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Departments of
*
Immunology and
Urology, Cleveland Clinic Foundation, Cleveland, OH 44195;
Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, OH 44106; and
§
¶
Immunology Department, Pfizer, Inc., Groton, CT 06340
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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following in vitro
stimulation with anti-CD3 mAb or with hapten-labeled cells were
present in the skin-draining lymph nodes of mice treated with
anti-SLC Ab during hapten sensitization. These T cells were unable,
however, to passively transfer CHS to naive recipients. Animals treated
with anti-SLC Ab during hapten sensitization were not tolerant to
subsequent sensitization and challenge with the hapten. In addition,
anti-SLC Ab did not inhibit CHS responses when given at the time of
hapten challenge. These results indicate an important role for SLC
during sensitization for CHS and suggest a strategy to circumvent
functional T cell priming for inflammatory responses through
administration of an Ab inhibiting dendritic cell
trafficking. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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-producing cells induced by
epicutaneous sensitization with DNFB or Ox are hapten-specific
CD8+ T cells (9). In contrast, most
of the hapten-specific CD4+ T cells develop to
IL-4/IL-10-producing cells. After these T cell populations are primed
during sensitization, hapten challenge induces recruitment of the
primed T cells to the challenge site and production of the
proinflammatory cytokines, resulting in the characteristic edema of the
response (10, 11). During sensitization of the epidermis, the reactive hapten is covalently coupled to cell surface proteins. Langerhans cells (LC), the dendritic cells (DC) of the epidermis, capture the hapten, enter lymphatic vessels, and migrate to skin-draining lymph nodes where hapten-MHC complexes are presented to the T cell (12, 13, 14). The important role of LC has been demonstrated by the absence of CHS responses observed when epidermal areas lacking LC are sensitized with hapten (15, 16). In addition, exposure of skin to UV B irradiation before hapten sensitization inhibits LC migration to the lymph nodes and T cell priming for CHS (17, 18). These results suggest that inhibition of DC migration might be an effective strategy to inhibit T cell-mediated responses such as CHS.
The chemotactic factors directing LC trafficking from hapten sensitization sites to draining lymph nodes are unclear. Chemokines are a superfamily of cytokines with chemoattractant properties for leukocytes (19, 20). Neutralization of specific chemokines has been shown to inhibit leukocyte infiltration and pathology in several experimental models of inflammation, demonstrating their critical role in directing cell infiltration into inflammatory sites (21, 22, 23, 24). Recent studies have also indicated the role of chemokines in directing cellular traffic into and within lymphoid tissues. Secondary lymphoid chemokine (SLC), a member of the C-C chemokine family, is constitutively expressed at high levels in the endothelium of the afferent lymphatics and high endothelial venules of lymph nodes (25, 26). The in vivo importance of this chemokine has recently been suggested in plt mice where an absence of SLC production was associated with the inability of DC to migrate from peripheral to lymphoid tissues (27). In further support of the role of SLC in DC trafficking, treatment of wild-type mice with anti-SLC mAb has been shown to inhibit DC migration to lymphoid organs (28). However, the functional consequences of this inhibition on an immune response have not been tested. Since effector T cell priming in CHS is dependent on LC migration from the skin to draining lymph nodes, we hypothesized that treatment with anti-SLC mAb may be an effective method of inhibiting the T cell-mediated response.
In this study, we examined the effect of anti-SLC Ab given at the time of hapten sensitization on the elicitation of the subsequent CHS response. The results demonstrate that administration of anti-SLC Ab during hapten sensitization inhibits the development of effector T cells and CHS responses, suggesting a novel strategy directed at SLC for inhibition of T cell-mediated skin allergies.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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BALB/c mice were obtained through Dr. Clarence Reeder (National Cancer Institute, Frederick, MD). Adult females of 6–10 wk of age were used throughout this study.
Abs and cytokines
Ab to mouse SLC and control Ig were obtained from R&D Systems
(Minneapolis, MN). mAb from the culture supernatant of the
IgG-producing hybridomas N418 (anti-mouse CD11c), 145.2C11
(anti-CD3
), and KJ23a (anti-Vß17a) were purified by
protein G chromatography. Capture and detecting Abs for
cytokine-specific ELISA and recombinant IFN- and IL-4 for
standardization of assays were purchased from PharMingen (San
Diego, CA).
