J H Frederik Falkenburg

Leiden University Medical Centre, Leyden, South Holland, Netherlands

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Publications (317)1923.72 Total impact

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    ABSTRACT: This is the sixth special report that the European Society for Blood and Marrow Transplantation regularly publishes on the current practice and indications for haematopoietic SCT for haematological diseases, solid tumours and immune disorders in Europe. Major changes have occurred in the field of haematopoietic SCT over the last years. Cord blood units as well as haploidentical donors have been increasingly used as stem cell sources for allo-SCT, thus, augmenting the possibility of finding a suitable donor for a patient. Continuous refinement of conditioning strategies has also expanded not only the number of potential indications but also has permitted consideration of older patients or those with co-morbidity for a transplant. There is accumulating evidence of the role of haematopoietic SCT in non-haematological disorders such as autoimmune diseases. On the other hand, the advent of new drugs and very effective targeted therapy has challenged the role of SCT in some instances or at least, modified its position in the treatment armamentarium of a given patient. An updated report with revised tables and operating definitions is presented.Bone Marrow Transplantation advance online publication, 23 March 2015; doi:10.1038/bmt.2015.6.
    Bone marrow transplantation 03/2015; DOI:10.1038/bmt.2015.6 · 3.47 Impact Factor
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    ABSTRACT: BH3 mimetic drugs may be useful to treat acute lymphoblastic leukemia (ALL) but the sensitivity of primary tumor cells has not been fully evaluated. Here B-lineage ALL cell cultures derived form a set of primary tumors were studied with respect to sensitivity to the BH3 mimetics ABT-263 and ABT-199 and to Bcl-2 dependence and function. These ALL cells each expressed high levels of Bcl-2 and exhibited great sensitivity to ABT-263 and ABT-199, which induced rapid apoptotic cell death. BH3 profiling indicated that the ALL cultures were Bcl-2 dependent. Co-immunoprecipitation studies revealed a multifaceted role for Bcl-2 in binding pro-apoptotic partners including Bax, Bak, Bik and Bim. ABT-263 disrupted Bcl-2:Bim interaction in cells. Mcl-1 overexpression rendered ALL cells resistant to ABT-263 and ABT-199 with Mcl-1 assuming the role of Bcl-2 in binding Bim. Freshly isolated pediatric ALL blasts also expressed high levels of Bcl-2 and exhibited high sensitivity to Bcl-2 inhibition by the BH3 mimetic compounds. Overall, our results showed that primary ALL cultures were both more sensitive to BH3 mimetics and more uniform in their response than established ALL cell lines which have been evaluated previously. Further, the primary cell model characterized here offers a powerful system for preclinical testing of novel drug and drug combinations to treat ALL. Copyright © 2015, American Association for Cancer Research.
    Cancer Research 02/2015; 75(7). DOI:10.1158/0008-5472.CAN-14-1849 · 9.28 Impact Factor
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    ABSTRACT: A record number of 39 209 HSCT in 34 809 patients (14 950 allogeneic (43%) and 19 859 autologous (57%)) were reported by 658 centers in 48 countries to the 2013 survey. Trends include: more growth in allogeneic than in autologous HSCT, increasing use of sibling and unrelated donors and a pronounced increase in haploidentical family donors when compared with cord blood donors for those patients without a matched related or unrelated donor. Main indications were leukemias, 11 190 (32%; 96% allogeneic); lymphoid neoplasias, 19 958 (57%; 11% allogeneic); solid tumors, 1543 (4%; 4% allogeneic); and nonmalignant disorders, 1975 (6%; 91% allogeneic). In patients without a matched sibling or unrelated donor, alternative donors are used. Since 2010 there has been a marked increase of 96% in the number of transplants performed from haploidentical relatives (802 in 2010 to 1571 in 2013), whereas the number of unrelated cord blood transplants has slightly decreased (789 in 2010 to 666 in 2013). The use of donor type varies greatly throughout Europe.Bone Marrow Transplantation advance online publication, 2 February 2015; doi:10.1038/bmt.2014.312.
