bluuebunny/crossref_metadata_2025_split huggingface dataset:
Novel Approach for NK Cell Therapy for Cancer
Authors: Parameswaran Reshmi , Wald David N. , De Lima Marcos , Lee Dean A. , Moreton Stephen 
Published: October 2019

Abstract
Novel therapeutic approaches are urgently needed for many malignancies such as Acute Myeloid Leukemia (AML). We have developed a new therapeutic strategy based upon NK cell immunotherapy that exhibits high clinical potential based upon cell and animal studies. While the harnessing of NK cells for cellular therapy against malignancies has been a topic of interest for several decades, our approach overcomes a major hurdle of insufficient NK cell cytotoxic activity. We have identified that targeting the kinase GSK3 through pharmacologic and genetic approaches leads to the hyperactivation of human blood derived NK cells and a significant improvement in efficacy as compared to traditionally used activated NK cells or chemotherapy in our mouse AML model systems. Importantly this GSK3 inhibition can be achieved through a short ex-vivo incubation of NK cells with a GSK3 inhibitor paving the way for a rapid implementation into a clinical trial. Utilizing both in vitro studies with AML cell lines (ex. OCI-AML3 and HL-60)) and primary human AML cells we observe approximately a 50% increase in efficacy with GSK3 inhibited NK cells as compared to untreated NK cells. Further, we demonstrate significant efficacy of GSK3 inhibited NK cells in a mouse model of circulating human AML. After 4 weekly injections of human NK cells, there is a 50% greater reduction in human AML cells present in the mouse bone marrow with GSK3 inhibited NK cells as compared to vehicle treated NK cells.
Besides efficacy studies, our work has led a model of how GSK3-inhibition enhances NK cell activity as depicted in figure 1. GSK3 inhibition leads to a dramatic increase in adhesion of NK cells to target cells as demonstrated by a flow cytometric adhesion assay (49% vs 83% after 20 min incubation) as well as live cell imaging. Consistent with the increased adhesion, GSK3 inhibited NK cells as well as target cells (after co-incubation) exhibit increased expression of essential NK cell-target adhesion molecules including L-selectin (on NK cells) and ICAM (on target cells). The induction of ICAM on target cells is due to a marked induction in TNFa production from the NK cells upon incubation with target cells (>7 fold increase in TNFa production). TNFa neutralization impairs the NK activity of the GSK3 inhibited NK cells (~30%) but not vehicle treated cells. Finally, GSK3 inhibition also leads to changes in the NK cells that enhance activity such as increased expression of granzyme and perforin and secretion of IFNg. Overall, our work has a revealed a novel strategy for NK cell therapy that holds high clinical potential.
Figure 1. Model of how GSK3 inhibition leads to hyperactive NK cells. GSK3I - GSK3 inhibitor Figure 1. Model of how GSK3 inhibition leads to hyperactive NK cells. GSK3I - GSK3 inhibitor

Disclosures
No relevant conflicts of interest to declare.
Outcome of Allogeneic Hematopoietic Stem Cell Transplantation in Aplastic Anemia: A Comparison of 3 Conditioning Regimens in 202 Consecutive Cases
Authors: Ahmed Parvez , Chaudhry Qamar Un Nisa , Mahmood Syed Karman , Satti Tariq Mahmood , Khan Mehreen Ali , Shahbaz Nighat , Satti Humayoon Shafique 
Published: October 2019