Sensitization and elicitation of contact hypersensitivity
Mice were sensitized and challenged to elicit CHS responses to DNFB and to Ox as described previously (8, 9). For the induction of CHS to DNFB, groups of three to four mice were either sensitized by two daily paintings (days 0 and +1) with 25 µl of 0.25% DNFB (Sigma-Aldrich, St. Louis, MO) on the shaved abdomen and 5 µl on the footpads or were sensitized by a single painting with 50 µl of 1% DNFB on the shaved abdomen and 5 µl on each footpad. For the induction of CHS to Ox, mice were sensitized by a single painting (day 0) of 25 µl of 3% Ox on the shaved abdomen and 5 µl on the footpads. For sensitization to FITC, mice were painted once (day 0) with 100 µl of 1% FITC (isomer I; Sigma-Aldrich) on the shaved abdomen and 5 µl on each footpad. On day +5, the ear thickness of sensitized and unsensitized control animals was measured with an engineer’s micrometer (Mitutoyo, Elk Grove Village, IL) and each animal was challenged by applying 10 µl of 0.2% DNFB, 0.6% Ox, or 0.5% FITC to each side of both ears. Increase in ear swelling was measured in a blinded manner 24 h after challenge and expressed in units of 10-4 inches. The magnitude of ear swelling is given as the mean increase of each group of three to four individual animals (i.e., six to eight ears) ± SEM. The change in ear thickness is calculated as ear thickness at the indicated time after challenge minus ear thickness before challenge. The statistical significance of ear-swelling responses between experimental groups of mice was determined using Student’s t test.
Flow cytometry
Two-color flow cytometry analyses to detect LC in the skin-draining lymph nodes of FITC-sensitized mice was performed as described previously (29). One to 3 days after FITC sensitization, lymph node cells (LNC) from two animals per group were pooled, and 5 x 105 cell aliquots were washed twice with staining buffer (Dulbecco’s PBS with 2% FCS/0.2% NaN3) and then incubated on ice in 100 µl rat serum (Rockland, Gilbertsville, PA) diluted 1/1000 in the staining buffer. After 20 min, the cells were washed twice, resuspended in 200 µl of staining buffer, and stained with biotinylated N418 Ab for 30 min on ice. The cells were washed five times and then incubated for 20 min with streptavidin-PE. After 30 min, the cells were washed five times, resuspended in staining buffer, and analyzed by two-color flow cytometry using a FACScan (Becton Dickinson, San Jose, CA). Sample data were collected on 20,000 cells. Each experiment was performed three times, and the results from a single representative experiment are shown.
Cell culture to stimulate cytokine production and ELISA
Cells were stimulated to produce cytokines by culture on
anti-CD3 Ab-coated tissue culture wells. The 96-well U-bottom
tissue culture plates were precoated with 30 µl/well of
anti-CD3 Ab or, as a negative control, anti-Vß17a Ab at 25
µg/ml for 90 min at 37°C and then washed extensively. LNC cells
were obtained from nonsensitized and hapten-sensitized mice on day +4,
suspended in complete medium, and 2 x 105
LNC from each group were delivered to each well in 200 µl complete
medium. After 48 h, culture supernatants were harvested and
assayed for cytokine production by ELISA. Each experiment was performed
three times, and a single representative experiment is shown in
Results. Previous results have demonstrated that lymph node
T cells from hapten-sensitized, but not from naive, mice produce
cytokines during anti-CD3 Ab-mediated stimulation similar to
cytokine production during culture with isolated hapten-presenting LC
(9).
Cytokine ELISAs for determining quantity of IFN- and IL-4 were
performed as described previously (9). Polyvinylchloride
ELISA plates were coated with capture Ab in 0.1 M bicarbonate buffer
(pH 8.6) overnight at 4°C and then blocked with 5% FCS/0.5% gelatin
in PBS. Duplicate aliquots of all supernatant dilutions were tested
undiluted and in at least two dilutions. Each plate also included
titrated recombinant cytokine as a positive control and to obtain a
standard curve for quantitation. Following incubation overnight at
4°C, each plate was washed and the biotin-labeled anti-cytokine-
detecting mAb was added. The plate was incubated for 2 h at
37°C, washed, and alkaline phosphatase-conjugated streptavidin
(Fisher Scientific, Pittsburgh, PA) added. Following a final incubation
for 1 h at 37°C, the plate was washed and the assay developed by
addition of the substrate p-dinitrophenylphosphate
(Sigma-Aldrich). Results were read at 405 nm and mean values were
calculated. The amount of cytokine in each test supernatant was then
calculated using the standard curve on each plate.
