bluuebunny/crossref_metadata_2025_split huggingface dataset:
Toll-Like Receptor 2 and 4 Are Expressed on Transplanted T-Cell Subsets and Influence Outcome after Allogeneic Unrelated Peripheral Blood Stem Cell Transplantation.
Authors: Platzbecker Uwe , Stoehlmacher Jan , Goekkurt Eray , Pabst Caroline , Thiede Christian , Ehninger Gerhard , Bornhäuser Martin 
Published: October 2019

Abstract
Introduction: Recently, Toll-like receptor (TLR) 2 and 4 have been identified as the most important receptors for LPS, which is contained in the cell wall of gram-negative bacteria and is known to be a main inducer of graft versus host disease (GVHD). The role of TLR expressing T-cells within the graft for the induction of GVHD in patients after unrelated peripheral blood stem cell transplantation (PBSCT) is unknown.
Methods and patients: We therefore determined by flow cytometry expression of TLR 2 and 4 on T-cells within the graft of 63 patients receiving unrelated PBSCT after intensive conditioning followed by cyclosporine A and methotrexate as GVHD prophylaxis. Additionally, donor specific single nucleotide polymorphisms (SNP) for TLR2 (R753Q), TLR4 (D299G) and TLR4 (Y135A) were determined. The data were finally correlated with clinical endpoints.
Results: As expected, TLRs were not expressed on T-cells in peripheral blood of healthy donors (TLR 2: <1.0%, TLR4 <0.5% of T-cells, n=6). In contrast we detected a distinct up-regulation of these receptors on T-cells within the grafts. TLR2 and TLR4 expression on CD4+ T-cells ranged from 1.2%–12.3% (median 3.1%) for TLR2 and from 1.2%–12.0% (median 3.7%) for TLR4. Among the CD8+ T-cells 0.9%–16.1% (median 3.3%) expressed TLR2 and 0.7%–13.3% (median 3.5%) expressed TLR4. The SNP for TLR2 (R753Q) and TLR4 (D299G) was found in 8.6% and 10.3% of the allogeneic donors, respectively but did neither correlate with the expression levels of TLR on T-cells nor with clinical endpoints. Treatment-related mortality from infections was observed in 10 patients (16%). Interestingly, higher expression of TLR2 and 4 on CD4+ but not CD8+ T-cells was significantly associated with an increased cumulative incidence of fatal infections (38% vs. 8% p=0.03 for TLR2 and 37% vs. 9% p=0.007 for TLR4). Neither the overall CD3+, CD4+ and CD8+ cell dose nor the expression of TLR2 and 4 on CD4+ and CD8+ T-cells showed a significant association with the incidence of acute or chronic GVHD or relapse.
Conclusion: These data suggest a previously unrecognised up-regulation of TLR, on CD4+ and CD8+ T-cells contained in G-CSF mobilized apheresis products. Whether these phenotypic changes impact on T cell function or patient outcome warrants further investigation.
Acute Graft-Versus-Host Disease Following Umbilical Cord Blood Transplantation: Retrospective Survey Involving 2015 Japanese Patients.
Authors: Miyakoshi Shigesaburo , Yamaguchi Takuhiro , Kami Masahiro , Matsumura Tomoko , Yuji Koichiro , Narimatsu Hiroto , Kusumi Eiji , Sakamaki Hisashi , Kouzai Yasushi , Okada Masaya , Osugi Yuko , Kobayashi Ryuji , Inoue Masami , Takahashi Satoshi , Murata Makoto , Kai Shunro , Kato Koji , Nagamura Tokiko , Taniguchi Shuichi , Kato Shunichi K. 
Published: October 2019

Abstract
BACKGROUND Limited information is available on incidences and clinical features of acute graft-versus host disease (GVHD) after cord blood transplantation and large-sized researches have been awaited.
METHODS We investigated the incidences and clinical features of acute GVHD in 2,015 patients reported to the Japan Cord Blood Bank Network, who underwent cord blood transplantation between June 1997 and August 2006.