    Bone Marrow Transplantation 02/2015; 50(4). DOI:10.1038/bmt.2014.312 · 3.47 Impact Factor
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    ABSTRACT: Purpose: T-cell recognition of minor histocompatibility antigens (MiHA) plays an important role in the beneficial graft-versus-leukemia (GVL) effect of allogeneic stem cell transplantation (allo-SCT) but also mediates serious graft versus host disease (GVHD) complications associated with allo-SCT. Using a reverse immunology approach we aim to develop a method enabling the identification of T-cell responses directed against predefined antigens, with the goal to select those MiHAs that can be used clinically in combination with allo-SCT. Experimental design: In this study, we used a recently developed MiHA selection algorithm to select candidate MiHAs within the HLA-presented ligandome of transformed B-cells. From the HLA-presented ligandome that predominantly consisted of monomorphic peptides 25 polymorphic peptides with a clinically relevant allele frequency were selected. By high-throughput screening the availability of high-avidity T-cells specific for these MiHA-candidates in different healthy donors was analyzed. Results: With the use of MHC-multimer enrichment, analyses of expanded T-cells by combinatorial coding MHC-multimer flow cytometry, and subsequent single cell cloning, positive T-cell clones directed to 2 new MiHA: LB-CLYBL-1Y and LB-TEP1-1S could be demonstrated, indicating the immunogenicity of these 2 MiHAs. Conclusion: The biological relevance of MiHA LB-CLYBL-1Y was demonstrated by the detection of LB-CLYBL-1Y specific T cells in a patient suffering from acute myeloid leukemia (AML) that experienced an anti-leukemic response after treatment with allo-SCT. Copyright © 2015, American Association for Cancer Research.
    Clinical Cancer Research 01/2015; 21(9). DOI:10.1158/1078-0432.CCR-14-2188 · 8.19 Impact Factor
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    ABSTRACT: Human Leukocyte Antigen (HLA) class I molecules generally present peptides (p) of 8 to 11 amino acids (aa) in length. Although an increasing number of examples with lengthy (>11 aa) peptides, presented mostly by HLA-B alleles, have been reported. Here we characterise HLA-A*02:01 restricted, in addition to the HLA-B*0702 and HLA-B*4402 restricted, lengthy peptides (>11 aa) arising from the B-cell ligandome. We analysed a number of 15-mer peptides presented by HLA-A*02:01, and confirmed pHLA-I-formation by HLA-folding and thermal stability assays. Surprisingly the binding affinity and stability of the 15-mer epitopes in complex with HLA-A*02:01 were comparable to the values observed for canonical length (8 to 11 aa) HLA-A*02:01-restricted peptides. We solved the structures of two 15-mer epitopes in complex with HLA-A*02:01, within which the peptides adopted distinct super-bulged conformations. Moreover, we demonstrate that T-cells can recognize the 15-mer peptides in the context of HLA-A*02:01, indicating that these 15-mer peptides represent immunogenic ligands. Collectively, our data expand our understanding of longer epitopes in the context of HLA-I, highlighting that they are not limited to HLA-B family, but can bind the ubiquitous HLA-A*02:01 molecule, and play an important role in T-cell immunity. Copyright © 2014, The American Society for Biochemistry and Molecular Biology.
