bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sensitive NOD-scid-IL2γnull (NOG-scid) Assay for Human Precursor B Acute Lymphoblastic Leukemias (ALL).
Authors: Morisot Sebastien , Hildreth Richard , Kaplan Ian M. , Stetler-Stevenson Maryalice , Taylor Julie A. , Ahuja Yachna , Bloom Lauren , Wayne Alan S. , Civin Curt I. 
Published: October 2019

Abstract
Results of treatment for childhood ALL have improved considerably over the past few decades, yet still ∼20% of pediatric patients relapse. New in vivo models of human ALL might increase understanding of ALL biology and speed preclinical development of novel treatment regimens. Multiple strains of immunodeficient mice support growth of human ALL cells (Bachmann, Current Drugs Target 2007, review). The recently described highly immunodeficient NOD-scid-IL2γnull (NOG-scid) mouse strain offers a number of advantages over the widely-used NOD-scid strain: NOG-scid mice have a normal lifespan, do not develop spontaneous lymphomas, and require ∼10-fold fewer normal human hematopoietic stem-progenitor cells to generate human hematopoiesis (Hiramatsu, Blood 2003; Ishikawa, Blood 2005). In addition, NOG-scid mice support generation of human leukemia from transplanted human acute myeloid leukemia (AML) (Ninomiya, Leukemia 2007). Herein we report a sensitive in vivo model using transplant of human childhood precursor B ALL cells into sublethally irradiated NOG-scid mice. Transplanted mice developed fatal leukemia 4–5 weeks after intravenous injection of 5x104 REH or KOPN8 cells (established precursor B ALL cell lines derived from pediatric ALL patients). At necropsy, the mice had massive splenomegaly and hyperplastic bone marrow. Blood, spleen, and marrow contained abundant human cells with immunophenotype matching the transplanted ALL cell line. We went on to similarly transplant primary leukemic blast cells from pediatric ALL patients. In 3 of 5 tested primary ALL cases, fatal human leukemias developed in the NOG-scid mice. In titrations of 103-2.5x106 transplanted ALL cells, time to clinical leukemia was dose-dependent, and 1000 primary human cells was sufficient to initiate leukemia development. In addition, we detected human leukemic blast cells by flow cytometric immunophenotyping in the blood of transplanted NOG-scid mice as early as 10 days post-transplant, enabling objective leukemia diagnosis ∼4 weeks prior to clinical signs.
CONCLUSIONS: NOG-scid mice supported the generation of human precursor B ALL from cell lines and primary ALL cases, objectively detectable as early as 10 days post-transplant. Our dose titration results suggest that this model detects a high frequency of ALL-initiating cells. This in vivo model may provide a powerful assay both for fundamental questions regarding the biology of leukemia stem cells and for preclinical studies of novel anti-neoplastic agents and regimens.
Frequent Occurrence of Aberrant Pax5 Splice Variants in Philadelphia Chromosome-Positive Lymphoblastic Leukemia.
Authors: Okawara Akemi , Tojo Arinobu 
Published: October 2019

Abstract
Pax5 is a key regulator of B cell commitment and is indispensable throughout the early stages of B cell differentiation. B cell development is blocked at the pro-B cell stage in Pax5-null mice. On the other hand, several investigators have reported that aberrant Pax5 transcripts as well as forced expression of Pax5 result from genetic and/or epigenetic mechanisms especially in B cell malignancies, suggesting the involvement of Pax5 abnormality in development and progression of B cell malignancies. In the present study, we examined Pax5 expression in Philadelphia chromosome (Ph)-positive lymphoid malignancies including 17 cases of Ph-positive acute lymphoblastic leukemia (Ph-ALL) and 3 cases of lymphoid blast crisis of chronic myeloid leukemia (CML-BC). By the reverse transcription-polymerase chain reaction (RT-PCR) method, we detected 4 types of Pax5 isoforms among those. In 16 samples, full-length as well as truncated forms of Pax5 were expressed. Expression of Pax5 could not be detected in 3 cases, and was only faintly found in the remaining one. These 4 cases did not express or very weakly expressed EBF, early B cell transcription factor upstream of Pax5. Sequencing analysis revealed that all 4 truncated isoforms were derived from exon skipping as follows. Type I is lacking in exon 2 and exon 3 (D2/3), type II shows deletion of exons 2–6 (D2-6), type III; D2-7, and type IV; D2-8. Two isoforms resulted in frame-shift, and two isoforms were in-frame splice variants without exons coding DNA binding domain. Transactivation potential of these two in-frame variants was examined by transfection of 293T cells with a reporter plasmid containing high-affinity Pax5 binding sites from the CD19 gene linked to the minimal TATA-like promoter, indicating much lower transcriptional activity in comparison to wild-type Pax5. These results suggest the frequent occurrence in adult Ph-positive lymphoid malignancies of aberrant Pax5 splice variants with reduced transcriptional activity, leading to impairment of normal B cell differentiation.
Analysis of the Role of Wnt Signaling for the Interaction between Leukemia Cells and Stroma.
Authors: Abe Akihiro , Niwa Yosuke , Xu Jinglan , Suzuki Momoko , Hayakawa Fumihiko , Katsumi Akira , Kiyoi Hitoshi , Naoe Tomoki 
Published: October 2019

