bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD6-Depleted Mobilized Stem Cells for Modification of HVG and GVH Reactions after HLA-Haploidentical Marrow Transplantation.
Authors: Kolb Hans-Jochem , Bigalke Iris , Simoes Belinda , Falk Christine , Tischer Johanna , Hill Wolfgang , Ledderose Georg 
Published: October 2019

Abstract
Allogeneic stem cell transplantation is limited to patients with a histocompatible donor, but for patients with advanced acute leukemia and high-grade lymphoma HLA-haploidentical transplantation may be considered. Rejection of the transplant and graft-versus-host disease are major obstacles and T-cell depletion eliminates the graft-versus-leukemia effect and causes prolonged immune deficiency. Here we studied the use of CD6-depleted G-CSF mobilized blood cells (mbc) 6 days after transplantation of unmodified marrow in order to suppress host-versus-graft and graft-versus-host reactions (GVHD) retaining a graft-versus-leukemia effect and the capacity to reconstitute the immune system. CD6-depleted mbc contain a large proportion of NK and NK-T cells. 63 patients with advanced disease (AML 32, ALL 15, NHL 11, CLL 2, CML 2, SAA 1) were transplanted with marrow from family donors sharing one HLA-haplotype and differing in 0 – 4 HLA-antigens of the second haplotype. Conditioning consisted of total body irradiation (TBI), antithymocyte globulin (ATG) and cyclophosphamide (CY), post-grafting immunosuppression of cyclosporin A (CSA) and a short course of methotrexate (sMTX). A transfusion of donor leukocytes was given prior to CY. Complete engraftment was observed in 34 evaluable patients given 12 Gy TBI. The dose of TBI could be reduced to 4 Gy without rejection in 25 evaluable patients. GVHD was severe (grade III and IV) in 12 of 48 evaluable patients. An improved method of CD6-depletion was administered to mbc in 9 patients and severe GVHD did not develop. GVHD responded to corticosteroids in most patients. 15 patients survive disease free up to 6 years (median 784 days). Recurrent infections including PTLD were the major cause of transplant-related mortality. Absolute counts of lymphocytes, CD4 and CD8 subpopulations were not different from those of a contemporary group of 46 patients in advanced disease given HLA-identical sibling transplants. However naïve CD4 cells and TRECs were low. Rejection, GVHD and GVL have been controlled by this regimen, but immune reconstitution remains a problem that may be solved by early discontinuation of immunosuppression in this regimen.
Hemostatic Alterations in Patients Undergoing Hematopoietic Stem Cell Transplantation.
Authors: Buchholz Stefanie , Woywodt Alexander , Scheer Johanna , Hambach Lothar , Kamal Haytham , Haller Hermann , Haubitz Marion , Ganser Arnold , Hertenstein Bernd , von Depka Mario 
Published: October 2019

Abstract
Damage of endothelial cells (EC) is known to be involved in pathogenesis of microangiopathy, hepatic veno-occlusive disease, sepsis, capillary leak syndrome, and acute or chronic graft-versus-host disease (GvHD) as major causes of morbidity and mortality in patients after hematopoietic stem-cell transplantation (HSCT). We recently demonstrated in 39 patients undergoing allogeneic stem cell transplantation that numbers of circulating EC (CEC) increased significantly after conditioning regimens and that patients treated with reduced intensity conditioning (RIC) showed significantly lower cell numbers (Woywodt et al., Blood, 2004 May 1;103(9):3603-5).
Here we report on the measurement of plasma levels of von Willebrand factor (vWF), thrombomodulin (TM), PAI-I and TAFI in the course of HSCT.
Measurement of vWF, TM, PAI-I and TAFI was performed in the 39 patients (20 male, 19 female; n = 21: HLA-matched unrelated donor; n = 18: related donors). Blood samples were collected before starting conditioning regimen (day −7), the day before transplantation (day −1) and at day +7, +14 and +21 after transplantation. 28 patients received a conventional regimen with cyclophosphamide (120 mg/kg) and either total body irradiation with 12 Gy (n = 14) or busulfan (16 mg/kg, n = 14). 11 patients undergoing stem-cell transplantation were treated with reduced intensity conditioning with fludarabine (150 mg/m²) and busulfan (8 mg/kg body) or melphalan (120 mg/m²).
