|
|
 Previous Article | Table of Contents | Next Article 
Blood, Vol. 94 No. 4 (August 15), 1999:
pp. 1201-1208
Complete Remission of Accelerated Phase Chronic Myeloid Leukemia by
Treatment With Leukemia-Reactive Cytotoxic T Lymphocytes
By
J.H. Frederik Falkenburg,
Amon R. Wafelman,
Peter Joosten,
Willem
M. Smit,
Cornelis A.M. van Bergen,
Rian Bongaerts,
Ellie Lurvink,
Menno van der Hoorn,
Petra Kluck,
James E. Landegent,
Hanneke C. Kluin-Nelemans,
Willem E. Fibbe, and
Roel Willemze
From the Departments of Hematology and Pharmacy, Leiden University
Medical Center, Leiden, The Netherlands; and the Department of Internal
Medicine, Medical Center Leeuwarden, Leeuwarden, The Netherlands.
 |
ABSTRACT |
Relapse of chronic myeloid leukemia (CML) in chronic phase after
allogeneic stem cell transplantation (SCT) can be successfully treated
by donor lymphocyte infusion (DLI). However, relapse of accelerated
phase CML, blast crisis, or acute leukemia after allogeneic SCT are
resistant to DLI in the majority of cases. In vitro-selected and
expanded leukemia-reactive T-cell lines may be more effective in
inducing an antileukemic response in vivo. To treat a patient with
accelerated phase CML after allogeneic SCT, leukemia-reactive cytotoxic
T-lymphocyte (CTL) lines were generated from her HLA-identical donor.
Using a modification of a limiting dilution assay, T cells were
isolated from the donor, selected based on their ability to inhibit the
in vitro growth of CML progenitor cells, and subsequently expanded in
vitro to generate CTL lines. Three CTL lines were generated that lysed
the leukemic cells from the patient and inhibited the growth of
leukemic progenitor cells. The CTL did not react with lymphocytes from
donor or recipient and did not affect donor hematopoietic progenitor
cells. The 3 leukemia-reactive CTL lines were infused at 5-week
intervals at a cumulative dose of 3.2 × 109 CTL. Shortly
after the third infusion, complete eradication of the leukemic cells
was observed, as shown by cytogenetic analysis, fluorescence in situ
hybridization, molecular analysis of BCR/ABL-mRNA, and
chimerism studies. These results show that in vitro cultured leukemia-reactive CTL lines selected on their ability to inhibit the
proliferation of leukemic progenitor cells in vitro can be successfully
applied to treat accelerated phase CML after allogeneic SCT.
© 1999 by The American Society of Hematology.
| |
INTRODUCTION |
ALLOGENEIC STEM CELL transplantation
(SCT) has been successfully applied in the treatment of hematological
malignancies, including chronic myeloid leukemia (CML).1
Depletion from an allogeneic stem cell graft of mature T lymphocytes,
which mediate graft-versus-host disease (GVHD), results in an increased
incidence of recurrence of leukemia after transplantation, especially
for CML.2,3 Thus, the antileukemic effect of allogeneic SCT
is not merely due to the cytoreductive treatment during the
conditioning regimen before transplantation, but is also due to a
graft-versus-leukemia (GVL) effect mediated by donor T
cells.4,5 Patients with recurrence of leukemia after
transplantation have been treated with donor lymphocyte infusions (DLI)
from their stem cell donor. In patients with relapsed CML in chronic
phase, 70% to 80% complete remissions have been reported after
DLI.6-13 In CML accelerated phase, blast crisis, acute
myeloid leukemia, and acute lymphoblastic leukemia, remission rates of
only 10% to 30% have been reported.11-13 Despite a
correlation between the occurrence of acute GVHD following DLI and the
remission rate, disappearance of the leukemic cells can be observed in
the absence of GVHD, suggesting that GVL and GVHD reactivity are not
always mediated by the same effector cells.
The induction of tolerance to donor cells is the immunological basis
for the successful treatment of relapsed leukemia after allogeneic SCT
with DLI. Donor T cells recognizing the recipient-derived leukemic
cells only and alloreactive donor T cells recognizing polymorphic
antigens on all hematopoietic cells from the recipient may result in
eradication of the leukemic cells without affecting donor hematopoiesis
in the patient.14
Previously, we have shown that primary cytotoxic T lymphocyte (CTL)
responses can be generated from donor lymphocytes against the leukemic
cells from the patient in HLA-identical donor-recipient pairs.15,16 Using a newly developed assay, the progenitor
cell inhibitor lymphocyte precursor (PCILp) frequency assay, we
recently showed that T cells recognizing CML precursor cells may be
responsible for the clinical response to DLI.17 We
hypothesized that, if T-cell lines recognizing the CML precursor cells
could be generated in vitro from peripheral blood of the stem cell
donor, these leukemia-reactive CTL lines may be capable of eradicating
the leukemic cells from the patient in vivo with a limited risk of GVHD.
In this report, we describe a patient with relapsed accelerated phase
CML after allogeneic stem cell transplantation who was successfully
treated with donor-derived leukemia-reactive CTL recognizing CML
precursor cells.
| |
MATERIALS AND METHODS |
Case history.
Patient KG, a 43-year-old white female, was diagnosed as
Ph+ CML in chronic phase in September 1991. Because no
HLA-identical sibling was available, treatment was started with
hydroxyurea and interferon- -2b at 3 × 106 IU/d for
5 days per week.18 In 1992 and 1993, a hematological response, but no cytogenetic response was observed. In 1993, cytogenetic analysis showed 46XX, t(9;22), 7q , 10, 12p+,
13, +2 marker chromosomes in 50% of the metaphases. Extended
family typing showed an HLA-identical niece (HS) as a potential
donor.19 The HLA-type of both donor and recipient was A1,
A2, B7, B8, Cw7, DR15, DR17, DQ6, DQ2. Mixed lymphocyte cultures were
negative in both directions. In 1994, allogeneic bone marrow
transplantation was performed. The conditioning regimen consisted of
Campath-1G intravenously (5 mg on days 8 through 3),
cyclophosphamide at 60 mg/kg of body weight on days 6 and
5, and total body irradiation (6 Gy) on days 1 and 0. T-cell depletion of the graft was performed by incubation with
Campath-1G (10 mg), as described.20 GVHD prophylaxis
consisted of cyclosporin at 3 mg/kg/d. Normal engraftment was observed.
At day 17, acute GVHD grade II of the skin developed that was treated
with methylprednisolone. Cyclosporin toxicity included hypertension,
fluid retention, and renal failure. Subsequent chronic GVHD of the skin
required treatment with low-dose cyclosporin.
