| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Regular Article |
From the Department of Dermatology, University of Würzburg Medical School, Würzburg, Germany
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
In a mouse CHS model Grabbe and
colleagues3
demonstrated that nonspecific effects of
epicutaneously applied haptens contribute to the elicitation of
CHS. Proinflammatory irritative rather than antigen-specific properties
of the hapten are furthermore responsible for the strict
concentration-dependence of the effector phase of CHS. Therefore,
it is tempting to speculate that such irritative properties of haptens
promote inflammatory skin reactions via induction of proinflammatory
cytokines, adhesion molecules, and chemoattractants. Accordingly, some
contact allergens such as urushiol, the relevant hapten in poison ivy,
and nickel sulfate have been demonstrated to directly induce
inflammatory activation of keratinocytes resulting in expression of
ICAM-1, interleukin (IL)-8, and/or tumor necrosis factor
(TNF)-
.4-6
In recent years, in particular chemokines
have emerged as potent stimulators of effector cell accumulation and
activation and are likely candidates to mediate leukocyte recruitment
during elicitation of CHS. Since the description of IL-8 more than a
decade ago, the supergene family of chemokines has increased enormously
and comprises a set of more than 30 different species.7-9
For many of them, the detailed understanding of their in
vivo role is fragmentary or even lacking. This is mainly because
of the limited availability of in situ data obtained under
pathological conditions. The individual role of chemokines during CHS
has been studied only for single species (eg, IP-10, MCP-1, Eotaxin)
and mainly in the murine system.10-13
Because mouse skin
exhibits quite a different morphology and physiology as compared to
human skin (eg, only few keratinocyte layers, high abundance of hair
follicles) and because chemokine homologues for both the human and
murine system are not always established, inflammatory disorders such
as CHS reactions should be studied, despite experimental restrictions,
in the human system itself. As most chemokines are regarded to be
redundant in their action on target cells, promiscuous in receptor
usage, and produced by both resident and passenger cells (leukocyte
subtypes) of the skin,14
understanding of the inflammatory
response in CHS requires the in vivo study of an extended
set of chemokines. Furthermore, diapedesis and migration from the
dermal into the epidermal compartment may be regulated by a sequential
and spatial different set of chemokines as recently demonstrated for
other inflammatory conditions of the skin.15,16
In
particular, the elegant in vitro studies of neutrophil
locomotion through complex fields of overlapping chemoattractant
gradients by Foxman and colleagues17
have strengthened the
still unproven concept that the spatial guidance of cells in tissue may
not be a linear event of a single chemokine gradient but rather
requires a complex network of sequential and combinatorial
chemoattractant effects. To prove this concept of effector cell
recruitment and positioning in vivo we applied in
situ hybridization to study a large panel of radiolabeled
chemokine anti-sense probes on serial tissue sections of biopsies
obtained at various time intervals after allergen exposure to the skin
of sensitized volunteers. We particularly focused our attention on
chemokines that are chemoattractive for monocytes and/or lymphocytes
such as MCP-1, MCP-3, macrophage inflammatory protein-1 (MIP-1)
and {beta}, RANTES, MDC, I309, IP-10, MIG, liver and
activationregulated chemokine (LARC), (synonymous with
Exodus-1/MIP-3), PARC (synonymous with DC-CK1/MIP4), TARC,
lymphotactin, stromal cell-derived factor-1 (SDF-1) and {beta}, and
interferon-inducible T cell alpha chemoattractant (I-TAC). In addition,
we studied expression of IL-8, growth-related oncogene
(Gro)/MGSA, granulocyte chemoattractant protein-2 (GCP-2), and
endothelial neutrophil-attracting protein 78 (ENA-78) that primarily
attract neutrophils as well as of hemofiltrate C-C chemokine (HCC-1).
