bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Use of Clinical Grade Magnetic Beads to Deplete Immune Suppressive Elements from Autologous Peripheral Blood Hematopoietic Stem Cell Grafts Enhances T Cell Immune Responses.
Authors: Al-Kadhimi Zaid S , Mahdi Haider , Abdalamir Moshrik , Paul Elyse , Ratanatharathorn Voravit , Uberti Joseph , Lum Lawrence G. 
Published: April 2020

Abstract
Background: Despite the demonstrated efficacy of high dose chemotherapy and autologous hematopoietic stem cell transplantation in Myeloma and relapsed Lymphoma, relapse continues to be a problem. Improving post transplant immune reconstitution in general and tumor specific immune response can reduce relapse rates. Pre-clinical and clinical data suggest that the abundance of CD4+/CD25+ regulatory T-cells (T-regs) and CD14+ monocytes in the autologous graft can contribute to the well described immune suppression state in the early months post transplant. We hypothesized that it is feasible to deplete both of these cellular inhibitory elements from the autologous grafts using clinical grade CD25 and CD14 magnetic beads from Miltenyi. We also hypothesized that the depletion of these populations would improve the proliferative capacity of T-cells in these grafts.
Methods: Samples of peripheral blood hematopoietic stem cell grafts from 9 patients (4 lymphomas and 5 myelomas) undergoing autologous stem cell transplantation, with G-CSF mobilized grafts. We depleted T regs and monocytes, using CD25 and CD14 beads from these graft samples. Then we compared T cell reactivity between the manipulated graft versus the unmanipulated graft samples, in a mixed lymphocyte reaction.
Results: We show that it is feasible to deplete both populations based on phenotype analysis that confirmed selective depletion of CD4+/CD25+/CD127-T reg population (70–80%) and CD14 population (74%.). The positively selected population was enriched for T regs CD4+CD25+CD127-(50–85%) and monocytes CD14+ (75%+). Furthermore in 9 out of 9 patients T cell proliferation measured by 3H thymidine uptake was close to two fold or higher in the double depleted grafts compared to the unmanipulated grafts (P=0.004 Wilcoxon Signed Rank Test).
Conclusion: Double depletion of T regs and monocytes from autologous grafts using clinical grade magnetic beads, leads to enhanced T cell proliferation. This work will lay the ground for a subsequent clinical trial of regulatory T-cell/Monocyte depleted autologous transplantation using the same strategy, to enhance immune recovery post transplant and achieve better anti-lymphoma, or anti-myeloma immune response.
Figure Figure
Ethnic Variations in Diagnosis, Treatment, Socioeconomic Status and Survival in a Large Population-Based Cohort of Elderly Patients with Non-Hodgkin’s Lymphoma
Authors: Song Shen , Burau Keith D , Fang Shenying , Wang Haijun , Du Xianglin L , Wang Luhua 
Published: April 2020

Abstract
Background: Little is known about ethnic disparities in survival among patients with Non-Hodgkin’s Lymphoma (NHL), although these disparities have been documented for patients with many solid tumors.
Methods: We studied a retrospective cohort of 13,321 patients diagnosed with incident NHL at age &gt;=65 in 1992–1999, identified from the SEER (Surveillance, Epidemiology and End Results)-Medicare linked database with 11 years of follow-up. Of these patients, 11,868 were Caucasians, 533 were African-Americans, and 920 were other ethnicities. A time-to-event Cox regression model was used to examine the relative risk of all-cause and disease-specific mortality.
Results: A larger proportion (72.2%) of African-Americans were in the poorest quartile of socioeconomic status as measured by the poverty level compared to 21.8% of Caucasians, and 43.2% of African-Americans received chemotherapy compared to 52.4% of Caucasians (p&lt;0.01).
Hazard ratio of all-cause and NHL-specific mortality increased significantly with age, advanced tumor stage, higher comorbidity scores, and poorer socioeconomic status. Patients receiving either chemotherapy or radiotherapy or both were significantly less likely to die. Compared to Caucasians, African-Americans were not statistically significant at risk of all-cause (Hazard ratio = 0.97, 95% confidence interval: 0.88–1.08) and disease-specific mortality (1.07, 0.92–1.25), after adjusting for treatment, comorbidity, and socioeconomic status.