In 1995, a cytogenetic relapse was found with 46XX, t(2;20)(p12;q13),
t(9;22)(q34;q11), t(8;15)(p10;q10) in all of 30 metaphases analyzed,
followed by a hematological relapse. Platelet counts exceeded 1,000 × 109/L and required treatment with increasing doses
of hydroxyurea. Discontinuation of the cyclosporin treatment did not
result in a hematological or cytogenetic response. In November 1995, interferon--2b (3 × 106 IU/d) was initiated, but
no improvement was observed. Cytogenetic analysis showed 46XX,
t(2;20)(p12;q13), inv(2)(p12;q37), t(8;15)(p10;q10), t(9;22)(q34;q11)
in all metaphases analyzed. Basophil counts were 5% to 10%. In April
1996, hydroxyurea treatment was discontinued and she received a DLI
from her donor at a dose of 0.6 × 107 lymphocytes/kg
of body weight. Eight weeks later, she developed acute GVHD of the skin
that was treated by a short course of methylprednisolone (30 mg/d),
whereas interferon treatment was stopped. Bone marrow cytogenetics
showed 20% normal metaphases of 30 metaphases tested. In August 1996, 16 weeks after DLI, moderately severe chronic GVHD of the skin required
treatment with cyclosporin and continuous treatment with low-dose
corticosteroids. Karyotypic analysis of the bone marrow showed again
100% abnormal metaphases. Thrombocytosis (1,600 × 109/L) necessitated reinstallment of hydroxyurea therapy.
Because reversal of the cytogenetic response was observed in the
presence of chronic GVHD, dose-escalation of DLI was not applied.
In February, March, and April 1997, she was treated with 3 leukemia-reactive CTL lines at 5-week intervals at the doses of 0.2 × 109, 1.3 × 109, and 1.7 × 109 CTL, respectively. Side effects included chills 2 hours
after the infusion and fever that resolved in 6 hours. In May 1997, she
developed severe pancytopenia and pneumococcal pneumonia. While the
chronic skin GVHD persisted, transient circumscript lesions were
observed on her lower legs. Skin biopsies were compatible with erythema
exsudativa multiforme. No acute GVHD was observed. A bone marrow biopsy
showed almost complete aplasia, with scattered small foci of myeloid
cells. Subsequently, a gradual increase in leukocytes and platelets was
observed, resulting in normal white blood cell counts at day 105 after
the first CTL infusion, with platelet counts of 30 × 109/L. Chimerism studies showed complete donor
hematopoiesis. At days 140 and 200, bone marrow and peripheral blood
cytogenetic analysis showed the absence of the Ph chromosome, which was
confirmed by fluorescence in situ hybridization (FISH) on interphase
cells using BCR- and ABL-specific probes and absence of
BCR/ABL mRNA transcripts using reverse transcriptase-polymerase
chain reaction (RT-PCR). In October 1997, she was treated
again for the persistant chronic GVHD of the skin with cyclosporin and
corticosteroids. This resulted in a hypertensive crisis with renal
failure, liver dysfunction, and signs of capillary leakage. After
discontinuation of the cyclosporin, the bilirubin normalized and the
oedema disappeared, but the renal insufficiency persisted with a
creatinin clearance of 10 mL/min. One year after the treatment, she
additionally developed angina pectoris. Two years after the treatment,
the patient died of ischemic heart disease, being still in complete
remission from the leukemia.
Patient and donor materials.
After informed consent was obtained, bone marrow and peripheral blood
(PBL) were obtained from patient KG and her donor HS. After Ficoll
Isopaque density cell separation, mononuclear cells (MNC) were
harvested from the interphase and polymorphic mononuclear cells (PMN)
were recovered from the pellet. MNC were cryopreserved under good
manufacturing practice (GMP) conditions in Isove's modified
Dulbecco's medium (IMDM; Biowhittaker, Verviers, Belgium) with 20 g/L
clinical grade human albumin (Central Laboratory of the Blood
Transfusion Service [CLB], Amsterdam, The Netherlands) and 10%
dimethyl sulphoxide (DMSO). Thawed cells were washed twice and
resuspended in IMDM containing 10% heat-inactivated heparin plasma
from the bone marrow donor.
CML PCILp assay.17
Precursor cell frequency analysis of T lymphocytes recognizing the CML
precursor cells was performed on PBL from the donor or the patient
before and after treatment. MNC were plated into 96-well U-bottom
plates using a pipetting robot (Biomek; Beckman, Mijdrecht, The
Netherlands) at 2-fold dilutions from 40,000 cells/well down to 625 cells/well in 24 replicates per concentration. Each well was stimulated
with 20,000 irradiated (25 Gy) leukemic cells from the patient obtained
before transplantation or at relapse when 100% of the metaphases were
of CML origin. Twenty-four wells contained irradiated stimulator cells
only and were used as a reference. After 6 days of culture, 120 IU/mL
of interleukin-2 (IL-2; Chiron, Amsterdam, The Netherlands) was added,
and at day 9 all wells were restimulated with 10,000 irradiated
leukemic MNC (CML-MNC) from the patient. Twice weekly, half of the
medium was refreshed. After 21 days of culture, cells from each
individual well were irradiated (15 Gy) and cocultured with 10,000 patient CML cells/well in medium consisting of IMDM with 15% donor
plasma, 50 ng/mL stem cell factor (SCF; Amgen, Thousand Oaks, CA), 25 ng/mL IL-3 (Sandoz, Basel, Switzerland), 100 ng/mL
granulocyte-macrophage colony-stimulating factor (GM-CSF; Sandoz), 100 ng/mL granulocyte colony-stimulating factor (G-CSF; Amgen), 2 U/mL
erythropoietin (EPO; Cilag Ag, Zug, Switzerland), 0.47 g/L human
transferrin, and 5 × 10 5 mol/L
 -mercaptoethanol. After 7 days of culture, 1 µCi
3H-thymidine deoxyribose was added to each well; after
another 6 hours of culture, 3H-thymidine incorporation was
measured. Previous studies have shown that proliferation of the cells
at day 7 reflected the progenitor cell growth of CD34+ CML
cells.17 Inhibition of CML progenitor cell proliferation was calculated in comparison to the reference wells. An individual well
was scored positive if the 3H-thymidine incorporation in
that well was lower than the mean minus 3× SD of the reference
wells. The percentage of growth inhibition of each well was determined
by the following formula: (1 [experimental CPM/mean CPM of
reference wells]) × 100%. PCILp frequencies were calculated
using a statistical computer program as described
previously.21
Generation of leukemia-reactive CTL lines.