The microanatomical location of chemokine expression and the
longitudinal profile was semiquantitatively evaluated and compared with
the infiltration pattern of leukocyte subtypes. We demonstrate that
during elicitation of CHS a highly diversified repertoire of chemokines
is expressed at distinct sites of the skin that spatially and
temporally correlates with the recruitment of macrophages and T
cells.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Thirteen patients referred to the Allergy Section of the Department of Dermatology, University of Würzburg, for evaluation of contact allergies by patch testing were included in this study after having given informed written consent. The study was approved by the Ethics committee at the University of Würzburg. Separate epicutaneous patch tests were performed on the volar forearm using occlusive Finn Chambers (Hermal, Reinbek, Germany). Standard concentrations of allergens prepared in soft paraffin (Hermal) were used to reproduce CHS reactions. All patch-test chambers were removed at the time of biopsy or after a maximum of 48 hours of contact. After local anesthesia with mepivacaine, 5-mm-punch skin biopsies (four to five biopsies from each individual) were taken from the patch-test sites before and 6, 12, 24, 48, 72, and/or 96 hours after allergen application. Evaluation of quantitative data is based on results obtained from at least four biopsies per time point. CHS reactions were elicited in five patients by 5% nickel sulfate; in three patients by 20% colophony; and in one patient by each isocillin, 2% nystatin, 0.25% sodium thiosulfatoaurate, 1% paraphenylenediamine, or 20% neomycin sulfate, respectively. Tissue samples were embedded in OCT compound (Tissue Tek, Diatec, Nürnberg, Germany) immediately after punch biopsy, frozen, and stored at -80°C. Cryostat sections (5 µm) were prepared on gelatin-coated slides (Merck, Darmstadt, Germany) for immunohistology and on poly-L-lysine-coated slides (Sigma, Deissenhofen, Germany) for in situ hybridization. After air-drying, sections were fixed in acetone (10 minutes, 4°C) for immunohistology or, for in situ hybridization, in 4% paraformaldehyde/phosphate-buffered saline (PBS) (20 minutes, room temperature; Sigma). Alternatively, specimens were fixed in 4% formaldehyde and embedded in paraffin wax for a superior microanatomical preservation.
Immunohistology
A three-step streptavidin-biotin-complex (StreptAB-Complex)/peroxidase method was used as previously described.18 The following monoclonal antibodies (mAbs) were used: anti-CD3 (at 1:500; BD, Sunnyvale, CA), reacting with the T cell receptor-associated CD3 antigen; anti-CD4, anti-CD8 (both at 1:200; BD); anti-HLA-DR (at 1:500; BD); anti-CD68 (clone KP-1, at 1:1,000; DAKO, Copenhagen, Denmark), recognizing macrophages; anti-neutrophil elastase (at 1:200; DAKO), specifically labeling neutrophils; anti-CXCR1 (clone 5A12 at 1:1,000; PharMingen, Hamburg, Germany), anti-CXCR2 (clone 6C6 at 1:1,000, PharMingen) and anti-CXCR3 (clone 44716.111 at 1:500; R&D Systems, Wiesbaden, Germany) against human chemokine receptors. After blocking Fc receptors with 20% heat-inactivated sheep serum in PBS, sections were incubated with primary mAb at 4°C overnight, followed by incubation with biotin-conjugated sheep anti-mouse IgG (Amersham Pharmacia Biotech, Freiburg, Germany) at 1:200 and preformed StreptAB-Complex/Peroxidase (DAKO) at room temperature for 1 hour. Sections were washed between each step and the reaction cascade was visualized by incubation with 3-amino-9-ethylcarbazole (Sigma) as substrate. For control purposes the primary mAb was replaced by isotype-matched IgG of nonrelevant specificity.
In Situ Hybridization
Preparation of 35S-Labeled RNA Probes
cDNA probes were provided by J. M. Farber (NIH, Bethesda,
MD), MIG; T. Yoshimura (NCI, Frederick, MA), MCP-1 and Eotaxin-1;
Genetics Institute (Cambridge, MA), MIP-1; T. Schall (DNAX Research
Institute, Palo Alto, CA), MIP-1{beta} and RANTES; J. A. Hedrick
(DNAX Research Institute), lymphotactin; A. Anisowicz (Dana Farber
Cancer Institute, Boston, MA), GRO; C. Weissmann (University of
Zürich, Zürich, Switzerland), IL-8; C. Müller
(University of Bern, Bern, Switzerland), MCP-3; R. Kulke (University of
Kiel, Kiel, Germany), ENA-78; M. Krangel (Duke University Medical
Center, Durham, NC), I309; W. G. Forssmann (Lower Saxony Institute
for Peptide Research, Hannover, Germany), HCC-1; J. B. Smith (UCLA
Medical Center, Los Angeles, CA), GCP-2; H. Nomiyama (Kumamoto
University, Kumamoto, Japan), PARC; O. Yoshie (Shionogi Institute for
Medical Science, Osaka, Japan), LARC; R. Godiska and P. Gray (ICOS,
Bothell, WA), MDC; and T. Honjo (Kyoto University Faculty of Medicine,
Kyoto, Japan), SDF and {beta}.
