|
|||||||||
|
|
|
|
|
|
|
|
|
|
|
Prepublished online as a Blood First Edition Paper on April 17, 2002; DOI 10.1182/blood-2002-01-0024.
IMMUNOBIOLOGY
From the Department of Immunohematology and Blood
Transfusion and the Department of Hematology, Leiden University Medical
Center, The Netherlands.
Successful stem cell transplantation (SCT) across HLA barriers can
be performed with cord blood, megadoses of stem cells, or with
nonmyeloablative conditioning strategies. Because the HLA-mismatched
transplants are often T-cell depleted, leukemia relapse rates are high.
Treatment of relapsed leukemia after HLA-mismatched SCT is difficult. A
novel potential strategy to treat relapsed leukemia after
HLA-mismatched SCT is the use of patients' mismatched HLA molecules as
antigen-presenting molecules to generate hematopoietic system-specific
cytotoxic T cells (CTLs) from the stem cell donor. Adoptive transfer of
these hematopoietic system-specific CTLs that are restricted by
nonself HLA molecules may eliminate leukemia without affecting the
patient's nonhematopoietic cells or donor hematopoietic cells. We
investigated the feasibility of this strategy using the hematopoietic
system-specific minor histocompatibility antigen HA-1, which
is known to induce HLA-A2-restricted CTLs. HLA-A2 Allogeneic stem cell transplantation (SCT) is
the treatment of choice for various hematologic
malignancies.1 The curative graft-versus-leukemia (GvL)
effect of SCT is mainly mediated by donor-derived alloreactive T cells.
The potent immunotherapeutic effect of donor-derived alloreactive T
cells is also illustrated by the successful application of donor
lymphocyte infusions to relapsed chronic myelogenous
leukemia.2 However, if the donor T cells are not selected
for preferential reactivity against leukemic cells, the GvL is often
associated with graft-versus-host disease (GvHD). Hence, the ideal
strategy for immunotherapy without GvHD is to target donor-derived T
cells to antigens specifically expressed by the patient's
hematopoietic system, including the leukemic cells. In HLA-matched,
minor histocompatibility antigen (mHag)-mismatched SCT settings, this
strategy is feasible by targeting the donor-derived cytotoxic T cells
(CTLs) to the polymorphic hematopoietic system-specific mHags HA-1 and
HA-2.3 Because of hematopoietic system-restricted and
patient-specific target-cell damage, the ex vivo-generated HA-1/HA-2-specific CTLs can be used as immunotherapeutic agents to
eliminate relapsed leukemia with a low risk of GvHD.
Because not every patient has an HLA-identical family member, a
substantial proportion of leukemia patients currently undergo transplantation with stem cells of unrelated, often partially HLA-matched donors.4 Successful marrow engraftment
across an HLA mismatch is possible by the use of cord
blood,5 megadoses of CD34+ stem
cells,6 and by nonmyeloablative
conditioning7,8 combined with intensive T-cell
depletion.8 Because T-cell-depleted grafts lack the GvL
effect,1,9 leukemia relapse rates in HLA-mismatched SCT
are high. As yet, there is no adequate therapy for leukemia relapses
after HLA-mismatched SCT. However, a novel approach for the treatment
of relapsed leukemia after HLA-mismatched SCT is to take advantage of
the HLA mismatch of the recipient for the generation of
immunotherapeutic T cells. Here, the recipients' mismatched HLA
molecules are applied as antigen-presenting molecules for hematopoietic
system-specific antigens to generate CTLs from the stem cell donor.
These CTLs have 2 characteristics: they are hematopoietic system
specific and they are "nonself" HLA restricted. Earlier studies
showed the existence of T cells restricted by nonself HLA molecules.
