bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pegfilgrastim Shows Safety and Efficacy Similar to Filgrastim in Patients Undergoing Autologous Stem Cell Transplantation for Hematological Malignancies.
Authors: Neumeister Peter , Holub Robert , Staber Phillipe B. , Linkesch Werner 
Published: October 2019

Abstract
Infectious complications are frequent events in patients undergoing high dose cytotoxic chemotherapy with subsequent autologous hematopoietic stem cell transplantation. In order to determine whether a single subcutaneous injection of pegfilgrastim (6mg) is as safe and effective as daily filgrastim (5mcg/kg/d) for reducing duration of grade 4 neutropenia and infectious complications, 60 consecutive autologous stem cell transplantations (ASCT) performed for various hematological malignancies have been analyzed retrospectively at our institution. 24 patients undergoing 30 consecutive ASCT received a single subcutaneous injection of 6 mg pegfilgrastim on day 5 after transplantation and were compared retrospectively to 30 patients receiving 5mcg/kg/d of filgrastim starting from day 7 post-transplantation. Both patient cohorts were comparable with respect to baseline patient and transplant characteristics. Duration of grade 4 neutropenia, infectious complications including incidence of febrile neutropenia, microbiological surveillance data and safety parameters were assessed. The mean duration of grade 4 neutropenia in the pegfilgrastim and filgrastim groups was 8.3 and 9.5 days, respectively (p=0,058). The results of the two groups were also not significantly different for febrile neutropenia, time to neutrophil recovery and toxicity profile, including bone pain episodes. However, a higher incidence of infectious complications associated with the filgrastim vs the pegfilgrastim group could be observed (50% vs 20%) (p=0,011). Pegfilgrastim was safe and well tolerated in this patient population. A single injection of pegfilgrastim administered at day 5 post-transplant shows comparable safety and efficacy profiles to that provided by daily injections of filgrastim. A fixed-dose pegfilgrastim facilitates the management of aplasia following autologous stem cell transplantation and may offer significant benefits for patients and health care providers. Consequently, it may serve as an attractive alternative to a repetitive single G-CSF administration modus. However, prospective randomized studies are warranted to corroborate the results obtained by this retrospective analysis.
Increased Bone Marrow Iron Stores, Severe Mucositis and Large Area under the Curve (AUC) of Profound Neutropenia Predict Severe Infection in 382 Myeloma Patients Undergoing Melphalan - Autologous Stem Cell Transplantation (MEL-ASCT).
Authors: Anaissie Elias J. , Miceli Marisa H. , Dong Li , Grazziutti Monica L. , Rajan Sabitha , Talamo Giampaolo , Thertulien Raymond , Fassas Anasthasios , Tricot Guido , Barlogie Bart 
Published: October 2019

Abstract
Background: MEL-ASCT is standard therapy for multiple myeloma (MM) but is associated with severe infections, at times life-threatening.
Objective: To determine the risk factors for severe infection (bacteremia, septic shock, colitis, pneumonia) following MEL - ASCT for MM.
Materials and Methods: 382 consecutive MM patients (pts) enrolled in our Total Therapy 2 protocol and who received their first MEL - ASCT between 10/1998 and 12/2002 were included. Variables evaluated included age, sex and MM remission status, severity of mucositis and others. Because of the known association between increased body iron stores and infection, pre-ASCT bone marrow (BM) iron stores were also evaluated. The AUC for severe neutropenia (<100 absolute neutrophils (ANC) / mL) was used as a single variable accounting for both the depth and duration of neutropenia.
Results: Median age was 56 years (range: 30–76) and 235 pts (62 %) were males. Severe infections developed in 77 pts (20%) including pneumonia (42 pts). Pre-ASCT risk factors for infection by univariate analysis were increased BM iron stores (OR= 3.601; 95%CI 1.795–7.222; p<0.0007) and low platelets counts (OR for -1000 platelets/μL = 0.997; 95% CI 0.994 – 1; p=0.0381). Increased BM iron stores remained significant by multivariate analysis (OR= 3.601; 95% CI 1.795–7.222; p<0.0007). Post-ASCT risk factors that were significant by both univariate and multivariate analysis were severe mucositis (Grades 3–4 by NCI Common Toxicity Criteria) (OR=1.916; 95% 1.093–3.36; p=0.02) and AUC of severe neutropenia (OR= 1.001/unit; 95% 1–1.002; p=0.03). Neither the duration (days with ANC <1000 / mL) nor the depth of moderate neutropenia (AUC < 1000 neutrophils / mL) predicted infection.
Conclusion: MM pts scheduled to undergo MEL - ASCT and who have increased BM iron stores, and those who develop severe mucositis and / or prolonged and profound neutropenia following ASCT should be considered at greater risk for developing severe infection. AUC of severe neutropenia is a useful single marker of both depth and duration of neutropenia and should be included in studies evaluating risk for infection in neutropenic pts.
NK Cell Recognition of Hepatocellular Carcinoma by Specific Induction of NKG2D Ligands Following Treatment with the Histone Deacetylase Inhibitor Sodium Valproate.
Authors: Salih Helmut R. , Armeanu Sorin , Lauer Ulrich M. , Venturelli Sascha , Pathil Anita , Smirnow Irina , Wagner Annika , Kanz Lothar , Steinle Alexander L. , Bitzer Michael 
Published: October 2019