RESULTS Of 2,015 patients, 1481 patients (73%) achieved neutrophil engraftment at a median of day 22 (range, 6–81). Cumulative incidence of neutrophil recovery at day 100 was 0.74 (95%CI, 0.73–0.76). Of 2015 patients, 708 patients developed grade II-IV acute GVHD: grade II (n=423), grade III (n=237), and grade IV (n=48). The median onset was day 19 (range, 4–190). The cumulative incidences of grade II-IV and III-IV acute GVHD at day 100 were 0.35 (95% CI, 0.33–0.37) and 0.14 (95% CI, 0.12–0.15), respectively. Skin and gastrointestinal acute GVHD was documented in 1,006 and 405 patients, respectively, whereas liver GVHD was diagnosed in 149 patients. Multivariate analysis identified the following predictors of grade II-IV acute GVHD: the number of infused nucleated cells, transplantation from female donors to female recipients, TBI-containing preparative regimens, methotrexate-containing GVHD prophylaxis, and tacrolimus-based GVHD prophylaxis. Overall survival rates at three years of patients with grade 0-I, II and III-IV GVHD who survived 100 days or longer were 0.58 (95%CI, 0.53–0.63), 0.61 (95% CI, 0.54–0.67) and 0.40 (95%CI, 0.31–0.49), respectively.
CONCLUSIONS Acute GVHD following cord blood transplantation is mild and has graft-versus malignancy effects.
Probability of event free survival after cord blood tranplantation in the patients with grade 0-I, II and III-IV who survived 100 days or longer Event - free survival of the patients with grade 0-I, II and III-IV GVHD who survived 100 days or longer was 0.54 (95% CI, 0.49–0.59), 0.58 (95%CI, 0.52–0.65) and 0.41 (95%CI, 0.32–0.49),respectively,3 years after transplantation. Probability of event free survival after cord blood tranplantation in the patients with grade 0-I, II and III-IV who survived 100 days or longer Event - free survival of the patients with grade 0-I, II and III-IV GVHD who survived 100 days or longer was 0.54 (95% CI, 0.49–0.59), 0.58 (95%CI, 0.52–0.65) and 0.41 (95%CI, 0.32–0.49),respectively,3 years after transplantation.
Is Splenectomy Necessary in Patients with Myelofibrosis and Extensive Splenomegaly Prior to Allogeneic Stem Cell Transplantation with Reduced Intensity Conditioning Regimens?.
Authors: Ciurea Stefan O. , Sadegi Barry , Wilbur Andrew , Alagiozian-Angelova Victoria , Gaitonde Sujata , Dobogai Lisa C. , Akard Luke P. , Hoffman Ronald , Rondelli Damiano 
Published: October 2019

Abstract
Allogeneic stem cell transplantation has been shown to restore normal hematopoiesis in patients with intermediate or high risk primary myelofibrosis (PMF) or myelofibrosis preceeded by polycythemia vera or essential thrombocythemia. However, in patients with PMF and extensive splenomegaly it is not clear whether transplant is associated with an unacceptable risk or if splenectomy should be performed prior to transplant. In this study, ten consecutive patients with myelofibrosis who were not splenectomized received an allogeneic hematopoietic stem cell transplantation (HSCT) from related or unrelated donors and were periodically monitored by ultrasound or CT scan for 12 months after transplant to assess the kinetics of the reduction of splenomegaly. These findings were correlated with the time to resolution of marrow fibrosis, time to engraftment and clinical outcome. Over a 12 month period a progressive reduction in spleen size was observed in all the patients and paralleled the reduction in marrow fibrosis (Figure 1). Of 10 patients, 5 with a splenic longitudinal diameter >30 cm showed a more prolonged time to engraftment ANC> 0.5 × 109/L (d 19±5 vs 13±2, p=0.05) and platelet > 20 × 109/L (d 75±104 vs 11±2, p=0.06). However, of the 5 patients with more extensive splenomegaly only 2 experienced delayed engraftment of platelets (d 77 and 256, respectively). Full donor chimerism was observed in all patients within 60 days after transplantation regardless of the size of the spleen. At a median follow-up of 54 months (range: 2–80) all the patients are alive but 1 who died of a TTP like syndrome 2 months after transplant and had a small spleen at the time of transplant. A clinical CR has been documented in 7 patients and 2 have achieved clinical improvement as assessed by IWG response criteria. These findings suggest that allogeneic HSCT with RIC regimen can be safely performed in patients with extensive splenomegaly. Although in these patients the time to engraftment may be prolonged, the significant risk of morbidity and mortality associated with splenectomy may be avoided.
Figure Figure
The Importance of Harmonizing Transplantation Procedures in Children with ALL: ALL-SCT-BFM 2003, a Prospective Multicenter Trial.