Sadovnikova et al10,11 have generated mdm-2- and
cyclin-D1-specific nonself major histocompatibility complex
(MHC)-restricted T cells using peptide-loaded transporter associated with antigen presentation (TAP)-deficient cells as stimulator cells for MHC-incompatible T cells. Similarly, Munz et
al12 showed the generation of nonself HLA-restricted,
peptide-specific T-cell responses by stimulating HLA-A2 In this study, we have used the mHag HA-1 as a model antigen to verify
the feasibility of generation of hematopoietic system-specific CTLs
restricted by nonself HLA molecules. The mHag HA-1 is an ideal antigen
to test the feasibility of this strategy because its several relevant
characteristics are known. HA-1 is an immunogenic T-cell epitope that
induces HLA-A2-restricted CTLs in vivo in recipients of HLA-matched,
HA-1-mismatched stem cell donors. Immunogenetic analyses demonstrated
mendelian segregation; the HA-1 phenotype frequency in the
HLA-A2+ population is 69%. The tissue distribution of HA-1
is restricted to the hematopoietic system; HA-1-specific CTLs lyse
leukemic cells and their progenitors but not nonhematopoietic
cells.15-18 The HA-1 T-cell epitope is a nonameric peptide
(ie, HA-1H) encoded by a diallelic locus that differs by a
single amino acid (ie, HA-1R) from its allelic
counterpart.19 The HLA-A2/HA-1R ligand is not
immunogenic (for a review, see Goulmy20).
The mHag HA-1 is recognized in association with the HLA-A2 molecule.
Therefore, throughout this study, PBMCs from HLA-A2 HA-1 peptide
Tetrameric HLA-A2/HA-1 peptide complexes
Induction of HA-1-specific CTLs restricted by nonself HLA-A2 molecules Responder cells.
The mHag HA-1 is recognized in the context of HLA-A2. Therefore,
HA-1-specific CTLs restricted by nonself HLA molecules were generated
using PBMCs from 2 HLA-A2 Stimulator cells. T2 cells.
TAP-defective T2 cells were cultured in RPMI 1640 medium (Life
Sciences, Breda, The Netherlands) with 15% human serum. T2 cells synthesize HLA-A2 and -B5 but do not express them because of the
TAP defect. T2 cells were pulsed with HA-1 peptide (10 µg/mL) for 16 hours at 37°C in the presence of 1 µg/mL
C1R-A2 cells. The HLA-A2 transfectant of C1R Epstein-Barr virus (EBV)-transformed B-cell line (EBV-LCL) was described elsewhere.11 Untransfected C1R cells do not express HLA-A or HLA-B, but show a low expression of Cw4. C1R-A2 cells were cultured in RPMI medium with 15% human serum. C1R2-A2 cells were pulsed with HA-1 peptide (10 µg/mL) for 16 hours at 37°C and irradiated (30 Gy) before coculture with responder cells at a stimulator cell-to-responder-cell ratio of 1:10. DCs transduced with HA-1 cDNA. The transduction of CD34+ progenitor cell-derived DCs with the HA-1-coding cDNA using a retroviral gene-delivery system is described elsewhere.25 The HLA typing of the HA-1-transduced DCs was as follows: HLA-A2, -A24, -B7, -B13, -Cw6, -Cw7, -DRB1*0701, -DRB1*11, -DQB1*0202, -DQB1*0301, -DPB1*0301, -DPB1*02012. The transduction efficiency of the DCs was 20%. HA-1 gene-transduced DCs were irradiated (30 Gy) before coculture with responder cells at a stimulator cell-to-responder cell ratio of 1:5. Generation of HA-1-specific CTLs restricted by nonself HLA-A2 molecules using T2 cells as stimulator cells The protocol described by Sadovnikova et al11 was used with slight modifications. Briefly, 20 × 106 responder cells from donor no. 1 were cocultured with T2 cells as stimulator cells in the presence of RPMI 1640 with 15% human serum (culture medium). At day 7, the CD8+ cells were sorted using immunomagnetic beads (MACS; Miltenyi Biotech, Bergisch Gladbach, Germany) and restimulated in culture medium by the addition of peptide-pulsed C1R-A2 cells (2 × 105 cells/mL), 30 Gy irradiated autologous PBMCs (2 × 106 cells/mL), and HA-1 peptide (0.5 µg/mL). The T-cell line