IP-10, TARC, and I-TAC were cloned by polymerase chain reaction using
the following primers: IP-10,
5'CGC-AAG-CTT-CGG-GAG-ACA-TTC-CTC-AAT-TGC-3' and
5'CGC-GGA-TCC-AGG-AGA-TCT-TTT-AGA-CAT-TTC-3' (with
HindIII and BamHI restriction sites); TARC,
5'ATG-GCC-CCA-CTG-AAG-ATG-3' and TCA-AGA-CCT-CTC-AAG-GCT-3'; and I-TAC,
5'CGG-GAT-CCC-GAT-GAG-TGT-GAA-GGG-CAT-3' and
5'CCG-CTC-GAG-CGG-TTA-AAA-ATT-CTT-TCT-TTC-3'. For polymerase chain
reaction amplification of TARC, PHA-stimulated peripheral blood
mononuclear cells were used whereas I-TAC and IP-10 were generated from
interferon (IFN)-
-stimulated buffy-coat monocytes. Total RNA was
then isolated and cDNA prepared using the primers described above
according to standard protocols. In vitro transcription of
sense and anti-sense probes was performed as described
earlier.15,16
Radiolabeled probes were obtained by
incubation of linearized plasmids with either T7, T3, or SP6 RNA
polymerases and ATP, GTP, CTP (all obtained from Roche Molecular
Biochemicals, Mannheim, Germany) and 35S-UTP
(Amersham Pharmacia Biotech) as substrates. The original template cDNA
was eliminated by DNase treatment and protein components were removed
by several phenol extraction steps. To facilitate the intracellular
accessibility of labeled probes, alkaline hydrolysis was performed to
get an average length of 50 to 150 bp. The radioactive probes
were adjusted to a specific activity of 2 x
106
cpm/µl in 0.01 mol/L Tris-HCl, pH 7.5,
containing 1 mmol/L ethylenediaminetetraacetic acid.
Hybridization Procedure
In situ hybridization was performed as previously described.15,16 Cryostat sections were fixed in 4% paraformaldehyde and treated with 1 µg/ml proteinase K (Roche Molecular Biochemicals). Sections were acetylated with acetic anhydride in 0.1 mol/L triethanolamine (pH 8.0, 10 minutes), dehydrated in alcohol, and air-dried. Sections were then overlaid with 20 µl of hybridization solution (50% formamide, 300 mmol/L NaCl, 20 mmol/L Tris-HCl, pH 8.0, 5 mmol/L ethylenediaminetetraacetic acid, 1x Denhardt’s solution, 10% dextran sulfate, 100 mmol/L dithiothreitol, and 2 x 105 cpm/µl heat-denaturated radioactive probe). All sense (control) and anti-sense probes were hybridized to at least three sections from the same biopsy. RNase treatment was used as a further control and consistently abrogated specific hybridization signals. After hybridization, sections were washed with a solution containing 50% formamide, 2x standard saline citrate (Sigma), and 5 mmol/L ethylenediaminetetraacetic acid at high stringency (54 to 57°C). To remove nonhybridized probes, slides were treated with RNase A (20 µl/ml) and RNase T1 (1 U/ml; Roche Molecular Biochemicals) for 30 minutes at 37°C. To visualize the hybridization reaction, slides were dipped in NTB solution (Kodak) and exposed for 1 to 4 weeks at 4°C.