Authors: Schrauder André , Stackelberg Arend , Schrappe Martin , Klingebiel Thomas , Holter Wolfgang , Sykora Karl-Walter , Handgretinger Rupert , Ebell Wolfram , Pötschger Ulrike , Zimmermann Martin , Karlhuber Susanne , Peters Christina 
Published: October 2019

Abstract
As results of frontline and relapse protocols for children with acute lymphoblastic leukaemia (ALL) are improving over time, there is a strong need for prospective haematopietic stem cell transplantation (SCT) trials, ensuring well standardized procedures for all relevant components which are potentially responsible for heterogeneity in post-SCT outcome. Therefore, in 2003 the Berlin-Frankfurt-Münster (BFM)-Study Group initiated a prospective international multicenter trial (ALL-SCT-BFM 2003). Main goals are the standardization and harmonisation of the SCT procedure and the question, if nowadays SCT from a matched sibling donor (MSD) is equivalent to SCT from a very well matched donor (MD). Further, we want to determine the efficacy of a SCT from HLA-mismatched donors (MMD) compared to SCT from MD/MSD as well as the incidence of acute and chronic graft versus host disease (GvHD) after SCT. Between September 2003 and June 2007, 381 patients (185 in CR1, 196 in CR2 or after subsequent relapse) were recruited by 27 participating SCT centers in Austria, Germany and Switzerland. Indications for SCT were poor response to induction treatment, either detected by morphology on day 33 or by minimal residual disease load measured after 11 weeks of treatment, cytogenetic aberrations [t(9;22), t(4;11)], early bone marrow relapses of ALL, or any subsequent ALL relapse. 76 patients had an indication for a MSD, 131 patients for a MD, and 174 patients for a MMD. Compliance with HLA-typing procedures according to the trial guidelines (high resolution typing of HLA A, B, C, DRB1 and DQB1) improved over time: 54% in 2004, 83% in 2005, 90% in 2006, and so far 100% in 2007. Unmanipulated bone marrow was used in 73% of MSD- and MD-SCT as recommended by the protocol. The recommended conditioning regimens (TBI+VP16 for MSD, TBI+VP16+ATG for MD, and TBI+Flu+VP16+ATG for MMD; in children younger than 2 years, TBI was substituted by BU+Cy) were used in 80% of all patients. GvHD prophylaxis (cyclosporine A for MSD, and cyclosporine A plus “short methotrexate” for MD) was given in 83% of all patients according to the trial guidelines. In most cases, deviations were due to clinical reasons. As the trial ALL-SCT-BFM 2003 is ongoing till 2009, interim results regarding outcome will not be presented. Nevertheless, treatment-related mortality is still very low (7%) and has improved over time since the very early beginning. Final results will be a very stable basis for subsequent ALL-SCT trials, focussing on controlled modifications and interventions in those patients being at highest risk for relapse subsequent to SCT.
The Transfusion Iron Overload Score Is a Potential Predictor of Survival in Stem Cell Transplant Patients.
Authors: Storey Jonathan A. , Connor Rebecca , Lewis Zachary T. , Lovato James , Hurd David , Torti Frank M. , Molnar Istvan 
Published: October 2019

Abstract
Iron overload at the time of bone marrow transplantation has recently been associated with increased mortality. The mechanism by which iron overload negatively impacts survival is unclear. In an effort to better define iron overload and its potential role in the transplant setting, we examined 78 consecutive patients at risk for transfusion-related iron overload (AML, ALL, MDS, and aplastic anemia) undergoing either autologous or allogeneic stem cell transplant at our institution between 1999 and 2004. To estimate transfusion-related iron overload, we devised a novel scoring system based on routinely available laboratory variables obtained prior to transplant. In our scoring system, a single point was awarded for each of the following:
number of red cell units transfused prior to transplantation > 25; serum ferritin > 1000 ng/ml; and a semi-quantitative bone marrow iron stain of 6+.
The Transfusion Iron Overload Score (range of 0 to 3) predicted for overall survival in our patient sample (Fig. 1, p = 0.0003 by Kaplan-Meier log rank trend) and was more powerful than using serum ferritin alone (p = 0.02). The calculated score retained its significant association with overall survival while controlling for traditional risk factors (age, sex, diagnosis, transplant type, donor relation, and remission status), inflammation (c-reactive protein), and measures of organ damage (transaminases, total bilirubin, INR, and cardiac ejection fraction) (p = 0.04 by Cox proportional-hazards regression). The 27 patients (35%) with a high score (≥ 2) had a dramatically lower median survival of 5.0 months compared to 29.3 months in those with an iron score of less than two (unadjusted hazard ratio of 2.5; 95% CI of 1.6 to 6.3). The observed mortality seen in those with a high score was primarily attributable to treatment-related mortality (p = 0.018) rather than disease-related mortality (p = 0.48). This finding likely accounts for the significant survival impact of iron overload seen in the allogeneic (p = 0.006) but not the autologous (p = 0.48) subgroup. In conclusion, the Transfusion Iron Overload Score served as a strong independent predictor of survival in our stem cell transplant population, identifying patients at significant risk for early treatment-related mortality. Our results lend further support to the notion that iron overload is detrimental in the transplant setting, and for the first time suggest that transfusion-related iron overload increases mortality via a mechanism independent of pre-existing inflammation and end-organ dysfunction.