was restimulated with peptide-pulsed T2 cells at day 14 and with peptide-pulsed C1R-A2 cells at day 21. The T-cell line was subcloned at day 28 by limiting dilution at 1000, 750, 500, 250, and 100 cells per well in 96-well, round-bottom tissue-culture plates in 50 µL/well culture medium supplemented with peptide-pulsed T2 cells (1 × 105 cells/mL), 30 Gy irradiated autologous PBMCs (1 × 106 cells/mL), and interleukin-2 (IL-2; 12 IU/mL). The subclones that showed more than 30% HA-1 peptide-specific lysis were pooled and restimulated in 24-well plates in the presence of peptide-pulsed C1R-A2 cells (2 × 105 cells/mL), autologous PBMCs (2 × 106 cells/mL), and IL-2 (60 IU/mL). The pooled subclones were stained with HA-1A2 tetramers and sorted by flow cytometry. The sorted cells were either restimulated as bulk or cloned by limiting dilution at 0.3 cell per well in 50 µL/well culture medium supplemented with a feeder cell/cytokine mix containing 60 IU/mL IL-2, 1% Leuco-agglutinin (Sigma, St Louis, MO), 30 Gy irradiated PBMCs from 6 random donors (1 × 106 cells/mL), and 50 Gy irradiated HA-1+ EBV-LCLs (1 × 105 cells/mL). The clones were expanded with IL-2-containing (60 IU/mL) culture medium. The HA-1-specific CTL clones restricted to nonself HLA-A2 molecules could be cultured for 4 to 6 weeks in vitro by weekly stimulations with the feeder cell/cytokine mix.Generation of HA-1-specific CTLs restricted by nonself HLA-A2 molecules using DCs transduced with HA-1 cDNA as stimulator cells HLA-A2 PBMCs (15 × 106) from donor
no. 2 were stimulated with HLA-A2+, HA-1-transduced DCs in
Iscoves modified Dulbecco medium (IMDM; Life Sciences)
supplemented with 10% heat-inactivated human plasma, 6 IU/mL IL-2, and
1 U/mL IL-12. At day 7, the CD8+ cells were positively
selected by immunomagnetic beads (MACS; Miltenyi Biotech). The
CD8+ T cells were restimulated with the original
HA-1-transduced DCs at days 7, 14, and 21. IL-2-containing (120 IU/mL) IMDM plus 10% pooled human plasma was added 24 hours after each
restimulation and every other 3 days. At day 28, the CD8+ T
cells that showed bright staining with HA-1A2 tetramers
were sorted by flow cytometry and cloned as described above.
Cell-mediated lympholysis assays Standard 4-hour chromium (51Cr)-release assays were performed as previously described.3 51Cr-labeled target cells were pulsed with 1 µg/mL peptide for 60 minutes before the addition of effector cells. For cold target inhibition assays, unlabeled target cells were added in the wells at various cold target cell-to-hot target cell ratios. The percentage specific lysis was calculated using the following formula: 100 × (cpm experimental release cpm spontaneous release)/(cpm
maximal release cpm spontaneous release).
Generation of HA-1-specific CTLs restricted by nonself HLA-A2 molecules using T2 cells pulsed with HA-1 peptide The alloreactive T-cell line induced with HA-1 peptide-pulsed T2 cells was subcloned by limiting dilution, and the proliferating subclones were analyzed for cytotoxic activity against unpulsed and HA-1 peptide-pulsed T2 cells. Whereas many (182 of 211) subclones lysed both peptide-pulsed and unpulsed T2 cells, 17 of 211 subclones showed preferential lysis against HA-1 peptide- pulsed T2 cells. The mean cytotoxic activities of these latter subclones against HA-1 peptide-pulsed and unpulsed T2 cells were 68% and 35%, respectively (Figure 1A). These "HA-1-specific" subclones were pooled and stained with HA-1A2 tetramers. A total of 7% lymphocytes showed bright staining with HA-1A2 tetramers (Figure 1B). To verify the specificity of the HA-1A2 tetramer staining, we analyzed 15 allo-HLA-A2-specific CTL clones. HA-1A2 tetramers stained solely HA-1-specific CTL clones. None of the 15 allo-HLA-A2-specific CTLs showed staining with HA-1A2 tetramers (Figure 1C).