Assessment of Leukocyte Subsets and Chemokine mRNA-Expressing Cells
Slides processed for in situ hybridization or immunohistochemistry were evaluated and documented with an Axiophot microscope (Zeiss, Oberkochen, Germany) equipped with interference contrast, epipolarization, and dark field illumination. For quantification of leukocyte subsets, positive cells in the dermis were counted using an ocular grid. They were related to the total number of cells in dermal areas whereas resident epidermal cells and cells infiltrating the epidermis were not being considered. Biopsy specimens from four to seven individual CHS reactions for each time point were examined. At least three randomly selected fields were evaluated in each case. Data are expressed as percentage of positive cells ±SEM. In addition, if applicable, the site of preferential expression was photographically documented.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
In a first series of experiments, sections of biopsies taken at
distinct time intervals (0, 6, 12, 24, 48, 72, and 96 hours) after
epicutaneous application of contact allergens were histologically
evaluated. After 6 hours of allergen application, the histological
picture still resembled that of normal skin whereas after 12 hours a
slight intercellular edema in the epidermis and some CD3+ T lymphocytes
and CD68+ macrophages scattered around superficial and deeper dermal
vessels as well as single macrophages immediately below the epidermis
could be observed (data not shown). After 24 hours, multifocal
infiltration of T cells into the epidermis with concomitant formation
of epidermal microvesicles occurred whereas monocytes/macrophages
almost exclusively resided in the subepidermal region. Later, at 48 to
72 hours, mononuclear cells increasingly infiltrated the upper dermal
compartment and pronounced epidermal edema with vesiculation was
visible. At this stage, neutrophils were detectable within the
edematous or blistered epidermis. Despite a variable degree of
infiltration and vesiculation between biopsies obtained from different
patients at the same time point after allergen exposure, the relative
composition of infiltrates was comparable with a clear predominance of
CD3+ T cells and CD68+ macrophages reaching maximum densities at 48 to
72 hours. Composition and time course of infiltrate formation during
elicitation of CHS are summarized in Figure 1
.
|
To assess the time course and the expression patterns of
chemokines attracting monocytes and macrophages, serial sections of
biopsies taken before or 6, 12, 24, 48, 72, and 96 hours after allergen
exposure were hybridized with anti-sense probes of I309, MCP-1, MCP-3,
RANTES, MIP-1, and MIP-1{beta}. In parallel, serial sections were
studied for localization of leukocyte subsets.
Before application of allergens (ie, at 0 hours) significant mRNA
expression of the chemokines studied was not detectable. After 6 hours
of allergen exposure, multifocal expression of MCP-1 was already
visible in single basal keratinocytes as shown in Figure 2A
. The observation that MCP-1 was the
first chemokine found to be up-regulated after elicitation of CHS was
made regardless of the kind of allergen applied. Notably, at this early
time point the dermal compartment still appeared to be unchanged and an
increased number of inflammatory cells could not be detected. At 12
hours, MCP-1 mRNA expression was further up-regulated with an even
and strong expression in the basal epidermal layer and a weak
expression in single vascular and perivascular cells (Figure 2B)
accompanied by a rather scarce perivascular infiltrate (data not
shown). MCP-1 mRNA expression further increased up to 48 hours as
demonstrated in Figure 2, C and D
, and declined after 72 hours. At
these time points, despite a massive expression in basal keratinocytes
overall expression of MCP-1 was higher in the dermal compartment
because of a larger number of strong MCP-1 mRNA expressing cells
(Figure 2, C and D)
. Expression of RANTES was first detected between 12
and 24 hours in keratinocytes of the basal layer in a pattern
resembling that of MCP-1. Later, RANTES expression in the basal layer
further increased and was observed in mononuclear cells immediately
below the epidermis (Figure 3, C and D)
.
Expression of most other monocyte-attractant chemokines was weak
(MIP-1 and MIP1-{beta}; Figure 3, E and F
) or undetectable (MCP-3,
I309; data not shown). A semiquantitative evaluation of chemokine
mRNA-expressing cells is shown in Figure 4
. In addition to the differential
expression patterns, the specificity of hybridization was further
confirmed using sense probes that did not result in cell-associated
hybridization signals (data not shown).
|
|
|
; A, B, and C). Accordingly,
monocytes/macrophages were only rarely detected in the epidermis beyond
the basal layer.
|
Because CHS is considered as a prototype of a T-cell-mediated
inflammatory reaction, it was mandatory to assess the expression
patterns of chemokines exhibiting lymphocyte-attractant properties.