Abstract

Objective: To compare outcome, disease-free survival and rejection in aplastic anaemia patients receiving: (A) ALG or ATG plus cyclophosphamide (CY) 200 mg/Kg ; (B) Fludarabine 120 mg/m2, ATG plus CY 300 mg/m2 and (C) Fludarabine 120 mg/m2, ATG plus CY 120 mg/Kg conditioning regimens.
Patients & Methods: The study included 202 consecutive aplastic anaemia patients undergoing hematopoietic stem cell transplantation from HLA matched sibling donors at this centre from July 2001 to April 2014. Overall 122 had very severe aplastic anemia, 76 severe and 4 non-severe aplastic anemia. Group A, B and C were compared for outcome, disease free survival and rejection. The stem cell source was bone marrow (39.6%); PBSC (7.9%) or both bone marrow plus PBSC (51.5%). GVHD prophylaxis consisted of cyclosporine (51% cases) and cyclosporine plus methotrexate (45.5% case). Chi-square test was used to compare categorical variables. Two way ANOVA was used to compare group means. Kaplan Meier survival curves with log rank test was applied to compare the groups for survival analysis.
Results: Group A included 99, group B 72 and group C 31 patients. Male to female ratio was 76/23 in group A; 56/16 in group B and 22/9 in group C. Median age of patients in group A, B and C were 16, 22 and 18 years respectively (p=0.001). At a median follow up of 1267 days the overall and disease free survival were 76.7% and 68.7% in group A, 70.8% and 69.4% in group B, 67.7% and 54.8% in group C (p=0.350, and p=0.412 respectively). The rejection rate was 11.1%, 9.7% and 35.5% in group A, B and C respectively (p=0.001). Frequency of chronic GVHD was 10%, 8% and 35.5 in respective groups (p<0.001). There was no significant difference between the groups for disease severity, gender and ABO mismatch, stem cell source and GVHD prophylaxis.
Conclusion: Conditioning regimens using ALG or ATG with Cy 200 mg/kg; and Fludarabine 120 mg/m2, ATG plus Cy 120 mg/kg give comparable results while the regimen using Fludarabine 120 mg/m2, ATG plus Cy 300 mg/m2 is associated with high rejection rate and inferior survival in aplastic anemia following allogeneic stem cell transplantation.

Disclosures
No relevant conflicts of interest to declare.
Development of an Alpha/Beta T-Cell Depleted Allogeneic Stem Cell Transplantation Protocol Form Matched Related and Unrelated Donor Grafts in Patients with Poor Risk Leukemia
Authors: DeWitte Moniek A , Boome Liane te , Wagen Lotte van der , Kuball Jurgen H 
Published: October 2019

Abstract

Introduction: The outcome of allo-SCT in patients with poor risk leukemia is still hampered by GVHD and relapse. The innate immune system has been reported to contribute to tumor control, with lower incidence of GVHD. Specific depletion of αβ T- cells – key players in the development of GVHD – will render NK cells and γδ T cells within the allograft. Recently reported results have shown the great promise of this approach in haploindentical transplantations. Within this study, we aim to extend αβT- cell depleted allo-SCT to patients with a MRD or MUD.
Methods: Patients with either ‘poor-risk’ or ‘very poor-risk’ leukemia were included in this phase I study. Either HLA matched siblings (MRD) or fully matched HLA matched (10/10) unrelated donors (MUD) were eligible. abT-cell reduction was performed by negative selection with anti-abTCR antibodies in combination with magnetic microbeads, using the automated CliniMACS device (Miltenyi Biotec, Bergisch Gladbach, Germany). The maximal contamination with αβT-cells for all dose levels was 5x105/kg. Three conditioning regimens have been investigated (I): fludarabine 120 mg/m2 + cyclophosfamide 4800 mg/m2, (II): fludarabine 120 mg/m2 + busilvex AUC=90 and (III): ATG (Genzyme®) 4 mg/m2 + fludarabine 120 mg/m2 + busilvex AUC=90 followed by αβT- cell depleted grafts from matched related or unrelated donors. Within cohort II and III, no additional immune suppression was given after allo-SCT.
Results: Products for 14 patients have been successfully processed and used for αβT-cell depleted allo-SCT between 2011 and 2013. A ~4 log depletion of αβT-cells has been observed in the product with a recovery of ~75% of CD34+ cells. In cohort I and cohort II, 60% and 25% primary graft failures were observed, whereas in cohort III primary engraftment (chimerism > 95%) was observed in all patients. The combination of ATG/fludarabine/busilvex was well tolerated with a hematological recovery of within 3 weeks. In all 14 patients immune reconstitution primarily consisted of innate cells (NK cells and γδ T cells) the first 6 months post transplantation. In addition, no increase in CMV or EBV reactivations has been observed so far under the profound “innate control”.
Conclusion: ATG Busulfan Fludarabine is a low toxicity platform for abTCR-depleted transplantations, resulting in a swift reconstitution of innate cells (NK cells and γδ T cells) the first 6 months post transplantation. This transplantation strategy can serve as a tool for future immunological interventions such as a low dose DLI or genetically modified T cells.

