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Blood, Vol. 92 No. 12 (December 15), 1998:
pp. 4539-4544
RAPID COMMUNICATION
Critical Requirement for Graft Passenger Leukocytes in Allograft
Tolerance Induced by Donor Blood Transfusion
By
Régis Josien,
Michèle Heslan,
Sophie Brouard,
Jean-Paul Soulillou, and
Maria-Cristina Cuturi
From the INSERM Unit 437 and ITERT "Institut de Transplantation et
de Recherche en Transplantation," Immeuble Jean Monnet, Nantes,
France.
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ABSTRACT |
Tolerance to a vascularized allograft can be induced in adult
animals by pregraft donor-specific blood transfusion (DST). Mechanisms
underlying this effect appear to depend on unresponsiveness of
alloreactive T-helper cells. In this study, we examined the roles of
DST and cellular components of the allograft that are important in
inducing T-cell unresponsiveness in a rat model. DST alone did not
tolerize alloreactive recipient T-helper cells, but the combination of
DST and heart allograft induced profound inhibition of the antidonor
proliferative response in spleen but not in lymph node cells. When
heart allografts were depleted of passenger leukocytes by pretreating
the donor with cyclophosphamide or by parking the graft for 2 months in
a tolerant recipient, tolerance induction in DST-treated recipients was
abrogated. Tolerance could then be restored in a majority of
DST-treated recipients of passenger leukocytes depleted grafts by
injecting them at the time of grafting with donor, but not third-party,
dendritic cells. This indicates that graft passenger leukocytes, most
likely dendritic cells, are required for DST-induced allograft
tolerance.
© 1998 by The American Society of Hematology.
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INTRODUCTION |
IT HAS LONG BEEN established that
pretransplant donor-specific blood transfusions (DST) result in the
enhancement or tolerance of vascularized allografts in
rodents.1-3 In addition to DST (reviewed in Anderson and
Brennan4), donor unrelated pregraft blood transfusions (BT)
have also been shown to either enhance kidney allograft survival or to
reduce the incidence of acute rejection in humans.5-7
Results obtained in experimental animal models suggest that the effects
of BT may be related to either fortuitous exposure to graft
donor-specific antigens or to T-cell cross-reactivity,2,8
suggesting that the immunomodulatory effect of DST and random BT may
share common mechanisms. At present, these mechanisms remain poorly
understood; however, studies in rodents have provided important
information. Unexpectedly, donor-specific cytotoxic T lymphocytes (CTL)
can be found in nonrejected cardiac or renal allografts from
DST-treated rats, indicating the absence of clonal deletion of
alloreactive CD8+ T cells.9-11 Moreover, the
fact that tolerance can be abrogated by the administration of exogenous
interleukin-2 (IL-2) or interferon- (IFN-) after grafting
suggests that alloreactive CD4+ T cells are also present,
but somehow silenced.11,12
T-helper (Th) cells play a pivotal role in acute allograft rejection,
and DST-induced allograft tolerance is likely due to a dramatic
downregulation of Th function as shown in rodent
models.11-13 Possible mechanisms underlying this defect are
not clear: anergy has been proposed,12 as has an immune
deviation towards a Th2-type reaction.14 However, we have
recently shown in a rat model that both Th1- and Th2-related cytokines
were downregulated in heart allografts from DST-treated animals, as
compared with untreated recipients.13
It is still unclear whether the DST by itself is tolerogenic. Although
the presence of donor-specific suppressor T cells in DST-treated rats
has been reported in vitro,15 such cells could be detected
in our model in DST-treated animals only after the transplantation.16 Moreover, results obtained in human have shown that fully major histocompatibility complex
(MHC)-incompatible BT resulted in an increase in both
donor-specific T-helper and CTL precursors,17,18 indicating
that allogenic BTs sensitize rather than tolerize. Based on these
results, we sought to define which kind of stimulus within the
allograft is responsible for turning off the T-helper response in
DST-treated recipients. We report that graft passenger leukocytes,
known to be highly immunogenic, are necessary for tolerance induction
in DST-treated rats. This effect can be substituted by purified donor
dendritic cells (DCs), indicating that these antigen-presenting cells
can govern the cellular events leading to both rejection and tolerance,
depending on the context in which they are involved.
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MATERIALS AND METHODS |
Animals and transplantation.
Eight- to 12-week-old male Lewis.1W (RT1.u), Lewis.1A (RT1.a), and
Lewis (RT1.l) rats were obtained from the Zentral Institut Für
Versuchstierzucht (Hannover, Germany) or kindly provided by E. Günther (Georg-August Universität Göttingen,
Göttingen, Germany). Heterotopic heart grafts were
performed using the Ono and Lindsey technique,19 and
function was monitored daily by palpation through the abdominal wall.