Median baseline vWF was elevated (262%, range 68–612, normal range: 50–150). Median baseline PAI-I (11.3 U/l; range 1.8–34.4; normal range: < 20) and median baseline TAFI (90%; range 46–126; normal range: 70–120) were within normal limits. TM level was lower than normal values (median 3.95 ng/ml; range 2.69–9.36; normal range: 6.6–10.6). Levels of vWF increased after conditioning regimen and remained elevated until day +21 (day −1: median 262; day+7: 268; day +14: 327; day +21: 374; p < 0.01). Median TM remained low at all time points (day −1: median 4.26; day +7: 3.86; day +14: 3.97; day +21: 4.52).
Levels of TAFI remained unchanged (day −1: median 82; day +7: median 91, day +14: median 88; day +21: median 89). There were also no differences in levels of PAI-I before or after conditioning regimen or after transplantation (day −1: median 11.4, day +7: median 10.8, day + 14: median 14, day + 21: median 11.8). There was no significant difference in vWF in patients undergoing reduced intensity conditioning compared to patients treated with conventional regimens.
Interestingly, there was no correlation between the endothelial markers as vWF and TM and numbers of CEC.
Our data indicate that significant alterations of the hemostatic system occur in patients undergoing HSCT. Further studies are warranted to define the clinical role of both, hemostatic alterations and CEC in the course of HSCT.
Molecular Targets of Glucocorticoids in Context of Apoptosis Induction in Childhood Precursor T-Cell ALL (T-ALL).
Authors: Karawajew Leonid , Rhein Peter , Kostka Dennis , Hoffmann Rosemarie , Schrappe Martin , Spang Rainer , Ludwig Wolf-Dieter 
Published: October 2019

Abstract
Glucocorticoids (GCs) are principal therapeutic agents in ALL. In spite of their prominent clinical significance, molecular targets of GCs and, consequently, the mechanisms of GC-resistance are still unclear. In the present study, we focussed on molecular mechanisms of GC cytotoxicity in T-ALL, and investigated GC-specific response in leukemic cells at transcriptional and cellular levels. To this end, T-ALL samples (n=14) were investigated for GC-specific gene expression changes before onset of apoptosis (10 μM dexamethasone, 6 hr of incubation). Gene expression data obtained using Affymetrix HG Focus microarrays were normalized by variance stabilization on probe level, and genes consistently changed in GC-treated vs -untreated cells were identified using a SAM-like analysis of log ratios in an unbalanced permutation test. By these procedures, a group of 195 genes (false discovery rate = 0.005) was identified. Furthermore, the T-ALL samples were investigated for GC-specific apoptosis (10 μM dexamethasone, 20 hr). Apoptotic response to dexamethasone was highly heterogeneous with an apoptosis rate varying from 0% to 56% and a mean value of 23% (assessed by annexin V/propidium iodide staining). To identify genes with the highest impact on GC-specific apoptosis, we investigated the correlation between GC-specific gene expression changes and apoptosis rate in T-ALL samples. Expression changes of 25 genes were found to correlate significantly with the rate of apoptosis (Spearman regression analysis; correlation coefficients rs=0.54–0.81, p<0.05). First, we found that the GC-specific apoptosis was associated with a downregulation of genes encoding for cell cycle-promoting proteins (PTTG1, CDC7) and an upregulation of antiproliferative factors (ID2, BTG1), thus indicating that antiproliferative and cytotoxic activities of GCs are coupled in T-ALL. Furthermore, the rate of apoptosis correlated with a suppression of the commonly known survival signaling pathways, as indicated by the upregulation of the NFkappaB inhibitor IkappaB-alpha and downregulation of c-myc. In addition, we observed a downregulation of Bcl11A, a survival factor in the T-cell development. Finally, GC-specific apoptosis correlated with an upregulation of genes encoding for an actin depolymerizing factor DSTN and catabolic enzymes ISG20 (exonuclease) and CLPP (protease). Taken together, investigation of gene expression changes in the context of apoptotic cellular response identified common and T-cell specific survival pathways as GC-targets in T-ALL. In addition, our data suggest a novel GC-specific mechanism of cytotoxicity via direct transcriptional regulation of genes encoding for cellular catabolic proteins.