Abstract
Histone deacetylase inhibitors (HDAC-I) are currently being evaluated as novel treatment for various malignancies. Recently, the HDAC-I sodium valproate (VPA) was shown to exert anti-neoplastic effects on human hepatoma cells while being well tolerated by primary human hepatocytes (PHH). However, the mechanisms underlying this differential mode of action are unclear. In this study, we used microarray analysis to unravel alterations of genes in hepatoma cells upon treatment with VPA. VPA induced the transcription of MICA and MICB, the ligands of the activating immunoreceptor NKG2D which is expressed on cytotoxic lymphocytes and has been shown to play a central role in the immune surveillance of tumors. Changes in MIC mRNA were mirrored by an increase of cell surface, soluble and total MIC protein as determined by immunofluorescense, flow cytometry and ELISA techniques. No significant changes of the NKG2D ligands (NKG2DL) ULBP1-3 were observed. Most importantly, VPA treatment did not induce MIC protein expression in PHH. The VPA-induced MIC expression profoundly triggered lysis of hepatoma cells by NK92 NK cells, which was abrogated by addition of a NKG2D antibody. Since the strength of an anti-tumor response by NK cells and CD8 T cells is critically depending on NKG2DL expression levels and the HDAC-I VPA mediates lysis of hepatoma cells via specific induction of NKG2DL, our results suggest a potential use of VPA in priming tumor cells for innate immune effector mechanisms.
MDR-1 Recognition by Cytotoxic T Cells.
Authors: Niethammer Andreas G. , Wodrich Harald , Loeffler Markus , Lode Holger , Krempien Robert , Huber Peter E. , Debus Juergen , Reisfeld Ralph A. 
Published: October 2019

Abstract
Acquired multidrug resistance (MDR) remains a major challenge in the treatment of cancer with chemotherapeutic drugs. It can be mediated by the upregulated expression of different proteins within the tumor cell membrane. Here we used murine MDR-1 as a target-antigen for the immunotherapy of cancer. We successfully demonstrated that peripheral T cell-tolerance can be broken by oral administration of a DNA vaccine encoding MDR-1 and carried by attenuated Salmonella typhimurium to secondary lymphoid organs. Thus, mice, immunized orally three times at 2-week intervals and challenged two weeks thereafter with either MDR-1 expressing CT-26 colon carcinoma cells or MDR-1 expressing Lewis lung carcinoma cells, revealed a significant increase in lifespan. This was evident, when compared to animals vaccinated with the empty control vaccine or to animals challenged with the maternal cell lines lacking overexpression of MDR-1. The immune response induced was antigen specific and CD8+ T cell-mediated. The presence of the target antigen led to upregulation of activation markers on CD8+ T cells and resulted in a strong cytotoxic T cell response as well as lysis of tumor target cells in vitro. We furthermore established the vaccine to be an effective treatment for established multidrug resistant tumor metastases resulting in a significantly increased lifespan of experimental animals. Absence of CD8+ T cells due to in vivo depletion led to abrogation of effectiveness. Taken together our results demonstrate, that T cell tolerance against the MDR-1 self antigen can be broken. It is anticipated that the combination of such an approach with chemotherapy could lead to more effective treatments of cancer.
Purification of c-Kit-Negative Hematopoietic Stem Cells from Runx1-GFP Knock-In Mice.
Authors: Lorsbach Robert B. , Ang Sonny O. , Sun Weili , Moore Jennifer , Downing James R. 
Published: October 2019