ELISA spot assays for enumeration of IFN--producing
cells
ELISA spot assays for IFN- were performed as described
previously (29, 30). Briefly, ELISA spot plates (Unifilter
350; Polyfiltronics, Rockland, MA) were coated with 4 µg/ml
IFN--specific mAb R46A2 and incubated overnight at 4°C. The plates
were blocked with 1% BSA in PBS for 1 h at room temperature and
then washed four times with PBS. LNC from unsensitized or
DNFB-sensitized mice treated with control or anti-SLC Ab were
prepared on day +4 after sensitization and used as responder cells.
Syngeneic spleen cells from naive BALB/c mice were labeled with 100
µg/ml DNBS as described previously (31), treated with 50
µg/ml mitomycin C, and used as stimulator cells. After washing in
RPMI 1640 medium, responder and stimulator cells were resuspended in
serum-free HL-1 medium (BioWhittaker, Walkersville, MD) supplemented
with 1 mM L-glutamine. Stimulator cells were plated at
5 x 105 cells/well with 2.5 x
105 responder cells/well. Responder cells plated
with unlabeled splenocytes were used as a negative (hapten-specificity)
control. After 24 h of cell culture at 37°C in 5%
CO2, cells were removed from the plate by
extensive washing with PBS/0.05% Tween 20 (PBS-T). Biotinylated
anti-IFN- mAb XMG1.2 (4 µg/ml) was added and the plate was
incubated overnight at 4°C. The following day the plate was washed
three times with PBS-T and conjugated streptavidin-alkaline phosphatase
was added to each well. After 2 h at room temperature, the plates
were washed with PBS-T and nitroblue
tetrazolium/5-bromo-4-chloro-3-indoyl substrate was added (Kirkegaard &
Perry Laboratories, Gaithersburg, MD). The resulting spots were counted
with an ELISA spot image analyzer (developed at Case Western Reserve
University) using Optimas (Optimas, Botheled, WA) that was designed to
detect ELISA spots with predetermined criteria on size, shape, and
colorimetric density.
Adoptive transfer of CHS
Hapten immune T cells were obtained from mice treated with 25-µg aliquots of anti-SLC mAb or control Ig the day of Ox sensitization (i.e., day 0). On day +4, LNC suspensions were prepared, washed, and resuspended at 125 x 106/ml in PBS and 400-µl aliquots (50 x 106 cells) were injected i.v. Immediately following cell transfer, ears were challenged with hapten and the increase in ear swelling was measured 24 h later.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Following hapten sensitization of the skin, LC migrate from the
epidermis to the skin-draining lymph nodes where they present
hapten-MHC complexes and prime the T cells mediating and regulating CHS
following hapten challenge. Application of the hapten FITC allows
detection of migrating epidermal LC as
FITC+/CD11c+ cells in the
skin-draining lymph nodes (12, 14, 18, 29). Previous
studies have demonstrated the decreased LC migration in response to
FITC application in plt mice that are defective in SLC
production (27). To test the ability of anti-SLC Ab to
inhibit LC migration from the skin to draining lymph nodes in wild-type
mice, groups of mice were treated with 100 µg of anti-SLC Ab or
control Ig i.v. and then sensitized with 0.5% FITC on the shaved
abdomen and each footpad. LNC were prepared from the mice at days +1,
+2, and +3 postsensitization and stained with anti-CD11c mAb, N418.
LNC from animals treated with the control Ig contained readily
detectable FITC-labeled CD11c+ cells, indicating
migration of the hapten-presenting LC from the epidermis. The presence
of FITC-labeled DC were considerably reduced in animals treated with
anti-SLC Ab, indicating that hapten-presenting LC migration to the
lymph nodes was inhibited by the Ab (Fig. 1
). Furthermore, the inhibitory effect of
the anti-SLC Ab treatment was maintained through at least day 3
after sensitization.