Using the computer-controlled pipetting robot in a biosafety cabinet
placed in a clean room facility, CTL lines were generated under GMP
conditions. MNC from donor HS were thawed, washed, resuspended in
culture medium, and plated at a concentration of 20,000 cell/well in 10 round-bottomed 96-well microtiter plates. To each well, 20,000 irradiated (25 Gy) CML-MNC derived from the patient at relapse when
100% of the metaphases were leukemic were added. The cells were
cultured in a sterile incubator at 37°C, 95% humidity, 5%
CO2. After 6 days, IL-2 was added (120 IU/mL), and at day
9, each well was restimulated with 10,000 irradiated CML-MNC from the
patient and the cells were expanded for 10 to 12 days. Growing wells
were split using the pipetting robot. From each well, one fourth was
used to analyze its potential to inhibit the CML progenitor cell growth
as described in the PCILp assay, and the remaining three fourths of the
cells from the wells were further expanded. From the positive wells
containing PCIL, the cells were collected and pooled to a T-cell line.
Using a 51Cr release assay, the T-cell line was analyzed
for its capacity to lyse phytohemagglutinin-stimulated T cells
(PHA-blasts) from the donor, leukemic cells from the patient, PHA
blasts from the patient that were derived from her peripheral blood
before transplantation, and stromal cells derived from bone marrow from
the patient before the transplantation. Stromal cells were generated by
culturing bone marrow MNC in the presence of 10% human serum. Two days
before harvesting and testing, the cells were incubated with 300 U/mL of interferon- to upregulate major histocompatibility complex (MHC) expression. To determine whether CD4+ or
CD8+ T cells were the main effector cells responsible for
the cytolytic activity, blocking experiments were performed using
antibodies against CD4, CD8, HLA-class I, or HLA-class II, as described
previously.15 In addition, the CTL line was analyzed for
its capacity to suppress the clonogenic erythroid (burst-forming
unit-erythroid [BFU-E]) or myeloid (colony-forming
unit-granulocyte-macrophage [CFU-GM]) precursor cell
growth from the normal bone marrow from the donor or the CML bone
marrow cells from the patient in a cell-mediated inhibition assay of
hematopoietic progenitor cells in semisolid medium cultures as
described previously.22,23
Molecular analysis of a clinical response.
Conventional cytogenetic analysis was performed on metaphases and FISH
was performed to detect the Ph chromosome on interphase nuclei using
BCR- and ABL-specific probes.24 The
presence of BCR/ABL mRNA transcripts was determined in PBL and
bone marrow MNC from the patient by RT-PCR.24 Chimerism
studies of MNC and/or PMN was performed after isolation of DNA from the
cell fractions, followed by PCR using as primers oligonucleotides
flanking the polymorphic regions of AFP and NGF,25 which
were informative for cells derived from the donor HS and patient KG, respectively.
| |
RESULTS |
DLI.
As determined by AFP polymorphism, before DLI, 5% of bone marrow MNC
and 20% of PBL MNC were of donor origin, whereas all PMN were of
recipient origin (Fig 1). Eight weeks after
DLI, when GVHD was present, donor chimerism increased to approximately
20% in bone marrow MNC, but no significant increase in donor-derived PMN was observed. Sixteen weeks after DLI, these chimerism parameters had returned to pre-DLI values, as shown in Fig 1. The transient increase in donor MNC coincided with a transient increase of normal donor metaphases in the bone marrow to 20%.
View larger version (38K):
[in this window]
[in a new window]
| Fig 1.
Chimerism analysis in bone marrow (BM) or peripheral
blood (PBL) from the patient after treatment with donor lymphocyte
infusion (DLI). Cells of donor (HS) origin could be detected by AFP
polymorphism (heterozygous 145 and 72 bp fragments). Temporary increase
of donor cells was observed in MNC in BM at 8 weeks after DLI, but
returned to pre-treatment levels at 16 weeks after DLI. PMN from the
patient remained of patient origin.
|
|
Generation of CML reactive T-cell lines from donor HS.
The PCILp frequency in PBL MNC from donor HS directed against CML cells
from the patient was 26 per 106 MNC. From these data, we
calculated that, if 20,000 donor MNC were plated per well, clonal
expansion of T cells capable of inhibiting the CML precursor cells
would be present in 40% to 50% of the wells. Three leukemia-reactive
CTL lines were generated with 5-week intervals. The T-cell lines showed
no bacterial or fungal contamination. For each CTL line, in a total of
960 wells, 20,000 donor MNC per well were stimulated with CML-MNC from
the patient, of which 30% expressed CD34. Each individual positive
well showing growth inhibition of 50% to 95% of the CML precursor
cells in the PCILp assay was harvested from the microtiter plates. The
numbers of positive wells harvested per line, the phenotypic analysis,
and cytotoxic activity against CML-MNC and PHA blast or bone marrow
stromal cells from patient KG as well as PHA blasts from donor HS are shown in Table 1. All CTL lines consisted
of CD3+ TCR T cells, with the majority of cells being
CD4+. The CTL lines did not react with PHA blasts of
patient origin and showed weak recognition of stromal cells derived
from the bone marrow of the patient, showing a relative specificity for the patient leukemic cells. As shown in Fig
2, the cytolytic activity against the leukemic cells could be inhibited
by the addition of anti-CD4 but not anti-CD8 antibodies to the effector
cells or by the addition of anti-HLA-DR, but not anti-HLA class I
antibodies. Control experiments using an anti-HLA-A2 specific CTL
clone showed that the leukemic cells were susceptable to HLA class
I-restricted recognizion. The CTL lines showed specific inhibition of
the CML hematopoietic progenitor cells from the patient, but did not
affect normal donor hematopoietic progenitor cells, as shown in
Fig 3.
View larger version (28K):
[in this window]
[in a new window]
| Fig 2.
Recognition of leukemic cells by leukemia-reactive CTL or
an anti-HLA-A2 CD8+ control CTL clone. CTL were
preincubated with anti-CD4 or anti-CD8, or leukemic target cells were
preincubated with anti-HLA class I or class II monoclonal antibodies
for 30 minutes. The percentage of lysis was then measured in a
51Cr-release assay at an effector:target ratio of 3:1.
Recognition of leukemic cells by the leukemia-reactive CTL was blocked
by anti-CD4 or anti-class II antibodies.
|
|
View larger version (31K):
[in this window]
[in a new window]
| Fig 3.
Suppression of leukemic but not normal donor BFU-E and
CFU-GM growth by leukemia-reactive CTL line 3. CTL line 3 was
irradiated (10 Gy) and incubated for 4 hours with bone marrow MNC from
donor HS or patient KG at various effector:target ratios. After 4 hours, the cells were resuspended in methylcellulose as a single-cell
suspension and cultured in the presence of multiple hematopoietic
growth factors. Progenitor cell growth is expressed as the percentage
of BFU-E or CFU-GM growth in the absence of CTL. All metaphases from
MNC of patient KG were t(9;22) positive.
|
|
Treatment of patient KG with leukemia-reactive CTL lines.
The patient was treated at days 0, 35, and 70 with CTL lines 1, 2, and
3, respectively. No clinical response was observed before infusion of
CTL line 3 (day 70). A rapid hematologic response was observed between
day 70 and day 90 (Fig 4A).
