CD3, CD14, CD16, and CD19 and FITC-conjugated anti-HLA-DR Ab. Blood
DCs were defined as lineage-negative and HLA-DR-positive cells (lower
right quadrant). Nuclei of freshly isolated DCs were stained with
hematoxylin and eosin, and surface markers were identified with
PE-conjugated CD11c, CD80, CD83, and CD86. (B) Relative mRNA levels of
chemokines expressed by freshly isolated blood DCs, CD11c-
DCs stimulated with IL-3 and CD40L, and CD11c+ DCs
stimulated with MCM. Each graph represents data from one representative
donor out of 3. (C) Sandwich ELISA for MDC and TARC produced by
activated CD11c- DCs and CD11c+ DCs (for 3
days).
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(Fig. 6
). The chemokines DC-CK1, SDF-1, and SDF-1ß were not detected.
Interestingly, LCs expressed chemokines at levels similar to those of
activated peripheral blood DCs.
 View larger version (14K): [in a new window] |
Figure 6. LCs express high levels of IL-8, MDC, and TARC. LCs were allowed to
migrate out of epidermal sheets for 2 days. Relative mRNA levels of
chemokines expressed by LCs were measured. One representative
experiment out of 3 is shown.
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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, SDF-1, and SDF-1ß (data not shown). Real-time quantitative PCR analysis revealed that monocyte-derived DCs express high levels of the chemokines MDC and TARC. A recent study using serial analysis of gene expression also demonstrated an abundance of messages for MDC and TARC in cultured DCs [27 ]. Additionally, we demonstrated by ELISA that cultured DCs secrete large amounts of MDC and TARC proteins. Both chemokines attract T helper 2 cells, which express the chemokine receptor CCR4 [21 ]. Recently, it has been shown that monocyte-derived DCs mediating the induction of a T helper 1 response produce a cytokine profile different from that of those mediating the induction of a T helper 2 response [22 23 ]. Interestingly, we showed that these two DC populations express similar levels of chemokines including MDC and TARC. This suggests that with regard to chemokine levels, monocyte-derived DCs do not discriminate between the attraction of T helper 1 and T helper 2 cells. Rather, it seems that monocyte-derived DCs are able to attract naïve T cells (DC-CK1 and ELC) as well as memory T cells (MDC and TARC), which have also been shown to express CCR4 [28 ]. Additionally, NK cells have been shown to express CCR4 and as a result respond to TARC and MDC [29 30 ]. Thereby, DCs may provide a link between the innate and acquired immune responses, since they can attract and activate NK cells [31 ].
Maturation of monocyte-derived DCs by either LPS or TNF- induced a
significant up-regulation of DC-CK1 (60-fold), ELC (7-fold), and TARC
(10-fold), while the high-level MDC expression remained constant. Upon
arrival of mature DCs in secondary lymphoid tissue, DCs can attract
naive T cells by secretion of the DC-specific chemokine DC-CK1
[32
]. MDC and TARC will attract subpopulations of
recently activated and memory T cells. In the case of an
antigen-specific interaction, the DCs will activate the T cells,
resulting in up-regulation of CD40L. CD40L in turn further activates
the DCs. Interestingly, we observed that ligation of the CD40 molecule
on cultured DCs induced a further up-regulation of ELC (25-fold). ELC
can attract additional mature DCs and subsets of T lymphocytes
expressing CCR7 [33
] and thereby help to sustain an
inflammatory reaction. Accordingly, expression of CCR7 gradually
increases during DC maturation [26
]. These findings
support an important role for chemokines produced by mature DCs in the
organization of secondary lymphoid structures.
Compared to the PBGD housekeeping gene, monocyte-derived DCs produce large amounts of chemokines. The analysis of immature DCs cultured in autologous serum showed that these chemokine levels were independent of the presence of FCS. The most significant difference between DCs cultured in medium containing autologous serum and those cultured in the presence of FCS was a higher level of expression of IL-8, which was up-regulated during maturation in the former. Monocytes produce high levels of IL-8 which decrease during the differentiation of these cells into DCs (data not shown). Therefore, immature DCs cultured in the presence of autologous serum seem less differentiated from monocytes than FCS-cultured DCs. This is supported by the finding that we were not able to maturate autologous–serum-cultured DCs by addition of a single stimulus. The combination of stimuli needed for the full maturation of these DCs could have contributed to the observed up-regulation of IL-8 expression.