Some DST-treated recipients received a heart allograft that was parked
for 2 months in another tolerant DST-treated recipient. The grafts were
harvested from the tolerant recipient's abdominal location and
regrafted using the Ono and Lindsey technique19 into fresh
DST-treated recipients. Skin grafts were performed using standard
procedures.20
Donor and recipient pretreatments.
Allograft recipients received either no pretreatment or DST. Blood (1 mL), collected from a Lewis.1W donor by cardiac puncture into a syringe
containing heparin (final concentration, 20 U/mL), was immediately
injected intravenously (IV) into Lewis.1A recipients 14 and 7 days
before the transplantation. This regimen induces long-term allograft
tolerance (see Fig 7 for allograft survivals), which is donor MHC
specific, because third-party blood transfusions do not prolong graft
survival3 and because long-term tolerant recipients accept
donor-type, but not third-party MHC expressing skin grafts
(Fig 1). Where indicated, donor rats received, 5 days before graft harvesting, a single intraperitoneal dose (300 mg/kg) of
cyclophosphamide (Sigma, St Louis, MO) to deplete heart graft leukocytes.21
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| Fig 1.
DST-induced allograft tolerance is donor-specific.
Long-term (>100 days) tolerant heart allograft recipients were
challenged with donor-type (Lewis.1W) and third-party-type (Lewis.
RT1.l) and recipient-MHC type-expressing skin grafts. Note the
rejection of third-party but not of donor-type skin graft.
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Preparation of DCs.
Highly enriched (75% to 85%) suspensions of mature spleen DCs were
prepared as we previously described.22 Bone marrow-derived DCs (>85% pure as assessed by class II staining and morphology) were
prepared as described by Talmor et al.23 Briefly, femoral bone marrow cells were cultured in 24-well plates in complete medium
containing mouse granulocyte-macrophage colony-stimulating factor
(GM-CSF; 1/1,000 dilution of surpernatant from murine
GM-CSF-transfected COS cells) and rat IL-4 (1/1,000 dilution of
supernatant from rat IL-4-transfected COS cells). Cells were fed on
days 2, 4, 6, and 8 and transferred to 100-mm culture dishes on day 9 to induce maturation and were used on day 11. After three washes, cells
were resuspended in RPMI at 5 × 105/mL and kept on
ice until injected.
Proliferation assays.
A standard one-way mixed lymphocyte culture (MLC) was performed. Spleen
and LN cell suspensions were prepared as previously described24 from DST-treated but ungrafted, as well as from grafted DST-treated or untreated rats 5 days after the transplantation. In some experiments, CD4+ and CD8+ cells were
depleted from the responding cell population using anti-CD4 or anti-CD8
monoclonal antibodies (MoAbs) followed by antimouse IgG-conjugated
Dynabeads (Dynal, Oslo, Norway) as previously described.22
Irradiated donor-type Lewis.1W (RT1.u), third-party Lewis
(RT1.l), or Buffalo (RT1.b) splenocytes served
as stimulator cells. Responder and stimulator cells (2 × 105 cells/well) were plated in 96-well round-bottom plates
in triplicate in a final volume of 200 µL of complete medium and
cultured at 37°C in 5% CO2. Proliferation of the
responder population was assessed from day 1 to 7 by measuring the
incorporation of [3H]TdR (0.5 µCi per well; Amersham,
Les Ulis, France) during the last 8 hours of culture. Cells were then
harvested on glass fiber filters and [3H]TdR
incorporation was measured by standard scintillation procedures (Packard Instruments, Meriden, CT).
Immunohistology.
Heart tissue samples removed from untreated or cyclophosphamide-treated
LEW.1W donors were embedded in Tissue Tek (OCT Compound; Bayer
Diagnostics, Puteau, France), snap-frozen in liquid
nitrogen, and stored at  70°C. Five-micrometer cryostat
sections were cut, air-dried, and fixed in acetone for 10 minutes at
room temperature. Sections were then labeled using a three-step
indirect immunoperoxidase technique using either OX1+OX30 (CD45, all
leukocytes) or OX6 (MHC class II antigen) MoAbs. Nonspecific staining
was controlled for by omission of the first antibody. The number of
positively stained cells was counted on each slide using a grid mounted
in the ocular eyepiece.
Statistical analysis.
Graft survivals were compared using the Kaplan Meier method.
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RESULTS AND DISCUSSION |
DST does not tolerize T-helper cells in the absence of an allograft.
Previous studies have shown that DST, with or without a subsequent
allograft in adult rat, did not tolerize CTL but instead prime them to
donor alloantigens.25 To analyze CD4+ T-cell
function in DST-treated rats, we performed MLCs with spleen and lymph
node (LN) responder cells from transfused animals killed 14 days after
the first DST (the time at which the heart transplantation is normally
performed in this model). After mixing with stimulator cells, Lewis.1W
(RT1.u), or third-party Buffalo (RT1.b)
irradiated splenocytes, proliferative responses were assessed daily
from day 1 to 7 of culture. As shown in Fig 1, both spleen and LN cells
mounted a potent proliferative response against both donor and
third-party alloantigens. The kinetic and level of proliferation of LN
cells against donor and third party stimulators were not significantly
modified by DST (Fig 2). In contrast,
splenocyte proliferation appeared greater for DST-treated than for
naive animals from day 3 to 7 of culture (Fig 2).