Abstract
The Runx1/Core Binding Factor- β (CBF β) transcriptional complex is required for the establishment of hematopoiesis during development. To permit the analysis of Runx1 expression in hematopoietic cell subsets, we have recently developed a novel murine line in which the expression of both full-length Runx1 and GFP is driven by the Runx1 promoter [Runx1-internal ribosomal entry site-green fluorescent protein (Runx1-GFP) knock-in mouse; Blood 103:2522]. Analysis of these mice has revealed that Runx1 is expressed in all hematopoietic lineages with the exception of erythroid cells. During our analysis, we identified in the bone marrow of these mice a cell population that expresses GFP at levels 2–5 fold higher than any other cell type (GFPhi). These cells have low forward and side scatter properties and do not express c-kit or several lineage-associated cell surface markers (lin−). In comparison to c-kit+lin− cells, these GFPhic-kit−lin− cells possess little colony forming activity in vitro. While they lack primary CFU-S activity in contrast to c-kit+lin− cells, GFPhic-kit−lin− cells possess secondary CFU-S activity. A c-kit−lin− hematopoietic stem cell (HSC) has been described by others that contributes to long-term, but not short-term, hematopoietic reconstitution in lethally irradiated recipients and may represent progenitors of c-kit+lin− HSCs in vivo (Ortiz et al. Immunity 10:173). The GFPhi bone marrow cells that we have identified share many of the properties of those c-kit−lin− cells identified by others; consequently, they likely represent the same cell population. Our ability to isolate these cells based on differential GFP expression should enable us to highly purify these GFPhic-kit−lin− cells and further characterize their immunophenotypic and biologic properties.
In Multiple Myeloma, Clonal Cells Are Unevenly Distributed among Peripheral Blood B Cell Subpopulations.
Authors: Papadhimitriou Stefanos I. , Gligori Ioanna , Kritikou-Griva Elpiniki , Gortzolidis Georgios , Davea Aggeliki , Paterakis Georgios S. , Goumakou Eleni , Savvidou Ioanna , Grigoraki Vassiliki , Anagnostopoulos Nikolaos I. , Georgiakaki Maria 
Published: October 2019

Abstract
BACKGROUND: Current evidence supports the existence of circulating clonal B cells in Multiple Myeloma (MM). However, attempts to enumerate and phenotypically characterise them have so far provided inconsistent results. Most investigators have studied unselected peripheral blood (PB) mononuclear cells, among which the clonal ones make only a small minority. Moreover, in most cases, numerical chromosomal changes were employed as a clonal marker, but aneuploidy is considered a late event in myelomagenesis. To overcome these difficulties, we have followed an alternative approach, by studying purified PB B cells and focusing on chromosomal translocations involving the immunoglobulin heavy chain gene (IGH) on region 14q32, a frequent, early and possibly crucial pathogenetic event in MM.
METHODS: The study included 33 MM patients with 14q32 rearrangements, detected by conventional cytogenetics or florescence in-situ hybridisation (FISH) in the bone marrow at diagnosis. PB CD19+ cells were immunomagnetically isolated (>99% purity) and cytocentrifuged on slides. The slides were studied with a FICTION technique, ie a combination of FISH using a “break-apart” IGH probe set (Vysis Inc, Downers Grove, Il, USA) and indirect immunofluorescence for CD34, CD5, CD10, CD23 and CD38. To avoid the possibility of contaminating plasma cells, isolates with >1% CD19+CD38+++ cells on flow cytometry were excluded from FICTION study.
RESULTS: “Positive” cells above the cutoff level of false positivity (4%) were detected in 25 cases (75.7%), ranging from 4% to 33% (median 9%) among the total CD19+ population. These cells were found to consistently express CD10 (83% to 100%, median 96%) and CD38 (79% to 100%, median 89%). They less commonly expressed CD23 (39% to 67%, median 46%) and very rarely CD34 (0% to 5%, median 0%) and CD5 (0% to 3%, median 0%).
CONCLUSIONS: Our data suggest that circulating cells bearing IGH rearrangements are the rule in MM, making a small but detectable fraction of CD19+ cells. There mmunophenotypic profile supports the concept that clonal B cells represent advanced ontogenetic rather than early stages in B lineage differentiation. Finally, the virtual absence of CD5+ clonal cells is in accord with the view that the high number of PB CD5+ B cells in MM reflects an immunoregulatory network and does not result from clonal expansion.
The Modulating Effect of Azacitidine (Aza C) on the Cytogenetically Tracked MDS Clone Impact Survival.
Authors: Najfeld Vesna , Scalise Angela , Odchimer-Reissig Rosalie , Silverman Lewis R. 
Published: October 2019

