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*
Department of Dermatology, University of Texas Southwestern Medical Center, Dallas, TX 75235; and
Department of Immunology, Juntendo University School of Medicine, Tokyo, Japan
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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and
MIP-1
, and DC-CK, which attract T cells (18, 19, 20, 21). In sum, DC possess
all phenotypic and functional properties required for the efficient
induction of cellular immunity. DC do not simultaneously exhibit all the properties described above; rather they acquire them sequentially, apparently when needed. For example, DC-lineage Langerhans cells (LC) in skin are relatively deficient in their capacity to activate naive T cells when tested immediately after isolation from skin, but they acquire this capacity during short term culture in vitro. This capacity is supported by an elevated expression of MHC class II and costimulatory molecules and by increased production of several cytokines (22). At the same time, cultured LC lose other properties, including pinocytosis, phagocytosis, and the ability to process complex protein Ag (1, 6, 9). These changes in LC function are thought to reflect their "maturation" from DC specialized for Ag uptake and processing into DC specialized for delivering T cell-stimulatory signals (9, 22). Importantly, LC undergo a similar maturation process in vivo following topical application of reactive haptens (23, 24, 25), the treatment that also triggers LC migration into draining lymph nodes (26). Therefore, when LC reach lymph nodes, they most likely have completed their transition into the T cell-stimulatory DC. This transition illustrates the dynamic nature of DC biology; DC change both phenotype and function in a life cycle, depending on their maturational states and location.
The ultimate question concerns the terminal fate of DC after Ag presentation has been accomplished. One scenario is that they reside in lymph nodes for relatively long periods of time, ensuring maximal activation of T cells. Perhaps they leave lymph nodes and return to their tissues of origin. Even more remote is the possibility that DC are killed by the very T cells that they activate. Ingulli et al. (27) developed a unique experimental system to follow the migration and the fate of DC. In this system, splenic DC were pulsed with OVA, labeled with a fluorescent dye, and then injected s.c. into syngeneic mice that had received an adoptive transfer of OVA-reactive, naive CD4+ T cells from transgenic animals. Ingulli et al. observed that the DC migrated into draining lymph nodes where they interacted closely with CD4+ T cells. Interestingly, these infused DC disappeared rapidly (by 48 h) from the lymph nodes at the time when clonal expansion of OVA-reactive T cells became detectable. However, these in vivo experiments failed to determine whether those DC had died or whether they had migrated out of the lymph nodes. In the present study, we developed in vitro experimental systems to determine the terminal fate of DC. Our results document that DC undergo rapid apoptosis upon Ag-specific interaction with a CD4+ T cell clone.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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BALB/c mice, B6.MRL-FASlpr
mice, and C57BL/6J mice (all 6–8 wk old, female) were purchased from
The Jackson Laboratory (Bar Harbor, ME). Splenic DC were isolated as
described previously (28). Briefly, spleen cell suspensions were
prepared by mechanical dissociation using fine forceps, followed by
collagenase treatment (1% collagenase, 1 h, 37°C). After lysis
of erythrocytes, splenic cells were subjected to gradient
centrifugation with Percoll (Pharmacia, Piscataway, NJ); cells
collected from the interface between 1.035 and 1.075 g/ml of Percoll
were then incubated on tissue culture plates. After a 90-min
incubation, nonadherent cells were removed by extensive pipeting, and
the adherent cells were cultured overnight. Cells released during the
second culture period were harvested and used as splenic DC
preparations; they contained 
40–50% CD11c+ DC,
30–40% B220+ B cells, 5–10% CD3+ T cells,
and <5% of pan-NK mAb DX-5+ NK cells assessed by FACS
analysis. XS52 is a long term DC line we established from the epidermis
of a newborn BALB/c mouse (29). XS52 cells were maintained and expanded
in complete RPMI in the presence of GM-CSF (2 ng/ml) and NS47
fibroblast supernatant (10%). Phenotypic and functional features of
this DC line are described elsewhere (29, 30, 31, 32, 33). As responder T cells, we
used the HDK-1 cells, which are a keyhole limpet hemocyanin
(KLH)-specific CD4+ Th1 clone (34); these cells were
stimulated every 4 wk with KLH-pulsed spleen cells and expanded by
repeated feeding with IL-2 (200 U/ml). In some experiments, we used a
short term-cultured KLH-reactive CD4+ T cell line. Briefly,
BALB/c mice were immunized by s.c. injection of 100 µg of KLH in PBS
emulsified in CFA into the foot pads. After 7 days, T cells were
isolated from the draining lymph nodes and cultured in the presence of
-irradiated syngeneic spleen cells plus KLH (100 µg/ml) and IL-2
(25 U/ml). The resulting populations were >95% CD4+, and
they exhibited significant proliferative responses to KLH-pulsed XS52
DC.