Disclosures
No relevant conflicts of interest to declare.
CD70/CD27 Signaling Mediates Resistance of Chronic Myeloid Leukemia Stem Cells to Tyrosine Kinase Inhibitors By Compensatory Activation of the Wnt Pathway
Authors: Riether Carsten , Schürch Christian M. , Radpour Ramin , Hinterbrandner Magdalena , Huguenin Anne-Laure , Ochsenbein Adrian F. 
Published: October 2019

Abstract
The introduction of BCR/ABL-specific tyrosine kinase inhibitors (TKIs) a decade ago revolutionized chronic myelogenous leukemia (CML) therapy. However, disease-initiating leukemia stem cells (LSCs) in CML are resistant to TKIs despite BCR/ABL inhibition. Therefore, CML will ultimately relapse upon drug discontinuation. We have previously shown that blocking CD70/CD27 signaling targets LSCs by inhibiting the activation of the Wnt pathway. Here, we investigated a combination therapy of TKIs and CD70/CD27 blocking monoclonal antibodies in human and murine CML. We demonstrate that TKI-mediated BCR/ABL inhibition down-regulates miR-29, leading to increased expression of specificity protein 1 (SP1), a transcription factor with binding site in the CD70 promoter. In addition, TKI treatment reduced the expression of DNA methyltransferases resulting in de-methylation of the CD70 promoter. These combined effects resulted in CD70 up-regulation on LSCs, enhanced CD70/CD27 signaling and compensatory Wnt pathway activation. Combined BCR/ABL and CD70/CD27 inhibition synergistically reduced Wnt signaling and eradicated leukemia cells in vitro. More importantly, combination therapy effectively eliminated CD34+ CML stem/progenitor cells in murine xenografts and LSCs in a murine CML model. Therefore, TKI-induced CD70 up-regulation triggers CD70/CD27 signaling leading to compensatory Wnt activation. These findings identify an important targetable TKI resistance mechanism of CML LSCs and may lead to new therapeutic strategies to directly target LSCs to overcome treatment resistance.

Disclosures
No relevant conflicts of interest to declare.
Sphingosine 1-Phosphate (S1P)/S1P Receptor 1 Pathway Has an Essential Role for Sickle Cell Disease
Authors: Zhang Yujin , Zhao Shushan , Wu Hongyu , Hu Xia , Luo Renna , Parchim Nicholas , Kellems Rodney E , Xia Yang 
Published: October 2019

