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ABSTRACT: Mesenchymal stromal cells (MSC) are an important component of the bone marrow microenvironment. Notch ligands expressed by MSC are known to play a regulatory role for hematopoietic stem and progenitor cells (HSPC) and support of bone marrow homeostasis. While the role of Notch signaling in HSPC, their progeny and MSC has been relatively well studied, little is known about the Notch-independent regulatory impact of Notch ligands on MSC themselves. In the present study, we used genetically engineered bone marrow-derived human MSC to study the function role of Jagged-1 and the Jagged-1 intracellular domain (JICD) with respect to the interaction with HSPC. We demonstrate that Jagged-1 in human MSC undergoes cleavage to produce an intracellular domain that translocates into the nucleus. JICD but not Jagged-1 overexpression was associated with an increased expression of stromal cell-derived factor-1 (SDF-1). Short-term co-culture (7 days) of HSPC with JICD-overexpressing MSC, but not with Jagged-1-overexpressing MSC, led to increased proliferation of CD34+ progenitors. In contrast, long-term co-culture of HSPC with Jagged-1-overexpressing MSC (up to 6 weeks) led to a significantly better support of cobblestone area-forming cells (CAFCs) and long-term culture-initiating cells (LTC-ICs) compared to JICD-overexpressing MSC. Taken together, results of this study indicate that full-length Jagged-1 and JICD have differential effects on MSC and on their interaction with HSPC ex-vivo. JICD-overexpressing MSC induce proliferation of HSPCs in short-term culture at the expense of immature precursors (LTC-ICs), whereas Jagged-1 overexpressing MSC favor LTC-IC formation.
Stem cells and development 06/2013; · 4.15 Impact Factor
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Ruben A Ferrer,
Manja Wobus,
Catrin List,
Rebekka Wehner,
Claudia Schönefeldt,
Barbara Brocard,
Brigitte Mohr,
Martina Rauner,
Marc Schmitz,
Maik Stiehler,
Gerhard Ehninger,
Lorenz C Hofbauer, Martin Bornhäuser,
Uwe Platzbecker
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ABSTRACT: The contribution of the bone marrow microenvironment in myelodysplastic syndrome is controversially discussed. Therefore, the functional properties of primary mesenchymal stromal cells from patients with myelodysplastic syndrome were analyzed in the presence or absence of lenalidomide. Compared to healthy controls, clonality and growth were reduced across all disease stages. Further, differentiation defects and particular expression of adhesion and cell surface molecules (e.g. CD166, CD29, CD146) were detected. Interestingly, the levels of stromal derived factor 1-alpha (SDF-1α) in patients' cells culture supernatants were almost 2-fold lower (p<0.01) compared to controls and this was paralleled by a reduced induction of migration of CD34+ hematopoietic cells. Cocultures of mesenchymal stromal cells from patients with CD34+ cells from healthy donors resulted in reduced numbers of cobblestone area forming-cells and fewer colony forming units. Exposure of stromal cells from patients and controls to lenalidomide led to a further reduction of SDF-1α secretion and cobblestone area formation, respectively. Moreover, lenalidomide pretreatment of mesenchymal stromal cells from low but not high-risk myelodysplastic syndrome was able to rescue impaired erythroid and myeloid colony formation of early hematopoietic progenitors. In conclusion, our analyses support the notion that the stromal microenvironment is involved in the pathophysiology of myelodysplastic syndrome thus representing a potential target for therapeutic interventions.
Haematologica 05/2013; · 6.42 Impact Factor
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Nona Shayegi,
Michael Kramer, Martin Bornhäuser,
Markus Schaich,
Johannes Schetelig,
Uwe Platzbecker,
Christoph Röllig,
Caroline Heiderich,
Olfert Landt,
Gerhard Ehninger,
Christian Thiede
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ABSTRACT: Mutations of the NPM1-gene (NPM1(mut)) are among the most common genetic alterations in acute myeloid leukemia (AML) and suitable for minimal residual disease (MRD) detection. Retrospectively, we investigated the prognostic impact of NPM1(mut) based MRD-detection from bone marrow for development of relapse by using a newly developed Real-Time PCR based on LNA-containing primers in 174 patients, 155 of whom were treated within prospective protocols. The prognostic value of five cut-off values after completion of treatment or after allogeneic transplantation was studied using cause specific hazard models. Subsequent validation using cross-validated partial likelihood analysis revealed that a rise over 1% NPM1(mut)/ABL1 was most prognostic for relapse after chemotherapy, whereas after allogeneic transplantation an increase over 10% NPM1mut/ABL1 was most prognostic for relapse. Univariate and multivariate analysis of disease free survival and overall survival revealed a significantly worse outcome in patients exceeding >1% NPM1(mut)/ABL1 and > 10% NPM1mut/ABL1 respectively, which remained significant after adjustment for FLT3-ITD status. Our results in a large data set define and optimize cut-off values for early diagnosis of molecular relapse. These results may be especially important to define triggers for early therapeutic intervention.