Apoptosis assays
XS52 DC (1 x 105 cells/ml) or splenic DC
(1 x 106 cells/ml) were cocultured with HDK-1 T cells
(4 x 105 or 1 x 106 cells/ml,
respectively) in the presence of KLH (100 µg/ml). Unless otherwise
mentioned, all incubations were conducted in 5-ml tissue culture tubes
using complete RPMI in the absence of added growth factors. In some
experiments, XS52 DC or HDK-1 T cells were prelabeled with FITC (100
µg/ml, 10 min, 4°C) before coculture (33). Alternatively, XS52 DC
and HDK-1 T cells were first incubated together and then postlabeled
with FITC-conjugated mAb against Iad, Thy-1.2, or CD11c
(PharMingen, San Diego, CA). At various intervals during coincubation,
cells were collected by centrifugation, labeled with propidium iodide
(PI), and analyzed by FACScan (Becton Dickinson, Mountain View, CA)
(35). To identify apoptotic DC visually, samples were postlabeled with
PE-conjugated anti-Iad mAb (PharMingen), fixed with 3%
paraformaldehyde, and then stained with 10 µg/ml Hoechst 33342
(Sigma, St. Louis, MO). We then counted the numbers of
Iahigh DC showing the condensed nuclei under fluorescence
microscopy at x400. To examine the role of CD95L, XS52 DC were
cocultured with HDK-1 T cells and KLH in the continuous presence of 10
µg/ml anti-CD95L mAb (MFL-1) (36), anti-CD48 mAb
(PharMingen), or control hamster IgG (PharMingen). To examine the role
of CD54, we added anti-CD54 mAb (PharMingen) or rat IgG2a control
(PharMingen). DNA degradation was examined as described previously with
slight modification (35, 37). Briefly, XS52 cells were labeled with
[3H]]thymidine (5 µCi/ml) for 48 h, washed
extensively, and then cocultured for 8 h in the presence or
absence of HDK-1 T cells and/or KLH. Subsequently, these cells were
collected by centrifugation and then lysed in a buffer consisting of 10
mM Tris-HCl, 10 mM EDTA and 0.2% Triton X-100 (pH 7.5). The
lysate was centrifuged (13,000 x g) for 10 min at
4°C, and low m.w. DNA was then purified from the supernatant by
phenol-chloroform extraction followed by ethanol precipitation. DNA was
separated in a 2% agarose gel and then transferred onto a 
-probe
membrane (Bio-Rad, Hercules, CA). The membrane was exposed to Kodak
x-ray film (Eastman Kodak, Rochester, NY) at -80°C for 10 days.
Assays for CD95 and CD95 ligand (CD95L) expression
XS52 DC and HDK-1 T cells were labeled with anti-CD95 mAb
(Jo2, PharMingen) or anti-CD95L mAb (MFL-1) and then incubated with
PE-conjugated goat anti-hamster IgG (Jackson ImmunoResearch, West
Grove, PE). In some experiments, the cells were stimulated with LPS
(100 ng/ml) or Con A (4 µg/ml) for 8 h before analysis. To
examine CD95 and CD95L mRNA expression, total RNA isolated from these
cells was subjected to RT-PCR using the following primers:
5'-ATGCACACTCTGCGATGAAG-3' and 5'-TTCAGGGTCATCCTGTCTCC-3' for CD95,
and 5'AGCTACCTGGGGGCAGTATT-3' and 5'-ATGCAGGCATTAAGGACCAC-3' for
CD95L. Primers for IL-1ß, IFN-, and ß-actin were
purchased from Clontech Laboratories (Palo Alto, CA). PCR
products were harvested within the linear range of amplification (30
cycles for CD95, CD95L, IFN-, and IL-1ß, and 25 cycles for
ß-actin) and then separated on 1% agarose gel containing ethidium
bromide. In some experiments, PCR products were analyzed further by
Southern blotting using the internal oligo probes
(5'-AAACAAATCGCACCCTGACC-3' for CD95 and 5'-TCATGGGCACCAGGAATATT-3' for
CD95L), as described previously (38).