Figure 1. Overall post-transplant survival by the Transfusion Iron Overload Score. Figure 1. Overall post-transplant survival by the Transfusion Iron Overload Score.
Prognostic Impact of Elevated Pretransplant Serum Ferritin in Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Mahindra Anuj , Bolwell Brian , Sobecks Ronald , Rybicki Lisa , Brown Stacey , Pohlman Brad , Dean Robert , Andresen Steve , Kalaycio Matt , Sweetenham John , Copelan Edward 
Published: October 2019

Abstract
Iron overload is a well-established adverse prognostic factor in patients who undergo hematopoietic stem-cell transplantation (HSCT) for thalassemia and appears to play a similar role in patients with acute leukemia and myelodysplastic syndrome. We report the results of a large study of consecutive patients undergoing myeloablative allogeneic transplantation to assess the influence of pretransplantation ferritin on treatment-related mortality (TRM), graft-versus-host disease (GVHD) and survival. Pretransplantation ferritin, obtained within 100 days preceding transplant, was available on 222 consecutive patients ≥18 years of age undergoing allogeneic HSCT from 10/20/2000 to 12/04/2006. Transplantation was performed for various disorders (AML n= 105, ALL= 44, CML= 25, MDS= 19, NHL=18, and other= 11) and mainly with busulfan-based preparation (n=153). One hundred twenty four donors were siblings and 98 unrelated. The median patient age was 43 years. Median ferritin level was 1433 ng/ml (range 7–17029). Seventy-five patients had ferritin levels >1910 ng/ml. Median follow-up for surviving patients was 2.5 years. Recursive partitioning analysis identified significantly lower survival for patients with serum ferritin >1910 ng/ml as shown: ESTIMATED SURVIVAL BY PRETRANSPLANTATION FERRITIN LEVEL
Figure Figure
Age, gender, disease, disease status at transplant, interval from diagnosis to transplant, number of prior chemotherapy regimens, prior radiation therapy, ferritin, albumin, AST, Alkaline phosphatase, LDH, preparative regimen and CD 34+ dose elevated were assessed. In a multivariable model for survival, elevated ferritin was a significant,independent,adverse prognostic factor for survival (p=0.011, HR=1.68). The addition of albumin, a negative acute phase reactant, did not change the prognostic impact of elevated ferritin. Patients with ferritin >1910 ng/ml had a higher incidence of TRM(p=0.046), as illustrated below: INCIDENCE OF TRM BY FERRITIN LEVELS
Figure Figure
This siginificantly higher incidence of TRM occured despite a significantly lower incidence of chronic GVHD (p=0.019) and a trend towards a lower incidence of acute GVHD (p=0.10) in patients with elevated ferritin levels. The incidences of hepatic venoocclusive disease and relapse mortality were not different between the two groups.
Conclusions: Elevated ferritin is an independent adverse risk factor in a large group of patients undergoing myeloablative transplantation for various diseases. Higher TRM occured despite the association of elevated ferritin with a significantly decreased incidence of chronic GVHD, consistent with an immunosuppressive effect demonstrated in other settings. Trials should be undertaken to analyze the benefit of iron chelation therapy prior to BMT in patients with iron overload.
SAGE Analysis Demonstrates Increased Expression of TOSO Contributing to Fas Mediated Resistance in CLL.
Authors: Proto-Siqueira Rodrigo , Panepucci Rodrigo A. , Careta Francisco P. , Rizzatti Edgar G. , Lee Abigail , Clear Andrew , Morris Kelly , Falcão Roberto P. , Zago Marco A. , Gribben John G. 