Generation of HA-1-specific CTLs restricted by nonself HLA-A2 molecules using DCs transduced with HA-1 cDNA The PBMCs of another HLA-A2 individual were
stimulated with HLA-A2+ DCs that were transduced with HA-1.
After selection of CD8+ T cells at day 7, the cells were
restimulated weekly. The development of HA-1-specific CTLs was
monitored with HA-1A2 tetramers. As illustrated in Figure
2, tetramer-positive T cells totaled only
0.5% at day 14. The frequency of HA-1A2 tetramer-positive
cells increased significantly upon antigen-specific restimulations and
totaled 2% and 7% at days 21 and 28, respectively (Figure
2).
Selection of HA-1-specific CTLs restricted by nonself HLA molecules using tetrameric HLA-A2/HA-1 peptide complexes The HA-1A2 tetramer-positive CD8+ cells in the polyclonal cultures were sorted by flow cytometry. In the CTL line induced by T2 cells, the HA-1A2 tetramer-positive T cells were enriched from 7% to 87% after a single round of sorting (Figure 3A). The sorted T cells did not cross-react with the control HA-2A2 tetramer (Figure 3A) and showed strong lysis of HA-1 peptide-pulsed T2 cells (Figure 3B). The H-Y peptide-pulsed T2 cells were lysed by 30%. The latter lysis on the control H-Y peptide-pulsed T2 cells could be due to allo-HLA-A2-specific T cells or natural killer (NK) cells contaminating the culture. Therefore, cold target inhibition studies were performed by the addition of cold (unlabeled) K562 cells and HLA-A2+ HA-1 EBV-LCLs during the
cell-mediated lympholysis assays. The lysis of HA-1 peptide-pulsed T2
cells was not significantly affected by the addition of cold K562 cells
or HLA-A2+ HA-1 EBV-LCLs. In contrast, the
lysis of H-Y peptide-pulsed T2 cells was abrogated by the addition of
both cold K562 cells and HLA-A2+ HA-1
EBV-LCLs. Thus, the T-cell line contained not only HA-1-specific CTLs,
but also allo-HLA-A2-specific T cells and NK cells (Figure 3B).
Addition of the HA-1 peptide-pulsed HLA-A2+ EBV-LCLs as
cold target cells abrogated the lysis of HA-1 peptide-pulsed targets,
indicating the specificity of the assay (data not shown). The
CTL line was subsequently tested against target cells that express the
natural HLA-A2/HA-1 ligand. Unlabeled K562 cells and HLA-A2+ HA-1 EBV-LCLs were used as cold
targets to inhibit the cytotoxic activity mediated by contaminating NK
cells and allo HLA-A2-specific CTLs. Effective lysis was demonstrated
against HA-1 peptide-pulsed target cells, HA-1+
phytohemagglutinin blasts, EBV-LCLs, and leukemic cells of AML and acute lymphocytic leukemia (ALL) origin. HA-1 or
HLA-A2 target cells were not lysed (Figure
4). The HA-1-specific CTLs induced by
transduced DCs could be enriched to 20% purity by FACS sorting (data
not shown). This CTL line was therefore directly cloned by
limiting dilution.
Generation of HA-1-specific CTL clones restricted by nonself HLA-A2 molecules Because the HA-1-specific CTLs induced by T2 cells and HA-1-transduced DCs contained undesired allo-HLA-A2-specific cells and NK cells, CTL clones were obtained by limiting dilution at 0.3 cell per well. Two CTL clones showed bright staining with HA-1A2 tetramer (Figure 5A), but no staining with the control HA-2A2 tetramer. As expected, both CTL clones showed strong and specific lysis of both HA-1 peptide-pulsed EBV-LCLs and EBV-LCLs that naturally express the HLA-A2/HA-1 ligand (Figure 5B).