Blood 05/2013; · 9.90 Impact Factor
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Markus Schaich,
Stefani Parmentier,
Michael Kramer,
Thomas Illmer,
Friedrich Stölzel,
Christoph Röllig,
Christian Thiede,
Mathias Hänel,
Kerstin Schäfer-Eckart,
Walter Aulitzky, [......],
Hubert Serve,
Wolfgang E Berdel,
Jiri Mayer,
Norbert Schmitz,
Stefan W Krause,
Andreas Neubauer,
Claudia D Baldus,
Johannes Schetelig, Martin Bornhäuser,
Gerhard Ehninger
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ABSTRACT: PURPOSETo assess the treatment outcome benefit of multiagent consolidation in young adults with acute myeloid leukemia (AML) in a prospective, randomized, multicenter trial. PATIENTS AND METHODS
Between December 2003 and November 2009, 1,179 patients (median age, 48 years; range, 16 to 60 years) with untreated AML were randomly assigned at diagnosis to receive either standard high-dose cytarabine consolidation with three cycles of 18 g/m(2) (3× HD-AraC) or multiagent consolidation with two cycles of mitoxantrone (30 mg/m(2)) plus cytarabine (12 g/m(2)) and one cycle of amsacrine (500 mg/m(2)) plus cytarabine (10 g/m(2); MAC/MAMAC/MAC). Allogeneic and autologous hematopoietic stem-cell transplantations were performed in a risk-adapted and priority-based manner.ResultsAfter double induction therapy using a 3 + 7 regimen including standard-dose cytarabine and daunorubicin, complete remission was achieved in 65% of patients. In the primary efficacy population of patients evaluable for consolidation outcomes, consolidation with either 3× HD-AraC or MAC/MAMC/MAC did not result in any significant difference in 3-year overall (69% v 64%; P = .18) or disease-free survival (46% v 48%; P = .99) according to the intention-to-treat analysis. Furthermore, MAC/MAMAC/MAC led to additional GI and hepatic toxicity and a higher rate of infection and bleeding, resulting in significantly shorter 3-year overall survival in the per-protocol analysis compared with 3× HD-AraC (63% v 72%; P = .04). CONCLUSION
In younger adults with AML, multiagent consolidation using mitoxantrone and amsacrine in combination with high-dose cytarabine does not improve treatment outcome and confers additional toxicity.
Journal of Clinical Oncology 04/2013; · 18.37 Impact Factor
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Brigitte Mohr,
Johannes Schetelig,
Kerstin Schäfer-Eckart,
Norbert Schmitz,
Mathias Hänel,
Wolf Rösler,
Norbert Frickhofen,
Hartmut Link,
Andreas Neubauer,
Ulrich Schuler,
Uwe Platzbecker,
Jan M Middeke,
Gerhard Ehninger, Martin Bornhäuser,
Markus Schaich,
Friedrich Stölzel
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ABSTRACT: The role of allogeneic stem cell transplantation (HSCT) as compared to chemotherapy in acute myeloid leukaemia (AML) patients with abnormalities of chromosome 17p [abnl(17p)] has not yet been defined. Therefore, we analysed 3530 AML patients treated in three randomized, prospective, controlled clinical trials and compared post-remission therapies using a multivariate Cox regression analysis to determine whether allogeneic HSCT is superior than chemotherapy in overcoming the detrimental impact of patients with abnl(17p) AML. One hundred and forty-three patients (4%) were identified with abnl(17p) AML. All patients had received intensive induction chemotherapy. Forty-seven patients with a median age of 54 years (18-69 years) proceeded to allogeneic HSCT in first or second remission. The 3-year overall survival (OS) rate for the entire cohort of patients was 4% [95% confidence interval (CI), 1-7%]. OS and event-free survival at 3 years, calculated from the day of HSCT, was 11% (95% CI, 2-20%) and 6% (95% CI, 0-13%), respectively. Multivariate Cox regression analysis showed no benefit of allogeneic HSCT compared to chemotherapy (Hazard Ratio 0·97, 95% CI 0·56-1·67, P = 0·9). In conclusion, allogeneic HSCT does not improve survival in patients with abnl(17p) AML as compared to other adverse cytogenetic risk abnormalities.