Abstract
Sickle cell disease (SCD) is a devastating hemolytic genetic disorder associated with high morbidity and mortality. In order to understand the pathogenesis of this disease, we have conducted non-biased metabolomic screening and found that circulating sphingosine-1-phosphate (S1P) was significantly elevated in mice and patients with SCD. S1P is an important bioactive lipid signaling molecule known to regulate inflammation. Our previous study demonstrated that reduced S1P level in plasma and erythrocytes by treatment with sphingosine kinase 1 (SPHK1) inhibitor, PF-543, significantly decreased sickling cells, hemolysis and inflammation in SCD mouse model, which indicated that S1P may play an important role in an SCD complication, especially in inflammation.
S1P engages five G-protein coupled receptors known as S1PR1-5. Targeting S1P signaling has been successfully applied in the treatment of the autoimmune disease-multiple sclerosis with the compound named FTY720. In order to understand the roles of S1P/S1PRs signal pathway in pathophysiology of SCD, we treated SCD mice with S1P receptors antagonist FTY720. The results showed that FTY720 successfully inhibited S1P receptors, especially S1P1 expression on immune cells from thymus and lymph node (P<0.05). Circulating white blood cells and inflammatory cytokines, such as CRP, IL-1β, TNF-α and IL-6, also decreased significantly measured by ELSIA kit. Additionally, FTY720 treatment significantly ameliorated organ damage. To investigate the roles of S1P1 receptor in SCD, we treated SCD mice with S1P1 specific antagonist, SEW2871. The results demonstrated that circulating white blood cells and inflammatory cytokines reduced significantly. Histologic studies revealed that the necrosis and congestion of multiple organs including kidney, lung and spleen were substantially reduced by SEW2841.Our studies demonstrate the elevated circulating S1P signaling via its receptor (likely S1PR1) directly contributes to inflammation and multiple tissue damage. Thus, it provides strong evidence that S1P/S1P1 pathway is likely a therapeutic target for SCD.

Disclosures
No relevant conflicts of interest to declare.
Bone Marrow Derived Mesenchymal Cells Secrete Granulopoietic Cytokines upon Danger Signaling
Authors: Wirths Stefan , Bugl Stefanie , Radsak Markus P. , Märklin Melanie , Müller Martin R. , Kanz Lothar , Kopp Hans-Georg 
Published: October 2019

Abstract
Granulopoietic homeostasis is regulated at steady-state to supply sufficient numbers of pooled and circulating neutrophils to maintain barrier function against commensal flora. In addition, upon pathogenic microbial challenge, an increased formation of neutrophils is induced, termed ‘emergency granulopoiesis’.
Antibody-mediated reduction of neutrophil numbers in steady-state induces a feedback loop leading to an increase of bone marrow granulopoiesis with expansion of hematopoetic stem and progenitor cells. This feedback loop was demonstrated to depend on TLR4 and TRIF, but not MyD88 signaling (Bugl et al. Blood 2013). In contrast, emergency granulopoiesis was shown to be dependent on MyD88 signaling in endothelial cells (Boettcher et al. Blood 2014).
Bone marrow mesenchymal stromal cells (MSC) are niche-forming cells, harboring and regulating hematopoiesis. Upon steady-state neutropenia an increase of niche size was observed. Here we investigated, whether niche-forming MSC act as sensors of pathogen-associated molecular patterns (PAMPs) and induce granulopoietic cytokines to stimulate expansion of adjacent hematopoietic stem and progenitor cells.
MSC of C57BL/6 and TLR4-KO mice were cultured in vitro and treated with LPS for 24 hours. Cells were harvested and qRT-PCR for G-CSF, TLR4, MyD88, TRIF, GM-CSF, IL-1β, IL-18 and Casp-1 was performed
After treatment with LPS, RNA of granulopoietic cytokines G-CSF and GM-CSF were massively up regulated in MSC of WT mice. Upstream regulating, inflammasome components IL-1ß and caspase-1 RNA levels increased as well, with little changes in IL-18, TLR4, MyD88 and TRIF.
Unexpectedly, TLR4-KO MSC up regulated transcription of IL-1β and G-CSF upon LPS stimulation as well, and caspase-1 was found to be strongly up-regulated in unstimulated TLR4-KO compared to WT MSC.
In summary, bone marrow stromal cells are found to be PAMP-sensing and secrete cytokines that regulate granulopoiesis. TLR4-independent sensing of LPS by MSC might correspond to the alternative noncanonical inflammasome pathway recently described (Kayagaki et al. Science 2013).