Abstract
Wnt signaling plays an important role in hematopoietic stem cell self-renewal and proliferation. Recent results also revealed that an aberrant activation of Wnt signaling has been related with hematological malignancies. We have previously reported a stroma-dependent human leukemia cell line, TRL-01, which could not be maintained by any cytokines without stroma or on fibronectin-coated plate although they periodically stimulated the proliferation. We here analyzed the role of Wnt signaling on the cell-adhesion, proliferation and survival of this cell line. TRL-01 was maintained on human bone marrow stroma cell line, hTERT. The apoptosis of TRL-01 on hTERT was suppressed by Wnt-receptor competitor, secreted Frizzled related protein (sFRP)-1, or Rho kinase inhibitor, Y29632 but not by specific inhibitor of canonical pathway, DKK-1. However, the apoptosis of TRL-01 without hTERT was not affected by sFRP-1, Y29632 or DKK-1. These results suggest that survival of TRL-01 on stroma was regulated by non-canonical pathway of Wnt signaling. Next, the effects of Wnt3A, 5A and 10B on TRL-01 were analyzed by establishing hTERT transduced with Wnt3A, 5A or 10B genes. Wnt3A is known as an important factor for the stimulation of self-renewal and proliferation of hematopoietic stem cells, and Wnt5A and 10B were reported to be expressed in B-ALL. When TRL-01 was cocultured with hTERT/Wnt3A or 10B, adhesion was reduced and β-catenin was significantly increased. The condition medium from hTERT/Wnt3A or 10B also stimulates the accumulation of β-catenin but inhibited the survival and proliferation of TRL-01. Wnt5A did not have any significant effects. These results suggest that Wnt3A and 10B stimulate canonical pathway with accumulation of β-catenin but negatively regulate the survival and proliferation of TRL-01. Comparing the expression pattern of Wnt transcripts of TRL-01 and hTERT by RT-PCR, Wnt5A, 5B and 14 were found to be dominantly expressed in hTERT. The role of Wnt5B and 14 for the interaction between TRL-01 and hTERT remains to be elucidated.
In Vitro Study of the Effect of Cyclosporine A, Tetrandrine and Their Combination on the Reversion of Multidrug Resistance in Leukemia Cell Line.
Authors: Chen Bao-An , Guo Jing-jing , Gao Feng , Gao Chong , Chen Jian , Sun Yun-Yu , Wang Jun , Zhao Gang , Ding Jia-Hua , Chen Ning-Na , Pei Xiao-Ping , Dai Yong-Yuan , Shun Qian 
Published: October 2019