Leukemia-reactive PCILp frequencies in peripheral blood of the patient
measured before and 1 or 2 days after the infusion of the CTL lines
showed a minor increase after the first infusion and a high increase of
after the infusion of CTL line 3 (Fig 4B). Chimerism studies of bone
marrow and PBL MNC and PMN as well as the detection of BCR/ABL
transcripts in bone marrow MNC are shown in
Fig 5. Disappearance of the
patient-specific polymorphic band was observed from the MNC fraction
between days 70 and 105, as shown by NGF- polymorphism (Fig 5A). In
addition, no patient-derived PMN could be determined after day 90 in
peripheral blood or bone marrow from the patient (Fig 5B). From day
105, all MNC and PMN from blood and bone marrow from the patient
were found to be of donor origin. As shown in Fig 5C, BCR/ABL
transcripts could not be detected from day 140 after the infusion of
CTL line 1 (detection limit, 1 per 104 cells). Similarly,
at days 140 and 200, the Ph chromosome could no longer be detected by
FISH and the karyotype normalized. These results illustrate the
complete disappearance of leukemic cells and the reconstitution of full
donor chimerism after treatment with the leukemia-reactive CTL lines.
View larger version (27K):
[in this window]
[in a new window]
| Fig 4.
Platelets, white blood cells (WBC), and PCILp before and
after treatment with leukemia-reactive CTL lines 1, 2, and 3. (A)
Between days 80 and 90, 10 to 20 days after the infusion of CTL line 3, a rapid decrease of ( ) platelets and ( ) WBC was
observed, followed by a gradual recovery. (B) A strong increase of
PCILp frequencies in PBL from the patient was observed between days 70 and 90.
|
|
View larger version (49K):
[in this window]
[in a new window]
| Fig 5.
Chimerism analysis and BCR/ABL minimal residual
disease detection in bone marrow (BM) or peripheral blood (PBL) from
the patient after treatment with leukemia-reactive CTL. Cells of donor
(HS) origin could be detected by AFP polymorphism (heterozygous 145- and 72-bp fragments); cells of patient (KG) origin could be detected by
NGF- polymorphism (heterozygous for 100- and 50-bp fragments). (A)
Between days 70 and 105 after the first CTL infusion, chimerism of MNC
in BM and PBL completely converted to donor origin. (B) PMN from the
patient were completely of donor origin from day 105 (5 weeks after the
last CTL infusion). (C) Complete molecular remission was documented at
day 140, 10 weeks after the last CTL infusion by disappearance of the
patient b2a2 BCR/ABL transcript. C1, control b3a2 cDNA; C2,
control b2a2 cDNA; N, negative control. RT-PCR of the HPRT gene served
as an internal control for cDNA synthesis.
|
|
| |
DISCUSSION |
We report here the first successful treatment with leukemia-reactive
CTL lines showing a relative specificity for the leukemic cells in a
patient with relapsed accelerated phase CML after allogeneic SCT. This
patient had been treated with low-dose DLI 10 months before treatment
with the leukemia-reactive CTL lines. After DLI, a temporary reduction
of the leukemic cells was observed, but she developed acute GVHD
followed by chronic GVHD of the skin, limiting further dose escalation
of DLI. Several reports have shown that relapsed CML in chronic phase
responds more frequently to DLI than CML in accelerated phase, blast
crisis, or acute leukemia.6-13 One hypothetical explanation
for this observation is that CML is a malignancy of hematopoietic
precursor cells giving rise in chronic phase to malignant professional
antigen-presenting cells such as dendritic cells that are capable of
inducing strong T-cell responses.24,26,27 In contrast, the
malignant cells present in accelerated phase CML, in blast crisis, or
in acute leukemia may be more inappropriate antigen-presenting cells,
which may lead to the induction of anergy rather than to a specific
antileukemic T-cell response in vivo. We hypothesized that in vitro
generation of leukemia-reactive CTL lines may bypass the induction
phase of the antileukemic immune response in vivo.
In vitro cultured T-cell lines or clones that recognize viral antigens
can be effective in suppressing Epstein-Barr virus (EBV)-associated lymphoproliferative disorders or
cytomegalovirus (CMV) disease after allogeneic SCT without
significant GVHD.28-32 Previously, we have shown that
leukemia-reactive CTL lines can be generated with relative specificity
for the leukemic cells.15,16 We have demonstrated that the
generation of such leukemia-reactive CTL lines is more efficient using
a modified limiting dilution assay.33 By pooling the T
cells from individual wells that were inhibitory to CML precursor
cells, CTL lines were generated showing cytolytic activity and
growth-inhibitory activity of CML precursor cells from patient KG, but
not of normal hematopoietic progenitor cells from the donor. In
addition, no reactivity was found with donor or recipient
PHA-stimulated lymphoblasts, whereas stromal cells derived from the
leukemic bone marrow before transplantation were only weakly
recognized, illustrating a relative specificity for the leukemic cells.
Despite this relative specificity of the CTL lines for the leukemic
cells, the antigen specificity of these CTL lines is unknown. Based on
the relatively high PCILp frequency in donor peripheral blood, it is
unlikely that these CTL recognized a BCR/ABL-specific
peptide.34,35 Because many non-HLA molecules may be
different between such HLA-matched, but not genotypically identical
individuals, it is more likely that the majority of the CTL were
directed against minor histocompatibility antigens (mHAg), with, at
least in part, a restricted tissue distribution. Recently, several of
such mHAg antigens restricted to the hematopoietic lineages have been
characterized that could be recognized in the context of HLA class I
molecules.36-38
Previously, we have shown that both CD4+ and
CD8+ mHAg-specific CTL are capable of strong antigen
specific growth inhibition of leukemic precursor
cells.22,23,39,40 As shown by these specific inhibition of
the recognition of leukemic cells by antibodies against CD4 and HLA
class II, we demonstrated that the CD4+ cells, which were
the majority of the cells present in the CTL lines, were responsible
for most of the cytolytic activity. This does not exclude reactivity by
the CD8+ cells as well, but these cells were present in the
CTL lines at a frequency too low to allow sufficient inhibition by
anti-CD8 and anti-class I antibodies. CD8+ T cells may
require the presence of helper CD4+ T cells to exhibit
their clinical effect. For an optimal effect, a direct cytolytic effect
of the effector cells infused is probably not sufficient. It is likely
that the infused cells have to proliferate and expand in vivo and
survive for several weeks to months to exhibit their full clinical
effect. Based on these previous findings, we hypothesized that
leukemia-reactive CTL lines composed of both CD4+ and
CD8+ T cells may be most efficient in exhibiting an
antileukemic effect. The strong clinical response to the CTL lines was
observed after infusion of the third CTL line. A total of 3.2 × 109 T cells was infused into the patient. We estimated the
leukemic burden of the patient in bone marrow, spleen, and blood to be approximately 1 to 3 × 1012 cells. This would result
in an initial effector target ratio of 1:1,000. Because the CML cells
will be dividing in vivo, it is unlikely that, without in vivo
expansion of the T cells, this treatment would have been effective. The presence of both CD4+ and CD8+ T cells in the
lines have probably supported the in vivo proliferation.