We isolated peripheral blood DCs and LCs to study chemokine expression
by DC subsets in vivo. Freshly isolated blood DCs expressed only very
low levels of IL-8, lymphotactin, and MIP-1, while activated blood
DCs expressed nearly all tested chemokines; ELC, MDC, and TARC were
up-regulated most strongly, while SDF1- and SDF-1ß were not
detected. Strikingly, the CD11c- lymphoid-like and
CD11c+ myeloid blood DC subsets had comparable chemokine
profiles. The DC-specific chemokine DC-CK1 was detected only in
activated CD11c+ blood DCs. The absence of DC-CK1
expression in activated CD11c- DCs could either represent
a lineage-dependent difference or result from the difference in
stimulation of the two subsets. The chemokine expression pattern of
activated blood DCs closely resembled that of monocyte-derived DCs.
Consequently, peripheral blood DCs are able to attract the same
population of cells as monocyte-derived DCs. The absolute chemokine
levels expressed by peripheral blood DCs were significantly lower than
in monocyte-derived DCs. However, in vivo, the chemokine concentration
will be dependent not only on the amount of protein produced but also
on chemokine stability and the capture of chemokines by extracellular
matrix proteins such as proteoglycans. This could allow chemokines
expressed at lower concentrations to be functionally active in the
appropriate microenvironment.
We (Fig. 5A) and others [15 24 ] have demonstrated that the CD11c- blood DC subsets can be clearly distinguished phenotypically from the CD11c+ blood DCs. However, at present it is still uncertain which cells in vivo correspond to the in vitro-stimulated blood DC subsets. The presence of lymphoid-like CD4+ CD11c- DCs, also referred to as plasmacytoid cells, surrounding the high endothelial venules (HEV) in secondary lymphoid organs [34 35 ] strongly suggests that lymphoid-like DCs have directly migrated from the blood through the HEV into the T-cell area of the lymph node. This migration pattern is functionally distinct from that of DCs which have migrated from peripheral tissues to secondary lymphoid tissues through the afferent lymphatic system. CD11+ blood DCs are also capable of migrating across HEV but seem to migrate into the B-cell areas, likely represented by the phenotypically similar CD4+ CD11c+ germinal-center DCs [15 34 ]. This is supported by specific expression of the B-lymphocyte chemoattractant CXCL13 in both germinal-center DCs and activated CD11c+ blood DCs [36 ]. The presence of high levels of costimulatory markers on the activated blood DCs used for our analysis indicates that these cells most likely correspond to these lymphoid-tissue DCs.
LCs are bone marrow-derived DCs from the myeloid lineage that localize in the epidermis of the skin and are involved in antigen uptake [25 ]. The LC population used for our PCR analysis had migrated out of the epidermis over a 2-day period, thereby inducing LC activation. These LCs mainly express IL-8, MDC, and TARC, which are also expressed at high levels by activated blood DCs. LCs do not express DC-CK1 and express only minor levels of ELC, although these chemokines are expressed by activated blood DCs (DC-CK1 only by CD11c+ blood DCs) and in vitro-cultured DCs. Further studies in our laboratory indicate that at least DC-CK1 is a lineage-specific chemokine expressed by DCs and is not expressed in LCs, supporting the theory that LCs form a separate myeloid DC lineage.
DCs attract different cell types by adjusting their production of chemokines. Chemokine expression can be influenced by signals from inflammatory agents, contact with T cells, and a changing microenvironment. Here we demonstrated that despite differences in chemokine expression levels, all DC subsets, in vitro and in vivo, are able to attract the same leukocyte subsets. Therefore, regulation at the level of chemokine receptor expression on responder cells, including DCs, is also important. In addition, high levels of chemokines will desensitize the receptor and render cells unresponsive to the appropriate chemokine(s), allowing responder cells to reside at particular sites despite their continued exposure to chemokines. Although various aspects of the chemokine system still have to be unraveled, chemokine regulation is now well appreciated to be essential for immune cells to find the proper partner at the right place and time during the immune response.