Abstract
MDS is a clonal hematopoietic stem cell disorder characterized by progressive bone marrow failure leading to death from infections and bleeding in the majority of patients. Transformation to acute leukemia occurs in 35-40%. Most treatments, other than allogeneic SCT, have been ineffective. Treatment with Aza C has been shown to be clinically effective and alters the natural history of disease (Silverman et al JCO 2002). We have previously demonstrated Aza C modulation of the MDS clone and established five different cytogenetic categories based on its modulating effect (Najfeld et al, ASH 2002). The goal of this extended study was to determine biological and clinical consequences of Aza C modulation on the five cytogenetic categories. Among 224 patients treated with AzaC, multiple sequential studies were performed in187 pts, while 41 pts without follow up studies, were excluded from this report. The initial karyotype prior to therapy with Aza C was normal in 99 pts (53%) and abnormal in 88 pts (47%). These latter patients were not further subdivided into good, intermediate or poor risk according to IPSS. The table below shows the Kaplan Meier median survival of the 5 subcategories based on the modulating effects of Aza C on the cytogenetically identified MDS clone.
Cyto Category No of Frequency of Hematological Survival Group Pts Emerging Response In No Chromosome Abnormalities Months 1 Normal->Normal 67 None CR:3, PR:10, IMP:31 29.3 2 Normal->Abnormal 32 +8:25%l+1q:25% CR:1, PR:6, IMP:18 28.1 +21:9% 3 Abnormal-> Abnormal 50 −7/del(7q):24%; CR:1, PR:4, IMP:30 11.7 −5/del(5q):20%,+1q:14% 4 Abnormal->Clonal 20 −7/del(7q):25%;+21:20% IMP:8 9.1 5 Abnormal->Normal 18 del(7q):33%,+8,28% CR:1, PR:1,IMP:8 22.7
When survival was compared between cytogenetic subgroups there was a statistical difference between some groups. Survival of patients with normal cytogenetics (group 1) was superior when compared to either group 3 (p=.0001) or group 4 (p=.00015). Even for pts who developed an abnormal clone while on AzaC (group 2) survival was significantly longer when compared to either group 3 (p=0004) or group 4 (p=00001). Moreover, prognostic significance was observed for pts who initially had an abnormal karyotype that was diminished or eradicated on AZA C therapy (group 5) when compared to groups 3 (p=0.009) and 4 (p-0.0006). The emergence of a cytogenetically abnormal clone while on treatment with Aza C is not associated with shortened survival compared to those who remain cytogenetically normal. Response to Aza C can occur even with the persistence of the MDS clone or with the emergence of a new clone. This suggest that AZA C can modulate the responsivness of MDS hematopoietic progenitors. Our findings demonstrate that AZA C modulation of the cytogenetically marked MDS clone is associated with distinct subgroups with differing survival characteristics.
The BCR-ABL1 Kinase Interferes with Pre-B Cell Receptor Signal Transduction in B Cell Precursor Leukemia Cells.
Authors: Klein Florian , Feldhahn Niklas , Wernet Peter , Müschen Markus 
Published: October 2019

Abstract
The BCR-ABL1 kinase expressed in B cell precursor leukemia drives malignant transformation of human pre-B cells. To identify novel target genes of BCR-ABL1-mediated transformation and to elucidate how the oncogenic BCR-ABL1 kinase may interfere with normal pre-B cell receptor signaling, we compared genome-wide gene expression of BCR-ABL1+ B cell precursor leukemia cells with their normal counterpart.
While pre-B cell receptor signals are critical for differentiation and survival of normal pre-B cells, nearly all components of pre-B cell receptor-dependent signal transduction were downregulated in the leukemia cells. Surprisingly, BCR-ABL1+ leukemia cells mostly even do not harbor a productively rearranged IGH allele. In these cases, we identified traces of secondary VH gene rearrangements, which may have rendered an initially productive VH gene rearrangement nonfunctional. However, even BCR-ABL1+ leukemia cells harboring a functional VH gene rearrangement are unresponsive to pre-B cell receptor engagement. Independently from engagement, BCR-ABL1 kinase activity induces activation of PLCg1, resulting in an autonomous oscillatory Ca2+ signaling. BCR-ABL1 kinase activity is also linked to the expression of a truncated isoform of the pre-B cell receptor-associated linker molecule SLP65, which acts as a tumor suppressor in mice. Conversely, leukemia subclones surviving inhibition of BCR-ABL1 show responsiveness to antigen receptor engagement and express only full-length SLP65. These cells differentiate into immature B cells expressing Ig light chains.
We conclude that BCR-ABL1 interferes with pre-B cell receptor signal transduction in B cell precursor leukemia cells, while its inhibition reconstitutes the selection for functional (pre-)B cell receptor signaling. Hence, a functional pre-B cell receptor signal-transduction cascade may provide these cells with a potential mechanism to escape apoptotic cell death induced by inhibition of BCR-ABL1.
Figure Legend: BCR-ABL1 + leukemia cells were incubated for 48 hours in the presence or absence of 10 μmol/l STI571. Differentiating subclones among the STI571-treated cells replace their pre-B cell receptor by a B cell receptor and were enriched for surface IgM-expression by MACS.
Figure Figure
Growth Potential of Human Leukemia Blast Cells in SCID Mice and Association with Prognosis of Human Acute Leukemias.
Authors: Yan Ying , Steinherz Peter , Guan Xiuqin , Jakubowski Ann , McGuirk Joesph P. , O’Reilly Richard J. 
Published: October 2019