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Since treatment with anti-SLC Ab in FITC-sensitized mice
inhibited LC migration to the lymph nodes, we next evaluated the
functional significance of this inhibition by testing whether the
anti-SLC Ab would inhibit CHS to FITC. Groups of mice were injected
with 100 µg of anti-SLC Ab or control Ig i.v. and then sensitized
with 1% FITC on day 0. On day +5, the sensitized animals and a group
of unsensitized animals were challenged by applying 0.5% FITC to each
side of both ears and the increase in ear thickness was measured
24 h later. Readily detectable ear-swelling responses to FITC were
observed in mice treated with control Ig during hapten sensitization
when compared with unsensitized mice (Fig. 2, group 1 vs group 3). Treating mice
with the anti-SLC Ab at the time of sensitization inhibited the
magnitude of the CHS response to near background levels (group
2).
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A, group 1) but not the
unsensitized control animals (group 5) had readily apparent
ear-swelling responses to challenge with Ox. Treatment of Ox-sensitized
animals with anti-SLC Ab inhibited the CHS response in a
dose-dependent manner (groups 2–4). Treatment with 25 µg of
anti-SLC Ab resulted in 74% inhibition of the CHS response. This
inhibitory effect was long-lasting as an ear-swelling response was not
observed in treated animals through 72 h after challenge (Fig. 3B). Inhibition of CHS to Ox was also observed when 100 µg
of anti-SLC Ab was administered i.p. at the time of sensitization
(data not shown).
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, group 1 vs group 2). In
contrast, CHS responses of mice sensitized with 0.25% DNFB on 2
consecutive days and challenged with the hapten on day +5 were not
inhibited by either single or dual administration of 75 µg of
anti-SLC Ab (group 3 vs groups 4 and 5).
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, groups 1
and 2). CHS responses to painting with the irritant followed by Ox or a
dual sensitization with Ox were not inhibited by administration of the
anti-SLC Ab (groups 3 and 4).
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We next tested the effect of anti-SLC Ab treatment on T cell
priming in the lymph nodes draining the skin sensitization site by
assessing cytokine production. Groups of mice were administered 10–100
µg of anti-SLC Ab or control Ig i.v. and sensitized with 3% Ox.
Four days later, LNC suspensions were prepared and cultured on
anti-CD3 Ab-coated wells to stimulate the hapten-primed T cells to
produce cytokines. The supernatants were collected 48 h later and
tested by ELISA for IFN- and IL-4 production. Supernatants of LNC
from naive mice cultured on anti-CD3 mAb-coated wells yielded low
to nondetectable levels of IFN- and IL-4, whereas supernatants of
LNC from hapten-immune mice treated with the control Ab produced
readily detectable amounts of IFN- and IL-4 (Fig. 6). LNC from mice treated with increasing
doses of anti-SLC Ab at Ox sensitization produced equivalent
amounts of IFN- and IL-4. These results suggested that although LC
migration to the lymph nodes was inhibited, anti-SLC Ab did not
inhibit priming for cytokine production in the lymph nodes of
hapten-sensitized mice.
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in a
hapten-specific manner. Mice were treated i.p. with 100 µg of
anti-SLC Ab or control Ig at the time of a single painting with
DNFB. Four days later, the LNC from each group of sensitized mice and a
group of unsensitized mice were prepared and cocultured with either
unlabeled or DNP-labeled spleen cells on anti-IFN--coated
filters. After 24 h, the cells were removed, the filters were
developed, and the number of IFN--producing cells in each group was
quantified using an enzyme-linked immunospot image analyzer. In
contrast to LNC from naive mice, LNC from DNFB-sensitized mice treated
with control Ig contained a readily detectable population of cells that
produced IFN- when cultured with the DNP-labeled, but not with the
unlabeled, spleen cells (Fig. 7).
Treatment with anti-SLC Ab did not affect the number of
IFN--producing cells in DNFB-sensitized mice. The results indicated
that sensitization of control and anti-SLC Ab-treated mice induced
equivalent hapten-specific IFN--producing cell responses.