Induction of DTH response
Splenic DC isolated from B6.MRL-FASlpr mice and from control C57BL/6J mice were pulsed with KLH (100 µg/ml) during the overnight incubation period. After extensive washing, the KLH-pulsed DC were suspended in PBS and injected s.c. into C57BL/6J mice. As a positive control, a different group of mice were immunized by s.c. injection of KLH plus CFA. Ten days later, these animals were challenged by s.c. injection of KLH (50 µg/20 µl PBS/mouse) into the left hind footpad. The same volume of PBS alone was injected into the right hind footpad as control. Footpad swelling responses were measured at 24 and 48 h using a caliper-type engineer’s micrometer (Mitsutoyo, Kawasaki, Japan).
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In the first set of experiments, we examined the fate of splenic
DC during Ag-specific interaction with CD4+ T cells. We
isolated DC from BALB/c mouse spleens with a standard protocol that
used centrifugation through Percoll gradient, removal of plastic
nonadherent cells, and overnight culture (28). As a responder T cell
population, we used a KLH-specific, CD4+ Th1 clone HDK-1.
Splenic DC preparations were cocultured with HDK-1 T cells for an
additional 20 h in the presence or absence of Ag (KLH), labeled
with FITC-conjugated anti-CD11c mAb, and then examined for
viability by PI uptake. As noted in Fig. 1
A, 50% of the
CD11c+ cells were judged to be PI positive after the second
(20-h) incubation even in the absence of either HDK-1 cells or KLH.
Cell viability as measured by trypan blue exclusion remained relatively
high during the isolation procedure, i.e., >95% in low density spleen
cells after gradient centrifugation and 80–85% in DC preparations
after the first overnight culture. On the other hand, 35–40% of
CD11c+ cells in the same DC preparations were judged to be
"dead" by the more sensitive FACS-based assay of PI uptake. We
interpreted these observations to indicate that CD11c+
splenic DC undergo spontaneous cell death during prolonged culture
periods, with 35–40% death during the isolation procedure and
additional 10–15% death during the second 20-h incubation.
Importantly, the frequency of PI-positive CD11c+ splenic DC
increased to 90% after coincubation with both HDK-1 T cells and KLH,
whereas incubation with either T cells or KLH alone had only modest
effects (Fig. 1A). In six independent experiments,
relatively consistent numbers of CD11c+ splenic DC
(85–91%, 87.3 ± 2.9%) were judged to be PI positive after
20 h of incubation with T cells and KLH; these values were
statistically higher (p < 0.01, Student’s
t test) than those observed in any of the control groups, DC
+ T cells, DC + KLH, or DC alone (Fig. 1B). These
observations indicate that splenic DC undergo apoptosis spontaneously
in culture and that Ag-specific interaction with CD4+ T
cells augments this cell death.
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). These
results suggest strongly that splenic DC undergo spontaneous apoptosis
in culture and that this process is augmented significantly by
Ag-specific interaction with CD4+ T cells.
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To study the process of DC apoptosis in detail and to identify its
mechanism(s), we developed a better defined experimental system using
the long term DC line XS52. We observed previously that XS52 DC are at
least as potent as splenic DC in their capacity to present KLH to the
HDK-1 T cells (29). As shown in Fig. 3A, XS52 cells showed only
modest (<10%) spontaneous cell death (assessed by PI uptake) after
16 h of incubation in the absence of added growth factors.
Likewise, the majority of HDK-1 T cells remained viable over the same
time period. When XS52 DC and HDK-1 T cells were incubated together in
the presence of KLH (i.e., "complete" coculture), substantial
numbers of these cells became PI positive. By contrast, they remained
mostly viable when incubated together in the absence of KLH or
incubated individually in the presence of KLH (i.e., "incomplete"
cocultures). To determine whether cell death occurred primarily among
XS52 DC (or HDK-1 T cells), XS52 cells were first labeled with FITC and
then incubated with HDK-1 T cells and KLH. As shown in Fig. 3B, relatively large numbers of the FITC-positive cells
(i.e., XS52 cells) showed significant PI uptake in the complete
coculture. In time course experiments, significant cell death became
detectable among XS52 cells as early as the 2-h point, with maximal
cell death between 8 and 24 h, illustrating rapid kinetics (Fig. 3C). In three independent experiments, the percentages of
dead XS52 cells at 16 h varied from 44 to 86% (67.0 ±
22.9%) in complete cocultures (Table I).
These values were significantly (p < 0.01)
higher than those observed for any of the incomplete cocultures.