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| Fig 2.
DST does not tolerize Th cells to donor alloantigen.
Spleen and LN cells were prepared from untreated naive or DST-treated
(1 mL on days 14 and 7) rats 14 days after the first transfusion.
Cells (2 × 105) were plated in triplicate with an equal
number of donor-specific Lew.1W (RT1.u) (A) or third-party Buffalo
(RT1.b) (B) irradiated spleen cells in 96-well round-bottom plates and
cultured for 1 to 7 days. Thymidine incorporation during the last 8 hours of each of these cultures was assessed. Representative data of
three independent experiments are shown.
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Some investigators have proposed that DST and random BT effects are
mediated by an anergy of donor-reactive T cells after encountering
alloantigens on resting donor B cells.26,27 However, our
results and those of others suggest that this may not always be the
case. Indeed, allogenic DST primes the recipient CTLs against donor
alloantigens,10,25 and the data presented here suggest that
donor-reactive Th cells are not anergized by DST (Fig 2). In humans,
MHC-mismatched BT resulted in an increase of donor-specific T-helper
cell precursors. Moreover, it has been shown that resting B cells can
induce tolerance to minor histocompatibility antigen, but not to MHC
antigen-mismatched allografts.28
Both DST and heart allograft are required to tolerize splenic Th
cells.
We previously showed that expression of both Th-1- and
Th-2-type cytokine mRNA was strongly diminished in heart grafts from DST-treated rats as compared with allografts in naive
recipients.13 This suggests that Th function is depressed.
To further assess donor-reactive Th function in these animals, we
performed MLC with spleen and LN cells from untreated and DST-treated
allograft recipients killed 5 days after transplantation. The results
of the antidonor proliferative response are described in
Fig 3. Both spleen and LN cells from
untreated animals exhibited a strong proliferative response against
irradiated donor-type spleen cells. Depleting CD4+ but not
CD8+ cells from the responding population abolished this
response indicating that, as previously described,29
proliferation in MLC is mediated by T-helper cell ( Fig
4). In DST-treated animals, LN cells proliferated in the presence of
donor antigen, whereas the response of spleen cells was dramatically
reduced. Although these cells proliferated normally in response to
Concanavalin A (Fig 5), their response against third
party stimulator cells from Buffalo (RT1.b) (Fig 5) or
Lewis (RT1.l) (data not shown) rats were also reduced,
indicating a nonspecific in vitro suppression. However, it is clear
that in vivo the effect of DST is donor-specific, because third-party
pregraft BT did not enhance Lewis.1W heart graft survival in Lewis.1A
recipients3 and because long-term tolerant animals accept
donor-type but not third-party-type skin grafts (Fig 1). It
was important to exclude that this difference between pregraft and
postgraft profiles could reflect kinetic variations in the pattern of
the proliferative response in DST-treated rats. To answer this
question, we performed the same experiments as described in the Fig 2
with DST-treated ungrafted animals killed 12 days after the second
transfusion (equivalent to day +5 after transplantation) or with
DST-treated animals that received on day 0 a third DST instead of an
allograft. As expected, in both cases the proliferative response was
similar to that described in Fig 2 (data not shown). Finally, these
results led us to conclude that both DST and allograft are required to tolerize splenic alloreactive T-helper cells. These results are consistent with our previous preliminary report showing that DST and
heart allograft, but not DST alone, induce suppressor T cells in
recipient spleen.16
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| Fig 3.
Both DST and heart allograft are required to tolerize
recipient splenic Th cells. Spleen and LN cells were prepared from
DST-treated and untreated heart graft recipient rats killed 5 days
after the transplantation. Proliferation against irradiated spleen from
LEW.1W (RT1.u) or third-party Buffalo (RT1.b) and Lewis (RT1.l) (data
not shown) was assessed every day as described in Fig 1. Representative
data of five independent experiments are shown.
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| Fig 4.
The proliferative response during MLC is mediated by
CD4+ cells. Spleen cells were prepared from DST-treated
and untreated heart graft recipients sacrificed 5 days after the
transplantation. Total, CD4-depleted, or CD8-depleted (Dynal) cells (2 × 105) were plated in triplicate with an equal number of
Lew.1W (RT1.u) irradiated spleen cells in 96-well round-bottom plates
and cultured for 5 days. Thymidine incorporation was assessed during
the last 8 hours. Representative data of three independent experiments
are shown.
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| Fig 5.
