Frank Preijers - Academia.edu (original) (raw)

Papers by Frank Preijers

$Research paper thumbnail of 088 A phase I/II study on the anti-CD3/CD7 immunotoxin combination for the treatment of steroid-refractory acute GVHD$

Journal of Investigative Dermatology, 2019

Skin aging is a continuous process that affects skin appearance and function manifesting by sever... more Skin aging is a continuous process that affects skin appearance and function manifesting by several outcomes such as wrinkling and sagging. Researchers have identified impacting environmental factors (sun exposure, smoking, etc.) and several molecular mechanisms leading to skin aging. Recently, genome wide association studies (GWAS) have led to new insights into the molecular mechanisms of skin aging. Since individual SNP associations in GWAS explain only a small fraction of the genetic impact in complex polygenic phenotypes, we applied the Gene Set Enrichment Analysis on the genotype data obtained from 502 verywell characterized women to identify biological pathways associated with four major outcomes of skin aging, namely photoaging, solar lentigines, wrinkling, and sagging. This analysis revealed new relevant pathways and genes, some likely specific of skin aging such as the WNT7B and PRKCA genes in the "melanogenesis" pathway, some likely involved in global aging such as the DDB1 gene in the "nucleotide excision repair" pathway, not picked up in the previously published GWAS. Overall, our results suggest that photoaging, solar lentigines, wrinkling, and sagging involve specific molecular mechanisms such as "proteasome" and "mTOR signaling pathway" but also shared molecular mechanisms such as "nucleotide excision repair".

$Research paper thumbnail of A Phase I/II Study on the Anti-CD3/CD7 Immunotoxin Combination (T-GuardTM) for the Treatment of Steroid-Refractory Acute Gvhd$

Blood, 2017

Background More effective therapies for steroid-refractory acute graft-versus-host disease (SR-aG... more Background More effective therapies for steroid-refractory acute graft-versus-host disease (SR-aGVHD) are urgently needed. Because infections and relapse of the hematological malignancy contribute to the dismal overall survival (OS) therapies that limit the duration of immune suppression after achieving a remission might be preferred. The immunotoxin (IT)-combination (T-GuardTM) consists of two antibody-drug conjugates (i.e. Ricin A) that target CD3 and CD7 on activated T lymphocytes and has shown efficacy as third-line therapy in SR-aGVHD while allowing fast immune reconstitution. Objectives We conducted a prospective phase I/II multicenter trial on the safety and efficacy of T-GuardTM for the treatment of SR-aGVHD (NCT02027805). Methods Adult patients with grade II-IV SR-aGVHD were eligible for inclusion. Exclusion criteria consisted of the presence of uncontrolled infections, signs of chronic GVHD, severe renal impairment and severe hypoalbuminemia. T-Guard was given as 4-hour in...

CHAPTER 1: Introduction. CHAPTER 2: Relationship between internalization and cytotoxicity of rici... more CHAPTER 1: Introduction. CHAPTER 2: Relationship between internalization and cytotoxicity of ricin Α-chain immunotoxins. CHAPTER 3: Different susceptibilities of normal Τ cells and Τ cell lines to immunotoxins. CHAPTER 4: Human Τ lymphocyte differentiation antigens as target for immunotoxins or complementmediated cytotoxicity. CHAPTER 5: Cytotoxic potential of anti-CD7 immunotoxin (WTl-ricin A) to purge ex vivo malignant Τ cells in bone marrow. CHAPTER 6: Autologous transplantation of bone marrow purged in vitro with anti-CD7-(WTl-) ricin A immunotoxin in Τ cell lymphoblastic leukemia and lymphoma.