Objective: The aim of this study was to investigate the mechanism to reverse the drug resistance of leukemia cells in Cyclosporine A(CsA), tetrandrine (Tet) alone or their combination by using empirical methods, and to provide the basic evidence for further study, and lay the theoretical basis for the clinical applications.
Method: By MTT assay, the IC50(the concentration causing 50% inhibitor of cell growth) of DNR was determined;By flow cytomet ry (FCM) assay, the intracellular DNR concentration and the expression of P-glyco-protein (P-gp) were examined, and the apoptotic changes were observed by Anexin-V. By fluorescent semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR), the mRNA expression of multidrug resistance (MDR) was measured.
Results:
The IC50 of DNR for K562 and K562/A02 cells were 0.26mg/l and 21.22mg/l respectively. Treating K562/A02 cells with CsA(1mg/L), TTD(0.1mg/L)and both for 48 hours partially restored the sensitivity of K562/A02 cells to DNR(IC50 were 7.83 mg/L,5.32 mg/L,2.33 mg/L respectively.) but had not effect on K562 cells; K562/A02 cells showed apoptotic characteristics after treated with CsA, and both for 48 hours(Apoptosis rate was 10.27%,17.64%,52.79% respectively); The fluorescence intensity of intracellelar DNR in K562 and K562/A02 cells treated with DNR(1mg/L)was 522,173 respectively. CsA and TTD (alone or combination) elevated the intracellular DNR concentration in K562/A02 cells(the fluorescence intensity of intracellelar DNR in K562/A02 cells was 188,339,515 respectively); The fluorescence intensity of P-gp in K562 and K562/A02 cells was 0.5,97.97. The P-gp expression was down after treated with CsA, TTD and both(73.7,65.05,58.1); mdr1 mRNA was also down regulated, and the effect of their combination was greater. Date was analyzed by SPSS 13.0 software and expressed as mean ± SD.
Conclusions:
High expression of P-gp may be involved in the mechanism of multidrug resistance (MDR) of K562/A02 cells line; MDR can be partially reversed by CsA or TTD, of which the combination shows a great synergistic reversal effect.
The Reduction of Leukemic Blasts in Bone Marrow Aspirate on Day 6 of Remission Induction Treatment Is Predictive for Complete Remission Rate and Survival in Adult Acute Myeloid Leukemia; the Results of Multicenter, Prospective Polish Adult Leukemia Group Trial.
Authors: Holowiecki Jerzy , Grosicki Sebastian , Kyrcz-Krzemien Slawomira , Kuliczkowski Kazimierz , Kielbinski Marek , Skotnicki Aleksander B. , Jakubas Beata , Hellmann Andrzej , Robak Tadeusz , Seweryn Marek J. , Stella-Holowiecka Beata , Sadus-Wojciechowska Maria 
Published: October 2019

Abstract
The goal of this study was to prove the significance of an early evaluation of leukemic blast reduction based on cytological examination of bone marrow aspirates obtained on day 6 of remission induction treatment. Patients. 90 adult AML patients aged 18–60 (median 47) registered to the PALG prospective trial evaluating the efficacy of 3 remission induction protocols:
DAF: daunorubicine (DNR) 60 mg/m2/d iv, d 1–3; cytarabine (AraC) 200 mg/m2/d ci, d 1–7, and fludarabine 25 mg/m2 2h inf. iv d 1–5), DAC (like DAF but cladribin is used at 5 mg/m2 instead of fludarabine) and the standard DA 3+7 regimen.
Bone marrow aspirates were performed before the treatment and on the 6-th day of the first remission induction course, and the MGG stained marrow smears were evaluated at each centre by two experienced hematologists.
Results. Based on the proportion of blasts in bone marrow specimen on day 6 patients were arranged into 2 groups:
“responders”with ≤5% of blasts (n=59) and “non responders” with >5% of myeloblasts in bone marrow (n=31).
The complete remission rate (CR) in the first group equalling 89% (51/59) was significantly higher in comparison to that obtained in the second group 29%(9/31), p=0,008. The probability of overall survival (OS) was also higher in the group of “responders” if compared to “non responders”; 65% versus 45% respectively, p=0,009. In multivariate analysis including bone marrow examination, cytogenetic risk group, age, leukocyte count at diagnosis and the induction arm only the persistence of leukemic blasts >5% in bone marrow specimen on 6-th day of induction was associated with higher risk of not achieving CR (HR = 52,6; p=0,00002), and with shorter OS (HR=3,13; p=0,02). Conclusion. This prospective, multicenter study demonstrates that a simple and commonly accessible evaluation of the leukemic blasts reduction in bone marrow smear on the 6-th day of remission induction treatment is a reliable and independent predictor for achieving CR and for overall survival. This offers a decision point for an early modification of the treatment strategy.
Partial Efficacy of Bortezomib on Acute Myeloid Leukemia (AML) Cells in a Patient with Concomitant Multiple Myeloma and AML.
Authors: Joly Bertrand , Quillet Patrick , Salanoubat Celia , Bouledroua Selwa , Petitdidier Charlotte , Devidas Alain 
Published: October 2019