The suppression of the proliferative response against
alloantigens in spleen cells from DST-treated and grafted animals is
not donor-specific. Spleen cells were prepared from DST-treated and
untreated heart graft recipients killed 5 days after the
transplantation. Cells (2 × 105) were plated in
triplicate alone with an equal number of Lew.1W (RT1.u) or third-party
Buffalo (RT1.b) irradiated spleen cells or in the presence of 5 µg/mL
of Concanavalin A (Con A) in 96-well round-bottom plates and cultured
for 5 days. Thymidine incorporation was assessed during the last 8 hours. Representative data of five independent experiments are shown.
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The disappearance of the proliferative response in splenocytes
from DST-treated rats after the transplantation is unlikely to be due
to trapping of alloreactive CD4+ T cells in heart graft,
because this response can be restored in vitro with a combination of
IL-2 (50 U/mL) and anti-transforming growth factor- (TGF-)
MoAb.29a Different T-cell response patterns between spleen
and LN compartments have been previously shown in other
models25,30 and could be related to differential migratory
properties of naive and activated T cells. Also, this discrepancy might
be due to differential access of alloantigen in these two compartments.
Indeed, it has been shown that, after IV infusion, allogenic leukocytes
are almost exclusively eliminated in spleen in normal euthymic
rats,31 and, moreover, after vascularized allografts in
rodents, graft interstitial DCs rapidly migrate into recipient spleen
and then associate with CD4+ T cells.32 Thus,
it is possible that alloantigens do not gain access to LN compartments
after BT and allograftment, and therefore LN T cells in vivo might be
naive for these antigens. A role for spleen as a critical site for the
induction of tolerance by DST is further supported by previous studies
showing that splenectomy abrogates the induction of unresponsiveness
after IV alloantigen injection.33,34
Graft passenger leukocytes are required for induction of tolerance in
DST-treated rats.
Having demonstrated the requirement for the heart allograft to induce
unresponsiveness of splenocytes from DST-treated rats to donor
alloantigens, we then sought to determine which cell type within the
graft could be responsible for turning off the Th response. It is well
known that organ allografts contain two distinct cell subsets
controlling tissue immunogenicity. Resident parenchymatous cells of the
heart themselves are thought to be antigenic but poorly immunogenic,
mainly because of their lack of expression of costimulatory molecules.
In contrast, bone marrow-derived passenger leukocytes and more
precisely interstitial DCs are potent stimulators of allogenic
responses both in vitro and in vivo and are suggested to account for
the majority of the allograft immunogenicity.35 Indeed,
depletion of passenger leukocytes from allografts before transplantation has been shown to enhance allograft survival in some
rodent combinations.21,36-38 Thus, to differentiate the
role of passenger leukocytes and heart graft tissue in the tolerizing effect of the graft in DST-treated rats, we pretreated heart donors with a single dose of cyclophosphamide (300 mg/kg) 5 days before organ
harvesting. As shown in Fig 6, this
treatment led to a greater than 95% reduction in the number of
interstitial CD45 and class II-positive cells in the donor hearts at
the time of transplantation. Previous studies have shown that these
cyclophosphamide-sensitive cells expressing class II molecules are
exclusively interstitial DCs.39,40 Interstitial DC-depleted
hearts were acutely rejected upon grafting into DST-treated recipients,
with a mean survival time (MST) ± SD of 11.7 ± 4.3 days (not
significant [NS] as compared with untreated recipients
of normal heart grafts, MST = 11.9± 3.4 days;
Fig 7). Moreover, depletion of passenger
leukocytes did not significantly prolong allograft survival in
untreated recipients (8.4 ± 1.5 days). This result suggest that the
mechanism of acute rejection occurring in these animals mostly depends
on the presentation of graft derived alloantigen by host
antigen-presenting cells (APC; indirect pathway of allorecognition)
instead of alloantigen recognition on donor APC (direct pathway). Heart
grafts from cyclophosphamide-treated donors into syngenic recipients
survived indefinitely (n = 5), indicating that abrogation of graft's
ability to induce tolerance in DST-treated recipients was not simply
due to myocardial toxicity of cyclophosphamide. Finally, 3 DST-treated
recipients received heart allografts that were parked in tolerant
DST-treated recipients for 2 months, a period by which donor
interstitial DCs are replaced by recipient interstitial
DCs.41 Rejection occurred in these animals in 10, 12, and
22 days. Therefore, we conclude that graft interstitial DCs are
required for the induction of tolerance in DST-treated rats.
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| Fig 6.
The effect of pretreatment of the donor with
cyclophosphamide on leukocyte content in heart. Cryostat section of
heart tissue samples from untreated ( ) or cyclophosphamide-treated
( ) donor rats were labeled with OX1+OX30 (CD45) and OX6 (class II
MHC) MoAbs using a three-step immunoperoxidase technique. Positive
cells were counted using a grid in ocular eyepiece and each bar
represents the mean ± SD of 4 animals in each group.
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| Fig 7.