Serial skin sections were hybridized with anti-sense probes of IP-10,
MIG, MDC, RANTES, LARC (Exodus-1), PARC (DC-CK1, MIP-4), TARC, MIP-1
and -{beta}, MCP-1 and -3, lymphotactin, I-TAC, HCC-1, IL-8, Gro, and
SDF and -{beta}. Among all chemokines studied MIG- and IP-10-specific
probes showed the highest levels of cell-associated signals. Expression
of these started between 12 and 24 hours after allergen application and
reached maximal levels after 72 hours (Figure 6)
. Notably, both chemokines were
preferentially expressed at identical sites of the epidermis and,
unlike MCP-1, could also be observed at suprabasal layers of the
epidermis after 48 hours (Figure 6)
. In serial sections, epidermal
MCP-1 mRNA expression was always restricted to the basal layer.
Increasing expression of IP-10 and particularly of MIG was additionally
detected in the subepidermal region and co-localized expression of both
chemokines correlated with dense infiltration of CD3+ and CXCR3+
lymphocytes in the dermis and exocytosis into the epidermis (Figure 5
;
D, E, and F; and data not shown). MCP-1 and RANTES, which attract both
monocytes and lymphocytes, exhibited a similar expression pattern,
albeit with another time course as described for IP-10 and MIG and were
not detected to be expressed in suprabasal layers. In particular, MCP-1
expression started much earlier than that of IP-10 and MIG. A
quantitative evaluation of IP-10 and MIG mRNA expression is given in
Figure 7
.
|
|
. MDC mRNA was
first found to be moderately expressed by perivascular mononuclear
cells around superficial vessels at 12 hours (data not shown). Maximal
expression levels after 48 to 72 hours were frequently detected in
areas of strong leukocyte infiltration and were sometimes much stronger
than expression of MCP-1 (Figure 9)
.
Single mononuclear cells directly below the epidermis or intermingled
between basal keratinocytes and thus identified as
monocytes/macrophages also expressed MDC message; however, as opposed
to MCP-1, MDC mRNA has never been detected in epidermal keratinocytes
(Figure 9, C and D)
. The time course and quantity of MDC, LARC, PARC,
and TARC mRNA expression is summarized in Figure 10
. All other chemokines studied were
only weakly expressed or not detectable (data not shown). IL-8 and
GRO which, besides their role as important neutrophil attractants,
have also been implicated in T cell trafficking,19,20
were
found to be weakly expressed in the epidermis (IL-8) or both the dermis
and epidermis (GRO; data not shown). Expression of IL-8 and GRO
was multifocal and especially seen in areas of epidermal edema and
blister formation that were infiltrated by single neutrophil elastase+,
CXCR1+, and CXCR2+ cells, ie, neutrophils (data not shown). These
observations suggest that, during elicitation of CHS, both CXC
chemokines influence neutrophil recruitment rather than T cell
migration. Message of neutrophil-attracting chemokines ENA-78 and GCP-2
could not be detected at all (data not shown).
|
|
|
. In summary, during elicitation of CHS the chemokines MIG,
IP-10, RANTES, and MCP-1 are strongly expressed in the epidermis and,
to a lesser extent, in the dermis. Furthermore, perivascular expression
of MDC, LARC, and PARC can be observed that is accompanied by
infiltration of lymphocytes. |
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
and -{beta},
GRO, IL-8, and lymphotactin) or undetectable (I309, LARC, Eotaxin-1,
I-TAC, GCP-2, ENA-78). Our study which we believe is the first that
analyzed the complexity of chemokine action in vivo
indicates that a restricted and timely changing set of chemokines
governs the migration of monocytes and lymphocytes during evolving CHS
in human skin. Notably, with the exception of IP-10, MIG, PARC, and
TARC as selective T cell attractants, most other chemokines detected in
CHS (MCP-1, RANTES, MDC) are more or less chemoattractant for both
monocytes and lymphocytes but not for neutrophils.7,9
This
may explain the predominance of mononuclear cells in CHS and the
simultaneous immigration of lymphocytes and macrophages into areas of
allergen-exposed skin. Moreover, the preponderance of lymphocytes among
the mononuclear cells in CHS (Figure 1)
may be explicable by a broader
spectrum of lymphocyte-attractant chemokines (IP-10, MIG, MCP-1, MDC,
PARC, RANTES, TARC) than of monocyte-attractant chemokines (see Figures 7 and 10
). Because, unlike for sensitization, epidermal Langerhans
cells and dermal dendritic cells do not seem to be necessary for
elicitation of CHS,2
trafficking of this cell type was not
investigated in the present study.