In this study, we show that the T-cell repertoire of
HLA-A2 To generate HA-1-specific CTLs restricted by nonself HLA-A2 molecules, we used 2 approaches. In the first approach, similar to that of Sadovnikova et al,11 we used T2 cells pulsed with HA-1 peptide as stimulator cells. Because the empty HLA-A2 molecules on the surface of T2 cells can be uniformly loaded with exogenous peptides, this approach could minimize the induction of undesired allo-HLA-A2 reactivities. Moreover, similar to Sadovnikova et al,11 we restimulated the responder cells with the alternative usage of C1R-A2 cells and T2 cells. Because C1R-A2 cells and T2 cells are genetically distinct, it was expected that different sets of "irrelevant" endogenous peptides would be presented by T2 cells and C1R-A2 cells. Thus, alternative restimulation of alloreactive T-cell lines with T2 and C1R-A2 cells could prevent the restimulation of undesired allo-HLA-A2 CTLs directed to "undesired" endogenous peptides. Our second approach involved the use of DCs transduced with the HA-1 cDNA as stimulator cells, which also could present the "irrelevant" endogenous peptides. Both approaches induced HA-1-specific CTLs restricted by nonself HLA-A2 molecules, but also induced similar levels of undesired allo-HLA-A2 reactivities. It is noteworthy that in both approaches, the responder cells were mHag HA-1H positive. Thus, endogenous expression of HA-1 by the responder does not affect the generation of HA-1-specific CTLs restricted by nonself HLA-A2 molecules. In both approaches, the responder and the stimulator cells were mismatched not only for HLA-A2, but also for other HLA class I molecules that could induce additional undesired alloresponses. The presence of these additional mismatches apparently did not hamper the generation of HA-1-specific CTLs in the responder/stimulator combinations we used. Nonetheless, because the HLA system is highly polymorphic, our data cannot rule out the possibility that mismatches for highly immunogenic HLA class I alleles could negatively influence the generation of nonself-restricted HA-1-specific CTLs. The negative effect of additional mismatches may be less relevant for the "T2" approach because T2 cells and C1R-A2 cells do not express HLA class I molecules on the cell surface. However, it may be preferable to select closely HLA-matched responder and stimulator cells for the generation of nonself-restricted CTLs, as is the case in the HLA-mismatched SCT setting. It was reported that undesired alloreactive T cells from donor PBMCs can be efficiently removed by depletion of CD69/CD25+ T cells triggered by short-term stimulation of HLA-mismatched patient cells.22 We indeed observed that CD69/CD25-depleted T cells showed no anti-HLA-A2 reactivity in the initial weeks of the cultures. However, strong anti-HLA-A2 alloreactivity developed upon restimulation with allo-HLA-A2 target cells in the long-term cultures of more than 40 days, indicating ineffective elimination of allo-HLA-A2-specific responses using these depletion strategies (data not shown). Earlier we reported that mHag-specific CTLs show specific staining with HLA class I/mHag peptide tetramers.21 The intensity of the tetramer staining correlates with the avidity of the mHag-specific CTLs to the natural mHag ligand.23 On the basis of these results, we used tetrameric HLA/mHag peptide complexes for the selection and enrichment of HA-1-specific CTLs restricted by nonself HLA-A2 molecules. A recent report showed that certain alloreactive T cells can stain with tetrameric HLA/peptide complexes independent of the peptide complexed with the HLA molecule.24 Thus, tetramer staining in alloreactive T-cell cultures may not always reflect the peptide specificity. In our study, we did not observe nonspecific staining of alloreactive T cells with HA-1A2 tetramers. Thus, in our protocols, tetrameric HLA class I/peptide complexes served as satisfactory tools for the monitoring and enrichment of HA-1-specific CTLs restricted by