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Blood, Vol. 92 No. 12 (December 15), 1998:
pp. 4539-4544
RAPID COMMUNICATION
By
From the INSERM Unit 437 and ITERT "Institut de Transplantation et
de Recherche en Transplantation," Immeuble Jean Monnet, Nantes,
France.
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.
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- 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.
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
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 Statistical analysis.
Graft survivals were compared using the Kaplan Meier method.
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).
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
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.
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.
The authors thank Dr Ralph Steinman and Brian Wong for critically
reading the manuscript.
Submitted August 20, 1998;
accepted October 5, 1998.
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.
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
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-
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
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
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
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-
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
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
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
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
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
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
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
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
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
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
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]
This article has been cited by other articles:
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Top
Abstract
Introduction
(IFN-
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.
(TGF-
) 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.
1 helix of the first domain of HLA-B7 01.
Transplantation
59:661, 1995