Conclusions: The risk of mortality in patients with NH-lymphoma was associated with socioeconomic status and was reduced in patients receiving chemotherapy. No significant differences in the risk of mortality were observed between African-American and Caucasian patients after controlling for factors such as treatment and socioeconomic status.
Disruption of Leukemia/Stroma Cell Interactions by CXCR4 Antagonist CTCE-9908 Enhances Chemotherapy-Induced Apoptosis in AML
Authors: Lu Hongbo , Zeng Zhihong , Shi Yuexi , Konoplev Sergej , Wong Donald , Konopleva Marina , Andreeff Michael 
Published: April 2020

Abstract
The chemokine receptor CXCR4 is critically involved in the migration of hematopoietic cells towards the stromal derived factor (SDF-1α)-producing bone marrow microenvironment. We and others have previously demonstrated that stroma/leukemia interactions mediate protection of leukemic cells from chemotherapy-induced apoptosis (Konopleva, Leukemia 2002). Using a peptide analog of SDF-1α designated CTCE-9908, we tested the hypothesis that CXCR4 inhibition interferes with stromal/leukemia cell interactions resulting in increased sensitivity to chemotherapy. Our results showed that CTCE-9908 significantly inhibits SDF-1α-induced migration of U937 (43% inhibition) and OCI-AML3 cells (40% inhibition) in a dose-dependent manner. In three of the four primary AML samples which expressed CXCR4 on cell surface and migrated in response to SDF-1α, 50 μg/ml CTCE-9908 reduced SDF-1α-induced migration of leukemic blasts (60%, 19% and 50% inhibition respectively). In in vitro co-culture systems, stromal cells significantly protected OCI-AML3 cells from chemotherapy induced apoptosis [no MS-5, 75.2±5.2% annexinV(+); with MS-5, 59±1.1% annexinV(+)]. Western blot analysis revealed that CTCE-9908 inhibits Akt and Erk phosphorylation in a dose-dependent manner in the OCI-AML3 cell line stimulated by SDF-1α. Blockade of CXCR4 expression with CTCE-9908 markedly abrogated the protective effects of stromal cells on OCI-AML3 [Ara-C, 59±1.1% annexinV(+); Ara-C + CTCE-9908, 76.9±1.35 annexinV(+)]. Most importantly, it decreased stroma-mediated protection from AraC-induced apoptosis in four out of five primary AML samples with surface expression of functional CXCR4 (mean increase, 25.1±9.3% compared to chemotherapy alone). In vivo, subcutaneous administration of 1.25mg CTCE-9908 induced mobilization of leukemic cells from primary AML patient transplanted into NOD/Scid-IL2Rγ-KO mice (from 15% to 27% circulating leukemic cells 1 hour post CTCE-9908 injection). Taken together, our data suggest that SDF-1α/CXCR4 interactions contribute to the resistance of leukemic cells to chemotherapy-induced apoptosis via retention of leukemic cells in the bone marrow microenvironment niches. Disruption of these interactions by the potent CXCR4 inhibitor CTCE-9908 represents a novel strategy for targeting leukemia cell/bone marrow microenvironment interaction. Based on these observations, in vivo experiments are ongoing to characterize the efficacy of chemotherapy combined with CTCE-9908.
Disruption of the Osteoblast Niche by G-CSF Is Associated with Hematopoietic Stem Cell Quiescence and Loss of Long-Term Repopulating Activity: Role of Cdkn1a
Authors: Gopalan Priya K. , Christopher Matthew J. , Greenbaum Adam M. , Link Daniel C. 