British Journal of Haematology 02/2013; · 4.94 Impact Factor
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Hanka Jähnisch,
Rebekka Wehner,
Antje Tunger,
Anja Kunze,
Stephanie Oehrl,
Knut Schäkel,
Jacques Rohayem, Martin Bornhäuser,
Torsten Tonn,
Michael Bachmann,
Marc Schmitz
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ABSTRACT: Imiquimod and resiquimod represent Toll-like receptor (TLR) 7 and 8 agonists, which emerged as attractive candidates for tumor therapy. To elucidate immune cells, which mainly contribute to TLR7/8-mediated antitumoral activity, we investigated the impact of imiquimod and resiquimod on native human 6-sulfo LacNAc (slan) dendritic cells (DCs). We found that both TLR7/8 agonists significantly improve the release of various proinflammatory cytokines by slanDCs and promote their tumor-directed cytotoxic activity. Furthermore, resiquimod efficiently augmented the ability of slanDCs to stimulate T cells and natural killer cells. These results indicate that imiquimod and resiquimod trigger various immunostimulatory properties of slanDCs, which may contribute to their antitumor effects.
Cancer letters 02/2013; · 4.86 Impact Factor
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Claudia Ortlepp,
Christine Steudel,
Caroline Heiderich,
Sina Koch,
Angela Jacobi,
Martin Ryser,
Sebastian Brenner, Martin Bornhäuser,
Benedikt Brors,
Wolf-Karsten Hofmann,
Gerhard Ehninger,
Christian Thiede
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ABSTRACT: Autotaxin (ATX) has been reported to act as a motility and growth factor in a variety of cancer cells. The ATX protein acts as a secreted lysophospholipase D (lysoPLD) by converting lysophosphatidylcholine (LPC) to lysophosphatidic acid (LPA), which signals via G-protein coupled receptors and has important functions in cell migration and proliferation. The current study demonstrates that ATX expression is specifically upregulated and functionally active in acute myeloid leukemia (AML) harboring an internal tandem duplication (ITD) mutation of the FLT3 receptor gene. Moreover, ATX expression was also found in normal human CD34+ progenitor cells and selected myeloid and lymphoid subpopulations. Enforced expression of mutant FLT3-ITD by retroviral vector transduction increased ATX mRNA in selected cell lines, whereas inhibition of FLT3-ITD signaling by sublethal doses of PKC412 or SU5614 led to a significant down-regulation of ATX mRNA and protein levels. In the presence of LPC, ATX expression significantly increased proliferation. LPA induced proliferation, regardless of ATX expression and induced chemotaxis in all tested human leukemic cell lines and human CD34+ progenitors. LPC increased chemotaxis only in cells with high expression of endogenous ATX by at least 80% demonstrating the autocrine action of ATX. Inhibition of ATX using a small molecule inhibitor selectively induced killing of ATX-expressing cell lines and reduced motility in these cells. Our data suggest that the production of bioactive LPA through ATX is involved in controlling proliferation and migration during hematopoiesis and that deregulation of ATX contributes to the pathogenesis of AML.
Experimental hematology 01/2013; · 3.11 Impact Factor
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Stefanie Koristka,
Marc Cartellieri,
Claudia Arndt,
Claudia C Bippes,
Anja Feldmann,
Irene Michalk,
Kristin Wiefel,
Slava Stamova,
Marc Schmitz,
Gerhard Ehninger, Martin Bornhäuser,
Michael Bachmann
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ABSTRACT: The nuclear autoantigen La can be detected on the surface of dying cells. Here we present an assay which enables us to show that La protein is not limited to the surface of dying cells but will be released upon stress-induced cell death. As released La protein tightly binds to the surface of neighboring intact cells we asked the question whether or not La protein could serve as a stress-inducible target e.g. for redirecting of regulatory T cells (Tregs) into damaged tissues to downregulate an immune response. In order to provide first proof of concept we developed a novel fully humanized single-chain bispecific antibody (bsAb) which on the one hand is directed to the La antigen and on the other hand to the CD3 complex of T cells. A cross-linkage of Tregs with La-decorated target cells mediated by this bsAb resulted indeed in the activation of the Tregs in a target-dependent manner. Moreover, such bsAb activated Tregs displayed a potent suppressive capacity and negatively influenced proliferation, expansion and cytokine production of autologous CD4(+) and CD8(+) Teff cells.