    Journal of Biological Chemistry 12/2014; 290(5). DOI:10.1074/jbc.M114.607028 · 4.60 Impact Factor
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    ABSTRACT: Immunotherapy of B-cell malignancies using CD19-targeted chimeric antigen receptor (CAR)-transduced T-cells or CD20-targeted therapeutic monoclonal antibodies has shown clinical efficacy. However, refractory disease and the emergence of antigen-loss tumor escape variants after treatment demonstrate the need to target additional antigens. Here, we aimed to target the B-cell receptor associated protein CD79b by a T-cell receptor (TCR)-based approach. Since thymic selection depletes high-avidity T-cells recognizing CD79b-derived peptides presented in self-HLA molecules, we aimed to isolate T-cells recognizing these peptides presented in allogeneic HLA. Peptide-HLA tetramers composed of CD79b peptides bound to either HLA-A2 or HLA-B7 were used to isolate T-cell clones from HLA-A*0201 and B*0702-negative individuals. For three distinct T-cell clones CD79b specificity was confirmed through CD79b gene transduction and CD79b-specific shRNA knockdown. The CD79b specific T cell clones were highly reactive against CD79b-expressing primary B-cell malignancies whereas no recognition of non-hematopoietic cells was observed. Although lacking CD79b cell surface expression, intermediate reactivity towards monocytes, hematopoietic progenitor cells and T-cells was observed. Quantitative RT-PCR revealed low CD79b gene expression in these cell types. Therefore, aberrant gene expression must be taken into consideration when selecting common, apparently lineage-specific self-antigens as targets for TCR-based immunotherapies. Copyright © 2014 American Society of Hematology.
    Blood 11/2014; 125(6). DOI:10.1182/blood-2014-07-587840 · 9.78 Impact Factor
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    ABSTRACT: Effective T cell therapy against cancer is dependent on the formation of long-lived, stem cell-like T cells with the ability to self-renew and differentiate into potent effector cells. Here, we investigated the in vivo existence of stem cell-like antigen-specific T cells in allogeneic stem cell transplantation (allo-SCT) patients, and their ex vivo-generation for additive treatment post-transplant. Early after allo-SCT, CD8(+) Tscm cells targeting minor histocompatibility antigens (MiHAs) expressed by recipient tumor cells were non-detectable, emphasizing the need for improved additive MiHA-specific T cell therapy. Importantly, MiHA-specific CD8(+) T cells with an early CCR7(+)CD62L(+)CD45RO(+)CD27(+)CD28(+)CD95(+) memory-like phenotype and gene signature could be expanded from naive precursors by inhibiting Akt-signaling during ex vivo-priming and expansion. This resulted in a MiHA-specific CD8(+) T cell population containing a high proportion of stem cell-like T cells, compared to terminal differentiated effector T cells in control cultures. Importantly, these Akt-inhibited MiHA-specific CD8(+) T cells showed a superior expansion capacity both in vitro as well as in immunodeficient mice, and induced a superior anti-tumor effect in intrafemural multiple myeloma-bearing mice. These findings provide a rationale for clinical exploitation of ex vivo generated Akt-inhibited MiHA-specific CD8(+) T cells in additive immunotherapy to prevent or treat relapse in allo-SCT patients.
    Blood 10/2014; 124(23). DOI:10.1182/blood-2014-05-578583 · 9.78 Impact Factor
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    ABSTRACT: Knowledge of the accurate copy number of HLA class I presented ligands is important in fundamental and clinical immunology. Currently, the best copy number determinations are based on mass spectrometry, employing single reaction monitoring (SRM) in combination with a known amount of isotopically labeled peptide. The major drawback of this approach is that the losses during sample pretreatment, i.e. immunopurification and filtration steps, are not well defined and must, therefore, be estimated. In addition, such losses can vary for individual peptides. Therefore, we developed a new approach in which isotopically labeled peptide-MHC monomers (hpMHC) are prepared and added directly after cell lysis, i.e. before the usual sample processing. Using this approach, all losses during sample processing can be accounted for and allows accurate determination of specific MHC class I-presented ligands. Our study pinpoints the immunopurification step as the origin of the rather extreme losses during sample pretreatment and offers a solution to account for these losses. Obviously, this has important implications for accurate HLA-ligand quantitation. The strategy presented here can be used to obtain a reliable view of epitope copy number and thus allows improvement of vaccine design and strategies for immunotherapy.