Obesity and Body Weight Independently Predict Relapse and Survival in Preadolescents and Teenagers with Acute Lymphoblastic Leukemia (ALL). A Retrospective Analysis of Five Children Cancer Group (CCG) Studies.
Authors: Butturini Anna , Dorey Fred , Gaynon Paul , Fu Cecilia , Franklin Janet , Siegel Stuart , Sacks Nancy , Siebel Nita , Henry David , Rogers Paul , Lange Beverly , Sather Harland , Trigg Michael , Bleyer Archie 
Published: October 2019

Abstract
We analyzed data from 4356 patients with ALL diagnosed from 1988 to 1995 at age 2–20 years and enrolled in CCG studies 1881, 1891, 1922, 1882 or 1901. Patients with Down Syndrome and those with CNS disease at diagnosis were excluded. Obesity was defined as a body mass index (BMI)>95th percentile by the Center for Disease Control growth charts (www.cdc.gov/growthcharts). Obesity at diagnosis did not affect outcome in patients younger than 10 years but independently predicted 5y risk of leukemia relapse and event free survival (EFS) in the 1026 patients aged 10 years or older. Multivariate analysis in patients aged 10 years or older showed that obese patients had increased risk of leukemia relapse (hazard ratio-HR- 1.5, p=0.013) and of all the events (HR 1.5, p=0.009). Other independent predictors of poor outcome were high WBC at diagnosis and failure to achieve bone marrow M1 status by day 7 of therapy. Age had borderline significance; sex, race and type of therapy were not significant. To assess whether the effect of obesity on outcome was due to absolute body weight, we studied the effect of weight in Kg at diagnosis in patients aged 10 years or older. Patients >60kg had increased risk of leukemia relapse (HR 1.4, p=0.002) and of all the events (HR 1.4, p=0.006). High WBC at diagnosis and bone marrow status at day 7 maintained their significance; age, sex, race and therapy were not significant. The effects of obesity and weight on outcome only partially overlapped: obesity still predicted outcome when only patients <60 kg were analyzed as well as weight >60 kg predicted outcome when obese patients were excluded from the analysis. Obesity and weight >60 kg at diagnosis were not associated with differences in early toxic deaths, lenght of initial phases of therapy or days of hospitalization. In conclusion, preadolescents and teenagers with ALL who were either obese or >60 kg of weight at diagnosis had higher risk of leukemia relapse and poorer EFS.
von Willebrand Factor Protects Ca2+-Dependent Structure of the Factor VIII Light Chain.