We thank Dr. Ewald Mensink and Louis van de Locht (Department of Hematology, UMC Nijmegen St. Radboud, Nijmegen, The Netherlands) for giving us advice on the ABI/PRISM 7700 Sequence Detector System. We thank Maaike Looman, Dagmar Trancikova, and Linda Engelen for technical assistance. We thank Dr. Jolanda de Vries for stimulating discussions and critical reading of the manuscript.
Received October 17, 2000; revised December 28, 2000; accepted December 29, 2000.
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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A W T van Lieshout, P Barrera, R L Smeets, G J Pesman, P L C M van Riel, W B van den Berg, and T R D J Radstake Inhibition of TNF{alpha} during maturation of dendritic cells results in the development of semi-mature cells: a potential mechanism for the beneficial effects of TNF{alpha} blockade in rheumatoid arthritis Ann Rheum Dis, March 1, 2005; 64(3): 408 - 414. [Abstract] [Full Text] [PDF]
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T R D J Radstake, R van der Voort, M ten Brummelhuis, M de Waal Malefijt, M Looman, C G Figdor, W B van den Berg, P Barrera, and G J Adema Increased expression of CCL18, CCL19, and CCL17 by dendritic cells from patients with rheumatoid arthritis, and regulation by Fc gamma receptors Ann Rheum Dis, March 1, 2005; 64(3): 359 - 367. [Abstract] [Full Text] [PDF]
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E. Real, A. Kaiser, G. Raposo, A. Amara, A. Nardin, A. Trautmann, and E. Donnadieu Immature Dendritic Cells (DCs) Use Chemokines and Intercellular Adhesion Molecule (ICAM)-1, But Not DC-Specific ICAM-3-Grabbing Nonintegrin, to Stimulate CD4+ T Cells in the Absence of Exogenous Antigen J. Immunol., July 1, 2004; 173(1): 50 - 60. [Abstract] [Full Text] [PDF]
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B. L. Colvin, A. E. Morelli, A. J. Logar, A. H. Lau, and A. W. Thomson Comparative evaluation of CC chemokine-induced migration of murine CD8{alpha}+ and CD8{alpha}- dendritic cells and their in vivo trafficking J. Leukoc. Biol., February 1, 2004; 75(2): 275 - 285. [Abstract] [Full Text] [PDF]
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L. Stievano, V. Tosello, N. Marcato, A. Rosato, A. Sebelin, L. Chieco-Bianchi, and A. Amadori CD8+{alpha}{beta}+ T Cells That Lack Surface CD5 Antigen Expression Are a Major Lymphotactin (XCL1) Source in Peripheral Blood Lymphocytes J. Immunol., November 1, 2003; 171(9): 4528 - 4538. [Abstract] [Full Text] [PDF]
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I. J. M. de Vries, W. J. Lesterhuis, N. M. Scharenborg, L. P. H. Engelen, D. J. Ruiter, M.-J. P. Gerritsen, S. Croockewit, C. M. Britten, R. Torensma, G. J. Adema, et al. Maturation of Dendritic Cells Is a Prerequisite for Inducing Immune Responses in Advanced Melanoma Patients Clin. Cancer Res., November 1, 2003; 9(14): 5091 - 5100. [Abstract] [Full Text] [PDF]
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L. Skelton, M. Cooper, M. Murphy, and A. Platt Human Immature Monocyte-Derived Dendritic Cells Express the G Protein-Coupled Receptor GPR105 (KIAA0001, P2Y14) and Increase Intracellular Calcium in Response to its Agonist, Uridine Diphosphoglucose J. Immunol., August 15, 2003; 171(4): 1941 - 1949. [Abstract] [Full Text] [PDF]
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C. M. van Herpen, R. Huijbens, M. Looman, J. de Vries, H. Marres, J. van de Ven, R. Hermsen, G. J. Adema, and P. H. De Mulder Pharmacokinetics and Immunological Aspects of a Phase Ib Study with Intratumoral Administration of Recombinant Human Interleukin-12 in Patients with Head and Neck Squamous Cell Carcinoma: A Decrease of T-bet in Peripheral Blood Mononuclear Cells Clin. Cancer Res., August 1, 2003; 9(8): 2950 - 2956. [Abstract] [Full Text] [PDF]