    Journal of Proteomics 07/2014; 109. DOI:10.1016/j.jprot.2014.07.009 · 3.93 Impact Factor
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    ABSTRACT: Patients after allogeneic stem cell transplantation are at risk for invasive aspergillosis, especially during neutropenia. Recent data suggest that an impaired T-cell immune reconstitution post-transplantation plays an important role in this increased risk. In this study we investigated whether Aspergillus-specific T-cells are involved in the recovery from invasive aspergillosis by analyzing the Aspergillus-specific T-cell response in patients with invasive aspergillosis. In 9 patients with improvement of their aspergillus infection, we identified Crf1- or Catalase1-specific T-cells on the basis of CD154 expression and IFNγ production by stimulating with overlapping peptides of the A.fumigatus proteins Crf1 and Catalase1. These Aspergillus-specific T-cells were induced at the moment of regression of the aspergillus lesions. Crf1- and Catalase1-specific T-cells, sorted on the basis of CD154 expression at the peak of the immune response, had a Th1 phenotype and recognized a variety of T-cell epitopes. In contrast, in 2 patients with progressive invasive aspergillosis we were not able to identify any Crf1- or Catalase1-specific T-cells. These data indicate that the presence of Aspergillus-specific T-cells correlates with the clearance of aspergillus infection and suggest that adoptive immunotherapy may be beneficial in patients with invasive aspergillosis after stem cell transplantation.
    Haematologica 04/2014; 99(7). DOI:10.3324/haematol.2013.098830 · 5.87 Impact Factor
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    ABSTRACT: In all, 661 of 680 centers in 48 countries reported 37 818 hematopoietic SCT (HSCT) in 33 678 patients (14 165 allogeneic (42%), 19 513 autologous (58%)) in the 2012 survey. Main indications were leukemias, 10 641 (32%; 95% allogeneic); lymphoid neoplasias, 19 336 (57%; 11% allogeneic); solid tumors, 1630 (5%; 3% allogeneic); and nonmalignant disorders, 1953 (6%; 90% allogeneic). There were more unrelated donors than HLA-identical sibling donors (54% versus 38% (8% being mismatched related donor HSCT)). Cord blood was almost exclusive in allogeneic transplants (5% of total). Since 2011, the highest increases in allogeneic HSCT were for AML in CR1 (12%) and for myeloproliferative neoplasm (15%). For autologous HSCT the main increases were for plasma cell disorders (7%), non-Hodgkin lymphoma (4%) and autoimmune disease (50%). There were 4097 pediatric patients <18 years of age receiving HSCT, 2902 received an allogeneic and 1195 an autologous HSCT. Overall, 69% of allogeneic and 64% of autologous HSCT were performed in dedicated pediatric centers and the remainder in combined adult and pediatric centers. Distributions of diseases, donor types and stem cell source for all patients and pediatric patients in particular are shown. A percentage of centers fulfilling the annual required criteria for patient numbers for JACIE accreditation are provided.Bone Marrow Transplantation advance online publication, 17 March 2014; doi:10.1038/bmt.2014.55.
    Bone marrow transplantation 03/2014; DOI:10.1038/bmt.2014.55 · 3.47 Impact Factor
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    ABSTRACT: Following allogeneic stem cell transplantation (alloSCT) donor T-cells may recognize minor histocompatibility antigens (MiHA) specifically expressed on cells of the recipient. It has been hypothesized that T-cells recognizing hematopoiesis-restricted MiHA mediate specific graft-versus-leukemia (GvL) activity without inducing graft-versus-host-disease (GvHD), whereas T-cells recognizing ubiquitously expressed MiHA induce both GvL and GvHD-reactivity. Furthermore, alloreactive CD4 T-cells are hypothesized to be able to mediate specific GvL-reactivity due to the hematopoiesis-restricted expression of HLA-class-II. However, clinical observations suggest that an overt GvL-response, associated with expansion of T-cells specific for hematopoiesis-restricted antigens, is often associated with GvHD-reactivity. Therefore, we developed in-vitro models to investigate whether alloreactive T-cells recognizing hematopoiesis-restricted antigens induce collateral damage to surrounding non-hematopoietic tissues. We demonstrated that collateral damage to MiHA-negative fibroblasts was induced by misdirection of cytotoxic granules released from MiHA-specific T-cells activated by MiHA-positive hematopoietic cells resulting in granzyme-B-mediated activation of apoptosis in the surrounding fibroblasts. We demonstrated that direct contact between the activated T-cell and the fibroblast was a prerequisite for this collateral damage to occur. In conclusion, these data suggest that hematopoiesis-restricted T-cells actively participating in an overt GvL-response may contribute to GvHD via induction of collateral damage to non-hematopoietic targets.
    Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation 03/2014; 20(6). DOI:10.1016/j.bbmt.2014.03.002 · 3.35 Impact Factor
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    ABSTRACT: Hematological malignancies often express surface HLA-class-II, making them attractive targets for CD4+ T-cell therapy. We previously demonstrated that HLA-class-II ligands can be divided into DM-resistant and DM-sensitive antigens. In contrast to DM-resistant antigens, presentation of DM-sensitive antigens is suppressed by HLA-DM, but can be rescued by HLA-DO. We also showed that HLA-DO expression remains low in non-hematopoietic cells under inflammatory conditions, suggesting that DM-sensitive antigens may be ideal T-cell targets with a low risk for GvHD. Here, we demonstrated that B-cell malignancies often express HLA-DO and that levels are in particular high in chronic lymphocytic leukemia. Moreover, we showed that surface presentation of DM-sensitive antigens is regulated by HLA-DO, and that DM-sensitive antigens are relevant T-cell targets for B-cell malignancies and especially chronic lymphocytic leukemia. These data open the perspective to target HLA-class-II ligands with specific processing and presentation behavior for CD4+ T-cell therapy of hematological malignancies.
    Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation 02/2014; 20(5). DOI:10.1016/j.bbmt.2014.02.005 · 3.35 Impact Factor
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    ABSTRACT: Allogeneic stem cell transplantation (alloSCT) followed by donor lymphocyte infusion (DLI) can be applied as immunotherapeutic intervention to treat malignant diseases. Here, we describe a patient with progressive metastatic clear cell renal cell carcinoma (RCC) who was treated with T cell depleted non-myeloablative alloSCT and DLI resulting in disease regression accompanied by extensive graft versus host disease (GVHD). We characterized the specificity of this immune response, and detected a dominant T cell population recognizing a novel minor histocompatibility antigen (MiHA) designated LB-FUCA2-1V. T cells specific for LB-FUCA2-1V were shown to recognize RCC cell lines, supporting a dominant role in the graft versus tumor (GVT) reaction. However, coinciding with the gradual disappearance of chronic GVHD, the anti-tumor effect declined and 3 years after alloSCT the metastases became progressive again. To re-initiate the GVT reaction, escalating doses of DLI were given, but no immune response could be induced and the patient died of progressive disease 8.5 years after alloSCT. Gene expression studies illustrated that only a minimal number of genes shared expression between RCC and professional antigen presenting cells but were not expressed by non-malignant healthy tissues, indicating that in patients suffering from RCC, GVT reactivity after alloSCT may be unavoidably linked to GVHD.