Disclosures
No relevant conflicts of interest to declare.
Further Research on the Activity of Ribonucleases in the Blood and Urine of Patients Suffering from Proliferative Hemocytopathia
Authors: ALEKSANDROWICZ J. , URBANCZYK J. , OSTROWSKA ALEKSANDRA , SIERKO J. 
Published: October 2019

Abstract
1. A simple quantitative method (based on McCarty’s method) of measuring the RN-ase activity in urine and blood is presented.
2. The RN-ase activity was examined in the blood and urine of patients suffering from chronic granulocytic leukemia, myeloblastic leukemia, lymphatic leukemia, granulocytopenia, lymphogranulomatosis, and in cases of infectious disease with an increased leukocytosis.
3. The RN-ase activity in the urine is increased in chronic granulocytic leukemia, while the RN-ase activity in the blood serum remains unchanged. In other diseases, in spite of an increased or decreased leukocytosis, no essential changes in the RN-ase activity in the urine and blood serum have been observed, which indicates that there is no close relationship between the number of leukocytes in the peripheral blood and the RN-ase activity in the 24-hour sample of urine.
Effect of Chemical Protection and Bone Marrow Treatment on Radiation Injury in Mice
Authors: URSO PAUL , CONGDON C. C. , DOHERTY D. G. , SHAPIRA RAYMOND 
Published: October 2019

Abstract
MEG (prepared from 9.0 mg. of AET) significantly modified the response of the bone marrow, peripheral blood leukocytes, spleen, thymus, body weight, hematocrit, and histology of the hematopoietic organs to lethal (900 r) and sublethal (450 r) x-irradiation in CAF1 mice.
MEG reduced the effect of 900 r on the bone marrow, granulocytes of the blood, hematocrit, spleen, thymus, and body weight by a factor of approximately two.
Combined treatment (MEG and isologous bone marrow) of mice exposed to 900 r of x-rays demonstrated that MEG is primarily responsible for preventing the early destruction of the bone marrow, but bone marrow injection was primarily responsible for causing a more rapid recovery of the bone marrow.
In mice receiving combined treatment, recovery of the leukocytes and spleen was primarily influenced by the bone marrow injection; whereas recovery of the thymus and body weight was primarily influenced by MEG. The hematocrit values were normal after combined treatment.
A Rapid Method for Estimation of the Total Leukocyte Count
Authors: BENJAMIN NAOMI R. 
Published: October 2019

Abstract
A technic has been devised for the estimation of the white blood cell count by examination of a stained drop of blood of standard size. The drop is measured by immersing a pinhead in the blood and then touching onto a microscope slide the blood which adheres to the surface of the pinhead. The drop is dried and stained and the count is estimated by comparing its microscopic appearance with photographs or "standard" preparations. It was developed to be used as a screening procedure in case of atomic war, but it may prove useful in epidemics or as a routine screening method in the hematology laboratories of large hospitals.
A New Hereditary Hemoglobinopathy (the Lepore Trait) and Its Interaction with Thalassemia Trait
Authors: GERALD PARK S. , DIAMOND LOUIS K. 
Published: October 2019

Abstract
A new hemoglobinopathy, termed the "Lepore trait," is described. The Lepore trait is characterized by an altered erythrocyte morphology, resembling classical thalassemia trait,[unknown] and by the presence of a hitherto unreported abnormal hemoglobin occurring in low concentration (10-12% of the total hemoglobin). The trait is transmitted as if due to a single gene defect. Simultaneous inheritance of the Lepore trait and classical thalassemia trait occurred in one member of the pedigree studied and resulted in a severe hemolytic anemia clinically indistinguishable from thalassemia major.
On the basis of the erythrocyte morphology, the clinical findings, and the electrophoretic studies, it is conjectured that the Lepore trait may be more closely related to thalassemia than to the other abnormal hemoglobin syndromes.
The Anemia of Human Riboflavin Deficiency
Authors: LANE MONTAGUE , ALFREY CLARENCE P. 
Published: October 2019