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M. Vulcano, S. Struyf, P. Scapini, M. Cassatella, S. Bernasconi, R. Bonecchi, A. Calleri, G. Penna, L. Adorini, W. Luini, et al. Unique Regulation of CCL18 Production by Maturing Dendritic Cells J. Immunol., April 1, 2003; 170(7): 3843 - 3849. [Abstract] [Full Text] [PDF]
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O. Bruna-Romero, J. Schmieg, M. Del Val, M. Buschle, and M. Tsuji The Dendritic Cell-Specific Chemokine, Dendritic Cell-Derived CC Chemokine 1, Enhances Protective Cell-Mediated Immunity to Murine Malaria J. Immunol., March 15, 2003; 170(6): 3195 - 3203. [Abstract] [Full Text] [PDF]
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A. la Sala, D. Ferrari, F. Di Virgilio, M. Idzko, J. Norgauer, and G. Girolomoni Alerting and tuning the immune response by extracellular nucleotides J. Leukoc. Biol., March 1, 2003; 73(3): 339 - 343. [Abstract] [Full Text] [PDF]
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Y. K. Choi, B. A. Fallert, M. A. Murphey-Corb, and T. A. Reinhart Simian immunodeficiency virus dramatically alters expression of homeostatic chemokines and dendritic cell markers during infection in vivo Blood, March 1, 2003; 101(5): 1684 - 1691. [Abstract] [Full Text] [PDF]
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G. Penna, M. Vulcano, A. Roncari, F. Facchetti, S. Sozzani, and L. Adorini Cutting Edge: Differential Chemokine Production by Myeloid and Plasmacytoid Dendritic Cells J. Immunol., December 15, 2002; 169(12): 6673 - 6676. [Abstract] [Full Text] [PDF]
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J. H. Ahn, Y. Lee, C. Jeon, S.-J. Lee, B.-H. Lee, K. D. Choi, and Y.-S. Bae Identification of the genes differentially expressed in human dendritic cell subsets by cDNA subtraction and microarray analysis Blood, August 13, 2002; 100(5): 1742 - 1754. [Abstract] [Full Text] [PDF]
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M. K. Park, D. Amichay, P. Love, E. Wick, F. Liao, A. Grinberg, R. L. Rabin, H. H. Zhang, S. Gebeyehu, T. M. Wright, et al. The CXC Chemokine Murine Monokine Induced by IFN-{gamma} (CXC Chemokine Ligand 9) Is Made by APCs, Targets Lymphocytes Including Activated B Cells, and Supports Antibody Responses to a Bacterial Pathogen In Vivo J. Immunol., August 1, 2002; 169(3): 1433 - 1443. [Abstract] [Full Text] [PDF]
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S. A. Luther, A. Bidgol, D. C. Hargreaves, A. Schmidt, Y. Xu, J. Paniyadi, M. Matloubian, and J. G. Cyster Differing Activities of Homeostatic Chemokines CCL19, CCL21, and CXCL12 in Lymphocyte and Dendritic Cell Recruitment and Lymphoid Neogenesis J. Immunol., July 1, 2002; 169(1): 424 - 433. [Abstract] [Full Text] [PDF]
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H. Ni, J. Capodici, G. Cannon, D. Communi, J.-M. Boeynaems, K. Kariko, and D. Weissman Extracellular mRNA Induces Dendritic Cell Activation by Stimulating Tumor Necrosis Factor-alpha Secretion and Signaling through a Nucleotide Receptor J. Biol. Chem., April 5, 2002; 277(15): 12689 - 12696. [Abstract] [Full Text] [PDF]
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A. la Sala, S. Sebastiani, D. Ferrari, F. Di Virgilio, M. Idzko, J. Norgauer, and G. Girolomoni Dendritic cells exposed to extracellular adenosine triphosphate acquire the migratory properties of mature cells and show a reduced capacity to attract type 1 T lymphocytes Blood, March 1, 2002; 99(5): 1715 - 1722. [Abstract] [Full Text] [PDF]
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