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Since LNC from Ox-sensitized mice treated with anti-SLC mAb
and control Ig produced comparable levels of IFN-, we wanted to
determine whether the anti-SLC Ab treatment affected the function
of the T cells in CHS. Therefore, we tested the ability of the T cells
to mediate CHS responses following passive transfer to naive
recipients. Donor mice were injected with 25 µg of anti-SLC Ab or
control Ig i.v. and were sensitized with Ox. Four days later, pooled
LNC from each group of sensitized mice and from unsensitized mice were
transferred to naive recipients. Immediately after transfer, the
recipients were challenged on the ears with 1% Ox. Delivery of LNC
from control Ig-treated mice sensitized with Ox, but not from
unsensitized donors, elicited an ear-swelling response (Fig. 8, group1 vs group 3). Delivery of LNC
from anti-SLC Ab-treated mice sensitized with Ox, however, did not
induce an ear-swelling response (group 2). Therefore, T cells were
induced in the lymph nodes of anti-SLC Ab-treated animals that
produced IFN- in response to anti-CD3 mAb or to DNP-labeled
spleen cells in vitro, but these cells could not elicit a CHS response
to hapten challenge in vivo.
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Since T cells from mice given anti-SLC Ab at the time of
hapten sensitization could not mediate a CHS response, we next tested
whether these mice had developed immunological tolerance to the hapten.
Groups of mice were injected with 25 µg of anti-SLC or control Ig
i.v. and were sensitized with 3% Ox. Five days later, the mice were
challenged with 1% Ox and the ear thickness was measured 24 h
later. As previously observed, the primary CHS response to Ox was
inhibited in mice treated with anti-SLC Ab during hapten
sensitization (data not shown). Five weeks later, these anti-SLC
Ab-treated/Ox-sensitized and challenged mice were repainted with either
3% Ox or 0.25% DNFB and were challenged 5 days later (Fig. 9, groups 1–3). Mice treated with the
control Ig before primary Ox sensitization had an increased response
following resensitization and challenge with Ox (group 3) when compared
with the primary response in mice sensitized and challenged a single
time (group 4). Mice treated with anti-SLC Ab during primary
sensitization and then resensitized with Ox had less of an ear-swelling
response to Ox challenge than mice that were treated with control Ig
during primary sensitization and resensitized 5 wk later (group 1 vs
group 3). This response was equivalent, however, to the primary
response in control mice sensitized and challenged a single time (group
1 vs group 4). Ear-swelling responses were also observed in response to
DNFB sensitization and challenge of mice previously treated with
anti-SLC Ab and sensitized with Ox (group 2). In addition, control
groups of naive mice were treated with anti-SLC Ab or control Ig
and were sensitized and challenged with Ox (groups 4 and 5). Again,
treatment with anti-SLC Ab inhibited the primary CHS response when
compared with control Ig-treated mice. These results indicated that the
inhibition observed with anti-SLC Ab treatment during primary CHS
responses was not sustained during subsequent sensitization and
challenge. Thus, treatment with anti-SLC mAb during sensitization
did not induce tolerance to the hapten.
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Finally, we tested whether hapten-immune T cell recruitment and
function was inhibited by treatment with anti-SLC Ab given to
sensitized animals at the time of challenge. Groups of mice were
injected with 25 µg of anti-SLC Ab or control Ig i.v. at the time
of sensitization with 3% Ox or were sensitized with Ox and given the
anti-SLC Ab or control Ig at the time of hapten challenge with
0.6% Ox. As previously observed, treatment with anti-SLC Ab at the
time of sensitization inhibited the ear-swelling response to Ox
challenge (Fig. 10, group 1 vs group
2). Giving the anti-SLC Ab at the time of challenge, however, had
no effect on the elicitation of CHS when compared with control
Ig-treated immune animals (group 3 vs group 1).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In this study, we have extended these observations and demonstrated
that the CHS response to Ox is inhibited in a dose-dependent manner by
treating mice with anti-SLC Ab at the time of hapten sensitization.
The effect of this single dose was not transient because inhibition of
the ear-swelling response continued for at least 3–4 days after Ox
challenge, which is the time that the response in control Ig-treated
mice nears resolution. Consistent with these results, we have also
shown that migration of FITC-presenting LC from skin sensitization
sites to the draining lymph nodes is inhibited in mice treated with
anti-SLC Ab for at least 3 days after epicutaneous application of
the FITC. On the basis of these results, we predicted that T cells in
the lymph nodes would not be primed to react to the hapten and
therefore not be able to elicit an immune response to hapten challenge.