nonself HLA molecules. However, depletion of all undesired alloreactivity by the sole use of HA-1A2 tetramers was not possible, and limiting dilution of the sorted T cells was necessary for the effective selection of HA-1-specific CTLs. In conclusion, our results provide proof of the principle that specific CTLs can be generated against a nonself HLA-A2/HA-1 peptide ligand. Naturally, further optimization of the in vitro protocols will be indispensable for effective elimination of the undesired alloreactivity. The successful induction of both WT-1- and HA-1-specific CTLs restricted by nonself HLA molecules clearly underlines the feasibility of adoptive immunotherapy of hematologic malignancies with hematopoietic system-specific CTLs restricted by nonself HLA molecules after HLA-mismatched SCT. This novel strategy also introduces the possibility of a variety of other hematopoietic system-specific antigens to be used as target antigens for the generation of CTLs restricted by nonself HLA molecules. The candidate antigens include leukemia-associated antigens with known immunogenicity, hematopoietic system-specific transcription factors with high expression on leukemic cells (GATA-1, PU-1, IKAROS), and hematopoietic system-specific differentiation antigens (CD45, CD19, and CD20). Naturally, future studies are necessary to test the immunogenicity of peptides of these candidate proteins in the context of various HLA alleles. These studies will expand the arsenal of target antigens for the generation of hematopoietic system-specific CTLs restricted by nonself HLA molecules and provide a basis for broader application of this strategy for the treatment of relapsed leukemia after HLA-mismatched SCT.
We thank Prof Dr A. Brand and Dr M. Oudshoorn for reading the manuscript.
Submitted January 3, 2002; accepted March 5, 2002.
Prepublished online as Blood First Edition Paper, April 17, 2002; DOI 10.1182/blood-2002-01-0024.
Supported in part by grants from the Leiden University Medical Center, the J.A. Cohen Institute for Radiopathology and Radiation Protection, the Dutch Cancer Society, and the Leukemia & Lymphoma Society. T.M. is a Leukemia & Lymphoma Society Translational Research Fellow.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Tuna Mutis, Department of Immunohematology and Blood Transfusion, Bldg 1 E3-Q, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands; e-mail: t.mutis{at}lumc.nl.
1.
Horowitz MM, Gale RP, Sondel PM, et al.
Graft-versus-leukemia reactions after bone marrow transplantation.
Blood.
1990;75:555-562
2.
Kolb HJ, Schattenberg A, Goldman JM, et al.
Graft-versus-leukemia effect of donor lymphocyte transfusions in marrow grafted patients. European Group for Blood and Marrow Transplantation Working Party Chronic Leukemia.
Blood.
1995;86:2041-2050
3.
Mutis T, Verdijk R, Schrama E, Esendam B, Brand A, Goulmy E.
Feasibility of immunotherapy of relapsed leukemia with ex vivo-generated cytotoxic T lymphocytes specific for hematopoietic system-restricted minor histocompatibility antigens.
Blood.
1999;93:2336-2341
4.
Scott I, O'Shea J, Bunce M, et al.
Molecular typing shows a high level of HLA class I incompatibility in serologically well matched donor/patient pairs: implications for unrelated bone marrow donor selection.
Blood.
1998;92:4864-4871
5.
Gluckman E, Rocha V, Boyer-Chammard A, et al.
Outcome of cord-blood transplantation from related and unrelated donors. Eurocord Transplant Group and the European Blood and Marrow Transplantation Group.
N Engl J Med.
1997;337:373-381
6.
Aversa F, Tabilio A, Velardi A, et al.
Treatment of high-risk acute leukemia with T-cell-depleted stem cells from related donors with one fully mismatched HLA haplotype.
N Engl J Med.
1998;339:1186-1193 7. Sykes M, Preffer F, McAfee S, et al. Mixed lymphohaemopoietic chimerism and graft-versus-lymphoma effects after non-myeloablative therapy and HLA-mismatched bone-marrow transplantation. Lancet. 1999;353:1755-1759[CrossRef][Medline] [Order article via Infotrieve].