Authors: Takeyama Masahiro , Shima Midori , Nogami Keiji , Okuda Masahiro , Sakurai Yoshihiko , Tatsumi Kohei , Takeda Tomohiro , Tanaka Ichiro , Yoshioka Akira 
Published: October 2019

Abstract
Factor VIII (FVIII) cofactor function is dependent on the presence of divalent cation, since its active form is metal-linked heterotrimer as well as factor V (FV). The activity of FVIII (FVIII:C) and FV in normal plasma were completely and selectively inactivated by the treatment with divalent cation exchange resin, imidinoacetate. However, FVIII antigen (FVIII:Ag) was preserved, suggesting that deprivation of FVIII:C by the resin was not due to absorption of FVIII but due to inactivation. Both FVIII:Ag and FVIII:C of recombinant FVIII were decreased by treatment with the resin. However, FVIII:Ag but not FVIII:C, was preserved in FVIII and von Willebrand factor complex (FVIII/VWF). Sandwich ELISA using anti-A3 and anti-A2 monoclonal antibodies revealed that association with heavy and the light chains were impaired in the resin-treated FVIII. These results suggested that FVIII was inactivated by deprivation of the metal ion by the resin and that VWF protected cation-dependent FVIII structure. Therefore, we tested if the resin deprives the metal ion such as Ca2+ from FVIII or FVIII/VWF. [Ca2+] in FVIII preparation was decreased from 1.30 to 0.07 mM by the resin. Furthermore, [Ca2+] was decreased from 0.39 to 0.01 mM in FVIII/VWF preparation. [Ca2+] was recovered completely by elution with 1 N HCl from the resins used in both preparations. FVIII:C of the resin-treated FVIII/VWF was partially recovered by addition of Ca2+, whilst FVIII:C of the resin-treated FVIII was not recovered. When FVIII was treated with the resin after addition of [Ca2+], the inactivation of FVIII was dose-dependently inhibited by ~20 % (at [Ca2+]: ~75 mM). On the other hand, the inactivation of FVIII was inhibited by ~60 % (at [Ca2+]: ~25 mM) in FVIII/VWF preparation. Present results demonstrated that FVIII was selectively inactivated by the cation exchange resin due to deprivation of Ca2+. Kinetic experiments by surface plasmon resonance using BIAcore demonstrated that the resin-treated FVIII as well as treated C2 didn’t interact with phospholipid and VWF. Furthermore, immunoblot analysis using the resin-treated FVIII revealed that anti-A2 monoclonal antibody reacted with the heavy chain, whilst anti-A3 and anti-C2 antibodies failed to react with the light chain. On the other hand, these antibodies reacted with the light chain in the experiment using FVIII/VWF, indicating that VWF protects antigenic conformation of the FVIII light chain. Present findings suggest another protection mechanism of VWF on FVIII through stabilization of Ca2+-dependent structure of the FVIII light chain.
C6PS Regulates the Inactivation of Factor Va by Activated Protein C.
Authors: Majumder Rinku , Wang JinMing , Lentz Barry R. 
Published: October 2019

Abstract
Protein C is a vitamin K-dependent, anti-thrombotic protein that is proteolytically cleaved by thrombin to produce the active serine protease, activated protein C (APC). APC inactivates co-factors Va and VIIIa, leading to down-regulation of thrombin generation. Factor Va requires phosphatidylserine (PS) for full cofactor activity. APC inactivates bovine factor Va by catalyzing cleavage in its heavy-chain at Arg505, Arg 662 and Arg306. The cleavage at Arg 306 is stimulated by PS-containing membranes. In this paper, we use water-soluble 2-dicaproyl-sn-glycero-3-phospho-L-serine (C6PS) to ask whether a membrane or molecular PS regulates inactivation of factor Va by APC. Synthetic substrate titration suggested that APC has two Ca2+-requiring binding sites for C6PS: one site increased the APC amidolytic activity (Kdeff ~ 1.3 μM) while the other site decreased it (Kdeff ~ 2 μM) in the presence of 2 mM Ca2+. The effect of C6PS on APC amidolytic activity was PS-specific, with C6PE having no effect. However, titration of both intrinsic fluorescence and DEGR ([5-(dimethylamino)-l-naphthalenesulfonyl] glutamylycylarginyl chloromethyl )-labeled APC fluorescence showed only one Ca2+-requiring C6PS binding site (Kdeff ~ 0.8 μM). The fluorescence anisotropy of DEGR-APC in the presence of 200 μM C6PS revealed C6PS-dependent 1:1 binding to both factor Va isoforms (Va1or Va2) with Kdeff of 1.13x10-9 M (Va1) and 0.3x10-9 M (Va2). The inactivation of factors Va1/Va2 by APC was also promoted by C6PS. In the presence of 200 μM C6PS to saturate sites on both APC and factor Va, both factors Va1/Va2 were fully inactivated by APC (indicating cleavage at Arg 306). However, in the absence of C6PS or presence of only 4 μM C6PS (sufficient to saturate sites only on APC), only partial inactivation (48–52%) of factor Va1/Va2 was observed. These results suggest that PS binding to APC may have some effect on cleavage at Arg505 or Arg 662 but that PS binding to factor Va was needed to promote cleavage at Arg 306. Supported by USPHS grant HL072827 to BRL.