    PLoS ONE 01/2014; 9(1):e85198. DOI:10.1371/journal.pone.0085198 · 3.53 Impact Factor
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    ABSTRACT: A two-step procedure in which T-cell depleted allogeneic stem cell transplantation is followed by sequential treatment with donor lymphocyte infusion at 6 months can significantly reduce the risk and severity of graft-versus-host disease, with postponed induction of the beneficial graft-versus-leukemia effect. However, patients with high-risk leukemia have a substantial risk of relapse early after transplantation, at a time when administration of donor lymphocyte infusion has a high likelihood of resulting in graft-versus-host disease, disturbing a favorable balance between graft-versus-leukemia effect and graft-versus-host disease. Therefore, new therapeutic modalities are required to allow early administration of T-cells capable of exerting graft-versus-leukemia effect without graft-versus-host disease. Here, we describe the isolation of virus-specific T-cells using the Streptamer-based isolation technology and subsequent transfer of the minor histocompatibility antigen HA-1-specific T-cell receptor using retroviral vectors. Isolation of virus-specific T-cells and subsequent transduction with HA-1-T-cell receptor resulted in rapid in vitro generation of highly pure dual-specific T-cells with potent anti-leukemic reactivity. Due to the short production procedure of only 10-14 days and the defined specificity of the T-cells, administration of virus-specific T-cells transduced with the HA-1-T-cell receptor as early as 8 weeks after allogeneic stem cell transplantation is feasible. (This clinical trial is registered at www.clinicaltrialsregister.eu as EudraCT number 2010-024625-20).
    Haematologica 12/2013; DOI:10.3324/haematol.2013.093690 · 5.87 Impact Factor
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    ABSTRACT: The prognosis of patients with relapsed acute myeloid leukemia after allogeneic transplant is poor. We hypothesized that initial disease control by effective cytoreduction, followed by rapid induction of a profound allo-immune response by donor-lymphocyte infusion during the neutropenic phase is essential for long-term survival. Additional interferon-α was administered in case no acute graft-versus-host-disease occurred within 3 weeks after donor-lymphocyte infusion. Overall, 44 patients with relapsed acute myeloid leukemia were assessed; 26 relapsed after myeloablative, and 18 after reduced-intensity conditioning. Of these, 7 patients were not eligible due to poor performance status (n=3) or severe graft-versus-host-disease (n=4) at time of relapse. Patients with smoldering relapses (n=5) received donor-lymphocyte infusion only. Thirty-two patients received cytoreductive treatment, followed by donor-lymphocyte infusion in 22 patients. Reasons for not receiving donor-lymphocyte infusion were chemotherapy-related death (n=1) and chemotherapy-refractory disease (n=9). Two-year overall survival after donor-lymphocyte infusion was 36% (95% confidence-interval: 16-57%). The impact of acute graft-versus-host-disease on survival was calculated with a Cox-regression model including onset of acute graft-versus-host-disease as a time-dependent variable. Development of grade 1-3, but not grade 4, acute graft-versus-host-disease, was associated with superior survival as compared to absence of graft-versus-host-disease (hazard ratio 0.22, p=0.03). In conclusion, efficient cytoreduction followed by donor-lymphocyte infusion and subsequent interferon-α leading to limited acute graft-versus-host-disease represents a potentially curative option for relapsed acute myeloid leukemia after allogeneic transplant.
    Haematologica 11/2013; DOI:10.3324/haematol.2013.089565 · 5.87 Impact Factor
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    ABSTRACT: Aberrant PI3K/AKT/mTOR signaling has been linked to oncogenesis and therapy resistance in various malignancies including leukemias. In Philadelphia chromosome (Ph) positive leukemias, activation of PI3K by dysregulated BCR-ABL tyrosine kinase (TK) contributes to the pathogenesis and development of resistance to ABL-TK inhibitors (TKI). The PI3K pathway thus is an attractive therapeutic target in BCR-ABL positive leukemias, but its role in BCR-ABL negative ALL is conjectural. Moreover, the functional contribution of individual components of the PI3K pathway in ALL has not been established. We compared the activity of the ATP-competitive pan-PI3K inhibitor NVP-BKM120, the allosteric mTORC1 inhibitor RAD001, the ATP-competitive dual PI3K/mTORC1/C2 inhibitors NVP-BEZ235 and NVP-BGT226 and the combined mTORC1 and mTORC2 inhibitors Torin 1, PP242 and KU-0063794 using long-term cultures of ALL cells (ALL-LTC) from patients with B-precursor ALL that expressed the BCR-ABL or TEL-ABL oncoproteins or were BCR-ABL negative. Dual PI3K/mTOR inhibitors profoundly inhibited growth and survival of ALL cells irrespective of their genetic subtype and their responsiveness to ABL-TKI. Combined suppression of PI3K, mTORC1 and mTORC2 displayed greater antileukemic activity than selective inhibitors of PI3K, mTORC1 or mTORC1 and mTORC2. Inhibition of the PI3K/mTOR pathway is a promising therapeutic approach in patients with ALL. Greater antileukemic activity of dual PI3K/mTORC1/C2 inhibitors appears to be due to the redundant function of PI3K and mTOR. Clinical trials examining dual PI3K/mTORC1/C2 inhibitors in patients with B-precursor ALL are warranted, and should not be restricted to particular genetic subtypes.