Treatment of the accelerated phase CML with the leukemia-reactive CTL
lines was effective in our patient. However, we cannot exclude that
these cells have contributed to the persistence of the chronic GVHD
that developed after DLI and was still present after the CTL treatment.
We observed an increase in the skin GVHD around day 200 after the first
CTL line infusion. This chronic GVHD was treated again with cyclosporin
and prednisone. The immunosuppressive treatment 4 months after the
clinical response did not result in a relapse of the leukemia,
suggesting a sustained effect of the treatment of leukemia-reactive CTL.
In conclusion, treatment of relapsed chronic phase CML after allogeneic
stem cell transplantation with DLI appears to be successful in most of
the patients, without lethal GVHD in the majority of cases. Our results
show that leukemia after allogeneic SCT can respond to treatment with
leukemia-reactive CTL lines generated in vitro. These CTL lines may be
more specific to the leukemic cells than to the GVHD target organs,
leading to a relative specific antileukemic effect. However, because
the treatment with leukemia-reactive CTL requires the presence of
significant quantities of (cryopreserved) leukemic cells to be used as
stimulator cells and a relatively complex infrastructure to generate
such CTL lines under GMP conditions, the general applicability of this
treatment may be limited at present. Future characterization of the
target peptides that can be recognized by CTL on leukemic cells is
essential. Adoptive immunotherapy of relapsed leukemia after allogeneic
transplantation with in vitro-expanded CTL lines may significantly
contribute to the curative potential of allogeneic stem cell
transplantation for hematological malignancies.
| |
ACKNOWLEDGMENT |
The authors gratefully acknowledge Roel de Paus and Jacqueline Bergsma
for their technical assistance, Dr S.L. Bhola for cytogenetic analysis,
Dr Ronald van Soest for his help in preparing the figures, and Clary
Wiarda for preparing the manuscript.
| |
FOOTNOTES |
Submitted October 20, 1998; accepted April 14, 1999.
Supported by grants from the Dutch Cancer Society and the J.A. Cohen
Institute for Radiopathology and Radiation Protection.
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 J.H. Frederik Falkenburg, MD, PhD,
Department of Hematology, C2-R, Leiden University Medical Center, PO
Box 9600, 2300 RC Leiden, The Netherlands; e-mail:
falkenburg{at}hematology.azl.nl.
| |
REFERENCES |
1.
O'Reilly RJ:
Allogeneic bone marrow transplantation: Current status and future directions.
Blood
62:941, 1983[Free Full Text]
2.
Goldman JM, Gale RP, Horowitz MM, Biggs JC, Champlin RE, Gluckman E, Hoffmann RG, Jacobsen SJ, Marmont AM, McGlave PB, Messner HA, Rimm AA, Rozman C, Speck B, Tura S, Weiner RS, Bortin MM:
Bone marrow transplantation for chronic myelogenous leukemia in chronic phase. Increased risk for relapse associated with T-cell depletion.
Ann Intern Med
108:806, 1988
3.
Apperley JF, Mauro FR, Goldman JM, Gregory W, Arthur CK, Hows J, Arcese W, Papa G, Mandelli F, Wardle D, Gravett P, Franklin M, Bandini G, Ricci P, Tura S, Iacone A, Torlontano G, Heit W, Champlin R, Gale RP:
Bone marrow transplantation for chronic myeloid leukaemia in first chronic phase: Importance of a graft-versus-leukaemia effect.
Br J Haematol
69:239, 1988[Medline]
[Order article via Infotrieve]
4.
Horowitz MM, Gale RP, Sondel PM, Goldman JM, Kersey J, Kolb HJ, Rimm AA, Ringden O, Rozman C, Speck B, Truitt RL, Zwaan FE, Bortin MM:
Graft-versus-leukemia reactions after bone marrow transplantation.
Blood
75:555, 1990[Abstract/Free Full Text]
5.
Antin JH:
Graft-versus-leukemia: No longer an epiphenomenon.
Blood
82:2273, 1993[Free Full Text]
6.
Kolb HJ, Mittermüller J, Clemm Ch, Holler E, Ledderiose G, Brehm G, Heim M, Wilmanns W:
Donor leukocyte transfusions for treatment of recurrent chronic myelogenous leukemia in marrow transplant patients.
Blood
76:2462, 1990[Abstract/Free Full Text]
7.
Cullis JO, Jiang YZ, Schwarer AP, Hughes TP, Barrett AJ, Goldman JM:
Donor leukocyte infusions for chronic myeloid leukemia in relapse after allogeneic bone marrow transplantation.
Blood
79:1379, 1992[Free Full Text]
8.
Bär BMAM, Schattenberg A, Mensink EJBM, Geurts van Kessel A, Smetsers TFCM, Knops GHJN, Linders EHP, de Witte T:
Donor leukocyte infusions for chronic myeloid leukemia relapsed after allogeneic bone marrow transplantation.
J Clin Oncol
11:513, 1993[Abstract/Free Full Text]
9.
Porter DL, Roth MS, McGarigle C, Ferrara JLM, Antin JH:
Induction of graft-versus-host disease as immunotherapy for relapsed chronic myeloid leukemia.
N Engl J Med
330:100, 1994[Abstract/Free Full Text]
10.
van Rhee F, Lin F, Cullis JO, Spencer A, Cross NCP, Chase A, Garicochea B, Bungey J, Barrett J, Goldman JM:
Relapse of chronic myeloid leukemia after allogeneic bone marrow transplant: The case of giving donor leukocyte transfusions before the onset of hematologic relapse.
Blood
83:3377, 1994[Abstract/Free Full Text]
11.
Kolb HJ, Schattenberg A, Goldman JM, Hertenstein B, Jacobsen N, Arcese W, Ljungman P, Ferrant A, Verdonck L, Niederwieser D, van Rhee F, Mittermueller J, de Witte T, Holler E, Ansari H:
Graft-versus-leukemia effect of donor lymphocyte transfusions in marrow grafted patients.
Blood
86:2041, 1995[Abstract/Free Full Text]
12.
Mackinnon S, Papadopoulos EB, Carabasi MH, Reich L, Collins NH, Boulad F, Castro-Malaspina H, Childs BH, Gillio AP, Kernan NA, Small TN, Young JW, O'Reilly RJ:
Adoptive immunotherapy evaluating escalating doses of donor leukocytes for relapse of chronic myeloid leukemia after bone marrow transplantation: Separation of graft-versus-leukemia from graft-versus-host disease.