Similar observations were made in a parallel experimental system in
which HDK-1 T cells, instead of the XS52 cells, were prelabeled with
FITC (Table I). Considering the possibility that FITC might have acted
as a reactive hapten, thereby triggering DC maturation (and possibly
death), we examined XS52 cell death under hapten-free conditions; XS52
cells and HDK-1 cells were first cocultured in the presence of KLH,
postlabeled with anti-Ia mAb (or anti-Thy-1 mAb), and then
examined for PI uptake. With this postlabeling method, 65% of the
Ia+ cells (i.e., XS52 cells) were judged to be PI positive
(Table I). In each experimental system, 25% of HDK-1 T cells died
in complete cocultures, corroborating previous reports by others that T
cells undergo apoptosis on activation in the absence of added growth
factors (39, 40, 41). In sum, these observations indicate that XS52 DC,
like splenic DC, undergo rapid and prominent death after Ag-specific
interaction with HDK-1 T cells.
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The observations of chromatin condensation among splenic DC and
the rapid onset of XS52 DC death both supported the hypothesis that DC
die by an apoptotic mechanism. To test this hypothesis more directly,
we examined DNA degradation in XS52 DC. XS52 cells were first labeled
with [3H]thymidine, washed extensively, and then
incubated for 8 h in the presence or absence of HDK-1 T cells
and/or KLH. Low m.w. DNA isolated from complete cocultures showed
characteristic DNA laddering in autoradiography, whereas DNA remained
relatively intact when the XS52 cells were incubated by themselves or
with either HDK-1 cells or KLH alone (Fig. 4). Taken as a whole, our observations
indicate that DC do undergo apoptosis after Ag-specific interaction
with CD4+ T cells.
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secretion by HDK-1 T cells and IL-1ß
secretion by XS52 cells (32). To determine whether such signaling was
also required to induce XS52 cell death, we blocked Ag-dependent
interaction with anti-Ia mAb. As shown in Fig. 5B, anti-Ia mAb inhibited
the secretion of IFN- and IL-1ß in the complete cocultures,
corroborating our previous observations (32). Importantly, anti-Ia
mAb also inhibited XS52 cell death significantly in the same complete
coculture, whereas control IgG showed minimal effects (Fig. 5A and B). We interpret these observations to
indicate that MHC class II-dependent DC-T cell interaction is required
for delivering an apoptotic signal.
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A). Treatment with LPS, which
activated XS52 cells as judged by up-regulated IL-1ß mRNA expression,
failed to significantly modulate mRNA expression for either CD95 or
CD95L. On the other hand, HDK-1 T cells expressed constitutively both
CD95 and CD95L mRNA. Treatment with Con A, which activated HDK-1 T
cells as judged by up-regulated IFN- mRNA expression, showed minimal
effects on CD95L mRNA expression, but it induced up-regulation of CD95
mRNA expression (Fig. 6A). These observations were confirmed
by Southern blot analysis of the PCR products (Fig. 6B). By
FACS analyses, we observed that XS52 DC express CD95, but not CD95L,
regardless of their activation states, whereas HDK-1 T cells express
CD95L only after activation (Fig. 7).
Thus, it appeared that surface expression of CD95 and CD95L is highly
regulated by posttranscriptional mechanisms.
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A, those T
cells expressed CD95 on their surface after Con A activation. When
incubated with these T cells in the presence of KLH, XS52 DC underwent
significant cell death. Once again, only modest cell death was observed
with XS52 DC after incubation with T cells alone or KLH alone,
duplicating our observations with the HDK-1 T cell clone (Fig. 8B).
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, anti-CD95 mAb Jo2 induced
significant death of thymocytes within 24 h, and thymocyte death
was further augmented by cycloheximide (CHX), an inhibitor of protein
synthesis, corroborating the reports of others (52, 53). The same
anti-CD95 mAb induced significant XS52 cell death only in the
presence of CHX. This is also consistent with previous reports that CHX
augments, or is even required, for anti-CD95-triggered apoptosis
(52, 54, 55). These results suggest that thymocytes and DC undergo
apoptosis by CD95-dependent, but perhaps different, mechanisms. To
determine the extent to which T cell-dependent DC apoptosis depends on
the CD95/CD95L pathway, we added neutralizing mAb against CD95L in
complete cocultures. As shown in Fig. 10, this mAb inhibited significantly,
albeit partially (50%), HDK-1 T cell-induced XS52 cell death. The
extent of inhibition remained at 50%, even with higher
concentrations (up to 30 µg/ml) of anti-CD95L (data not shown).