Allograft survivals. Donors were pretreated with a single
intraperitoneal injection of cyclophosphamide (cyp) at 300 mg/kg on day
5. Recipients were pretreated with two DSTs (1 mL of fresh blood) on
days 14 and 7. Recipients were posttreated where indicated by IV
injection of 2.5 × 105 donor or third-party splenic or
bone marrow-derived DCs in 100 µL RPMI at the end of the surgical
procedure.
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In a previous study, Heineman et al,42 using rats from a
different genetic background than the one used in this study, showed that donor irradiation before graft harvesting also abrogated the
induction of allograft tolerance in a DST-treated recipient, indicating
that this effect is not restricted to particular strains. Although
cyclophosphamide could theoretically interfere with activity of
DST-induced regulatory cells, it is unlikely that sufficient amount of
this drug could be carried over into the recipient with the graft,
because hearts were extensively washed before engraftment and because
the heart does not efficiently accumulate
cyclophosphamide.43
The injection of donor-type, but not third-party, DCs can
restore tolerance to a passenger leukocyte-depleted heart allograft in
a majority of DST-treated animals.
To further assess the role of donor interstitial DCs in inducing
tolerance in DST-treated rats, we injected DST-treated recipients of an
interstitial DCs-depleted heart allograft with donor and third-party
(Lewis, RT1.l) DCs at the time of transplantation. Because it is
difficult to obtain a sufficient number of donor DCs from heart, they
were prepared from spleen or bone marrow. The results in Fig 7 show
that the IV injection of 2.5 × 105 (the average
number of interstitial DCs in a rat heart; see McKenzie et
al21) donor spleen or bone marrow-derived DCs restored
indefinite allograft survival in 56% (5/9) and 63% (5/8),
respectively, of DST-treated recipients of interstitial DCs-depleted
heart graft (P < .001 as compared with noninjected
recipients). In contrast, the injection of a similar number of
third-party (RT1.l) DCs did not significantly prolong allograft
survival (Fig 7). Increasing the number of DC injected to 1 × 106 cell per recipient did not further increase the
percentage of surviving allografts.
Previous studies in rats have shown that interstitial DCs play a
critical role in inducing acute allograft rejection by directly stimulating recipient CD4+ and CD8+ T cells
(direct pathway of allorecognition).35,41,44 We showed here
that the direct pathway of allorecognition could also elicit tolerance
in CD4+ T cells from DST-treated rats. Because, in the
absence of graft interstitial DCs, the hearts are rejected in these
animals, this rejection is probably due to indirect recognition of
donor alloantigens. Interestingly, interstitial DCs have been shown to
disappear more rapidly from allografts in DST-treated than in untreated
recipients.9 However, it is not known whether this results
from in situ cell death or migration. Based on the present results, we
hypothesize that heart interstitial DCs rapidly migrate to the spleen
of a DST-treated host. This migration would result in an inactivation rather than a stimulation of recipient splenic CD4+ T
cells. The absence of suppression in the lymph nodes compared with
spleen could be explained by the inability of DCs to access to the
lymph nodes from the blood.45 We recently showed that the
bystander suppression of allogenic proliferative response observed in
spleen from DST-treated and grafted rats is related to
TGF-.29a Moreover, TGF-1 was overexpressed in
nonrejected allografts from DST-treated recipient, and allograft
tolerance was abrogated by anti-TGF- MoAb treatment. This strongly
suggests that induction of allograft tolerance in this model is
dependent on TGF--producing cells. It is possible that donor Ags
present on graft interstitial DCs are required for activation of, and
TGF- production by, these putative regulatory cells.
Hart and Fabre46 have showed that the presence of
interstitial DCs in kidney graft was also required for tolerance to be induced in passively enhanced recipient rats. They suggested that the
mechanism of induction of passive enhancement was related to an
interaction of enhancing antibodies with donor interstitial DCs.46 We have previously reported, in our model, that a
high level of anti-class II IgG2a could be detected at the time of transplantation in the DST-treated rat.47 It is likely that these antibodies interact with graft interstitial DCs during the first
2 to 3 days after grafting, because they are the only
cells expressing class II antigen in normal rat heart.40
However, the fact that interstitial DC-depleted allografts were
rejected in unmodified recipients indicates that anti-class II
antibodies do not simply act through opsonisation and destruction of
interstitial DCs. Nevertheless, anti-class II Ab could modify
interstitial DCs function and mask class II molecules that would
prevent CD4+ T-cell activation. On the other hand, rapid
opsonisation of interstitial DCs could favor their processing by
recipient spleen APC and then would allow indirect alloantigen
presentation to T cells in a tolerogenic fashion.