    PLoS ONE 11/2013; 8(11):e80070. DOI:10.1371/journal.pone.0080070 · 3.53 Impact Factor
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    ABSTRACT: The low frequency of antigen-specific naïve T cells has challenged numerous laboratories to develop various techniques to study the naïve T-cell repertoire. Here we combine the generation of naïve repertoire-derived antigen-specific T-cell lines based on MHC-tetramer staining and magnetic-bead enrichment with in-depth functional assessment of the isolated T cells. Cytomegalovirus (CMV) specific T-cell lines were generated from seronegative individuals. Generated T-cell lines consisted of a variety of immunodominant CMV-epitope specific oligoclonal T-cell populations restricted to various HLA-molecules (HLA-A1, A2, B7, B8 and B40), and the functional and structural avidity of the CMV-specific T cells was studied. Although all CMV-specific T cells were isolated based on their reactivity towards a specific peptide-MHC complex, we observed a large variation in the functional avidity of the MHC-tetramer positive T-cell populations, which correlated with the structural avidity measured by the recently developed Streptamer koff -rate assay. Our data demonstrate that MHC-tetramer staining is not always predictive for the sensitivity of specific T-cell reactivity, and challenge the sole use of MHC-tetramers as an indication of the peripheral T-cell repertoire, independent of the analysis of functional activity or structural avidity parameters. This article is protected by copyright. All rights reserved.
    European Journal of Immunology 11/2013; 43(11). DOI:10.1002/eji.201343397 · 4.52 Impact Factor
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    ABSTRACT: It is well known that allo-reactive T-cells play a crucial role in graft-versus-leukemia and graft-versus-host disease after allogeneic hematopoietic stem cell transplantation. Allo-reactive CD4+ T-cells can mediate direct cytolysis, but may also stimulate production of IgG antibodies as helper cells. Immune complexes may subsequently be processed and presented by professional antigen presenting cells and stimulate induction of specific CD8+ T-cells. As such, proteins targeted in coordinated T- and B-cell responses may represent a class of immunodominant antigens in clinical responses after alloSCT. We previously identified LB-PTK2B-1T as HLA class II restricted polymorphic antigen in a patient treated with donor lymphocyte infusion for relapsed chronic myeloid leukemia after HLA-matched alloSCT. Since PTK2B has also been described as antibody target, we here investigated whether a coordinated T- and B-cell response against PTK2B was induced. Patient serum before and after alloSCT and DLI was screened for antibodies, and we indeed observed development of a humoral immune response against PTK2B. Antibodies against PTK2B were only found after DLI and, in contrast to the CD4+ T-cells, recognized a monomorphic region of the protein. To our knowledge this is the first description of a coordinated allo-reactive CD4+ T-cell and auto-reactive antibody response against an autosomal antigen.