The specificity of inhibition with anti-CD95L mAb was verified by
the observations that neither a conventional hamster IgG control nor
anti-CD48 mAb (hamster IgG) that bound to the XS52 cell surface
(data not shown) blocked XS52 cell death significantly (Fig. 10). We
also tested whether the same anti-CD95L mAb would inhibit T
cell-induced apoptosis of splenic DC. As shown in Fig. 11, anti-CD95L mAb, but not
anti-CD48 mAb, prevented apoptosis of CD11c+ splenic DC
triggered by Ag-specific interaction with HDK-1 T cells. Thus, we
conclude that ligation of CD95 on DC with CD95L on activated T cells is
one of the mechanisms leading to DC apoptosis.
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A, CD54
and its ligand LFA-1 (CD11a/CD18) are both expressed by XS52 DC and by
HDK-1 T cells, suggesting that these adhesion molecules may play a
functional role in T cell-induced DC apoptosis. In fact, anti-CD54
mAb inhibited XS52 DC apoptosis in a dose-dependent manner, with 40%
inhibition achieved at 10 µg/ml. An isotype-matched control IgG had
minimal effects even at the highest concentration tested. Not only do
these observations indicate the involvement of CD54-CD11a/CD18
interaction in T cell-induced DC apoptosis, they also suggest that one
may be able to manipulate DC apoptosis experimentally using
anti-CD54, anti-CD95L, and/or other mAb.
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Our observations that DC undergo apoptosis upon Ag-specific
interaction with T cells imply that this may be a mechanism that limits
the extent of DC-induced T cell activation in vivo. To test this
hypothesis, we used CD95-deficient animals
(B6.MRL-FASlpr mice). Splenic DC
isolated from the CD95-deficient mice and from wild-type mice
(C57BL/6J) were pulsed with KLH and then injected s.c. into otherwise
untreated C57BL/6J mice. These animals were then challenged and
examined for footpad swelling responses to KLH. When relatively large
numbers (10–50 x 103 cells/mouse) of DC were
injected, CD95-deficient DC and control DC induced marked DTH reactions
at comparable levels (Fig. 13). The
extent of swelling response inducible by a single injection of
KLH-pulsed DC (5 x 104 cells/mouse) was roughly 70%
of that induced by immunization with KLH plus CFA. Splenic DC isolated
from control mice induced only minimal DTH responses at smaller numbers
(0.4–2 x 103 cells/mouse). By contrast, splenic DC
isolated from the CD95-deficient mice induced maximal responses even at
the smallest number (0.4 x 103 cells/mouse). Thus,
CD95-deficient DC do differ from control DC in their abilities to
induce DTH responses in vivo. We interpreted these results to support
our hypothesis that CD95/CD95L-mediated, T cell-induced apoptosis of DC
is a unique mechanism that prevents the interminable activation of T
cells by Ag-bearing DC.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Working with DC isolated from B6.MRL-FASlpr mice, we have observed that CD95-deficient DC are more potent than wild-type DC in their efficiency to induce DTH responses, especially at lower cell numbers. These results imply that CD95 molecules on DC (either expressed constitutively or induced on activation) have a negative impact on DC-dependent T cell activation in vivo. We have also observed in a series of in vitro experiments that CD95/CD95L interaction plays a functional role in T cell-induced DC apoptosis. Taken together, our observations suggest unique bidirectional signaling pathways during Ag presentation, in which DC deliver activation signals to T cells, whereas the same T cells deliver apoptotic signals to DC. The latter signaling pathway may serve as a mechanism that limits the extent of T cell activation inducible by a given number of Ag-pulsed DC.
In the present study we observed that Ag-specific interaction with the CD4+ T cell clone HDK-1 augments significantly apoptotic cell death of CD11c+ splenic DC in vitro. This corroborates the in vivo observations by Ingulli et al. (27) that OVA-pulsed, fluorescent dye-labeled DC initially interacted closely with OVA-reactive, CD4+ naive T cells in the lymph node but disappeared from this location at later time points. These two sets of observations suggest that DC die after completing the task of Ag presentation to CD4+ T cells. Using a long term DC line XS52, we have addressed the mechanisms of this event: 1) XS52 DC express CD95 on their surface; 2) CD95L expression becomes detectable on HDK-1 T cells after stimulation; 3) ligation of surface CD95 with anti-CD95 mAb triggers apoptosis of XS52 DC only in the presence of cycloheximide; and 4) neutralizing mAb against CD95L blocks significantly, albeit partially, HDK-1 T cell-induced XS52 DC apoptosis. These observations imply that coupling of CD95 (on DC) with CD95L (on T cells activated by DC) serves as one, but likely not the only, mechanism by which DC undergo apoptosis during Ag presentation. Obviously, further studies, especially at animal levels, will be required to determine the relative contribution of the CD95/CD95L system to T cell-mediated apoptosis of DC.