In addition to their major role in initiating primary T-cell
response both in vitro and in vivo,48 DCs could also exert an inhibitory action on the immune response. The strongest support for
this theory is evidenced by the critical role of DCs in the negative
selection process occurring in thymus. In the periphery, DCs may also
be involved in maintaining tolerance to peripheral tissue
antigen.49 Süss and Shortman50 have shown
that a subset of CD8+ splenic DCs in mice have the capacity
to delete CD4+ T cells that interact with them, by
FasL/Fas-mediated apoptosis, suggesting that this DC subset could have
a regulatory function. In addition, we recently showed that rat spleen
DCs exhibit a Ca2+-dependent NK-like cytotoxic
function.22 Bone marrow-derived DC progenitors in mouse
have been shown to induce T-cell hyporeponsiveness in vitro and to
prolong heart allograft survival when injected 7 days before
grafting.51 Finally, donor DCs are thought to play an
important role in establishing spontaneous tolerance to liver
allografts in rodent.51 Together with our present study, these data suggest that, in addition to their role in triggering allograft rejection, DCs might also be involved in allograft tolerance under certain circumstances.
In conclusion, we have shown that the presence of interstitial
DCs is required for tolerance to be induced in heart allograft recipient rats pretreated by DST. Thus, direct recognition of alloantigen does not always evoke acute allograft rejection but could
also, in DST-primed recipients, tolerize T-helper cells. Our results
suggest that the pregraft immunologic status of allograft recipients
and, in particular, whether they have received pregraft BTs should be
carefully considered in studies analyzing the effect of organ DC
depletion on allograft survival in humans.
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ACKNOWLEDGMENT |
The authors thank Dr Ralph Steinman and Brian Wong for critically
reading the manuscript.
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FOOTNOTES |
Submitted August 20, 1998;
accepted October 5, 1998.
The publication costs of this
article were defrayed in part by
page charge payment. This article
must therefore be hereby marked
"advertisement"
in accordance with 18 U.S.C. section
1734 solely to indicate this fact.
Address reprint requests to Régis Josien, MD, PhD, INSERM UNIT
437, Immeuble Jean Monet, 30, bd Jean Monet, 44035 Nantes Cédex
01, France; e-mail: rjosien{at}nantes.inserm.fr.
| |
REFERENCES |
1.
Fabre JW, Morris PJ:
The effect of donor strain blood pre-treatment on renal allograft rejection in rat.
Transplantation
14:608, 1972[Medline]
[Order article via Infotrieve]
2.
Peugh WN, Wood KJ, Morris PJ:
Genetic aspects of the blood transfusion effect.
Transplantation
46:438, 1988[Medline]
[Order article via Infotrieve]
3.
Soulillou J, Blandin F, Günther E, Lemoine V:
Genetics of the blood transfusion effect on heart allografts in rats.
Transplantation
38:63, 1984[Medline]
[Order article via Infotrieve]
4.
Anderson C, Brennan D:
A sanguine outlook: The role of donor-specific transfusion in renal transplantation and tolerance.
Transplant Rev
9:49, 1995
5.
Opelz G, Terasaki P:
Improvment of kidney graft survival with increased numbers of blood transfusions.
N Engl J Med
299:799, 1978[Abstract]
6.
Opelz G, Sengar DPS, Mickey MR, Terasaki PI:
Effect of blood transfusions on subsequent kidney transplants.
Transplant Proc
5:253, 1973[Medline]
[Order article via Infotrieve]
7.
Baatard R, Dantal J, Cantarovich D, Cesbron A, Bignon J, Soulillou J:
Effect of the number of pregraft blood transfusions in kidney graft recipients treated with bioreagents and cyclosporine A.
Transplant Int
4:235, 1991[Medline]
[Order article via Infotrieve]
8.
Bushell A, Morris PJ, Wood KJ:
Induction of operational tolerance by random blood transfusion combined with anti-CD4 antibody therapy. A protocol with significant clinical potential.
Transplantation
58:133, 1994[Medline]
[Order article via Infotrieve]
9.
Armstrong H, Bolton E, McMillan S, Spencer S, Bradley J:
Prolonged survival of actively enhanced rat renal allograft despite accelerated cellular infiltration and rapid induction of both class I and class II MHC antigens.
J Exp Med
169:891, 1987
10.
Dallman M, Wood K, Morris P:
Specific cytotoxic T cells are found in the non-rejected kidneys of blood transfused rats.
J Exp Med
165:566, 1987[Abstract/Free Full Text]
11.
Bugeon L, Cuturi M, Hallet M, Paineau J, Chabannes D, Soulillou J:
Peripheral tolerance of an allograft in adults rats characterized by low interleukin-2 and IFN- mRNA levels and by strong accumulation of major histocompatibility complex transcripts within the graft.
Transplantation
54:219, 1992[Medline]
[Order article via Infotrieve]
12.
Dallman MJ, Shiho O, Page TH, Wood KJ, Morris PJ:
Peripheral tolerance to alloantigen results from altered regulation of the interleukin 2 pathway.
J Exp Med
173:79, 1991[Abstract/Free Full Text]
13.