Surprisingly, although anti-SLC Ab inhibited the CHS response, T
cells from hapten-sensitized mice treated with the anti-SLC Ab were
still capable of producing IFN- when stimulated by culture on
anti-CD3 Ab-coated wells. T cells from naive mice did not produce
IFN- during these cultures, indicating that the cytokine production
observed is the result of hapten application. Similarly, equivalent
numbers of hapten-specific IFN--producing cells were observed in
sensitized mice treated with control and anti-SLC Ab. An obvious
interpretation of this result is that CD8+ T cell
production of IFN- is not critical for the elicitation of CHS and
the response is mediated by other proinflammatory cytokines produced by
the hapten-primed T cells. However, recent observations in our
laboratory have indicated a direct correlation between hapten-primed
CD8+ T cell IFN- production and the magnitude
of the CHS response observed following challenge, including the absence
of CHS responses in IFN--deficient mice (our unpublished
observations).
To explore the paradoxical induction of IFN--producing T cells with
the lack of CHS responses in mice treated with anti-SLC Ab during
hapten sensitization, we passively transferred LNC from sensitized mice
treated with control or anti-SLC Ab. These experiments demonstrated
that even though there were hapten-specific IFN--producing T cells
in the lymph nodes of anti-SLC Ab-treated mice, these cells were
unable to mediate CHS in naive mice following transfer and hapten
challenge. This argues that treatment with anti-SLC Ab inhibited
the development of a critical immune function during hapten priming of
these T cells. This result raised two important questions regarding the
priming of T cells in anti-SLC Ab-treated mice during hapten
sensitization: 1) If the LC do not migrate to the lymph nodes, how are
the T cells stimulated to produce cytokines? and 2) If the T cells can
produce IFN-, why are they unable to mediate CHS in response to
hapten challenge? It may be possible that hapten-protein complexes
drain from the epidermal sensitization site into the lymph nodes
through the blood and lymphatics and are picked up by resident DC in
the lymphoid tissue which subsequently prime hapten-specific T cells.
Such presentation may be sufficient to allow T cell priming for
production of proinflammatory cytokines such as IFN- but not in a
manner which renders the cells capable of functioning in response to
epicutaneous challenge with the hapten.
One result of priming by alternative hapten-presenting cells may be the lack of T cell programming for circulation to challenge sites in the skin. Several studies have indicated the requirement for expression of specific adhesion markers on T cells that enable the cells to enter cutaneous inflammatory sites during CHS (34, 35). Recent studies in other laboratories have indicated that a specific homing receptor/endothelial ligand pair, the cutaneous lymphocyte-associated Ag and E-selectin on endothelial cells, is associated with the infiltration of human T cells into inflammatory sites in the skin (36, 37). Similar studies have been performed in mice and have suggested that only CD4+ T cells expressing P- and E-selectin-binding proteins are recruited to inflammatory sites in the skin (38, 39). As such, treatment with anti-SLC mAb might inhibit critical priming factors that induce the expression of skin-homing receptors on the T cells mediating the CHS response. LC may provide such a critical component, inducing the expression of these skin-homing receptors during hapten priming. This possibility is currently being explored.
The results of this study have demonstrated the ability of a single dose of Ab to SLC to inhibit functional T cell priming for elicitation of CHS to Ox. In contrast, treatment with the Ab did not inhibit the response when more than one application of hapten or when an additional irritant were used to sensitize mice. Two applications of hapten may cause more irritation of the skin, which may increase inflammation at the site and stimulate LC or dermal DC migration to draining lymph nodes despite the anti-SLC Ab treatment. This is demonstrated by the lack of anti-SLC Ab-mediated inhibition of CHS when an irritant is applied to the skin before hapten sensitization. Consistent with this proposal, an increased dose of anti-SLC Ab was required to inhibit the CHS response to a single sensitizing application of DNFB when compared with the amount required to inhibit the response to Ox. This indicates an important difference between these two haptens during skin sensitization. A possible explanation is that the increased inflammation induced by DNFB application may either stimulate the production of higher levels of SLC or other chemokines that direct LC migration to the lymph nodes. Alternatively, this increased inflammation may stimulate a higher density of CCR7 on the LC surface, which renders this migration less sensitive to the Ab treatment.
Although a good deal is known about the role of LC in the induction of CHS, very little is known about the role these DC play in the elicitation of the response. Results from some laboratories have suggested that the presence of LC at the challenge site during elicitation of CHS enhances the magnitude of the response (40). In contrast, a recent report indicated the presence of LC at the hapten challenge site resulted in decreased CHS responses (41). Results from the current report indicate that treatment with anti-SLC Ab inhibits LC migration and CHS but does not have an effect on the elicitation of the response when administered at the time of challenge. These results suggest that inhibition of LC migration from the challenge site does not affect the magnitude of the elicited response in a positive or negative manner.