Abstract
1. Riboflavin deficiency was induced in 8 adult males with a riboflavin deficient diet and a riboflavin antagonist, galactoflavin.
2. Each patient developed a normochromic normocytic anemia and reticulocytopenia.
3. The leukocyte counts and platelet counts remained within the normal range throughout the period of deficiency.
4. The bone marrow changes and ferrokinetic characteristics of this anemia have been described.
5. The anemia was reversed by riboflavin administration.
Intranuclear inclusions in Bence Jones lambda plasma cell myeloma
Authors: Cohen HJ , Lefer LG 
Published: October 2019

Abstract
A patient with plasma cell myeloma producing only Bence Jones lambda protein was found to have pale intranuclear inclusions in the majority of the bone marrow plasma cells. These inclusions, previously undescribed in myeloma patients producing only Bence Jones protein, contained Bence Jones lambda protein, were non-electron dense, bound by a single membrane,and contained no cytoplasmic structures. Intracytoplasmic inclusions were not present, and the perinuclear cistern was not dilated. Thus, the inclusions may represent intranuclear protein synthesis with anomalous release in the abnormal cells.
Effect of blood in the gut on measurements of endogenous carbon monoxide production
Authors: Brouillard RP , Conrad ME , Bensinger TA 
Published: October 2019

Abstract
Changes induced in measurements of endogenous carbon monoxide (CO) production by blood in the lumen of the gut were studied in five normal volunteers. The study was undertaken because exogenous heme is absorbed by intestinal mucosal cells where the porphyrin ring is split with the release of CO that could contribute to blood CO levels and lead to a fallacious diagnosis of hemolytic disease. Volunteers who consumed 200 ml of their own blood doubled their endogenous production of CO (0.69 versus 0.34 mumoles/kg/hr). This suggested that at least 3% of the ingested heme was degraded and recovered as CO within 2 1/2 hr. Measurements of serum bilirubin also showed a significant increase after ingestion of blood. These data indicate that blood in the gastrointestinal tract can interfere with quantification of heme and bilirubin turnover from measurements of either endogenous CO production or bilirubin and suggest that this might occur with the ingestion of meat.
Dibromomannitol in the treatment of chronic granulocytic leukemia: a prospective randomized comparison with busulfan
Authors: Canellos GP , Young RC , Neiman PE , DeVita VT Jr 
Published: October 2019

Abstract
Dibromomannitol (DBM) is a new agent for the treatment of chronic granulocytic leukemia. A propsective evaluation of the drug was undertaken in a randomized comparison with busulfan. Forty previously untreated, Philadelphia chromosome-positive cases were treated, with 20 patients in each treatment group. The protocol provided for continuous maintenance therapy after remission induction, with a crossover to the opposite drug in patients who became refractory to the primary agent but are without evidence of blastic tranformation. There were 14 remissions in the DBM group and 15 in those treated with busulfan. The rate of decrease of the elevated leukocyte count was more rapid with DBM, but prolonged disease control off treatment occurred in only three of 14 cases as opposed to nine of fifteen busulfan-treated patients who required a median delay of 12 mo before maintenance could be initiated. Hypoplasia occurred in one DBM patient and two busulfan cases. Following recovery, crossover to the opposite drug in two cases again resulted in hypopllasia. Increased skin pigmentation, amenorrhea, pulmonary fibrosis, and cytologic dysplasia, commonly associated with busulfan adminstration, were also noted with DBM. The median duration of disease control with busulfan was 34 mo and 26 mo with DBM. There was no signigicant difference in the incidence of blastic transformation, and median survival for both groups was 44 mo. DBM appears to be as effective as busulfan in the treatment of the chronic phase of CGL but with a more predictable myelosuppressive action. The principal advantage of busulfan over DBM is the fact that more than half the busulfan-treated patients experienced prolonged disease control off treatment.