CD34+ Selection before Autologous Peripheral Blood Stem Cell Transplantation: Comparison of the Nexell Isolex 300i® and Miltenyi Biotec CliniMACS® Systems.
Authors: Mantha Simon , Savage David G. , Nichols Gwen L. , Skerrett Donna , Hesdorffer Charles S. 
Published: October 2019

Abstract
Autologous stem cell transplant has been used with variable success rates in the treatment of different malignancies. One of the potential causes of relapse is contamination of the stem cell collection by neoplastic cells. Purging the stem cells by CD34+ selection has been used to reduce such contamination. In this study two different devices were compared in their efficiency to achieve CD34+ selection of peripheral blood stem cells collected in individuals undergoing autologous transplant for breast cancer, neuroblastoma, plasma cell dyscrasia, lymphoma (both Hodgkin and non Hodgkin), rhabdomyosarcoma and amyloidosis. A total of 28 patients were randomized to CD34 selection on the Nexell Isolex 300i® system (NIS) or the Miltenyi Biotec CliniMACS® (MCMS) system. The prevalence of plasma cell dyscrasia was higher in the MCMS group (57%) than in the NIS group (22%). On the other hand, there were 2 cases of Hodgkin lymphoma and 2 cases of breast cancer in the NIS group, in comparison to none in the MCMS group. Average values for parameters of selection efficacy for the 19 patients transplanted are shown in table 1 (recovery = number of cells post / number of cells pre-selection X 100, viability measured with the trypan blue exclusion test). Transplant was done as per local protocols according to the type of malignancy. CD34+ cell dose was determined by institutional guidelines. Average values for engraftment are shown in table 2 (two patients who died before engraftment in the MCMS group were excluded from calculation of the mean values). One patient died of graft failure in the MCMS group in contrast to none in the NIS group. Although the numbers of patients are relatively low, this is the first randomized study in which the selection efficiency of the NIS and MCMS devices have been compared in patients receiving CD34-selected autografts. CFU-GM recovery and cell viability were lower, while neutrophil engraftment was slower in the MCMS group, but these differences were not statistically significant. Our data do not show any clinically significant advantage for either the NIS or MCMS selection device.
Table 1 Device CD34+ recovery (%,range) CFU-GEMM recovery (%,range) CFU-GM recovery (%,range) Viability pre (%,range) Viability post (%,range) P-value calculated with Student’s t-Test. NIS 52 (16–76) 5 (1–14) 9 (0–42) 95 (91–99) 95 (90–99) MCMS 66 (48–75) 4 (1–5) 5 (0–9) 94 (80–99) 88 (60–100) P 0.17 0.71 0.57 0.69 0.16 Table 2 Device CD34+ infused (X 106/kg, range) Time to ANC ≥500 (days, range) Time to plt ≥20 (days, range) # Units plt P-value calculated with Student’s t-Test. NIS 3.6 (2.0–5.2) 12.2 (9–20) 15.9 (11–38) 19.4 (0–30) MCMS 3.6 (1.5–5.1) 15.1 (11- ∞) 16.8 (9-∞) 40.2 (6–150) P - 0.22 0.79 0.23
The Critical Role of Survivin in the Survival and Proliferation of Human Myeloma Cells.