This is not the first report to describe the potential of DC to undergo apoptosis or to document CD95 expression by DC. Ludewig et al. (62) reported previously that DC undergo spontaneous apoptosis in culture and that this process is augmented by IL-10. We observed previously that ultraviolet B radiation sensitizes DC to become highly susceptible to apoptotic signals delivered by LPS treatment (35). More recently, Chambers et al. (63) found that DC serve as a sensitive target of NK cell-mediated, perforin-dependent cytolysis. Thus, DC undergo apoptosis in response to several different stimuli and by different mechanisms. With respect to CD95 expression, Winzler et al. (64) reported surface expression of CD95 by a long term DC line derived from mouse spleen. CD95L expression was also detected on this splenic DC line, corroborating the report by Süss and Shortman (60) that a subpopulation of splenic DC (i.e., CD8+ lymphoid DC) express CD95L and trigger apoptosis of alloreactive T cells. Thus, it is reasonable to propose that the CD95/CD95L system, which is now known to be involved in several immunological phenomena (e.g., CTL-mediated killing of target cells, suicidal cell death of T cells, and induction of immunological tolerance) (42), plays two distinct roles in DC-T cell interaction to induce apoptotic cell death of Ag-reactive T cells and Ag-pulsed DC.
Other APC populations (e.g., B cells and macrophages) are known to be killed by CD4+ T cells in Ag-specific and CD95/CD95L-dependent manners (43, 44, 50). Likewise, CD54-CD11a/CD18 interaction has been reported to be involved in T cell-mediated apoptosis of B cells (65). Thus, our observations made with DC are in complete agreement with the previous reports with other APC populations. It is to be emphasized that this is the first report formally documenting that DC, the most potent APC population, undergo apoptosis during Ag-specific interaction with CD4+ T cells. Taken together, one may propose that apoptosis occurs commonly in all APC populations after interaction with CD4+ T cells, thereby serving as a mechanism to down-regulate cellular immune responses.
In an attempt to develop strategies to experimentally control T cell-mediated DC apoptosis, we have identified that DC apoptosis is prevented by each of anti-Ia, anti-CD95L, and anti-CD54 mAb. Studies are under way to test the in vivo relevance of these observations. We anticipate that this line of investigation may ultimately lead to the development of new therapeutic reagents to prevent and even treat various human diseases that are caused by excessive or deficient responsiveness to environmental Ag, microbial Ag, tumor-associated Ag, or to self Ag.
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Footnotes |
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2 H.M. is a visiting scientist supported by the Ministry of Education, Science and Culture, Japan.
3 Address correspondence and reprint requests to Dr. Akira Takashima, Department of Dermatology, the University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75235-9069. E-mail address:
4 Abbreviations used in this paper: DC, dendritic cells; LC, Langerhans cells; PI, propidium iodide; CHX, cycloheximide; DTH, delayed type hypersensitivity; MIP, macrophage-inflammatory protein; KLH, keyhole limpet hemocyanin; CD95L, CD95 ligand.
Received for publication March 10, 1998. Accepted for publication February 11, 1999.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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B. Drenou, L. Amiot, N. Setterblad, S. Taque, V. Guilloux, D. Charron, R. Fauchet, and N. Mooney MHC class II signaling function is regulated during maturation of plasmacytoid dendritic cells J. Leukoc. Biol., April 1, 2005; 77(4): 560 - 567. [Abstract] [Full Text] [PDF]
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N. Setterblad, V. Blancheteau, A. Delaguillaumie, F. Michel, S. Becart, G. Lombardi, O. Acuto, D. Charron, and N. Mooney Cognate MHC-TCR interaction leads to apoptosis of antigen-presenting cells J. Leukoc. Biol., June 1, 2004; 75(6): 1036 - 1044. [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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W. Janssens, V. Carlier, B. Wu, L. VanderElst, M. G. Jacquemin, and J.-M. R. Saint-Remy CD4+CD25+ T Cells Lyse Antigen-Presenting B Cells by Fas-Fas Ligand Interaction in an Epitope-Specific Manner J. Immunol., November 1, 2003; 171(9): 4604 - 4612. [Abstract] [Full Text] [PDF]