8.
Slavin S, Nagler A, Naparstek E, et al.
Nonmyeloablative stem cell transplantation and cell therapy as an alternative to conventional bone marrow transplantation with lethal cytoreduction for the treatment of malignant and nonmalignant hematologic diseases.
Blood.
1998;91:756-763 9. Champlin R. T-cell depletion for allogeneic bone marrow transplantation: impact on graft-versus-host disease, engraftment, and graft-versus-leukemia. J Hematother. 1993;2:27-42[Medline] [Order article via Infotrieve].
10.
Sadovnikova E, Stauss HJ.
Peptide-specific cytotoxic T lymphocytes restricted by nonself major histocompatibility complex class I molecules: reagents for tumor immunotherapy.
Proc Natl Acad Sci U S A.
1996;93:13114-13118 11. Sadovnikova E, Jopling LA, Soo KS, Stauss HJ. Generation of human tumor-reactive cytotoxic T cells against peptides presented by non-self HLA class I molecules. Eur J Immunol. 1998;28:193-200[CrossRef][Medline] [Order article via Infotrieve].
12.
Munz C, Obst R, Osen W, Stevanovic S, Rammensee HG.
Alloreactivity as a source of high avidity peptide-specific human CTL.
J Immunol.
1999;162:25-34
13.
Mutis T, Schrama E, Luxemburg-Heijs SA, Falkenburg JH, Melief CJ, Goulmy E.
HLA class II restricted T-cell reactivity to a developmentally regulated antigen shared by leukemic cells and CD34+ early progenitor cells.
Blood.
1997;90:1083-1090
14.
Gao L, Bellantuono I, Elsasser A, et al.
Selective elimination of leukemic CD34+ progenitor cells by cytotoxic T lymphocytes specific for WT1.
Blood.
2000;95:2198-2203 15. de Bueger M, Bakker A, van Rood JJ, Goulmy E. Minor histocompatibility antigens, defined by graft-vs.-host disease-derived cytotoxic T lymphocytes, show variable expression on human skin cells. Eur J Immunol. 1991;21:2839-2844[Medline] [Order article via Infotrieve].
16.
van der Harst D, Goulmy E, Falkenburg JH, et al.
Recognition of minor histocompatibility antigens on lymphocytic and myeloid leukemic cells by cytotoxic T-cell clones.
Blood.
1994;83:1060-1066
17.
Marijt WA, Veenhof WF, Goulmy E, Willemze R, van Rood JJ, Falkenburg JH.
Minor histocompatibility antigens HA-1-, -2-, and -4-, and HY-specific cytotoxic T-cell clones inhibit human hematopoietic progenitor cell growth by a mechanism that is dependent on direct cell-cell contact.
Blood.
1993;82:3778-3785
18.
Falkenburg JH, Goselink HM, van der Harst D, et al.
Growth inhibition of clonogenic leukemic precursor cells by minor histocompatibility antigen-specific cytotoxic T lymphocytes.
J Exp Med.
1991;174:27-33
19.
den Haan JM, Meadows LM, Wang W, et al.
The minor histocompatibility antigen HA-1: a diallelic gene with a single amino acid polymorphism.
Science.
1998;279:1054-1057 20. Goulmy E. Human minor histocompatibility antigens: new concepts for marrow transplantation and adoptive immunotherapy. Immunol Rev. 1997;157:125-140[CrossRef][Medline] [Order article via Infotrieve]. 21. Mutis T, Gillespie G, Schrama E, Falkenburg JH, Moss P, Goulmy E. Tetrameric HLA class I-minor histocompatibility antigen peptide complexes demonstrate minor histocompatibility antigen-specific cytotoxic T lymphocytes in patients with graft-versus-host disease. Nat Med. 1999;5:839-842[CrossRef][Medline] [Order article via Infotrieve]. 22. Koh MB, Prentice HG, Lowdell MW. Selective removal of alloreactive cells from haematopoietic stem cell grafts: graft engineering for GVHD prophylaxis. Bone Marrow Transplant. 1999;23:1071-1079[CrossRef][Medline] [Order article via Infotrieve]. 23. Gillespie G, Mutis T, Schrama E, et al. HLA class I-minor histocompatibility antigen tetramers select cytotoxic T cells with high avidity to the natural ligand. Hematol J. 2000;1:403-410[CrossRef][Medline] [Order article via Infotrieve].