Blood
86:1261, 1995[Abstract/Free Full Text]
13.
Collins RH, Shpilberg O, Drobyski WR, Porter DL, Giralt S, Champlin R, Goodman SA, Wolff SN, Hu W, Verfaille C, List A, Dalton W, Ognoskie N, Chetrit A, Antin JH, Nemunaitis J:
Donor leukocyte infusions in 140 patients with relapsed malignancy after allogeneic bone marrow transplantation.
J Clin Oncol
15:433, 1997[Abstract/Free Full Text]
14.
Falkenburg JHF, Smit WM, Willemze R:
Cytotoxic T-lymphocyte (CTL) responses against acute or chronic myeloid leukemia.
Immunol Rev
157:223, 1997[Medline]
[Order article via Infotrieve]
15.
Faber LM, van Luxemburg-Heys SAP, Willemze R, Falkenburg JHF:
Generation of leukemia-reactive cytotoxic T lymphocyte clones from the HLA-identical donor of a patient with leukemia.
J Exp Med
176:1283, 1992[Abstract/Free Full Text]
16.
Falkenburg JHF, Faber LM, van den Elshout M, van Luxemburg-Heys SAP, Hooftman-den Otter A, Smit WM, Voogt PJ, Willemze R:
Generation of donor-derived antileukemic cytotoxic T lymphocyte responses for treatment of relapsed leukemia after allogeneic HLA-identical bone marrow transplantation.
J Immunother
14:305, 1993
17.
Smit WM, Rijnbeek M, Van Bergen CAM, Fibbe WE, Willemze R, Falkenburg JHF:
T cells recognizing leukemic CD34+ progenitor cells mediate the antileukemic effect of donor lymphocyte infusions for relapsed chronic myeloid leukemia after allogeneic stem cell transplantation.
Proc Natl Acad Sci USA
95:10152, 1998[Abstract/Free Full Text]
18.
Kluin-Nelemans JC, for the Benelux CML Study Group:
Randomized study on hydroxyurea alone versus hydroxyurea combined with low-dose interferon-alpha2b for chronic myeloid leukemia.
Blood
91:2713, 1998[Abstract/Free Full Text]
19.
Oudshoorn M, Lie JLWT, Schreuder GMTh, Fibbe WE, van Rood JJ, Schipper RF, Vossen JMJJ, Claas FHJ:
Extended family studies for the identification of bone marrow donors: How far?
Bone Marrow Transplant
19:S3, 1997
20.
Fibbe WE, Brouwer R, Hale G, Velders GJ, Planken EV, Visser HPJ, Posthuma W, Pruijt JFM, Falkenburg JHF, Barge R, Waldmann H, Willemze R:
Accelerated marrow reconstitution and earlier CMV infections in patients receiving T cell-depleted allogeneic blood cell grafts using "Campath-1G in the bag".
Bone Marrow Transplant
2:S14, 1998
21.
Strijbosch LWG, Buurman WA, Does RJMM, Zinken PH, Groenewegen G:
Limiting dilution assays: Experimental design and statisti cal analysis.
J Immunol Methods
97:133, 1987[Medline]
[Order article via Infotrieve]
22.
Falkenburg JHF, Goselink HM, Van der Harst D, Van Luxemburg-Heijs SA, Kooy-Winkelaar YM, Faber LM, De Kroon J, Brand A, Fibbe WE, Willemze R, Goulmy E:
Growth inhibition of clonogenic leukemic precursor cells by minor histocompatibility antigen-specific cytotoxic T lymphocytes.
J Exp Med
174:27, 1991[Abstract/Free Full Text]
23.
Faber LM, Van der Hoeven J, Goulmy E, Hooftman-den Otter AL, Van Luxemburg-Heijs SAP, Willemze R, Falkenburg JHF:
Recognition of clonogenic leukemic cells, remission bone marrow and HLA-identical donor bone marrow by CD8+ or CD4+ minor histocompatibility antigen specific cytotoxic T lymphocytes.
J Clin Invest
96:877, 1995
24.
Smit WM, Rijnbeek M, Van Bergen CAM, De Paus RA, Vervenne HAW, Van de Keur M, Willemze R, Falkenburg JHF:
Generation of dendritic cells expressing bcr/abl from CD34-positive chronic myeloid leukemia precursor cells.
Hum Immunol
53:216, 1997[Medline]
[Order article via Infotrieve]
25.
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlick HA:
Primer-directed enzymatic amplification of DNA with thermostable DNA polymerase.
Science
239:487, 1988[Abstract/Free Full Text]
26.
Choudhury A, Gajewski JL, Liang JC, Popat U, Claxton DF, Kliche KO, Andreeff M, Champlin RE:
Use of leukemic dendritic cells for the generation of antileukemic cellular cytotoxicity against Philadelphia chromosome positive chronic myelogenous leukemia.
Blood
89:1133, 1997[Abstract/Free Full Text]
27.
Eibl B, Ebner S, Duba C, Böck G, Romani N, Erdel M, Gächter A, Niederwieser D, Schuler G:
Dendritic cells generated from blood precursors of chronic myeloid leukemia patients carry the Philadelphia translocation and can induce a CML-specific primary cytotoxic T-cell response.
Genes Chromosomes Cancer
20:215, 1997[Medline]
[Order article via Infotrieve]
28.
Heslop E, Brenner MK, Rooney CM:
Donor T cells as therapy for EBV lymphoproliferation post bone marrow transplant.
N Engl J Med
331:679, 1994[Free Full Text]
29.
Rooney CM, Smith CA, Ng CYC, Loftin S, Li C, Krance RA, Brenner MK, Heslop HE:
Use of gene-modified virus-specific T lymphocytes to control Epstein-Barr-virus-related lymphoproli feration.
Lancet
345:9, 1995[Medline]
[Order article via Infotrieve]
30.
Walter EA, Greenberg PD, Gilbert MJ, Finch RJ, Watanabe KS, Thomas ED, Riddell SR:
Reconstitution of cellular immunity against CMV in recipients of allogeneic bone marrow transplantation by adoptive transfer of T cell clones from the donor.
N Engl J Med
333:1038, 1995[Abstract/Free Full Text]
31.
Riddell SR, Elloitt M, Lewinsohn DA, Gilbert MJ, Wilson L, Manley SA, Lupton SD, Overell RW, Reynolds TC, Corey L, Greenberg PD:
T cell mediated rejection of gene-modified HIV-specific cytotoxic T lymphocytes in HIV-infected patients.
Nat Med
2:216, 1996[Medline]
[Order article via Infotrieve]
32.
Yee C, Riddell SR, Greenberg PD:
Prospects for adoptive T cell therapy.
Curr Opin Immunol
9:702, 1997[Medline]
[Order article via Infotrieve]
33.