eLife, 2020

Atherosclerosis is the major cause of cardiovascular disease (CVD). Monocyte-derived macrophages ... more Atherosclerosis is the major cause of cardiovascular disease (CVD). Monocyte-derived macrophages are the most abundant immune cells in atherosclerotic plaques. In patients with atherosclerotic CVD, leukocytes have a hyperinflammatory phenotype. We hypothesize that immune cell reprogramming in these patients occurs at the level of myeloid progenitors. We included 13 patients with coronary artery disease due to severe atherosclerosis and 13 subjects without atherosclerosis in an exploratory study. Cytokine production capacity after ex vivo stimulation of peripheral blood mononuclear cells (MNCs) and bone marrow MNCs was higher in patients with atherosclerosis. In BM-MNCs this was associated with increased glycolysis and oxidative phosphorylation. The BM composition was skewed towards myelopoiesis and transcriptome analysis of HSC/GMP cell populations revealed enrichment of neutrophil- and monocyte-related pathways. These results show that in patients with atherosclerosis, activation o...

Clinical cancer research : an official journal of the American Association for Cancer Research, Jan 9, 2017

Older acute myeloid leukemia (AML) patients have a poor prognosis, therefore novel therapies are ... more Older acute myeloid leukemia (AML) patients have a poor prognosis, therefore novel therapies are needed. Allogeneic natural killer (NK) cells have been adoptively transferred with promising clinical results. Here, we report the first-in-human study exploiting an unique scalable NK cell product generated ex vivo from CD34+ hematopoietic stem and progenitor cells (HSPCs) from partially HLA-matched umbilical cord blood units. Ten older AML patients in morphologic complete remission received an escalating HSPC-NK cell dose (between 3 and 30x10^6/kg body weight) after lymphodepleting chemotherapy without cytokine boosting. HSPC-NK cell products contained a median of 75% highly activated NK cells, with <1x10^4 T cells/kg and <3x10^5 B cells/kg body weight. HSPC-NK cells were well tolerated and no graft-versus-host disease nor toxicity was observed. Despite no cytokine boosting was given, transient HSPC-NK cell persistence was clearly found in peripheral blood up to 21% until day 8, ...

Bone marrow transplantation, 1996

After lymphocyte-depleted BMT, CD8+ T cells have been expanded to or above normal levels in 45% o... more After lymphocyte-depleted BMT, CD8+ T cells have been expanded to or above normal levels in 45% of the recipients within 3 months. The mechanisms underlying proliferation of donor-derived CD8+ T cells after BMT are still unclear. We investigated whether these CD8+ T cells proliferate in response to specific antigens by determination of TCR clonality and whether these cells exert specific cytotoxicity. PCR analysis of TCR-gamma gene rearrangements showed a marked clonal predominance in CD8+ T cells of recipients with a high number of these cells. Strong association between expansion of CD8+ T cells and CMV infection suggests involvement of CMV antigens. Therefore, we examined CMV-specific cytotoxicity of freshly isolated CD8+ T cells of two BMT recipients with clonal expansion after the onset of CMV infection. CD8+ T cells exerted HLA-restricted cytotoxicity directed against CMV-infected fibroblasts indicating that CMV stimulates proliferation. The majority of CD8+ T cells in these r...

Cancer research, 1994

We analyzed the effect of isotype variation on effectiveness of B-cell CD19 immunotoxins (IT) by ... more We analyzed the effect of isotype variation on effectiveness of B-cell CD19 immunotoxins (IT) by using class switch variants (CLB-B4-IgG1 and CLB-B4-IgG2a) conjugated to ricin A. Notably, IgG1-IT appeared to be approximately 100-fold more potent than IgG2a-IT toward B-cell lines Daudi and KM3. Binding and internalization studies with 125I-labeled monoclonal antibodies (mAbs) revealed a higher cellular uptake of IgG1 compared to IgG2a, despite similar binding affinities. Following removal of the Fc part, both mAbs internalized at the same rate as IgG2a, indicating that the Fc part of IgG1 is involved in enhanced cellular uptake. The involvement of Fc gamma RII (CD32) in this process was demonstrated by a decreased cytotoxicity of IgG1-IT (and not IgG2a-IT) in the presence of Fc gamma RII-blocking mAbs AT10 or IV.3. To identify the isoform responsible for this phenomenon, internalization of IgG1 and IgG2a in 11 B-cell lines and malignant B-cells of 8 patients was compared with express...