Journal of Autoimmunity 01/2013; · 7.37 Impact Factor
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Felix Bahr,
Rebekka Wehner,
Uwe Platzbecker,
Martin Wermke,
Nona Shayegi,
Jan Moritz Middeke,
Christoph Röllig,
Johannes Schetelig,
Gerhard Ehninger,
Marc Schmitz, Martin Bornhäuser,
Sebastian Tuve
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ABSTRACT: Interleukin 17A (IL-17)-producing CD4(+) T helper type 17 (Th17) cells have recently drawn attention as possible effector cells of acute graft-versus-host disease (GvHD) following allogeneic hematopoietic cell transplantation (HCT) in murine models. Their role following allogeneic HCT in humans is unknown. In this prospective study Th17, Th1/17 and Th1 cells were quantified in the peripheral blood of 80 patients within the first 3 months following allogeneic HCT using intracellular cytokine staining and flow cytometry. Within the observation period Th1, Th1/17 and Th17 cells did not reconstitute to levels of healthy controls. In contrast to Th1 cells, no further expansion of Th1/17 and Th17 cells was observed following the 1(st) month after HCT. ATG during conditioning significantly reduced the frequency of Th1/17 and Th17 cells, but not of Th1 cells. Acute GvHD was not associated with significant changes in the size of the Th1, Th1/17 or Th17 cell subsets. CMV reactivation triggered the expansion of all T helper subsets and Th1 cells showed the strongest increase. In contrast, no significant changes were found in the T helper cell compartment of patients with bacterial infections compared to time matched controls. In conclusion, quantitative reconstitution of Th1, Th1/17 and Th17 cells is impaired within the first 3 months after HCT, especially when ATG is administered during conditioning. CMV reactivation, but not acute GvHD or bacterial infection, triggered the absolute expansion of these T cell subsets.
Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation 11/2012; · 3.15 Impact Factor
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Martin Bornhäuser,
Joachim Kienast,
Rudolf Trenschel,
Andreas Burchert,
Ute Hegenbart,
Michael Stadler,
Herrad Baurmann,
Kerstin Schäfer-Eckart,
Ernst Holler,
Nicolaus Kröger, [......],
Wolfgang Hiddemann,
Rainer Schwerdtfeger,
Stefanie Buchholz,
Peter Dreger,
Andreas Neubauer,
Wolfgang E Berdel,
Gerhard Ehninger,
Dietrich W Beelen,
Johannes Schetelig,
Matthias Stelljes
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ABSTRACT: Reduced-intensity conditioning regimens have been developed to minimise early toxic effects and deaths after allogeneic haemopoietic cell transplantation. However, the efficacy of these regimens before this procedure has not been investigated in a randomised trial. In this prospective, open-label randomised phase 3 trial we compared a reduced-intensity fludarabine-based conditioning regimen with a standard regimen in patients with acute myeloid leukaemia in first complete remission.
Patients were aged 18-60 years and had intermediate-risk or high-risk acute myeloid leukaemia (defined by cytogenetics) in first complete remission; an available HLA-matched sibling donor or an unrelated donor with at least nine of ten HLA alleles; and adequate renal, cardiac, pulmonary, and neurological function. Between Nov 15, 2004, and Dec 31, 2009, patients were randomly assigned (1:1, by a computer-based minimisation procedure that balanced patients for age, cytogenetic risk, induction therapy, and donor type) to receive either reduced-intensity conditioning of four doses of 2 Gy of total-body irradiation and 150 mg/m(2) fludarabine or standard conditioning of six doses of 2 Gy of total-body irradiation and 120 mg/kg cyclophosphamide. All patients were given ciclosporin and methotrexate as prophylaxis against graft-versus-host disease. Neither investigators nor patients were blinded to study treatment. Our primary endpoint was the incidence of non-relapse mortality, analysed in the intention-to-treat population. The trial is registered with ClinicalTrials.gov, number NCT00150878.