There is an obvious redundancy in expression of monocyte- and
lymphocyte-specific chemokines during elicitation of CHS that has
previously not been described. At first sight this seems to be a
redundancy for robust output of the chemokine system as recently
suggested by Mantovani.14
However, individual infiltrating
lymphocytes and monocytes bear more than one receptor and many of the
chemokines detected in CHS bind to different
receptors,24,25
thus enabling a strong persistent
stimulation of migrating effector cells despite a timely partial
unresponsiveness of one or two receptor pathways after ligand binding.
Therefore, one may speculate that the presence of a plethora of
different chemokines with different receptor specificities may not only
create robustness but also guarantees a rapid, nondelayed recruitment
of cells to sites of evolving inflammatory challenge. Therefore, our
data support the concept by Foxman and colleagues17
who
have shown in vitro that sequential migration through two
spatially distinct attractant fields can target leukocytes in a unique
manner that is determined by both the nature and the sequence of
attractants. The situation as exemplified in our study of inflamed skin
is even more complex because leukocytes have to cross different
compartments. Therefore, we propose a concept of multi
chemokine-mediated recruitment of monocytes and T lymphocytes during
elicitation of a CHS reaction as schematically illustrated in Figure 11
.
|
The relatively few neutrophils detected in CHS are possibly recruited
on moderate expression of IL-8 and GRO. Recent data obtained in the
murine system suggested that Gro expression and subsequent
neutrophil infiltration are prerequisites for T cell recruitment and
elicitation of the CHS reaction.26
In the human CHS
reaction, we could not only detect exclusive dermal Gro expression
but also simultaneous epidermal IL-8 and Gro expression. This may
explain why in humans neutrophils also infiltrate the epidermal
compartment after 48 hours. Whether neutrophils recruited to the skin
are a prerequisite for subsequent T cell trafficking as proposed by
Dilulio and colleagues26
remains to be elucidated for the
human system.
Our data further provide evidence that in the initial phase of CHS
basal keratinocytes as resident cells at the interface between dermis
and epidermis are the main contributors to chemokine expression. At
this stage, inflammatory cells have not been recruited to the skin that
argues, in agreement to earlier observations,4-6
that
keratinocytes are directly stimulated rather than activated by
cytokine-releasing infiltrating cells. The continuous expression of
MCP-1 in the basal layer after 12 hours also argues against
melanocytes, dendritic cells, or intraepidermal lymphocytes as a major
source of epidermal MCP-1 message. Moreover, antigen-specific cells by
chance patrolling through allergen-exposed skin areas are supposed to
be rare;2,27
therefore, a vigorous activation of
keratinocytes by those cells is very unlikely. The strong basal MCP-1
expression detected after 6 hours rather indicates that
nonimmunological stimuli, most likely the intrinsic proinflammatory
properties of the allergen itself, are responsible for such an early
expression by keratinocytes. Analysis of skin organ cultures obtained
from nonsensitized individuals revealed that epicutaneously applied
contact haptens such as nickel chloride induce chemokine and adhesion
molecule expression in the absence of specific T-cell-mediated immunity
(our own unpublished observation). Our data clearly support the concept
that allergens provide two distinct signals: beyond an antigen-specific
signal a defined nonspecific, proinflammatory signal is delivered that
is required to elicit a contact hypersensitivity
reaction.2
The hapten nickel chloride directly activates
the transcription factor nuclear factor-
B as well as p38 stress
kinase-dependent signaling pathways that finally results in expression
of adhesion molecules and chemokines.28-30
It is
conceivable that different types of contact allergens may activate
different cellular responses resulting in distinct chemokine expression
patterns. For example, urushiol, the active agent in poison ivy,
induces considerable IL-8 expression in keratinocytes.5,23
Further studies will have to focus on the heterogeneity of irritant
properties of different contact haptens and its impact on the
elicitation phase of CHS.