24.
Moris A, Teichgraber V, Gauthier L, Buhring HJ, Rammensee HG.
Cutting edge: characterization of allorestricted and peptide-selective alloreactive T cells using HLA-tetramer selection.
J Immunol.
2001;166:4818-4821 25. Mutis T, Ghoreschi K, Schrama E, et al. Efficient induction of minor histocompatibility antigen HA-1 specific cytotoxic T cells using dendritic cells retrovirally transduced with HA-1 coding cDNA. Biol Blood Marrow Transplant. 2002. In press.
© 2002 by The American Society of Hematology.
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
|
 |
|
  L. E. M. Oosten, D. Koppers-Lalic, E. Blokland, A. Mulder, M. E. Ressing, T. Mutis, A. G. S. van Halteren, E. J. H. J. Wiertz, and E. Goulmy TAP-inhibiting proteins US6, ICP47 and UL49.5 differentially affect minor and major histocompatibility antigen-specific recognition by cytotoxic T lymphocytes Int. Immunol., September 1, 2007; 19(9): 1115 - 1122. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Mullally and J. Ritz Beyond HLA: the significance of genomic variation for allogeneic hematopoietic stem cell transplantation Blood, February 15, 2007; 109(4): 1355 - 1362. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. M. E. Whitelegg, L. E. M. Oosten, S. Jordan, M. Kester, A. G. S. van Halteren, J. A. Madrigal, E. Goulmy, and L. D. Barber Investigation of Peptide Involvement in T Cell Allorecognition Using Recombinant HLA Class I Multimers J. Immunol., August 1, 2005; 175(3): 1706 - 1714. [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]
|
|
|
|
|
|
H. Fujiwara, F. El Ouriaghli, M. Grube, D. A. Price, K. Rezvani, E. Gostick, G. Sconocchia, J. Melenhorst, N. Hensel, D. C. Douek, et al. Identification and in vitro expansion of CD4+ and CD8+ T cells specific for human neutrophil elastase Blood, April 15, 2004; 103(8): 3076 - 3083. [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]
|
|
|
|
|
|
M. H. M. Heemskerk, M. Hoogeboom, R. A. de Paus, M. G. D. Kester, M. A. W. G. van der Hoorn, E. Goulmy, R. Willemze, and J. H. F. Falkenburg Redirection of antileukemic reactivity of peripheral T lymphocytes using gene transfer of minor histocompatibility antigen HA-2-specific T-cell receptor complexes expressing a conserved alpha joining region Blood, November 15, 2003; 102(10): 3530 - 3540. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Bachar-Lustig, S. Reich-Zeliger, and Y. Reisner Anti-third-party veto CTLs overcome rejection of hematopoietic allografts: synergism with rapamycin and BM cell dose Blood, September 15, 2003; 102(6): 1943 - 1950. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Orsini, R. Bellucci, E. P. Alyea, R. Schlossman, C. Canning, S. McLaughlin, P. Ghia, K. C. Anderson, and J. Ritz Expansion of Tumor-specific CD8+ T Cell Clones in Patients with Relapsed Myeloma after Donor Lymphocyte Infusion Cancer Res., May 15, 2003; 63(10): 2561 - 2568. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
 |
 |
|||||||
 |
 |
 |
 |
 |
 |
 |
 |
||
| Copyright © 2002 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
Top
Abstract
Introduction
2-microglobulin and irradiated (30 Gy) before coculture
with responder cells at a stimulator cell-to-responder-cell ratio of
1:10.