Blood Cancer Journal, 2013

Flow-cytometric detection of minimal residual disease (MRD) has proven in several single-institut... more Flow-cytometric detection of minimal residual disease (MRD) has proven in several single-institute studies to have an independent prognostic impact. We studied whether this relatively complex approach could be performed in a multicenter clinical setting. Five centers developed common protocols to accurately define leukemia-associated (immuno)phenotypes (LAPs) at diagnosis required to establish MRD during/after treatment. List mode data files were exchanged, and LAPs were designed by each center. One center, with extensive MRD experience, served as the reference center and coordinator. In quarterly meetings, consensus LAPs were defined, with the performance of centers compared with these. In a learning (29 patients) and a test phase (35 patients), a mean of 2.2 aberrancies/patient was detected, and only 1/63 patients (1.6%) had no consensus LAP(s). For the four centers without (extensive) MRD experience, clear improvement could be shown: in the learning phase, 39-63% of all consensus LAPs were missed, resulting in a median 30% of patients (range 21-33%) for whom no consensus LAP was reported; in the test phase, 27-40% missed consensus LAPs, resulting in a median 16% (range 7-18%) of 'missed' patients. The quality of LAPs was extensively described. Immunophenotypic MRD assessment in its current setting needs extensive experience and should be limited to experienced centers.

Leukemia, 2002

Using red cell phenotyping (RCP) and/or cytogenetics (CYT) we identified 19 patients with persist... more Using red cell phenotyping (RCP) and/or cytogenetics (CYT) we identified 19 patients with persisting mixed chimerism (MC) among 231 patients transplanted with partially T cell-depleted stem cell grafts from HLA-identical siblings. Persisting MC is defined as MC for more than 2 years in patients without any evidence of relapse. Median leukemia-free survival in these patients was 150 (range, 50-218) months. Diagnoses were ALL (n ‫؍‬ 10); AML (n ‫؍‬ 2); CML (n ‫؍‬ 2); NHL (n ‫؍‬ 2); MDS (n ‫؍‬ 1); MM (n ‫؍‬ 1) and SAA (n ‫؍‬ 1). Purpose of this study was the longterm follow-up of MC and definition of patterns of chimerism in the various subsets of PBMCs and granulocytes. Using a PCR-STR technique CD3 + /CD4 + (T4 lymphocytes), CD3 + /CD8 + (T8 lymphocytes), CD45 + /CD19 + (B lymphocytes), CD45 + /CD14 + (monocytes), CD45 + /CD15 + (granulocytes) and CD3 − /CD56 + (NKcells) were analyzed. The majority of patients with persisting MC were conditioned with a less intensive conditioning regimen and had little GVHD. Sequential monitoring of the chimerism resulted in a group of patients (n ‫؍‬ 7) with very slow transient mixed chimerism that resulted in complete DC after median 7 years. Another nine patients had a relatively high percentage of persisting autologous cells for a median of 12 years and in three patients we observed a stable low percentage of autologous cells. Only two out of 19 patients (AML-CR1, CML-CP1) relapsed during follow-up. Both patients had a relatively high percentage of autologous cells. Chimerism in granulocytes and PBMC subsets was analyzed at a median of 8 years after SCT in nine patients. In five patients mixed chimerism simultaneously detected by RCP and CYT was associated with MC in all subsets. Within each individual patient the percentages of donor and recipient cells were very different between the different subsets. Two CML-CP1 patients were mixed chimera in only two subsets and in one patient these subsets represented pending relapse. In another two patients mixed chimerism with a very low number of autologous red cells was not found in the PBMCs because of the different sensitivity level of the RCP and the PCR-STR technique. We conclude that in patients with persisting mixed chimerism after partially T cell-depleted SCT a remarkable number of patients had lymphoid malignancies, the majority of the patients were conditioned with less intensive conditioning regimens and the mixed chimerism was not correlated with relapse. Chimerism in granulocytes and PBMC subsets did show great intra-individual differences in the subsets and these data correlated well with RCP and CYT data with the exception of the NK cells.