In summary, the results of this report have indicated an important role for SLC in the CHS response. The inhibition of LC migration was associated with a selective inhibition of functional T cell priming to mediate contact allergy responses to challenge with the hapten. By examining the functional consequences of anti-SLC Ab treatment, we gain insight into DC trafficking and hapten priming of specific T cells which could potentially provide new therapies involving immune regulation in autoimmunity and transplantation.
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Robert L. Fairchild, Department of Immunology, NB3-79, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH 44195-0001.
3 Abbreviations used in this paper: CHS, contact hypersensitivity; DC, dendritic cell; DNFB, 2,4-dinitrofluorobenzene; LNC, lymph node cell; Ox, oxazolone; SLC, secondary lymphoid chemokine; LC, Langerhans cell.
Received for publication September 28, 1999. Accepted for publication March 8, 2000.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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M. Sugaya, L. Fang, A. R. Cardones, T. Kakinuma, S. H. Jaber, A. Blauvelt, and S. T. Hwang Oncostatin M Enhances CCL21 Expression by Microvascular Endothelial Cells and Increases the Efficiency of Dendritic Cell Trafficking to Lymph Nodes J. Immunol., December 1, 2006; 177(11): 7665 - 7672. [Abstract] [Full Text] [PDF]
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 E. Ziegler, F. Gueler, S. Rong, M. Mengel, O. Witzke, A. Kribben, H. Haller, U. Kunzendorf, and S. Krautwald CCL19-IgG Prevents Allograft Rejection by Impairment of Immune Cell Trafficking J. Am. Soc. Nephrol., September 1, 2006; 17(9): 2521 - 2532. [Abstract] [Full Text] [PDF]
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D. D. Kish, A. V. Gorbachev, and R. L. Fairchild CD8+ T cells produce IL-2, which is required for CD4+CD25+ T cell regulation of effector CD8+ T cell development for contact hypersensitivity responses J. Leukoc. Biol., September 1, 2005; 78(3): 725 - 735. [Abstract] [Full Text] [PDF]
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 K. R. Pilkington, I. Clark-Lewis, and S. R. McColl Inhibition of Generation of Cytotoxic T Lymphocyte Activity by a CCL19/Macrophage Inflammatory Protein (MIP)-3{beta} Antagonist J. Biol. Chem., September 24, 2004; 279(39): 40276 - 40282. [Abstract] [Full Text] [PDF]
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A. V. Gorbachev and R. L. Fairchild CD40 Engagement Enhances Antigen-Presenting Langerhans Cell Priming of IFN-{gamma}-Producing CD4+ and CD8+ T Cells Independently of IL-12 J. Immunol., August 15, 2004; 173(4): 2443 - 2452. [Abstract] [Full Text] [PDF]
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A. V. Gorbachev and R. L. Fairchild CD4+ T Cells Regulate CD8+ T Cell-Mediated Cutaneous Immune Responses by Restricting Effector T Cell Development through a Fas Ligand-Dependent Mechanism J. Immunol., February 15, 2004; 172(4): 2286 - 2295. [Abstract] [Full Text] [PDF]
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J. C. Dudda, J. C. Simon, and S. Martin Dendritic Cell Immunization Route Determines CD8+ T Cell Trafficking to Inflamed Skin: Role for Tissue Microenvironment and Dendritic Cells in Establishment of T Cell-Homing Subsets J. Immunol., January 15, 2004; 172(2): 857 - 863. [Abstract] [Full Text] [PDF]
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H. Guan, G. Zu, Y. Xie, H. Tang, M. Johnson, X. Xu, C. Kevil, W.-C. Xiong, C. Elmets, Y. Rao, et al. Neuronal Repellent Slit2 Inhibits Dendritic Cell Migration and the Development of Immune Responses J. Immunol., December 15, 2003; 171(12): 6519 - 6526. [Abstract] [Full Text] [PDF]