Authors: Romagnoli Mathilde , Bataille Regis , Barillé-Nion Sophie 
Published: October 2019

Abstract
Multiple myeloma (MM) is characterized by the accumulation of malignant plasma cells with an enhanced proliferation and survival capacity within the bone marrow. Myeloma cells often develop drug resistance leading to treatment failure in the patients. Survivin is a member of the inhibitors of apoptosis (IAP) gene family that has been implicated in both cell viability and cell cycle regulation and described as overexpressed in most cancers. We have observed that survivin is expressed in human myeloma cell lines (HMCL) from moderate level in both HMCL XG-6 and MM1S to strong level in U266. Survivin was also detectable in primary myeloma cells. Moreover, survivin expression peaked at G2/M phase and was induced by IL-6 and IGF-1 through JAK/STAT and PI3K/AKT signalling pathways. In order to elucidate the role of survivin in myeloma cells, MM1S and XG-6 HMCL were transfected with human survivin cDNA. First, The survivin transient transfectants MM1S were significantly less sensitive to different drugs (dexamethasone, melphalan, paclitaxel, vincristine, doxorubicin) than the control vector transfectants MM1S. Secondly survivin stable transfected clones established from the HMCL XG-6 that is dependent on IL-6 for its growth, could be maintained in culture without IL-6 longer than control stable clones. Moreover these survivin clones proliferated without exogenous IL-6 in contrast to control clones that kept their IL-6 dependent growth. Furthermore, in vivo preliminary data suggest that survivin overexpressing clones could develop tumor in the SCID-human MM model in contrast to control clones. These data tend to prove that survivin participates in drug sensitivity, escape from IL-6 dependence and tumor formation capacity of HMCL. In summary, our findings suggest that survivin plays an important role in the pathogenesis of MM. A more defined understanding of survivin biology should enhance the rational development of drugs to inhibit its function in myeloma cells.
Risk Factors for Second Malignancies after Transplantation Differ between Allogeneic and Autologous Transplantation.
Authors: Friedman Debra L. , Leisenring Wendy , Flowers Mary E. , Holmberg Leona , Schwartz Jeffrey , Deeg H. Joachim 
Published: October 2019

Abstract
Survivors of hematopoietic stem cell transplantation (HSCT) are at risk for second malignant neoplasms (SMN). We undertook this analysis to ascertain whether risk of SMN was comparable in survivors of allogeneic and autologous transplantation. Among 8662 transplant recipients treated at the Fred Hutchinson Cancer Research Center between November 1969 and April 2004, who survived at least 100 days post transplant, there were 1743 autologous and 6919 allogeneic HSCT recipients. Within this cohort, there were 56 SMNs among the autologous and 224 among the allogeneic recipients. Cumulative incidence of SMN at 10 years post-HSCT was 2.6% in the allogeneic and 4.2% in the autologous HSCT survivors. A multivariate Cox regression model adjusted for current age, TBI, gender and length of follow-up was fit and the hazard ratio (HR) for SMN for the allogeneic transplant survivors was 0.7 (95% confidence interval {CI} 0.5, 1.0) compared to the autologous transplant survivors (reference group), suggesting that the adjusted hazard of SMN is higher for autologous than for allogeneic HSCT recipients. Risk factors differed between the two groups. For survivors of autologous transplantation, in multivariate Cox regression models, only age > 18 years at transplant was associated with decreased risk of SMN (<18 years HR = 1.0; 18–39 years HR = 0.04; 40+ years HR = 0.004). Use of total body irradiation (TBI) was not significantly associated with risk among the autologous HSCT recipients. For allogeneic transplant survivors, increased risk of SMN was associated with TBI and the effect of TBI was stronger for younger (<18 years at HSCT: HR = 4.6; 95% CI 1.6, 13.5) than for older (≥18 years; HR = 1.5; 95% CI 1.0, 2.3) HSCT recipients (interaction p = 0.04) in multivariate Cox regression models. Risk was also increased after acute graft versus host disease (HR = 1.4; 95% CI 1.0, 1.9) and with ongoing follow-up time, with HRs of 1.7 (95% CI 1.1, 2.5) at 10 – 14 years, 2.2 (95% CI 1.3, 3.7) at 15–19 years and 2.6 (95% CI 1.2, 5.4) at 20+ years of follow-up. Unrelated HSCT also increased risk of SMN (HR = 1.4; 95% CI 1.0, 2.0). In adjusted analyses, risk of SMN thus appears slightly lower in allogeneic versus autologous transplant survivors, and risk factors appear different between the two groups. Attention should focus on modifiable risk factors in both groups. Particularly in autologous transplant survivors, the relative contribution of pre-transplant exposures must be established.