Proceedings of the National Academy of Sciences, 2012

Adaptive features of innate immunity, recently described as “trained immunity,” have been documen... more Adaptive features of innate immunity, recently described as “trained immunity,” have been documented in plants, invertebrate animals, and mice, but not yet in humans. Here we show that bacille Calmette-Guérin (BCG) vaccination in healthy volunteers led not only to a four- to sevenfold increase in the production of IFN-γ, but also to a twofold enhanced release of monocyte-derived cytokines, such as TNF and IL-1β, in response to unrelated bacterial and fungal pathogens. The enhanced function of circulating monocytes persisted for at least 3 mo after vaccination and was accompanied by increased expression of activation markers such as CD11b and Toll-like receptor 4. These training effects were induced through the NOD2 receptor and mediated by increased histone 3 lysine 4 trimethylation. In experimental studies, BCG vaccination induced T- and B-lymphocyte–independent protection of severe combined immunodeficiency SCID mice from disseminated candidiasis (100% survival in BCG-vaccinated m...

PLoS ONE, 2011

Natural killer (NK) cell-based adoptive immunotherapy is a promising treatment approach for many ... more Natural killer (NK) cell-based adoptive immunotherapy is a promising treatment approach for many cancers. However, development of protocols that provide large numbers of functional NK cells produced under GMP conditions are required to facilitate clinical studies. In this study, we translated our cytokine-based culture protocol for ex vivo expansion of NK cells from umbilical cord blood (UCB) hematopoietic stem cells into a fully closed, large-scale, cell culture bioprocess. We optimized enrichment of CD34 + cells from cryopreserved UCB units using the CliniMACS system followed by efficient expansion for 14 days in gas-permeable cell culture bags. Thereafter, expanded CD34 + UCB cells could be reproducibly amplified and differentiated into CD56 + CD3 2 NK cell products using bioreactors with a mean expansion of more than 2,000 fold and a purity of .90%. Moreover, expansion in the bioreactor yielded a clinically relevant dose of NK cells (mean: 2610 9 NK cells), which display high expression of activating NK receptors and cytolytic activity against K562. Finally, we established a versatile closed washing procedure resulting in optimal reduction of medium, serum and cytokines used in the cell culture process without changes in phenotype and cytotoxic activity. These results demonstrate that large numbers of UCB stem cell-derived NK cell products for adoptive immunotherapy can be produced in closed, large-scale bioreactors for the use in clinical trials.

PLoS ONE, 2008

A b s tra c t Background:Treg based immunotherapy is of great interest to facilitate tolerance in... more A b s tra c t Background:Treg based immunotherapy is of great interest to facilitate tolerance in autoimmunity and transplantation. For clinical trials, it is essential to have a clinical grade Treg isolation protocol in accordance with Good Manufacturing Practice (GMP) guidelines. To obtain sufficient Treg for immunotherapy, subsequent ex vivo expansion might be needed. M eth o d o lo g y/P rin c ip a l Findings: Treg were isolated from leukapheresis products by CliniMACS based GMP isolation strategies, using anti-CD25, anti-CD8 and anti-CD19 coated microbeads. CliniMACS isolation procedures led to 40-60% pure CD4posCD25highFoxP3pos Treg populations that were anergic and had moderate suppressive activity. Such CliniMACS isolated Treg populations could be expanded with maintenance of suppressive function. Alloantigen stimulated expansion caused an enrichment of alloantigen-specific Treg. Depletion o f unwanted CD19pos cells during CliniMACS Treg isolation proved necessary to prevent B-cell outgrowth during expansion. CD4posCD127pos conventional T cells were the major contaminating cell type in CliniMACS isolated Treg populations. Depletion o f CD127pos cells improved the purity of CD4posCD25highFoxP3pos Treg in CliniMACS isolated cell populations to approximately 90%. Expanded CD127neg CliniMACS isolated Treg populations showed very potent suppressive capacity and high FoxP3 expression. Furthermore, our data show that cryopreservation o f CliniMACS isolated Treg is feasible, but that activation after thawing is necessary to restore suppressive potential. C onclusions/Significance:The feasibility o f Treg based therapy is widely accepted, provided that tailor-made clinical grade procedures for isolation and ex vivo cell handling are available. We here provide further support for this approach by showing that a high Treg purity can be reached, and that isolated cells can be cryopreserved and expanded successfully.