Published: April 2020

Abstract
The bone marrow microenvironment plays a key role in regulating hematopoietic stem cell (HSC) function. In particular, bone marrow stromal signals contribute to the maintenance of HSC quiescence, a property that is thought to be associated with long-term repopulating activity. We previously reported that G-CSF treatment disrupts the osteoblast niche by inducing osteoblast apoptosis and inhibiting osteoblast differentiation. In this altered bone marrow microenvironment, we also showed that the number of HSCs in the bone marrow after G-CSF treatment (as defined by CD34− Kit+ Sca+ lineage-cells or CD150+ CD48− CD41− lineage-[SLAM] cells) was unchanged and that the HSCs were more quiescent than HSCs from untreated mice. However, despite the quiescent phenotype, there was a marked loss of HSC long-term repopulating activity. To define mechanisms for this phenotype, we first asked whether G-CSF acts directly on HSCs to inhibit their long-term repopulating activity. Bone marrow chimeras containing wild type and G-CSFR−/− cells were established and treated with G-CSF. The contribution of G-CSFR−/− cells to hematopoiesis remained stable for at least 3 months after G-CSF treatment, demonstrating that the effects of G-CSF on HSC function are not direct. We next performed RNA expression profiling on sorted SLAM cells, a cell population highly enriched for HSCs. These data showed that expression of Cdkn1a (p21cip1/waf1) was increased in HSCs harvested from G-CSF treated mice. To define the contribution of Cdkn1a to HSC quiescence and loss of repopulating activity following treatment with G-CSF, Cdkn1a−/− mice (inbred on a C57BL/6 background) were studied. Wild-type or Cdkn1a−/− mice were treated with G-CSF for 7 days and pulse labeled with bromo-deoxyuridine (BrdU), and the percentage of SLAM cells that labeled with BrdU was determined. Consistent with our previous observations, treatment of wild-type mice with G-CSF resulted in a significant decrease in the percentage of BrdU+ SLAM cells in the bone marrow. In contrast, in Cdkn1a−/− mice, no change in the percentage of BrdU+ SLAM cells after G-CSF treatment was observed [10.08 ± 2.26% (untreated); 10.96 ± 2.80% (G-CSF treated); p = NS]. To assess HSC function, competitive repopulation assays were performed using untreated or G-CSF treated bone marrow from wild type or Cdkn1a−/− mice. Surprisingly, G-CSF had a similar deleterious effect on HSC repopulating activity in both wild type and Cdkn1a−/− mice. Collectively, these data show G-CSF treatment, possibly through disruption of the osteoblast niche, induces HSC quiescence and loss of long-term repopulating activity. HSC quiescence, but not loss of repopulating activity, is dependent upon Cdkn1a−/−. The mechanisms by which G-CSF treatment results in a loss of HSC function are under investigation.
Red Blood Cell Scramblase Activation Is Induced by Phorbol Ester and Prevented by Inhibition of Protein Kinase C
Authors: Barber Latorya Arnold , Palascak Mary B , Joiner Clinton H. , Franco Robert S. 
Published: April 2020

Abstract
Phosphatidylserine (PS) is normally confined to the inner leaflet of the red blood cell (RBC) membrane, but is externalized in many sickle RBC. This external PS may contribute to decreased sickle RBC survival, increased thrombogenesis, and increased sickle RBC-endothelial interaction. Aminophospholipid Translocase (APLT) and Phospholipid Scramblase (PLSCR) are thought to regulate the distribution of PS between the inner and outer membrane leaflets. APLT maintains phospholipid (PL) asymmetry by returning externalized PS to the inner membrane leaflet. PLSCR disrupts PL asymmetry by causing nonspecific PL equilibration across the membrane. The mechanism(s) of PLSCR activation have not been defined, but previous studies have demonstrated PS externalization as a result of treatment with the potent PKC activator, PMA (phorbol-12-myristate-13-myristate). We examined the effect of PMA on PLSCR activation (measured by NBD-PC internalization) in normal RBC in the presence or absence of the PKC inhibitor, chelerythrine chloride. In response to PMA, PLSCR is significantly and rapidly activated as measured by the fraction of NBD-PC internalized after six minutes compared to a DMSO control (0.267±0.046 vs. 0.056±0.011, n=5, p&lt;0.001). There was also a significant increase in PS externalization (Annexin V binding) in response to PMA stimulation after thirty minutes compared to a DMSO control (14.76±4.80% vs. 1.50±1.55%, n=5, p=0.021). PMA-dependent NBD-PC internalization and PS externalization were significantly inhibited in the presence of chelerythrine chloride (0.180±0.031 vs. 0.267±0.046, n=5, p=0.016 for NBD-PC internalization at six minutes and 3.51±0.92% vs. 14.76±4.80%, n=5, p=0.011 for PS externalization at thirty minutes). These data provide evidence that PKC is involved in the regulation of scramblase activity and thus the distribution of PL across the RBC membrane. Future treatment strategies for sickle cell disease could include PKC inhibitors that would prevent PS externalization and potentially result in decreased sickle RBC-endothelial interactions, decreased thrombogenesis, and prolonged sickle RBC survival.