    Haematologica 10/2013; 99(2). DOI:10.3324/haematol.2013.086652 · 5.87 Impact Factor
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    ABSTRACT: The prognosis of adult patients with ALL remains unsatisfactory. AlloSCT is associated with a beneficial GVL response mediated by donor T cells. However, GVHD results in substantial mortality and long-term morbidity. T-cell depletion (TCD) of the graft reduces the severity of GVHD, but is associated with an increased relapse rate after alloSCT. Therefore, early sequential donor lymphocyte infusion (DLI) is likely to be necessary for a successful GVL reaction. Twenty-five adult ALL patients (10 Ph(+)ALL) were eligible for early DLI after initial disease control with myeloablative TCD-alloSCT in first CR (CR1), if active GVHD was absent at 3-6 months after alloSCT. Patients with a sibling donor or an unrelated donor were scheduled for 3.0 × 10(6) CD3(+) cells/kg or 1.5 × 10(6) CD3(+) cells/kg, respectively, at 6 months after alloSCT. Three patients died before evaluation (one early relapse). Five patients had active GVHD. Fourteen of the remaining seventeen patients received DLI (median time-to-DLI: 185 days). Overall, only 17% required long-term systemic immunosuppression for GVHD. With a median follow-up after TCD-alloSCT of 50 months, 2-year survival probability was 68% (95% confidence interval (CI) 49-87%). In conclusion, myeloablative TCD-alloSCT with early sequential DLI is an efficient and safe post-remission treatment for adult ALL patients in CR1.Bone Marrow Transplantation advance online publication, 12 August 2013; doi:10.1038/bmt.2013.111.
    Bone marrow transplantation 08/2013; DOI:10.1038/bmt.2013.111 · 3.47 Impact Factor
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    ABSTRACT: CD8+ T-cell depleted donor lymphocyte infusion (DLI) after T-cell depleted (TCD) allogeneic hematopoietic stem cell transplantation (alloSCT) has been associated with a reduced risk of Graft-versus-Host Disease (GvHD) while preserving conversion to donor hematopoiesis and anti-tumor immunity, providing a rationale for exploring CD4+ T-cell based immunotherapy for hematological malignancies. Here, we analyzed the clinical course and specificity of T-cell immune responses in two patients with acute myeloid leukemia who converted to full donor chimerism, but developed severe acute GvHD after prophylactic CD4+ DLI following 10/10-HLA-matched, but HLA-DPB1-mismatched TCD-alloSCT. Clonal analysis of activated T-cells isolated during GvHD demonstrated allo-reactivity exerted by CD4+ T-cells directed against patient mismatched HLA-DPB1 molecules on hematopoietic cells and skin-derived fibroblasts only when cultured under inflammatory conditions. At the time of CD4+ DLI, both patients contained residual patient-derived T-cells, including CMV-specific T-cells due to CMV-reactivations. Once activated by CMV-antigens, these CMV-specific T cells could stimulate HLA-DPB1-specific CD4+ T cells, which in turn could target non-hematopoietic tissues in GvHD. In conclusion, our data demonstrate that GvHD following HLA-DPB1-mismatched CD4+ DLI can be mediated by allo-reactive HLA-DPB1-directed CD4+ T-cells and that ongoing viral infections inducing HLA-class-II expression on non-hematopoietic cells may increase the likelihood of GvHD development. (ISRTCN registered: 51398568, http://www.controlled-trials.com/ISRCTN51398568/LUMC).
    Blood 06/2013; 122(11). DOI:10.1182/blood-2012-12-470872 · 9.78 Impact Factor

Publication Stats

8k Citations
1,923.72 Total Impact Points

Institutions

  • 1983–2015
    • Leiden University Medical Centre
      • • Department of Hematology
      • • Department of Clinical Pharmacy and Toxicology
      Leyden, South Holland, Netherlands
  • 1998–2014
    • Leiden University
      Leyden, South Holland, Netherlands
  • 1989–1991
    • Indiana University-Purdue University Indianapolis
      • • Division of Hematology/Oncology
      • • Department of Medicine
      Indianapolis, IN, United States