Published: October 2019

Abstract
The molecular pathogenesis of CLL remains unclear; therefore we sought to identify new genes involved in the pathogenesis or mechanisms of resistance in CLL and other B cell malignancies. To this end we performed SAGE in CLL cells and compared results with those from healthy B cells (nCD19+). Gene expression profiles were obtained by SAGE and individual gene expression was measured by real time PCR. Analysis revealed only 27 genes with significant (p< 0.001) differences between mutated and unmutated subtypes of CLL cells. In contrast to the high level of similarity between the CLL subtypes, comparison of the complete CLL SAGE libraries versus nCD19, revealed that 56 genes were over represented and 50 genes were down regulated in CLL (fold change higher than 20x, p<0.001 as a cut-off). A gene ontology analysis using the top 1,000 transcripts revealed some genes aberrantly expressed in CLL not previously reported. One of these is TOSO (regulator of Fas-induced apoptosis) of interest since CLL cells are resistant to Fas mediated killing. Real-time RT-PCR analysis of 78 cases of CLL and 12 nCD19+ confirmed that TOSO mRNA was over expressed (4.4 fold) in CLL (p<0.0001), and its expression was positively correlated with BCL-2 expression. There was no correlation between TOSO expression and prognostic factors, including IgVH mutational status, cytogenetics, evolution or previous treatment. In addition, using tissue microarray of 1mm cores arranged in triplicate followed by indirect immunohistochemistry we evaluated TOSO protein distribution and intensity in 44 CLL lymph nodes and 5 reactive tonsils and appendix. Scoring was performed by two histopathologists independently and demonstrated that more than 50% of CLL cases express TOSO strongly (2+, 3+) with a range of 10% to 95% of positive tumour cells (p=0.0057, Chi-Square test), whereas among control samples analyzed all were negative except the mantle cells from one tonsil and one appendix were scored as weakly positive (1+); the remainder were negative. Flow cytometry analysis in 12 CLL cases and 5 healthy subjects demonstrated a 5.6 fold increase of TOSO in peripheral CLL cells (p=0.013). SAGE results confirmed that both CLL subtypes share a similar expression profile and highlighted a novel gene, TOSO. This protein plays a functional role upstream of Fas in the extrinsic apoptosis pathway and ongoing studies are assessing the mechanisms whereby TOSO mediates resistance of CLL cells to anti-Fas treatment and ways to reverse this.
Mutual Regulatory Loop between miR-155 and PU.1 Is a Candidate Pathogenesis Factor in CLL.
Authors: Stopka Tomas , Vargova Karin , Kokavec Juraj , Pospisil Vit , Curik Nikola , Burda Pavel , Skoultchi Arthur I. , Zavadil Jiri 
Published: October 2019

Abstract
MicroRNAs are non-coding post-transcriptional regulators of gene expression often dysregulated in human cancers including hematological malignancies. MiR-155 is a putative oncogenic factor found overexpressed in chronic lymphocytic leukemia (CLL) cells. We have confirmed the increased levels of miR-155 in peripheral blood lymphocytes in 12 patients with chronic lymphocytic leukemia (CLL) compared to 11 normal controls, using quantitative PCR assay. Among the predicted high-scoring hematopoietic target genes of miR-155 is the ets-family transcription factor PU.1, a key regulator of myelo-lymphoid lineage development. Using gene expression profiling of CLL peripheral lymphocyte samples compared to normal controls, we have determined significant downregulation of the PU.1 mRNA and of additional numerous miR-155 targets previously reported to be required for myelo-lymphoid differentiation, and of a set of gene targets of PU.1-chromatin DNA interactions that we determined by chromatin immunoprecipitation analyses. Functional pathway analysis of the miR-155 targets downregulated in CLL patients suggest the suppression of T-cell receptor pathway, TGF-b/Bmp, MAPK, Jak/Stat and Wnt signaling and remodeling of actin cytoskeleton and cell-matrix interactions, as well as the regulation of numerous transcription regulators including c-FOS, c-MYC, MAFB and HDAC4. Recent reports suggested a role of miR-155 in lymphocytic differentiation, we have tested whether PU.1 may regulate expression of its own regulator miR-155. We used two cell systems expressing a transgene encoding PU.1 fused with the ligand binding domain of the estrogen receptor, ER):
murine erythroleukemia (MEL) cells (Rao 1997) and PU.1 null hematopoietic progenitor cells stably transfected with PU.1-ER (Walsh 2002).
Using a miRNA microarray assay we have identified a set of 21 miRNAs that are modulated by PU.1. Indeed, miR-155 was among the upregulated targets alongside a known PU.1 miR target miR-233. The direct interaction between miR-155 promoter and PU.1 protein was confirmed by chromatin immunoprecipitation in MEL cells. In summary, our data suggests that increased miR-155 levels may contribute to CLL by inhibiting PU.1 function and suppressing PU.1-mediated programs and that in two independent conditional PU.1 cell systems PU.1 directly binds miR-155 promoter and upregulates its expression suggesting thus a mutual regulatory mechanism between PU.1 and miR-155.