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P. Saint-Mezard, C. Chavagnac, S. Bosset, M. Ionescu, E. Peyron, D. Kaiserlian, J.-F. Nicolas, and F. Berard Psychological Stress Exerts an Adjuvant Effect on Skin Dendritic Cell Functions In Vivo J. Immunol., October 15, 2003; 171(8): 4073 - 4080. [Abstract] [Full Text] [PDF]
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 T. Murakami, W. Maki, A. R. Cardones, H. Fang, A. Tun Kyi, F. O. Nestle, and S. T. Hwang Expression of CXC Chemokine Receptor-4 Enhances the Pulmonary Metastatic Potential of Murine B16 Melanoma Cells Cancer Res., December 1, 2002; 62(24): 7328 - 7334. [Abstract] [Full Text] [PDF]
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H. Hammad, B. N. Lambrecht, P. Pochard, P. Gosset, P. Marquillies, A.-B. Tonnel, and J. Pestel Monocyte-Derived Dendritic Cells Induce a House Dust Mite-Specific Th2 Allergic Inflammation in the Lung of Humanized SCID Mice: Involvement of CCR7 J. Immunol., August 1, 2002; 169(3): 1524 - 1534. [Abstract] [Full Text] [PDF]
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T. Junt, H. Nakano, T. Dumrese, T. Kakiuchi, B. Odermatt, R. M. Zinkernagel, H. Hengartner, and B. Ludewig Antiviral Immune Responses in the Absence of Organized Lymphoid T Cell Zones in plt/plt Mice J. Immunol., June 15, 2002; 168(12): 6032 - 6040. [Abstract] [Full Text] [PDF]
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C. J. Kirk, D. Hartigan-O'Connor, and J. J. Mule The Dynamics of the T-Cell Antitumor Response: Chemokine-secreting Dendritic Cells Can Prime Tumor-reactive T Cells Extranodally Cancer Res., December 1, 2001; 61(24): 8794 - 8802. [Abstract] [Full Text] [PDF]
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 H. E. Wiley, E. B. Gonzalez, W. Maki, M.-t. Wu, and S. T. Hwang Expression of CC Chemokine Receptor-7 and Regional Lymph Node Metastasis of B16 Murine Melanoma J Natl Cancer Inst, November 7, 2001; 93(21): 1638 - 1643. [Abstract] [Full Text] [PDF]
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 E. Kriehuber, S. Breiteneder-Geleff, M. Groeger, A. Soleiman, S. F. Schoppmann, G. Stingl, D. Kerjaschki, and D. Maurer Isolation and Characterization of Dermal Lymphatic and Blood Endothelial Cells Reveal Stable and Functionally Specialized Cell Lineages J. Exp. Med., September 17, 2001; 194(6): 797 - 808. [Abstract] [Full Text] [PDF]
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A. V. Gorbachev, N. A. DiIulio, and R. L. Fairchild IL-12 Augments CD8+ T Cell Development for Contact Hypersensitivity Responses and Circumvents Anti-CD154 Antibody-Mediated Inhibition J. Immunol., July 1, 2001; 167(1): 156 - 162. [Abstract] [Full Text] [PDF]
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P. D. Rennert, P. S. Hochman, R. A. Flavell, D. D. Chaplin, S. Jayaraman, J. L. Browning, and Y.-X. Fu Essential Role of Lymph Nodes in Contact Hypersensitivity Revealed in Lymphotoxin-{alpha}-Deficient Mice J. Exp. Med., June 4, 2001; 193(11): 1227 - 1238. [Abstract] [Full Text] [PDF]
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P. Hjelmström Lymphoid neogenesis: de novo formation of lymphoid tissue in chronic inflammation through expression of homing chemokines J. Leukoc. Biol., March 1, 2001; 69(3): 331 - 339. [Abstract] [Full Text]
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A. V. Gorbachev, P. S. Heeger, and R. L. Fairchild CD4+ and CD8+ T Cell Priming for Contact Hypersensitivity Occurs Independently of CD40-CD154 Interactions J. Immunol., February 15, 2001; 166(4): 2323 - 2332. [Abstract] [Full Text] [PDF]
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M. Itakura, A. Tokuda, H. Kimura, S. Nagai, H. Yoneyama, N. Onai, S. Ishikawa, T. Kuriyama, and K. Matsushima Blockade of Secondary Lymphoid Tissue Chemokine Exacerbates Propionibacterium acnes-Induced Acute Lung Inflammation J. Immunol., February 1, 2001; 166(3): 2071 - 2079. [Abstract] [Full Text] [PDF]
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