The trial was stopped early on Dec 31, 2009, because of slow accrual of patients. 99 patients were randomly assigned to receive reduced-intensity conditioning and 96 to receive standard conditioning. The incidence of non-relapse mortality did not differ between the reduced-intensity and standard conditioning groups (cumulative incidence at 3 years 13% [95% CI 6-21] vs 18% [10-26]; HR 0·62 [95% CI 0·30-1·31]). Relapse incidence (cumulative incidence 3 years 28% [95% CI 19-38] vs 26% [17-36]; HR 1·10 [95% CI 0·63-1·90]), disease-free survival (3 year disease-free survival 58% [95% CI 49-70] vs 56% [46-67]; HR 0·85 [95% CI 0·55-1·32]), and overall survival (3 year overall survival 61% [95% CI 50-74] vs 58% [47-70]; HR 0·77 [95% CI 0·48-1·25]) did not differ significantly between groups. Grade 3-4 of oral mucositis was less common in the reduced-intensity group than in the standard conditioning group (50 patients in the reduced-intensity conditioning group vs 73 patients in the standard conditioning group); the frequency of other side-effects such as graft-versus-host disease and increased concentrations of bilirubin and creatinine did not differ significantly between groups.
Reduced-intensity conditioning results in a similar incidence of non-relapse mortality and reduced toxic effects compared with standard conditioning without affecting survival outcomes, and thus could be preferentially used in patients younger than 60 years with acute myeloid leukaemia transplanted in first complete remission.
Medical Faculty of Dresden University.
The lancet oncology 09/2012; 13(10):1035-44. · 14.47 Impact Factor
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Angela Scherwath,
Lena Schirmer,
Margitta Kruse,
Gundula Ernst,
Matthias Eder,
Andreas Dinkel,
Sabine Kunze,
Friedrich Balck, Martin Bornhäuser,
Gerhard Ehninger,
Karin Dolan,
Martin Gramatzki,
Hans-Jochem Kolb,
Pia Heußner,
Hans Wilhelm,
Dietrich W Beelen,
Frank Schulz-Kindermann,
Axel R Zander,
Uwe Koch,
Anja Mehnert
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ABSTRACT: BACKGROUND: Owing to its neurotoxicity, allogeneic hematopoietic stem cell transplantation (HSCT) carries risks for cognitive impairment. In this multicenter study, we prospectively evaluated cognitive functioning and its medical and demographic correlates in patients undergoing allogeneic HSCT. METHODS: A total of 102 patients were consecutively assessed prior to (T(0) ), 100 ± 20 days (T(1) ) after, and 12 ± 1 months (T(2) ) after HSCT (61% men, 41% acute myeloid leukemia). A comprehensive neuropsychological test battery was applied to evaluate attention, memory, executive function, and fine motor function, summing up into 14 test scores. RESULTS: Before and after HSCT, patients performed below test norms in up to 50% of the test scores. Patients were mostly impaired on word fluency (24%, T(0) ), fine motor function, and verbal delayed recall (19% each, T(2) ). Impairment on ≥1/5 cognitive domains occurred in 47% (T(0) ) and 41% (T(2) ) of the patients. Performance (mean z-scores) partially improved over time (i.e., visual span forward, verbal learning, and word fluency). However, from baseline to T(2) , 16% of the patients showed reliable decline on ≥3/14 test scores (reliable change index method). For the majority of neuropsychological subtests, no associations with conditioning intensity, total body irradiation, graft-versus-host disease, cyclosporine treatment, and length of hospital stay were found. Age and premorbid intelligence level were consistently associated with cognition. CONCLUSIONS: Below average cognitive performance is common in this patient group. In addition, a subgroup shows reliable cognitive decline after allogeneic HSCT. Healthcare professionals should be aware of these treatment-related cognitive side effects. Copyright © 2012 John Wiley & Sons, Ltd.
Psycho-Oncology 09/2012; · 3.34 Impact Factor
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Evgeny Klyuchnikov,
Ernst Holler, Martin Bornhäuser,
Guido Kobbe,
Arnon Nagler,
Avichai Shimoni,
Christian Könecke,
Christine Wolschke,
Ulrike Bacher,
Axel R Zander,
Nicolaus Kröger
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ABSTRACT: Thirty myelofibrosis patients (21 males, nine females) with relapse (n = 27) or graft-rejection (n = 3) after dose-reduced allografting underwent a salvage strategy including donor lymphocyte infusions (DLIs) and/or second allogeneic haematopoietic stem cell transplantation (HSCT). Twenty-six patients received a median number of three (range, 1-5) DLIs in a dose-escalated mode starting with a median dose of 1·2 × 10(6) (range, 0·003-8 × 10(6) ) up to median dose of 40 × 10(6) T-cells/kg (range, 10-130 × 10(6) ). 10/26 patients (39%) achieved complete response (CR) to DLIs. Acute (grade II-IV) and chronic graft-versus-host (GvHD) disease occurred in 12% and 36% cases. Thirteen non-responders to DLI and four patients who did not receive DLI due to graft-rejection or acute transformation of the blast phase underwent a second allogeneic HSCT from alternative (n = 15) or the same (n = 2) donor. One patient (6%) experienced primary graft-failure and died. Acute (II-IV) and chronic GvHD were observed in 47% and 46% of patients. Overall responses after second HSCT were seen in 12/15 patients (80%: CR: n = 9, partial response: n = 3). The 1-year cumulative incidence of non-relapse mortality for recipients of a second allograft was 6%, and the cumulative incidence of relapse was 24%. After a median follow-up of 27 months, the 2-year overall survival and progression-free survival for all 30 patients was 70% and 67%, respectively. In conclusion, our two-step strategy, including DLI and second HSCT for non-responding or ineligible patients, is an effective and well-tolerated salvage approach for patients relapsing after reduced-intensity allograft after myelofibrosis.