Deficient Apoptosis Signaling in Primary Pediatric BCP-ALL Xenograft Cells Determines the Kinetic of Engraftment in Vivo in a NOD/SCID Model and Patient Outcome
Authors: Queudeville Manon , Eckhoff Sarah M. , Stahnke Karsten , Debatin Klaus-Michael , Meyer Lueder H. 
Published: April 2020

Abstract
We previously identified the importance of intact apoptosis signaling for treatment response in primary pediatric ALL and AML by analysing two key apoptogenic events, caspase-3 activation and cytochrome c release. A correlation of both events was exclusively found in the groups of patients with good response to treatment, showing that intact apoptosis signaling is a critical feature for favorable outcome. Using a NOD/SCID mouse model for pediatric B cell precursor ALL we found that rapid in vivo leukemia growth (Time to leukemia (TTL) short) in transplanted recipients determines poor patient outcome. In this study we now investigated the importance of deficient apoptosis signaling for leukemia engraftment in this model. We analyzed a total of 17 BCP-ALL xenografts, 5 of which had a short time (TTLshort) to clinical manifestation of the disease (&lt; 10 weeks) and 12 with a significantly longer time (TTLlong) to leukemia (&gt; 10 weeks). The apoptosis signaling parameters cytochrome c release and caspase-3 activation were evaluated flowcytometrically in directly isolated cells from the leukemia bearing mice before and after induction of apoptosis by factor deprivation in culture and related to TTL. Comparing TTLshort and TTLlong a significant correlation of both apoptosis parameters was found in the TTLlong group only (P= .017). Importantly, patients whose blasts engrafted in NOD/SCID-mice with TTLlong showed a superior outcome compared to the TTLshort group. Additionally, proficient apoptosis signaling as indicated by the correlation of cytochrome c release and caspase-3 activation, was only found in blast cells from recipients where the corresponding patient had favorable prognostic factors such as TEL/AML1-positivity (P= .023), absence of hyperleucocytosis at diagnosis (P= .012), absence of relapse (P= .038) and no early relapse (P= .017). In contrast, the correlation of cytochrome c release and caspase-3 activation was absent in the groups with poor prognosis. We also found a direct positive correlation between TTL and cytochrome c (P= .022) and TTLlong and caspase-3 activation (P= .048), indicating that a longer engraftment time may be due to proficient apoptosis signaling in the leukemic blasts. Our finding in the NOD/SCID human-ALL model matches our retrospective results in pediatric ALL and AML to conclude that the functional integrity of a downstream apoptotic checkpoint (cytochrome-c related caspase activation) is an important feature regulating leukemia biology. Thus, deficient apoptosis signaling appears to determine rapid engraftment of leukemia cells in the NOD/SCID model in vivo and consequently poor patient outcome.
The Role of Activation-Induced C-Type Lectin (AICL) in NK Cell Reactivity against Acute Leukemia
Authors: Baessler Tina , Kuttfruff Sabrina , Krusch Matthias , Kanz Lothar , Steinle Alexander , Salih Helmut R 
Published: April 2020

Abstract
Studies of haploidentical stem cell transplantation (SCT) demonstrate that NK cells play an important role in immunosurveillance of leukemia and indicate that acute lymphoblastic leukemias (ALL) are less susceptible to NK cell reactivity than acute myeloid leukemia (AML). The molecular mechanisms mediating NK cell reactivity against leukemia cells, especially those responsible for the differential susceptibility of AML and ALL to NK cell immunosurveillance, are yet largely unclear. Recently we have identified AICL as a myeloid-specific ligand for the activating NK cell receptor NKp80 (Welte et al., Nat. Immunol.2006, 7:1334). Here we studied the role of AICL and NKp80 in NK cell reactivity against acute leukemia. Substantial levels of AICL were expressed on blasts in 23 cases (40%) of 57 AML patients studied, while no AICL expression was detected on leukemic cells of 21 investigated ALL patients. Healthy CD34+ cells did also not express AICL, which suggests that AICL may be a marker for a malignant phenotype of myeloid hematopoietic precursor cells. Expression in AML was not restricted to a specific French-American-British (FAB) subtype, but was significantly associated with myelomonocytic differentiation (FAB M4 and M5). No correlation with a particular cytogenetic abnormality or expression levels of HLA class I was observed. Inhibition of AICL-NKp80 interaction by blocking NKp80 (Fab’)2 fragments significantly diminished both cellular cytotoxicity (&gt;60% reduction, E:T ratio 40:1) and IFN-γ production (&gt;60% reduction) of NK cells in cocultures with AICL-positive AML cells demonstrating that AICL expression renders AML blasts susceptible to NK cell reactivity. On NK cells of AML patients, expression of NKp80 was found to be significantly reduced compared to healthy donors. Cocultures of AICL- and control-transfectants with NK cells revealed that NKp80 expression is downregulated after interacting with its ligand, which provides a functional explanation how leukemia cells may evade NKp80-mediated NK cell immunosurveillance. Taken together, our data indicate that AICL-NKp80 interaction substantially contributes to NK cell reactivity against AML but not against ALL, which may explain the differential susceptibility of AML and ALL to NK cell reactivity. Our data further support the notion that NK cells play an important role in the pathophysiology of acute leukemia and provide useful information for therapeutic approaches in AML which, like e.g. haploidentical stem cell transplantation, rely on a sufficient NK cell response.