Abstract
We have described a severe combined immunodeficiency (SCID) mouse model that permits the subcutaneous growth of primary human acute leukemia blast cells into a measurable subcutaneous nodule which may be followed by the development of disseminated disease. Utilizing the SCID mouse model, we examined the ability of patient-derived leukemic blasts to generate leukemic growth in the animals after subcutaneous inoculation without conditioning treatment. Leukemia blasts derived from 133 patients with acute leukemias, (67 acute lymphoblastic leukemia (ALL) and 66 acute myeloid leukemia (AML)), were inoculated into SCID mice. The leukemias displayed three distinct growth patterns: "aggressive", "indolent", or "no tumor growth". Out of 133 patients, leukemia samples 46 (34.5%) displayed an aggressive growth pattern, 14 (10.5%) indolent growth and 73 (55.0%) did not grow in SCID mice. The growth probability of leukemias from relapsed and/or refractory disease was 3 fold higher than that from patients with newly diagnosed disease. Serial observations found that leukemic blasts from the same individual, which did not initiate tumor growth at initial presentation and/or at early relapse, may engraft and grow in the later stages of disease, suggesting that the ability of leukemia cells for engraftment and proliferation was gradually acquired following the process of leukemia progression. Nine autonomous growing leukemia cell lines were established in vitro. These displayed an aggressive proliferation pattern, suggesting a possible correlation between the capacity of human leukemia cells for autonomous proliferation in vitro and an aggressive growth potential in SCID mice. It was demonstrated that patients whose leukemic blasts displayed an aggressive growth and dissemination pattern in SClD mice had a poor clinical outcome in patients with ALL as well as AML. Patients whose leukemic blasts grew indolently or whose leukemia cells failed to induce growth had a significantly longer DFS and more favorable clinical course.
Different Isoforms of BSAP Protein Regulate the Expression of Activation-Induced Cytidine Deaminase (AID) in Normal and CLL B-Cells.
Authors: Oppezzo Pablo J. , Dumas Gerard M. , Ines Lalanne Ana , Pritsch Otto F. , Vuillier Francoise M. , Dighiero Guillaume J. 
Published: October 2019

Abstract
Background: In B-cells somatic hypermutation (SHM) and class switch recombination (CSR) depend of AID expression, a typical event in germinal centers upon CD40 ligand (CD40L) stimulation. We recently showed, that in contrast to normal B-cells, AID is constitutively expressed in a majority of unmutated CLL patients (Oppezzo et al, Blood, 2003). Pax-5 gene, encodes for the B-specific activator protein (BSAP) an essential protein in B-cell development. BSAP has been shown to up-regulate AID gene expression, through binding to its promoter region, whereas inhibitor of differentiation Id-2 down-regulates it. To study the molecular mechanisms underlying AID regulation, we analyzed expression of BSAP and Id-2 transcripts in normal and CLL B-cells and correlated these results to expression of AID and CSR process.
Results: B-cells from 6 healthy donors and 40 CLL patients were analyzed for Pax-5, Id-2, AID and CSR. Results show that CLL and normal B-cells not expressing AID exhibit diminished expression of BSAP and express a spliced variant of the Pax-5 gene (Pax-5 ΔEx8) displaying a deletion in the C-terminal-active domain. Upon CD40-L and IL-4 stimulation expression of AID transcripts and disappearance of the Pax-5 ΔEx8 transcripts was observed. Translation of complete Pax-5 and Pax-5/ΔEx8 isoforms and their binding to AID gene promoter have been demonstrated by Western Blot and EMSA assays, respectively. In contrast, CLL B-cells with constitutive AID expression and active CSR, constantly show increased levels of BSAP and absence of Pax-5ΔEx8 isoform. High expression of AID and BSAP is also correlated with a decrease of Id-2 transcripts.
Conclusion: These results suggest that the presence of BSAP isoform deleted in exon 8 may interfere with the binding of the complete BSAP protein to AID promoter and down-regulate the expression of Pax-5a. Thus, we show for the first time a subtle regulation mechanism controlling the expression of AID protein through the differential splicing of a transcription factor as BSAP.