Josien R, Pannetier C, Douillard P, Cantarovich D, Menoret S, Bugeon L, Soulillou J-P, Cuturi M-C:
Graft infiltrating T helper cells, CD45RC phenotype, and Th1/Th2-related cytokines in donor specific transfusion-induced tolerance in adult rats.
Transplantation
60:1131, 1995[Medline]
[Order article via Infotrieve]
14.
Takeuchi T, Lowry RP, Konieczny B:
Heart allografts in murine systems: The differential activation of Th2-like effector cells in peripheral tolerance.
Transplantation
53:1281, 1992[Medline]
[Order article via Infotrieve]
15.
Quigley R, Wood K, Morris P:
Transfusion induces blood-donor specific suppressor cells.
J Immunol
142:463, 1989[Abstract]
16.
Chevalier S, Lacroix H, Moreau J, Soulillou J:
Blood transfusion plus allograft-but not blood transfusion alone-induce IL 2-producing suppressor cells in Lew-1A recipients of Lew-1W heart allograft.
Transplant Proc
19:544, 1987[Medline]
[Order article via Infotrieve]
17.
Vandekerckhove BA, van Bree S, Zhang L, Datema G, Zantvoort F, Claas FH:
Increase of donor-specific cytotoxic T lymphocyte precursors after transfusion.
Transplantation
48:672, 1989[Medline]
[Order article via Infotrieve]
18.
Vandekerckhove BA, Datema G, Zantvoort F, Claas FH:
An increase of donor-specific T helper precursors resulting from blood transfusions.
Transplantation
45:987, 1990
19.
Ono K, Lindsey ES:
Improved technique of heart transplantation in rats.
J Thorac Cardiovasc Surg
57:225, 1969[Medline]
[Order article via Infotrieve]
20.
Rosenberg A:
Skin allograft rejection, in
Coligan J,
Kruisbeek A,
Marguiles D,
Shevach E,
Strobe W
(eds):
Current Protocols in Immunology, vol 1. Bethesda, MD, Wiley, 1991, p 441.
21.
McKenzie J, Beard M, Hart D:
The effect of donor pretreatment on interstitial dendritic cell content and rat cardiac allograft survival.
Transplantation
38:371, 1984[Medline]
[Order article via Infotrieve]
22.
Josien R, Heslan M, Soulillou JP, Cuturi MC:
Rat spleen dendritic cells express natural killer cell receptor protein 1 (NKR-P1) and have cytotoxic activity to select targets via a Ca2+-dependent mechanism.
J Exp Med
186:467, 1997[Abstract/Free Full Text]
23.
Talmor M, Mirza A, Turley S, Mellman I, Hoffman LA, Steinman RM:
Generation or large numbers of immature and mature dendritic cells from rat bone marrow cultures.
Eur J Immunol
28:811, 1998[Medline]
[Order article via Infotrieve]
24.
Cuturi M, Josien R, Cantarovich D, Douillard P, Smit H, Menoret S, Pouletty P, Clayberger C, Soulillou J:
Prolongation of allogeneic heart graft survival in rats by administration of a peptide (a.a. 75-84) from the  1 helix of the first domain of HLA-B7 01.
Transplantation
59:661, 1995[Medline]
[Order article via Infotrieve]
25.
Quigley R, Wood K, Morris P:
Investigation of the mechanisms of active enhancement of renal allograft survival by blood transfusion.
Immunology
63:373, 1988[Medline]
[Order article via Infotrieve]
26. Fuchs E, Matzinger P: B cells turn off virgin but not memory T
cells. Science 258, 1992
27.
Matzinger P:
Tolerance, danger, and the extended family.
Annu Rev Immunol
12:991, 1994[Medline]
[Order article via Infotrieve]
28.
Niimi M, Roelen DL, Wong W, Hara M, Morris PJ, Wood KJ:
Resting B cells as tolerogens in vivo but only for minor histocompatibility antigens: Evidence for activation of resting B cells in vivo.
Transplantation
64:1330, 1997[Medline]
[Order article via Infotrieve]
29.
Mason D, Pugh C, Webb M:
The rat mixed lymphocyte reaction: Roles of a dendritic cell in intestinal lymph and T-cell subsets defined by monoclonal antibodies.
Immunology
44:75, 1981[Medline]
[Order article via Infotrieve]
29a. Josien R, Douillard P, Guillot C, Müschen M, Anegon I,
Chetritt J, Menoret S, Vignes C, Soulillou J-P, Cuturi, M-C: A critical
role for transforming growth factor- (TGF-) in
donor-transfusion-induced allograft tolerance. J Clin Invest (in press)
30.
Forsthuber T, Hualin C, Lehmann P:
Induction of Th1 and Th2 immunity in neonatal mice.
Science
271:1728, 1996[Abstract]
31.