Because CHS is regarded as a TH1 response with
TH1-related IFN--inducible chemokines MIG and
IP-10 appearing late after allergen contact (>48 hours), the
nonspecific inflammatory reaction induced by the irritative properties
of allergens possibly precedes the allergen-specific immunological
response by several hours and may furthermore accompany and amplify
antigen-specific CHS. Recently, Flier and colleagues31
demonstrated that at later time points (>48 hours) MIG, IP-10, and
IP-9 are essentially only detected in allergic but not in irritant
patch-test reactions thus supporting our concept that at early time
points the clinically invisible irritative inflammatory reaction is
dominating whereas at later time points the immunological response
evolves. During the later phase, strong expression of the
IFN--inducible chemokines IP-10 and MIG that is accompanied by a
moderate expression of MIP-1/{beta} may activate and preferentially
attract TH1 cells that express CXCR3 and CCR5,
respectively.32
In contrast, other T-cell-attracting
chemokines detected show less preference for TH1
subtypes or are rather specific for TH2 subtypes
such as TARC and MDC.33
Thus, the chemokine profile may
influence the pattern of T cell subsets recruited during inflammation
and supports the concept by D’Ambrosio and colleagues34
who hypothesize that different thresholds for TH1
and TH2 cells exist that differently localize
TH cell subsets in inflammation. Expression of
more promiscuous T cell attractants such as MCP-1 and RANTES may
explain the presence of bystander cells that are detected in
CHS.27
Taken together, our data provide evidence that 1) mononuclear cells in CHS encounter multiple chemoattractant signals in complex spatial and temporal patterns, 2) chemokines both mediate and integrate the irritant and allergen-specific pathways of CHS, and 3) a compartmentalized tissue such as the skin represents an ideal model to investigate the chemokine-mediated combinatorial control of leukocyte targeting in human tissue. Furthermore, our data provide evidence that inflammatory diseases such as CHS are characterized by their chemokine profile that may be a target for a specific anti-inflammatory intervention.
|
Acknowledgements |
|---|
|
Footnotes |
|---|
Supported by grants from the W. Sander-Stiftung (95.064.2) (to R. G.) and from the Deutsche Forschungsgemeinschaft (811/1-3) to M. G.
M. G. and A. Trautmann contributed equally to this work.
Accepted for publication October 27, 2000.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
mRNA in keratinocytes by different but potentially synergistic mechanisms. Int Immunol 1995, 7:343-352, MCP-1, IP-10, and Mig are sequentially and differentially expressed during phase-specific infiltration of leukocyte subsets in human wound healing. Am J Pathol 1998, 153:1849-1860 and IL-8 mRNA in psoriasis: A model for neutrophil migration and accumulation in vivo. J Invest Dermatol 1996, 107:778-782[Medline]
) in allergic contact dermatitis (rhus dermatitis). Br J Dermatol 1991, 124:519-526[Medline]
-mediated recruitment of neutrophils is required for elicitation of contact hypersensitivity. Eur J Immunol 1999, 29:3485-3495[Medline]
B and gene expression in human endothelial cells by the common haptens nickel and cobalt. J Immunol 1995, 155:2459-2467[Abstract]
B-dependent transcription of the monocyte chemoattractant protein-1 (MCP-1) gene in primary endothelial cells. Blood 2001, 790:46-55
This article has been cited by other articles:
 |
|
 |
|