New England Journal of Medicine, 2014

Journal of Translational Medicine, 2011

The Journal of Infectious Diseases, 2000

PloS one, 2012

A better understanding of human NK cell development in vivo is crucial to exploit NK cells for im... more A better understanding of human NK cell development in vivo is crucial to exploit NK cells for immunotherapy. Here, we identified seven distinctive NK cell developmental stages in bone marrow of single donors using 10-color flow cytometry and found that NK cell development is accompanied by early expression of stimulatory co-receptor CD244 in vivo. Further analysis of cord blood (CB), peripheral blood (PB), inguinal lymph node (inLN), liver lymph node (liLN) and spleen (SPL) samples showed diverse distributions of the NK cell developmental stages. In addition, distinctive expression profiles of early development marker CD33 and C-type lectin receptor NKG2A between the tissues, suggest that differential NK cell differentiation may take place at different anatomical locations. Differential expression of NKG2A and stimulatory receptors (e.g. NCR, NKG2D) within the different subsets of committed NK cells demonstrated the heterogeneity of the CD56(bright)CD16⁺/⁻ and CD56(dim)CD16⁺ subset...

Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K, 2012

Flow cytometry (FC) is increasingly recognized as an important tool in the diagnosis and prognosi... more Flow cytometry (FC) is increasingly recognized as an important tool in the diagnosis and prognosis of myelodysplastic syndromes (MDS). However, validation of current assays and agreement upon the techniques are prerequisites for its widespread acceptance and application in clinical practice. Therefore, a working group was initiated (Amsterdam, 2008) to discuss and propose standards for FC in MDS. In 2009 and 2010, representatives from 23, mainly European, institutes participated in the 2nd and 3rd European LeukemiaNet (ELN) MDS workshops. In the present report, minimal requirements to analyze dysplasia are refined. The proposed core markers should enable a categorization of FC results in cytopenic patients as "normal", "suggestive of" or "diagnostic of" MDS. A FC report should include a description of validated FC abnormalities such as aberrant marker expression on myeloid progenitors and, furthermore, dysgranulopoiesis and/or dysmonocytopoiesis if at l...

Japanese Journal of Cancer Research, 2002

An anchorage-dependent Wilms tumor cell line HFWT was found to stimulate selective and remarkable... more An anchorage-dependent Wilms tumor cell line HFWT was found to stimulate selective and remarkable expansion of human natural killer (NK) cells from human peripheral blood mononuclear cells (PBMC). After PBMC of healthy donors were cultured on irradiated HFWT cells for 10-21 days, the lymphocytes expanded 58-to 401-fold. This NK cell expansion required direct contact of PBMC with live, but not fixed, HFWT cells. The PBMC from an end-stage brain tumor patient also expanded 156-fold, whereas those cultured with irradiated NK-sensitive K562 grew only 30.5-fold. CD16 + CD56 + NK cells accounted for more than 70% of the population expanded on HFWT cells. No essential difference in expression of NK receptors was observed in the expanded NK cells on HFWT and K562 and without feeder cells. The expanded NK cells killed not only fresh HFWT cells but, unexpectedly, also MHC class I-expressing autologous brain tumor cells at an effector/target ratio of 4 for 24 h. These results will contribute to the development of a large-scale preparation method for human NK cells, which will aid studies of NK cell biology and possible treatment of brain tumors.