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 K. Vermaelen and R. Pauwels Accelerated Airway Dendritic Cell Maturation, Trafficking, and Elimination in a Mouse Model of Asthma Am. J. Respir. Cell Mol. Biol., September 1, 2003; 29(3): 405 - 409. [Abstract] [Full Text] [PDF]
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 M. Leverkus, A. D. McLellan, M. Heldmann, A. O. Eggert, E.-B. Brocker, N. Koch, and E. Kampgen MHC class II-mediated apoptosis in dendritic cells: a role for membrane-associated and mitochondrial signaling pathways Int. Immunol., August 1, 2003; 15(8): 993 - 1006. [Abstract] [Full Text] [PDF]
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A. M. Woltman, S. W. van der Kooij, P. J. Coffer, R. Offringa, M. R. Daha, and C. van Kooten Rapamycin specifically interferes with GM-CSF signaling in human dendritic cells, leading to apoptosis via increased p27KIP1 expression Blood, February 15, 2003; 101(4): 1439 - 1445. [Abstract] [Full Text] [PDF]
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 M. T. Cruz, C. B. Duarte, M. Goncalo, A. Figueiredo, A. P. Carvalho, and M. C. Lopes The Sensitizer 2,4-Dinitrofluorobenzene Activates Caspase-3 and Induces Cell Death in a Skin Dendritic Cell Line International Journal of Toxicology, January 1, 2003; 22(1): 43 - 48. [Abstract] [PDF]
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J.-G. Castaigne, W. Guo, C. Leveille, D. Charron, and R. Al-Daccak A CD18-dependent protein kinase C {beta}-mediated alternative cell death pathway of activated monocytes Int. Immunol., September 1, 2002; 14(9): 1003 - 1014. [Abstract] [Full Text] [PDF]
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A. E. Lokshin, P. Kalinski, R. R. Sassi, R. B. Mailliard, J. Muller-Berghaus, W. J. Storkus, X. Peng, A. M. Marrangoni, R. P. Edwards, and E. Gorelik Differential regulation of maturation and apoptosis of human monocyte-derived dendritic cells mediated by MHC class II Int. Immunol., September 1, 2002; 14(9): 1027 - 1037. [Abstract] [Full Text] [PDF]
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N. Bertho, V. M. Blancheteau, N. Setterblad, B. Laupeze, J. M. Lord, B. Drenou, L. Amiot, D. J. Charron, R. Fauchet, and N. Mooney MHC class II-mediated apoptosis of mature dendritic cells proceeds by activation of the protein kinase C-{delta} isoenzyme Int. Immunol., August 1, 2002; 14(8): 935 - 942. [Abstract] [Full Text] [PDF]
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L. Frasca, C. Scotta, G. Lombardi, and E. Piccolella Human Anergic CD4+ T Cells Can Act as Suppressor Cells by Affecting Autologous Dendritic Cell Conditioning and Survival J. Immunol., February 1, 2002; 168(3): 1060 - 1068. [Abstract] [Full Text] [PDF]
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K. Kawahata, Y. Misaki, M. Yamauchi, S. Tsunekawa, K. Setoguchi, J.-i. Miyazaki, and K. Yamamoto Peripheral Tolerance to a Nuclear Autoantigen: Dendritic Cells Expressing a Nuclear Autoantigen Lead to Persistent Anergic State of CD4+ Autoreactive T Cells After Proliferation J. Immunol., February 1, 2002; 168(3): 1103 - 1112. [Abstract] [Full Text] [PDF]
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M. Bonkobara, P. K. Zukas, S. Shikano, S. Nakamura, P. D. Cruz Jr., and K. Ariizumi Epidermal Langerhans Cell-Targeted Gene Expression by a Dectin-2 Promoter J. Immunol., December 15, 2001; 167(12): 6893 - 6900. [Abstract] [Full Text] [PDF]
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H. Matsue, K. Matsue, M. Kusuhara, T. Kumamoto, K. Okumura, H. Yagita, and A. Takashima Immunosuppressive properties of CD95L-transduced "killer" hybrids created by fusing donor- and recipient-derived dendritic cells Blood, December 1, 2001; 98(12): 3465 - 3472. [Abstract] [Full Text] [PDF]
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S. Latour, H. Tanaka, C. Demeure, V. Mateo, M. Rubio, E. J. Brown, C. Maliszewski, F. P. Lindberg, A. Oldenborg, A. Ullrich, et al. Bidirectional Negative Regulation of Human T and Dendritic Cells by CD47 and Its Cognate Receptor Signal-Regulator Protein-{alpha}: Down-Regulation of IL-12 Responsiveness and Inhibition of Dendritic Cell Activation J. Immunol., September 1, 2001; 167(5): 2547 - 2554. [Abstract] [Full Text] [PDF]