  V. Muller, D. Viemann, M. Schmidt, N. Endres, S. Ludwig, M. Leverkus, J. Roth, and M. Goebeler Candida albicans Triggers Activation of Distinct Signaling Pathways to Establish a Proinflammatory Gene Expression Program in Primary Human Endothelial Cells J. Immunol., December 15, 2007; 179(12): 8435 - 8445. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Frauenschuh, C. A. Power, M. Deruaz, B. R. Ferreira, J. S. Silva, M. M. Teixeira, J. M. Dias, T. Martin, T. N. C. Wells, and A. E. I. Proudfoot Molecular Cloning and Characterization of a Highly Selective Chemokine-binding Protein from the Tick Rhipicephalus sanguineus J. Biol. Chem., September 14, 2007; 282(37): 27250 - 27258. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Viemann, M. Schmidt, K. Tenbrock, S. Schmid, V. Muller, K. Klimmek, S. Ludwig, J. Roth, and M. Goebeler The Contact Allergen Nickel Triggers a Unique Inflammatory and Proangiogenic Gene Expression Pattern via Activation of NF-{kappa}B and Hypoxia-Inducible Factor-1{alpha} J. Immunol., March 1, 2007; 178(5): 3198 - 3207. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. d. Nadai, A.-S. Charbonnier, C. Chenivesse, S. Senechal, C. Fournier, J. Gilet, H. Vorng, Y. Chang, P. Gosset, B. Wallaert, et al. Involvement of CCL18 in Allergic Asthma J. Immunol., May 15, 2006; 176(10): 6286 - 6293. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Schutyser, A. Richmond, and J. Van Damme Involvement of CC chemokine ligand 18 (CCL18) in normal and pathological processes J. Leukoc. Biol., July 1, 2005; 78(1): 14 - 26. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Pastore, F. Mascia, F. Mariotti, C. Dattilo, V. Mariani, and G. Girolomoni ERK1/2 Regulates Epidermal Chemokine Expression and Skin Inflammation J. Immunol., April 15, 2005; 174(8): 5047 - 5056. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Gunther, C. Bello-Fernandez, T. Kopp, J. Kund, N. Carballido-Perrig, S. Hinteregger, S. Fassl, C. Schwarzler, G. Lametschwandtner, G. Stingl, et al. CCL18 Is Expressed in Atopic Dermatitis and Mediates Skin Homing of Human Memory T Cells J. Immunol., February 1, 2005; 174(3): 1723 - 1728. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. Hwang, J. Yamanouchi, P. Santamaria, and P. Kubes A Critical Temporal Window for Selectin-dependent CD4+ Lymphocyte Homing and Initiation of Late-Phase Inflammation in Contact Sensitivity J. Exp. Med., May 3, 2004; 199(9): 1223 - 1234. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. G. Boot, M. Verhoek, M. de Fost, C. E. M. Hollak, M. Maas, B. Bleijlevens, M. J. van Breemen, M. van Meurs, L. A. Boven, J. D. Laman, et al. Marked elevation of the chemokine CCL18/PARC in Gaucher disease: a novel surrogate marker for assessing therapeutic intervention Blood, January 1, 2004; 103(1): 33 - 39. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Struyf, E. Schutyser, M. Gouwy, K. Gijsbers, P. Proost, Y. Benoit, G. Opdenakker, J. Van Damme, and G. Laureys PARC/CCL18 Is a Plasma CC Chemokine with Increased Levels in Childhood Acute Lymphoblastic Leukemia Am. J. Pathol., November 1, 2003; 163(5): 2065 - 2075. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Klunker, A. Trautmann, M. Akdis, J. Verhagen, P. Schmid-Grendelmeier, K. Blaser, and C. A. Akdis A Second Step of Chemotaxis After Transendothelial Migration: Keratinocytes Undergoing Apoptosis Release IFN-{gamma}-Inducible Protein 10, Monokine Induced by IFN-{gamma}, and IFN-{gamma}-Inducible {alpha}-Chemoattractant for T Cell Chemotaxis Toward Epidermis in Atopic Dermatitis J. Immunol., July 15, 2003; 171(2): 1078 - 1084. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Senechal, P. de Nadai, N. Ralainirina, A. Scherpereel, H. Vorng, P. Lassalle, A.-B. Tonnel, A. Tsicopoulos, and B. Wallaert Effect of Diesel on Chemokines and Chemokine Receptors Involved in Helper T Cell Type 1/Type 2 Recruitment in Patients with Asthma Am. J. Respir. Crit. Care Med., July 15, 2003; 168(2): 215 - 221. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Mascia, V. Mariani, G. Girolomoni, and S. Pastore Blockade of the EGF Receptor Induces a Deranged Chemokine Expression in Keratinocytes Leading to Enhanced Skin Inflammation Am. J. Pathol., July 1, 2003; 163(1): 303 - 312. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
|
|
E. Schutyser, S. Struyf, P. Proost, G. Opdenakker, G. Laureys, B. Verhasselt, L. Peperstraete, I. Van de Putte, A. Saccani, P. Allavena, et al. Identification of Biologically Active Chemokine Isoforms from Ascitic Fluid and Elevated Levels of CCL18/Pulmonary and Activation-regulated Chemokine in Ovarian Carcinoma J. Biol. Chem., June 28, 2002; 277(27): 24584 - 24593. [Abstract] [Full Text] [PDF]
|