British Journal of Haematology 08/2012; 159(2):172-81. · 4.94 Impact Factor
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Anja Feldmann,
Claudia Arndt,
Katrin Töpfer,
Slava Stamova,
Franziska Krone,
Marc Cartellieri,
Stefanie Koristka,
Irene Michalk,
Dirk Lindemann,
Marc Schmitz,
Achim Temme, Martin Bornhäuser,
Gerhard Ehninger,
Michael Bachmann
[show abstract]
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ABSTRACT: Prostate cancer is the most common noncutaneous malignancy in men. The prostate stem cell Ag (PSCA) is a promising target for immunotherapy of advanced disease. Based on a novel mAb directed to PSCA, we established and compared a series of murine and humanized anti-CD3-anti-PSCA single-chain bispecific Abs. Their capability to redirect T cells for killing of tumor cells was analyzed. During these studies, we identified a novel bispecific humanized Ab that efficiently retargets T cells to tumor cells in a strictly Ag-dependent manner and at femtomolar concentrations. T cell activation, cytokine release, and lysis of target cells depend on a cross-linkage of redirected T cells with tumor cells, whereas binding of the anti-CD3 domain alone does not lead to an activation or cytokine release. Interestingly, both CD8(+) and CD4(+) T cells are activated in parallel and can efficiently mediate the lysis of tumor cells. However, the onset of killing via CD4(+) T cells is delayed. Furthermore, redirecting T cells via the novel humanized bispecific Abs results in a delay of tumor growth in xenografted nude mice.
The Journal of Immunology 08/2012; 189(6):3249-59. · 5.79 Impact Factor
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Jan M Middeke,
Dietrich Beelen,
Michael Stadler,
Gudrun Göhring,
Brigitte Schlegelberger,
Herrad Baurmann,
Gesine Bug,
Frauke Bellos,
Brigitte Mohr,
Stefanie Buchholz,
Rainer Schwerdtfeger,
Hans Martin,
Ute Hegenbart,
Gerhard Ehninger, Martin Bornhäuser,
Johannes Schetelig
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ABSTRACT: The European LeukemiaNet classification combines a heterogeneous group of aberrations as adverse-risk abnormalities. Our goal was to investigate the outcomes associated with distinct high-risk chromosomal abnormalities in acute myeloid leukemia (AML) after allogeneic hematopoietic stem cell transplantation (HSCT). We performed a retrospective cohort analysis in patients with high-risk AML who received first, HLA-compatible, allogeneic HSCT between January 2005 and December 2008. Data from 236 patients with a median age of 55 years were included. Because complex karyotype (CK), -5/5q-, and abnl(17p) are overlapping categories, a hierarchical classification system based on the presence or absence of abnl(17p) and -5/5q- was developed. Patients with abnl(17p) had a 2-year event-free survival (EFS) of 11% (95% confidence interval [CI], 0%-25%), patients with -5/5q- but no abnl(17p) a 2-year EFS of 29% (95% CI, 14%-44%), and patients with adverse-risk AML but neither of the 2 marker lesions a 2-year EFS of 49% (95% CI, 39%-59%). Notably, complex and monosomal karyotypes lost their prognostic value when these marker lesions were excluded. In conclusion, hierarchical classification of adverse-risk karyotypes by 2 marker lesions, abnl(17p) and -5/5q-, is effective in prognostication of the outcome of allogeneic HSCT in AML.