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M. L. Fields, C. L. Sokol, A. Eaton-Bassiri, S.-j. Seo, M. P. Madaio, and J. Erikson Fas/Fas Ligand Deficiency Results in Altered Localization of Anti-Double-Stranded DNA B Cells and Dendritic Cells J. Immunol., August 15, 2001; 167(4): 2370 - 2378. [Abstract] [Full Text] [PDF]
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 M. Rescigno, V. Piguet, B. Valzasina, S. Lens, R. Zubler, L. French, V. Kindler, J. Tschopp, and P. Ricciardi-Castagnoli FAS Engagement Induces the Maturation of Dendritic Cells (Dcs), the Release of Interleukin (Il)-1{beta}, and the Production of Interferon {gamma} in the Absence of IL-12 during Dc-T Cell Cognate Interaction: A New Role for FAS Ligand in Inflammatory Responses J. Exp. Med., December 4, 2000; 192(11): 1661 - 1668. [Abstract] [Full Text] [PDF]
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S. K. Bauman, K. L. Nichols, and J. W. Murphy Dendritic Cells in the Induction of Protective and Nonprotective Anticryptococcal Cell-Mediated Immune Responses J. Immunol., July 1, 2000; 165(1): 158 - 167. [Abstract] [Full Text] [PDF]
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F. Willems, Z. Amraoui, N. Vanderheyde, V. Verhasselt, E. Aksoy, C. Scaffidi, M. E. Peter, P. H. Krammer, and M. Goldman Expression of c-FLIPL and resistance to CD95-mediated apoptosis of monocyte-derived dendritic cells: inhibition by bisindolylmaleimide Blood, June 1, 2000; 95(11): 3478 - 3482. [Abstract] [Full Text] [PDF]
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 C. Servet-Delprat, P.-O. Vidalain, O. Azocar, F. Le Deist, A. Fischer, and C. Rabourdin-Combe Consequences of Fas-Mediated Human Dendritic Cell Apoptosis Induced by Measles Virus J. Virol., May 1, 2000; 74(9): 4387 - 4393. [Abstract] [Full Text]
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 K. Ariizumi, G.-L. Shen, S. Shikano, R. Ritter III, P. Zukas, D. Edelbaum, A. Morita, and A. Takashima Cloning of a Second Dendritic Cell-associated C-type Lectin (Dectin-2) and Its Alternatively Spliced Isoforms J. Biol. Chem., April 14, 2000; 275(16): 11957 - 11963. [Abstract] [Full Text] [PDF]
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N. Bertho, B. Drenou, B. Laupeze, C. L. Berre, L. Amiot, J.-M. Grosset, O. Fardel, D. Charron, N. Mooney, and R. Fauchet HLA-DR-Mediated Apoptosis Susceptibility Discriminates Differentiation Stages of Dendritic/Monocytic APC J. Immunol., March 1, 2000; 164(5): 2379 - 2385. [Abstract] [Full Text] [PDF]
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W.-P. Min, R. Gorczynski, X.-Y. Huang, M. Kushida, P. Kim, M. Obataki, J. Lei, R. M. Suri, and M. S. Cattral Dendritic Cells Genetically Engineered to Express Fas Ligand Induce Donor-Specific Hyporesponsiveness and Prolong Allograft Survival J. Immunol., January 1, 2000; 164(1): 161 - 167. [Abstract] [Full Text] [PDF]
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V. Loyer, P. Fontaine, S. Pion, F. Hetu, D.-C. Roy, and C. Perreault The In Vivo Fate of APCs Displaying Minor H Antigen and/or MHC Differences Is Regulated by CTLs Specific for Immunodominant Class I-Associated Epitopes J. Immunol., December 15, 1999; 163(12): 6462 - 6467. [Abstract] [Full Text] [PDF]
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S. Shikano, M. Bonkobara, P. K. Zukas, and K. Ariizumi Molecular Cloning of a Dendritic Cell-associated Transmembrane Protein, DC-HIL, That Promotes RGD-dependent Adhesion of Endothelial Cells through Recognition of Heparan Sulfate Proteoglycans J. Biol. Chem., March 9, 2001; 276(11): 8125 - 8134. [Abstract] [Full Text] [PDF]
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K. Ariizumi, G.-L. Shen, S. Shikano, S. Xu, R. Ritter III, T. Kumamoto, D. Edelbaum, A. Morita, P. R. Bergstresser, and A. Takashima Identification of a Novel, Dendritic Cell-associated Molecule, Dectin-1, by Subtractive cDNA Cloning J. Biol. Chem., June 23, 2000; 275(26): 20157 - 20167. [Abstract] [Full Text] [PDF]
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).
Samples were then examined for PI uptake. Data are the percentage of
inhibition of XS52 DC death (mean ± SD, n =
3) as compared with the control in the absence of added Abs.