J H Frederik Falkenburg

Leiden University, Leyden, South Holland, Netherlands

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Publications (326)2047.76 Total impact


  • No preview · Article · Feb 2016
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    ABSTRACT: In the field of hematopoietic stem cell transplantation, the common approach is to focus outcome analyses on time to relapse and death, without assessing the impact of post-transplant interventions. We investigated whether a multi-state model would give insight into the events after transplantation in a cohort of patients who were transplanted using a strategy including scheduled donor lymphocyte infusions. Seventy-eight consecutive patients who underwent myeloablative T-cell depleted allogeneic stem cell transplantation for acute myeloid leukemia or myelodysplastic syndrome were studied. We constructed a multi-state model to analyze the impact of donor lymphocyte infusion and graft-versus-host disease on the probabilities of relapse and non-relapse mortality over time. Based on this model we introduced a new measure for outcome after transplantation which we called "treatment success" being alive without relapse and immunosuppression for graft-versus-host disease. All relevant clinical events were implemented into the multi-state model and were denoted treatment success or failure (either transient or permanent). Both relapse and non-relapse mortality were causes of failure of comparable magnitude. Whereas relapse was the dominant cause of failure from the transplantation state, its rate was reduced after graft-versus-host disease and especially after donor lymphocyte infusion. The long-term probability of treatment success was approximately 40%. This probability was increased after donor lymphocyte infusion. Our multi-state model helps to interpret the impact of post-transplantation interventions and clinical events on failure and treatment success, thus extracting more information from observational data.
    Preview · Article · Jan 2016 · Haematologica
  • J.H.F. Falkenburg · I. Jedema
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    ABSTRACT: Several mechanisms can be responsible for control of hematological tumors by allo-reactive T cells. Following allogeneic stem cell transplantation (alloSCT) donor T cells recognizing genetic disparities presented on recipient cells and not on donor cells are main effectors of tumor control, but also of the detrimental graft versus host disease (GVHD). Since after transplantation normal hematopoiesis is of donor origin, any T cell response directed against polymorphic antigens expressed on hematopoietic recipient cells but not on donor cells will result in an anti-tumor response not affecting normal hematopoiesis. After fully HLA-matched alloSCT, T cells recognizing polymorphic peptides derived from proteins encoded by genes selectively expressed in hematopoietic lineages may result in anti-tumor responses without GVHD. Due to the high susceptibility of hematopoietic cells for T cell recognition, a low amplitude of the overall T cell response may also be in favor of the anti-tumor reactivity in hematological malignancies. A mismatch between donor and patient for specific HLA-alleles can also be exploited to induce a selective T cell response against patient (malignant) hematopoietic cells. If restricting HLA class II molecules are selectively expressed on hematopoietic cells under non-inflammatory circumstances, allo HLA class-II responses may control the tumor with limited risk of GVHD. Alternatively, T cells recognizing hematopoiesis-restricted antigens presented in the context of mismatched HLA alleles may be used to treat patients with hematological cancers. This review discusses various ways to manipulate the allo-immune response aiming to exploit the powerful ability of allo-reactive T-cells to control the malignancies without causing severe damage to non-hematopoietic tissues.
    No preview · Article · Oct 2015 · Molecular oncology
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    ABSTRACT: Allergic bronchopulmonary aspergillosis (ABPA) is characterized by an allergic immunological response to A.fumigatus. In this study we investigated whether certain Aspergillus antigens are more allergenic than others, as was postulated previously. We stimulated PBMC from patients with ABPA with the classically described A.fumigatus allergens Aspf1, Aspf2, Aspf3 and Aspf4, as well as 2 other Aspergillus antigens, Crf1 and Catalase1. Activated CD4+ T-cells displayed a Thelper2 phenotype with production of IL-4 in response to stimulation with several of these different antigens. Immune responses were not limited to the classically described A.fumigatus allergens. In healthy individuals we demonstrated a similar recognition profile to the different antigens, but in contrast the activated CD4+ T-cells exerted a Thelper1 phenotype and mainly produced IFNγ after stimulation with A.fumigatus antigens. In conclusion, irrespective of the A.fumigatus antigen the T-cell immune response in ABPA patients is skewed to a Thelper2 cytokine secretion profile. Allergic bronchopulmonary aspergillosis (ABPA) is a hypersensitivity pulmonary disease that occurs almost exclusively in patients with asthma or cystic fibrosis (1;2), and is characterized by an allergic immunological response to A.fumigatus. In patients with ABPA IgE, IgA and IgG anti-A.fumigatus antibodies are present (3-6), as well as Aspergillus-specific T-cells with a Th2 phenotype (7-9). The pathogenesis of ABPA is however not completely understood. Previously, it was postulated that certain A.fumigatus antigens, like Aspf1 or Aspf2 are more allergenic than others (6;8;9), and it was suggested this might play a role in the development of ABPA. In this study we studied T-cell responses against 4 classically described A.fumigatus allergens and 2 other A.fumigatus antigens in patients with ABPA, and compared these with healthy controls. This article is protected by copyright. All rights reserved.
    No preview · Article · Jul 2015 · Allergy
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    Full-text · Article · Jun 2015 · Haematologica
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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.
    Full-text · Article · Mar 2015 · Bone marrow transplantation
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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.
    Full-text · Article · Feb 2015 · Cancer Research
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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.
    Full-text · Article · Feb 2015 · Bone Marrow Transplantation
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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.
    No preview · Article · Jan 2015 · Clinical Cancer Research
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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.
    Full-text · Article · Dec 2014 · Journal of Biological Chemistry
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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.
    No preview · Article · Nov 2014 · Blood

  • No preview · Conference Paper · Nov 2014
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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.
    No preview · Article · Oct 2014 · Blood
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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.
    Full-text · Article · Jul 2014 · Journal of Proteomics
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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.
    Preview · Article · Apr 2014 · Haematologica
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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.
    Full-text · Article · Mar 2014 · Bone marrow transplantation
  • Boris van der Zouwen · Alwine B Kruisselbrink · J H Frederik Falkenburg · Inge Jedema
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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.
    No preview · Article · Mar 2014 · Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation
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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.
    Full-text · Article · Feb 2014 · Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation

  • No preview · Article · Feb 2014
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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.
    Full-text · Article · Jan 2014 · PLoS ONE

Publication Stats

9k Citations
2,047.76 Total Impact Points

Institutions

  • 1986-2015
    • Leiden University
      • Molecular Cell Biology Group
      Leyden, South Holland, Netherlands
  • 1983-2015
    • Leiden University Medical Centre
      • • Department of Hematology
      • • Department of Clinical Pharmacy and Toxicology
      • • Department of Immunhematology and Blood Transfusion
      Leyden, South Holland, Netherlands
  • 2002
    • Rijnland Hospital, Leiderdorp
      Лейдердорпе, South Holland, Netherlands
  • 1989-2002
    • Indiana University-Purdue University Indianapolis
      • • Department of Microbiology and Immunology
      • • Department of Medicine
      Indianapolis, Indiana, United States
  • 1998
    • Free University of Brussels
      • Physiology (FYSP)
      Bruxelles, Brussels Capital Region, Belgium