Rituximab in Combination with CHOP in Patients with Follicular Lymphoma: Analysis of Treatment Outcome of 552 Patients Treated in a Randomized Trial of the German Low Grade Lymphoma Study Group (GLSG) after a Follow up of 58 Months
Authors: Buske Christian , Hoster Eva , Dreyling Martin , Forstpointner Roswitha , Kneba Michael , Schmitz Norbert , Schmits Rudolf , Metzner Bernd , Reiser Marcel , Lengfelder Eva , Woermann Bernhard J , Harder Harriet , Hegewisch-Becker Susanna , Berdel Wolfgang E. , Hess Georg , Eimermacher Hartmut , Aldaoud Ali , Planker Manfred , Unterhalt Michael , Hiddemann Wolfgang 
Published: April 2020

Abstract
We have previously reported that compared to CHOP alone the addition of Rituximab (R) to CHOP significantly increases the response rate (RR), the time to treatment failure (TTF) and also the overall survival (OS) in patients with newly diagnosed advanced follicular lymphoma. However, in the previous report, the median observation time was short with 18 months and no data were reported on the outcome of different risk groups according to the FLIPI (Hiddemann et al., Blood 2005). We now report on the treatment outcome of 552 patients with advanced stage follicular lymphoma randomized between R-CHOP versus CHOP alone after a median follow up of 58 months. Responding patients &lt; 60 yrs. of age underwent a second randomization in first remission for Interferon alpha maintenance (IFN) versus myeloablative radio-chemotherapy with subsequent stem cell transplantation (ASCT), whereas all older pts received IFN maintenance.
R-CHOP induced a significantly higher RR (97% vs. 91%, p=0.005). Furthermore immunochemotherapy resulted in significantly longer TTF (median not reached vs. 35 months, 5-years TTF 65% vs. 32%, p&lt;0.0001), response duration (RD, 5-years RD 66% vs. 35%, p&lt;0.0001), and OS (5-years OS R-CHOP 90% vs. CHOP 84%, p = 0.0493). To analyse the impact of R-chemotherapy in different risk groups we performed subgroup analyses for patients with low (LR), intermediate (IR) or high risk (HR) according to the FLIPI: R-CHOP prolonged significantly TTF in all risk groups compared to CHOP alone (5-years TTF: 84% vs. 46% for LR (p=0.0021); 73% vs. 37% for IR (p&lt;0.0001); 49% vs. 23% for HR (p&lt;0.0001). The superiority of R-CHOP vs. CHOP could be documented in the patient group not treated with ASCT: the RD was significantly prolonged by the immunochemotherapy (5-years probability 62% vs. 26%, respectively; p&lt;0.0001). This could not be seen in the group of younger patients consolidated with ASCT (5-years probability 78% vs. 66%, p=0.43). However, the number of events was too low to draw any final conclusion.
These data with a mature follow-up demonstrate that first–line R-CHOP significantly improves response rate and TTF in patients with FL compared to the respective chemotherapy alone. However, they also demonstrate that R-chemotherapy might be not sufficient for a long-lasting improvement. This indicates the necessity to perform clinical trials that test whether adding consolidation and/or maintenance after initial cytoreduction further improves treatment outcome.