Blood 07/2012; 120(12):2521-2528. · 9.90 Impact Factor
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ABSTRACT: The concept that mesenchymal stromal cells (MSCs), a component of the hematopoietic microenvironment, can be a target for alloreactive effector cells in the context of graft-vs-host disease has not been investigated in detail. Mixed lymphocyte reaction (MLR) supernatant was used to mimic the inflammatory milieu induced by an allogeneic immune response in vitro. In addition to phenotype and proliferation, we monitored MSC differentiation, gene expression, and support of CD34(+) hematopoietic stem and progenitor cells after priming with MLR supernatant. Priming of MSCs with MLR supernatant led to an 11-fold decrease in cobblestone area-forming cells in the 4-week coculture (p < 0.05) and a threefold decrease of colony-forming unit macrophage in the colony-forming cell assay (p < 0.05). MSC proliferation over 8 days was increased 2.5-fold (p < 0.05). Osteogenic differentiation was enhanced, while adipogenesis was concurrently suppressed. In addition, the surface expression of HLA-DR and intercellular adhesion molecule-1 was increased 20-fold (p = 0.06) and 45-fold (p < 0.05), respectively. This was associated with increased adhesion of hematopoietic stem and progenitor cells to MLR-treated MSCs. In summary, our data shed light on the dysfunction of the stromal environment during graft-vs-host disease, possibly aggravating cytopenia and leading to an enhanced immunogenicity of MSCs.
Experimental hematology 07/2012; 40(11):934-44. · 3.11 Impact Factor
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ABSTRACT: Background. The melanoma cell adhesion molecule defines mesenchymal stromal cells in the human bone marrow that regenerate bone and establish a hematopoietic microenvironment in vivo.(1) The role of the melanoma cell adhesion molecule in primary human mesenchymal stromal cells and the maintenance of hematopoietic stem and progenitor cells during ex vivo culture has not yet been demonstrated.Design and Methods. We applied RNA interference or ectopic overexpression of the melanoma cell adhesion molecule in human mesenchymal stromal cells to evaluate the effect of the melanoma cell adhesion molecule on their proliferation and differentiation as well as its influence on co-cultivated hematopoietic stem and progenitor cells.Results. Knockdown and overexpression of the melanoma cell adhesion molecule affected several characteristics of human mesenchymal stromal cells related to osteogenic differentiation, proliferation, and migration. Furthermore, knockdown of the melanoma cell adhesion molecule in human mesenchymal stromal cells stimulated the proliferation of hematopoietic stem and progenitor cells and strongly reduced the formation of long-term culture-initiating cells. In contrast, melanoma cell adhesion molecule-overexpressing human mesenchymal stromal cells provided a supportive microenvironment for hematopoietic stem and progenitor cells. Expression of the melanoma cell adhesion molecule increased the adhesion of hematopoietic stem and progenitor cells to human mesenchymal stromal cells and their migration beneath the monolayer of human mesenchymal stromal cells. Conclusions. Our results demonstrate that the expression of the melanoma cell adhesion molecule in human mesenchymal stromal cells determines their fate and regulates the maintenance of hematopoietic stem and progenitor cells through direct cell - cell contact.
Haematologica 07/2012; · 6.42 Impact Factor
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Jürgen Finke,
Claudia Schmoor,
Wolfgang A Bethge,
Hellmut D Ottinger,
Matthias Stelljes,
Axel R Zander,
Liisa Volin,
Dominik A Heim,
Rainer Schwerdtfeger,
Karin Kolbe,
Jiri Mayer,
Johan A Maertens,
Werner Linkesch,
Ernst Holler,
Vladimir Koza, Martin Bornhäuser,
Hermann Einsele,
Hartmut Bertz,
Olga Grishina,
Gérard Socié
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ABSTRACT: Several prognostic factors for the outcome after allogeneic hematopoietic stem-cell transplant (HSCT) from matched unrelated donors have been postulated from registry data; however, data from randomized trials are lacking. We present analyses on the effects of patient-related, donor-related, and treatment-related prognostic factors on acute GVHD (aGVHD), chronic GVHD (cGVHD), relapse, nonrelapse mortality (NRM), disease-free survival (DFS), and overall survival (OS) in a randomized, multicenter, open-label, phase III trial comparing standard graft-versus-host-disease (GVHD) prophylaxis with and without pretransplantation ATG-Fresenius (ATG-F) in 201 adult patients receiving myeloablative conditioning before HSCT from HLA-A, HLA-B antigen, HLA-DRB1, HLA-DQB1 allele matched unrelated donors. High-resolution testing (allele) of HLA-A, HLA-B, and HLA-C were obtained after study closure, and the impact of an HLA 10/10 4-digit mismatch on outcome and on the treatment effect of ATG-F versus control investigated. Advanced disease was a negative factor for relapse, DFS, and OS. Donor age ≥40 adversely affected the risk of aGVHD III-IV, extensive cGVHD, and OS. Younger donors are to be preferred in unrelated donor transplantation. Advanced disease patients need special precautions to improve outcome. The degree of mismatch had no major influence on the positive effect of ATG-F on the reduction of aGVHD and cGVHD.
Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation 06/2012; 18(11):1716-26. · 3.15 Impact Factor
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ABSTRACT: Lenalidomide (LEN) has emerged as a promising therapeutic option for the management of various hematologic malignancies. Although its direct mechanisms of action on malignant cells have been studied intensively, its effects on the stromal compartment of bone marrow have not yet been analyzed systematically. Therefore, we investigated whether LEN alters the functional capacity of mesenchymal stromal cells (MSCs) as the main cellular component of the bone marrow microenvironment. In addition to their growth and differentiation characteristics, we focused on the ability of MSC to modulate T-cell function and support hematopoietic stem cells (HSCs).
Bone marrow-derived MSCs were exposed to LEN (10 μM), and differences in proliferation, phenotype, inhibition of T-cell proliferation, and differentiation capacity were analyzed. A Boyden chamber assay was used to test the migratory potential of HSC toward the conditioned medium of LEN-treated or untreated MSCs, and the stromal cell-derived factor-1 (SDF-1) concentrations in these supernatants were determined by enzyme-linked immunosorbent assay.
Treatment of MSCs with LEN did not affect their growth rate, proliferation, osteogenic and adipogenic differentiation potential, or capacity to inhibit T-cell proliferation. However, LEN treatment increased the average of mean fluorescence intensity of CD29 and CD73 by 15 and 22%, respectively. Interestingly, LEN reduced SDF-1 by MSCs by 32% compared to that of control cells. As a functional consequence, the serum-free supernatant of LEN-treated MSCs had a significantly lower potential to induce the directed migration of CD34(+) HSCs.
LEN can modulate the expression of cell surface molecules and the chemokine secretion of MSCs in vitro. These effects might contribute to the clinical effects of the compound in vivo for patients with hematological malignancies.
Experimental hematology 06/2012; 40(10):867-76. · 3.11 Impact Factor
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Konrad Mager,
Rebekka Wehner,
Felix Bahr,
Uta Oelschlägel,
Uwe Platzbecker,
Martin Wermke,
Nona Shayegi,
Jan Moritz Middeke,
Jörgen Radke,
Christoph Röllig,
Johannes Schetelig,
Christian Thiede,
Gerhard Ehninger,
Marc Schmitz, Martin Bornhäuser,
Sebastian Tuve
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ABSTRACT: BACKGROUND: Infections and acute graft-versus-host disease (GvHD) represent major complications of allogeneic stem-cell transplantation (SCT). Dendritic cells (DCs) display an extraordinary capacity to induce innate and adaptive immune responses. Therefore, they play a crucial role in the elimination of pathogens and in the pathogenesis of acute GvHD. 6-Sulfo LacNAc DCs (slanDCs) are a major subpopulation of human blood DCs with a high proinflammatory capacity. We investigated for the first time the reconstitution of slanDCs in the blood of patients after SCT and the modulation of their frequency by bacterial infection, cytomegalovirus (CMV) reactivation, and acute GvHD. METHODS: The frequency of slanDCs, CD1c myeloid DCs (mDCs), and plasmacytoid DCs (pDCs) in the peripheral blood was quantified by flow cytometry in 80 patients after SCT. To assess individual DC subsets, we used pregating of the HLADRLin subset and antibodies against slanDCs, blood DC antigen 1 (CD1c mDCs), and blood DC antigen 2 (pDCs). RESULTS: SlanDCs showed the slowest reconstitution in the first month after SCT compared with CD1c mDCs and pDCs. Interestingly, in the second and third months after SCT, their percentage steadily increased, and slanDCs were the most abundant DC subset. In addition, we observed a markedly reduced frequency of slanDCs in the blood of patients with bacterial infection, CMV reactivation, or severe acute GvHD. Furthermore, slanDCs showed the most prominent reduction after steroid treatment of acute GvHD. CONCLUSIONS: These results indicate that SCT-associated complications such as bacterial infection, CMV reactivation, and acute GvHD can significantly modulate the frequency of slanDCs.
Transplantation 05/2012; · 4.00 Impact Factor