McNeilage L, Heslop B:
Lymphocyte homing in syngeneic and unsensitized MHC compatible allogeneic hosts. I. Evidence for both syngeneic self-recognition and early killing allogeneic cells.
Cell Immunol
50:58, 1980[Medline]
[Order article via Infotrieve]
32.
Larsen C, Morris P, Austyn J:
Migration of dendritic leucocytes from cardiac allgrafts into host spleen. A novel pathway for initiation of rejection.
J Exp Med
171:307, 1990[Abstract/Free Full Text]
33.
Shelby J, Wakely E, Corry R:
Splenectomy abrogates the improved graft survival achieved by donor-specific transfusion.
Transplant Proc
17:1083, 1985
34.
Shoskes DA, Wood KJ:
Indirect presentation of MHC antigens in transplantation.
Immunol Today
15:32, 1994[Medline]
[Order article via Infotrieve]
35.
Roake JA:
Dendritic cells and the initiation of the immune response to organ transplants.
Transplant Rev
8:37, 1994
36.
Faustman D, Steinman R, Gebel H, Hauptfeld V, Davie J, Lacy P:
Prevention of rejection of murine islet allografts by pretreatment with anti-dendritic cell antibody.
Proc Natl Acad Sci USA
81:3864, 1984[Abstract/Free Full Text]
37.
Stegall M, Tezuka K, Oluwole S, Engelstad K, Jjing M, Andrew J, Hardy M:
Interstitial class II-positive cell depletion by donor pretreatment with gamma irradiation. Evidence of differential immunogenicity.
Transplantation
49:246, 1990[Medline]
[Order article via Infotrieve]
38.
Talmage D, Dart G, Radovich J, Lafferty K:
Activation of transplant immunity: Effect of donor leukocytes on thyroid allograft rejection.
Science
191:385, 1976[Abstract/Free Full Text]
39.
Spencer S, Fabre J:
Characterization of the tissue macrophage and the interstitial dendritic cells as distinct leucocytes normally resident in the connective tissue of rat heart.
J Exp Med
171:1841, 1990[Abstract/Free Full Text]
40.
Hart D, Fabre J:
Demonstration and characterization of Ia positive dendritic cells in the interstitial connnective tissues of rat heart and other tissues, but not brain.
J Exp Med
154:347, 1981[Abstract/Free Full Text]
41.
Lechler RI, Batchelor RJ:
Restoration of immunogenicity to passenger cell-depleted kidney allografts by the addition of donor strain dendritic cells.
J Exp Med
155:31, 1982[Abstract/Free Full Text]
42.
Heineman E, Bouwman E, de Bruin R, Marquet R, Jeekel J:
The role of passenger leukocytes in the manifestation of the donor-specific transfusion phenomenon.
Transplant Proc
17:821, 1985
43.
Bagley C, Bostick F, Devita V Jr:
Clinical pharmacology of cyclophosphamide.
Cancer Res
33:226, 1973[Abstract/Free Full Text]
44.
Braun M, McCormack A, Webb G, Batchelotr J:
Evidence for clonal anergy as a mechanism responsible for the maintenance of transplantation tolerance.
Eur J Immunol
23:1462, 1993[Medline]
[Order article via Infotrieve]
45.
Kupiec-Weglinski JW, Austyn JM, Morris PJ:
Migration patterns of dendritic cells in the mouse. Traffic from the blood, and T cell-dependent and -independent entry to lymphoid tissues.
J Exp Med
167:632, 1988[Abstract/Free Full Text]
46.
Hart D, Fabre J:
Mechanism of induction of passive enhancement. Evidence for an interaction of enhancing antibody with donor interstitial dendritic cells.
Transplantation
33:319, 1982[Medline]
[Order article via Infotrieve]
47.
Cuturi M, Josien R, Cantarovich D, Bugeon L, Anegon I, Menoret S, Smit H, Douillard P, Soulillou JP:
Decreased anti-donor major histocompatibility complex class I and increased class II alloantibody response in allograft tolerance in adult rats.
Eur J Immunol
24:1627, 1994[Medline]
[Order article via Infotrieve]
48.
Steinman RM:
The dendritic cell system and its role in immunogenicity.
Annu Rev Immunol
9:271, 1991[Medline]
[Order article via Infotrieve]
49.
Heath WR, Kurts C, Miller J, Carbone FR:
Cross-tolerance: A pathway for inducing tolerance to peripheral tissue antigens.
J Exp Med
187:1549, 1998[Free Full Text]
50.
Süss G, Shortman K:
A subclass of dendritic cells kill CD4 T cells via Fas/Fas-Ligand-induced apoptosis.
J Exp Med
183:1789, 1996[Abstract/Free Full Text]
51.
Thomson A, Lu L, Murase N, Demetris A, Rao AS, Starzl T:
Microchimerism, dendritic cell progenitors and transplantation tolerance.
Stem Cells
13:622, 1995[Abstract]
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Abstract
Introduction