Smit WM, Rijnbeek M, van Bergen CAM, Willemze R, Falkenburg JHF:
Generation of leukemia reactive cytotoxic T lymphocytes from HLA-identical donors of patients with chronic myeloid leukemia using modifications of a limiting dilution assay.
Bone Marrow Transplant
21:553, 1998[Medline]
[Order article via Infotrieve]
34.
Bocchia M, Korontsvit T, Xu Q, Mackinnon S, Yang SY, Sette A, Scheinberg DA:
Specific human cellular immunity to bcr-abl oncogene-derived peptides.
Blood
87:3587, 1996[Abstract/Free Full Text]
35.
ten Bosch GJA, Joosten AM, Kessler JH, Melief CJM, Leeksma OC:
Recognition of BCR-ABL positive leukemic blasts by human CD4+ T cells elicited by primary in vitro immunization with a BCR-ABL breakpoint peptide.
Blood
9:3522, 1996
36.
de Bueger M, Bakker A, van Rood JJ, van der Woude F, Goulmy E:
Tissue distribution of human minor histocompatibility antigens. Ubiquitous versus restricted tissue distribution indicates heterogeneity among human cytotoxic T-lymphocyte defined non-MHC antigens.
J Immunol
149:1788, 1992[Abstract]
37.
den Haan JMM, Sherman NE, Blokland E, Huczko E, Koning F, Drijfhout JW, Skipper J, Shabanowitz, Hunt DF, Engelhard VH, Goulmy E:
Identification of a graft versus host disease-associated human minor histocompatibility antigen.
Science
268:1476, 1995[Abstract/Free Full Text]
38.
Den Haan JMM, Meadows LM, Wang W, Pool J, Blokland E, Bishop TL, Reinhardus C, Shabanowitz J, Offringa R, Hunt DF, Engelhard VH, Goulmy E:
The human immunodominant minor histocompatibility antigen HA-1 represents a diallelic gene with a single amino acid polymorphism.
Science
279:1054, 1998[Abstract/Free Full Text]
39.
Faber LM, van Luxemburg-Heys SAP, Veenhof WFJ, Willemze R, Falkenburg JHF:
Generation of CD4+ cytotoxic T lymphocyte clones from a patient with severe graft-versus-host disease after allogeneic bone marrow transplantation: Implications for graft-versus-leukemia reactivity.
Blood
86:2821, 1995[Abstract/Free Full Text]
40.
Mutis T, Schrama E, van Luxemburg-Heijs SAP, Falkenburg JHF, Melief CJM, Goulmy E:
HLA class II restricted T-cell reactivity to a developmentally regulated antigen shared by leukemic cells and CD34+ early progenitor cells.
Blood
90:1083, 1997[Abstract/Free Full Text]
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
O. Bohana-Kashtan, S. Morisot, R. Hildreth, C. Brayton, H. I. Levitsky, and C. I. Civin
Selective Reduction of Graft-versus-Host Disease-Mediating Human T Cells by Ex Vivo Treatment with Soluble Fas Ligand
J. Immunol.,
July 1, 2009;
183(1):
696 - 705.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. S. M. Yong, K. Keyvanfar, N. Hensel, R. Eniafe, B. N. Savani, M. Berg, A. Lundqvist, S. Adams, E. M. Sloand, J. M. Goldman, et al.
Primitive quiescent CD34+ cells in chronic myeloid leukemia are targeted by in vitro expanded natural killer cells, which are functionally enhanced by bortezomib
Blood,
January 22, 2009;
113(4):
875 - 882.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Michalek, I. Kocak, V. Fait, J. Zaloudik, and R. Hajek
Detection and Long-Term In Vivo Monitoring of Individual Tumor-Specific T Cell Clones in Patients with Metastatic Melanoma
J. Immunol.,
June 1, 2007;
178(11):
6789 - 6795.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. Ciceri, C. Bonini, S. Marktel, E. Zappone, P. Servida, M. Bernardi, A. Pescarollo, A. Bondanza, J. Peccatori, S. Rossini, et al.
Antitumor effects of HSV-TK engineered donor lymphocytes after allogeneic stem-cell transplantation
Blood,
June 1, 2007;
109(11):
4698 - 4707.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
I. Jedema, P. Meij, E. Steeneveld, M. Hoogendoorn, B. A. Nijmeijer, M. van de Meent, S. A.P. van Luxemburg-Heijs, R. Willemze, and J.H. F. Falkenburg
Early Detection and Rapid Isolation of Leukemia-Reactive Donor T Cells for Adoptive Transfer Using the IFN-{gamma} Secretion Assay
Clin. Cancer Res.,
January 15, 2007;
13(2):
636 - 643.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Marijt, A. Wafelman, M. van der Hoorn, C. van Bergen, R. Bongaerts, S. van Luxemburg-Heijs, J. van den Muijsenberg, J. O. Wolbers, N. van der Werff, R. Willemze, et al.
Phase I/II feasibility study evaluating the generation of leukemia-reactive cytotoxic T lymphocyte lines for treatment of patients with relapsed leukemia after allogeneic stem cell transplantation
Haematologica,
January 1, 2007;
92(1):
72 - 80.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. H. Antin
Reduced-Intensity Stem Cell Transplantation: "...whereof a little More than a little is by much too much." King Henry IV, part 1, I, 2
Hematology,
January 1, 2007;
2007(1):
47 - 54.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. H. M. Heemskerk, R. S. Hagedoorn, M. A. W. G. van der Hoorn, L. T. van der Veken, M. Hoogeboom, M. G. D. Kester, R. Willemze, and J. H. F. Falkenburg
Efficiency of T-cell receptor expression in dual-specific T cells is controlled by the intrinsic qualities of the TCR chains within the TCR-CD3 complex
Blood,
January 1, 2007;
109(1):
235 - 243.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Greiner, L. Li, M. Ringhoffer, T. F. E. Barth, K. Giannopoulos, P. Guillaume, G. Ritter, M. Wiesneth, H. Dohner, and M. Schmitt
Identification and characterization of epitopes of the receptor for hyaluronic acid-mediated motility (RHAMM/CD168) recognized by CD8+ T cells of HLA-A2-positive patients with acute myeloid leukemia
Blood,
August 1, 2005;
106(3):
938 - 945.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Hoogendoorn, J. Olde Wolbers, W. M. Smit, M. R. Schaafsma, I. Jedema, R. M.Y. Barge, R. Willemze, and J.H. F. Falkenburg
Primary Allogeneic T-Cell Responses against Mantle Cell Lymphoma Antigen-Presenting Cells for Adoptive Immunotherapy after Stem Cell Transplantation
Clin. Cancer Res.,
July 15, 2005;
11(14):
5310 - 5318.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Kami, A. Makimoto, Y. Heike, and Y. Takaue
Reduced-intensity Hematopoietic Stem Cell Transplantation (RIST) for Solid Malignancies
Jpn. J. Clin. Oncol.,
December 1, 2004;
34(12):
707 - 716.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. S. Doubrovina, M. M. Doubrovin, S. Lee, J.-H. Shieh, G. Heller, E. Pamer, and R. J. O'Reilly
In vitro Stimulation with WT1 Peptide-Loaded Epstein-Barr Virus-Positive B Cells Elicits High Frequencies of WT1 Peptide-Specific T Cells with In vitro and In vivo Tumoricidal Activity
Clin. Cancer Res.,
November 1, 2004;
10(21):
7207 - 7219.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Yoshitake, T. Nakatsura, M. Monji, S. Senju, H. Matsuyoshi, H. Tsukamoto, S. Hosaka, H. Komori, D. Fukuma, Y. Ikuta, et al.
Proliferation Potential-Related Protein, an Ideal Esophageal Cancer Antigen for Immunotherapy, Identified Using Complementary DNA Microarray Analysis
Clin. Cancer Res.,
October 1, 2004;
10(19):
6437 - 6448.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. H.M. Heemskerk, M. Hoogeboom, R. Hagedoorn, M. G.D. Kester, R. Willemze, and J.H. F. Falkenburg
Reprogramming of Virus-specific T Cells into Leukemia-reactive T Cells Using T Cell Receptor Gene Transfer
J. Exp. Med.,
April 5, 2004;
199(7):
885 - 894.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
I. Jedema, N. M. van der Werff, R. M. Y. Barge, R. Willemze, and J. H. F. Falkenburg
New CFSE-based assay to determine susceptibility to lysis by cytotoxic T cells of leukemic precursor cells within a heterogeneous target cell population
Blood,
April 1, 2004;
103(7):
2677 - 2682.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H.-J. Kolb, C. Schmid, A. J. Barrett, and D. J. Schendel
Graft-versus-leukemia reactions in allogeneic chimeras
Blood,
February 1, 2004;
103(3):
767 - 776.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Renga, P. Pedrazzoli, and S. Siena
Present results and perspectives of allogeneic non-myeloablative hematopoietic stem cell transplantation for treatment of human solid tumors
Ann. Onc.,
August 1, 2003;
14(8):
1177 - 1184.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. R. Drobyski, M. Gendelman, S. Vodanovic-Jankovic, and J. Gorski
Elimination of Leukemia in the Absence of Lethal Graft-Versus-Host Disease After Allogenic Bone Marrow Transplantation
J. Immunol.,
March 15, 2003;
170(6):
3046 - 3053.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Or, M. Y. Shapira, I. Resnick, A. Amar, A. Ackerstein, S. Samuel, M. Aker, E. Naparstek, A. Nagler, and S. Slavin
Nonmyeloablative allogeneic stem cell transplantation for the treatment of chronic myeloid leukemia in first chronic phase
Blood,
January 15, 2003;
101(2):
441 - 445.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. V. Melo, T. P. Hughes, and J. F. Apperley
Chronic Myeloid Leukemia
Hematology,
January 1, 2003;
2003(1):
132 - 152.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. E. Heslop, F. K. Stevenson, and J. J. Molldrem
Immunotherapy of Hematologic Malignancy
Hematology,
January 1, 2003;
2003(1):
331 - 349.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. J. Barrett, K. Rezvani, S. Solomon, A. M. Dickinson, X. N. Wang, G. Stark, H. Cullup, M. Jarvis, P. G. Middleton, and N. Chao
New Developments in Allotransplant Immunology
Hematology,
January 1, 2003;
2003(1):
350 - 371.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Makita, A. Hiraki, T. Azuma, A. Tsuboi, Y. Oka, H. Sugiyama, S. Fujita, M. Tanimoto, M. Harada, and M. Yasukawa
Antilung Cancer Effect of WT1-specific Cytotoxic T Lymphocytes
Clin. Cancer Res.,
August 1, 2002;
8(8):
2626 - 2631.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. A. Nijmeijer, R. Willemze, and J. H. F. Falkenburg
An animal model for human cellular immunotherapy: specific eradication of human acute lymphoblastic leukemia by cytotoxic T lymphocytes in NOD/scid mice
Blood,
June 28, 2002;
100(2):
654 - 660.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
U. F. Hartwig, M. Robbers, C. Wickenhauser, and C. Huber
Murine acute graft-versus-host disease can be prevented by depletion of alloreactive T lymphocytes using activation-induced cell death
Blood,
April 15, 2002;
99(8):
3041 - 3049.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. A. Gruber, D. C. Skelton, and D. B. Kohn
Requirement for NK Cells in CD40 Ligand-Mediated Rejection of Philadelphia Chromosome-Positive Acute Lymphoblastic Leukemia Cells
J. Immunol.,
January 1, 2002;
168(1):
73 - 80.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Montagna, R. Maccario, F. Locatelli, V. Rosti, Y. Yang, P. Farness, A. Moretta, P. Comoli, E. Montini, and A. Vitiello
Ex vivo priming for long-term maintenance of antileukemia human cytotoxic T cells suggests a general procedure for adoptive immunotherapy
Blood,
December 1, 2001;
98(12):
3359 - 3366.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. A. Riemersma, E. S. Jordanova, R. F. J. Schop, K. Philippo, L. H. J. Looijenga, E. Schuuring, and P. M. Kluin
Extensive genetic alterations of the HLA region, including homozygous deletions of HLA class II genes in B-cell lymphomas arising in immune-privileged sites
Blood,
November 15, 2000;
96(10):
3569 - 3577.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. M. Lokhorst, A. Schattenberg, J. J. Cornelissen, M. H. J. van Oers, W. Fibbe, I. Russell, N. W. C. J. v. d. Donk, and L. F. Verdonck
Donor Lymphocyte Infusions for Relapsed Multiple Myeloma After Allogeneic Stem-Cell Transplantation: Predictive Factors for Response and Long-Term Outcome
J. Clin. Oncol.,
August 16, 2000;
18(16):
3031 - 3037.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
I. L. Weissman
Translating Stem and Progenitor Cell Biology to the Clinic: Barriers and Opportunities
Science,
February 25, 2000;
287(5457):
1442 - 1446.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
H. Kantarjian, J. V. Melo, S. Tura, S. Giralt, and M. Talpaz
Chronic Myelogenous Leukemia: Disease Biology and Current and Future Therapeutic Strategies
Hematology,
January 1, 2000;
2000(1):
90 - 109.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Brenner, C. Rossig, U. Sili, J. W. Young, and E. Goulmy
Transfusion Medicine: New Clinical Applications of Cellular Immunotherapy
Hematology,
January 1, 2000;
2000(1):
356 - 375.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
TOP
ABSTRACT
INTRODUCTION