Abstract
Bortezomib has a well known efficacy in multiple myeloma, but effects on AML cells is only observed in vitro and remain to be clinically proved. In a 78 year-old patient, we diagnosed two concomitant aggressive hematological malignancies: IgA Kappa multiple myeloma and de novo FAB-M5 AML. The patient was consulting for bone pain and progressive bicytopenia. The bone marrow was infiltrated with 30% dystrophic plasmocytes and 35% myeloïd monoblastes with CD13+, CD33+ phenotype. A serum M component of 5 g/dl IgA kappa was present at diagnosis with mild acute renal failure, elevated beta2microglobulinemia and IPSS score of 2. The cytogenetic study revealed poor prognosis factors for both malignancies with a complex caryotype including chromosome 13 abnormalities and MLL rearrangement. Standard treatments to cure both diseases one after the other could not be a good option, due to the presence of these poor prognosis factors. Thus, we decided to treat the patient with Bortezomib, a proteasome inhibitor known to induce clinical response in multiple myeloma and apoptosis of leukemia cell lines in vitro. After two cycles of Bortezomib (1.3 mg/m2 day 1,4,8,11) combined with steroids (dexamethasone, 40 mg day 1–4), the patient obtained a near complete myeloma remission with normal electrophoresis, but also disappearance of circulating AML blast and a 70% reduction of medullary AML blasts. Low-dose cytarabine (15 mg twice the day for 10 days) was introduced after the fourth cycle of bortezomib because of persistent circulating AML blasts. Bortezomib was stopped after 5 cycles because of neurological adverse event. After 5 cycles of cytarabine, the patient reached complete cytogenetic remission of both diseases. The patient still remains in complete remission after 13 months of follow-up. We conclude that bortezomib had a partial efficacy on AML cells in vivo and could have synergized with cytarabine to reach and maintain complete remission of both poor prognosis AML and multiple in this patient.
CD8+CD28− in Peripheral Blood of Patients with Cutaneus T-Cell Lymphoma (CTLC) in Different Clinical Stages.
Authors: Urbaniak-Kujda Donata M. , Kapelko-Slowik Katarzyna L. , Jazwiec Bozena , Maj Joanna , Dybko Jaroslaw , Slowik Miroslaw , Wolowiec Dariusz , Frydecka Irena , Kuliczkowski Kazimierz 
Published: October 2019

Abstract
Background: Patients with CTLC usually have a poor immune response. CD8+CD28− T cells are new types of immune suppressor cells. The study was to analyze the level and changes of them in peripheral blood with CTLC, their effects on immunosupression of CTLC, the influence factors to provide reference for treatment of CTLC patients.
Patients and Methods: 27 patients with CTLC were enrolled in the study: 8 women and 19 men aged 35–81, medium 57. According to Ann Arbor classification stage I was represented by 1 patient, stage II -6 patients, stage III - 5 patients, stage IV - 6 patients and stage early stage (premycoticus) was represented by 8 patients. Peripheral blood samples were assessed from all patients with CTLC before treatment and 8 normals. Percentage CD8+CD28− T cells was analyzed by flow cytometry.
Result: Compared with control group percentage of CD8+CD28− T cells was significantly higher in patients group p=0,005 (test U Mann Whitney). There were no significant differences of T-cells in among patients with different clinical stages. We revealed the correlation between CD8+CD28− cells and advanced clinical stage r=0,54, p=0,035 (Spearman correlation test).
Conclusion: Percentage of CD8+CD28− T cells is increased in peripheral blood in CTLC patients, and correlates with advanced stage.
Response of Hodgkin Lymphoma Patients of Different Ethnic Groups to Standard ABVD with or without Involved Field RT.
Authors: Ofori-Ntow Samuel , Ciobanu Bety , Telfer Margaret Clare , Nathan Sunita , Kovarik Paula , Kouz Rafid , Griza Decebal S. , Lad Thomas , Catchatourian R. 
Published: October 2019

Abstract
Background: The phenotypic expression of Hodgkin lymphoma among different ethnic groups is not well described. We conducted a study to look at the characteristics and the clinical response of Hodgkin lymphoma patients of different ethnicities to standard treatment.
Method: A retrospective analysis of 65 Hodgkin lymphoma patients diagnosed 2003 through 2006 was undertaken. Patients who have been compliant and completed assigned chemotherapy (ABVD) with or without Involved Field RT were evaluated for response to treatment using CT scan and PET scan. Data was analyzed using fisher exact test.
Results: Patients with limited staged disease received 4 to 6 cycles of ABVD +/− RT while those with advanced staged disease received 6 to 8 cycles of ABVD +/− RT. All 50 patients achieved objective response. Complete response rates were 52% Hispanic, 68% African American, and 75% Caucasians. (p=0.4)
Conclusion: Hodgkin lymphoma is as common in Hispanics as in African American. The mixed cellularity subtype is more common in the Hispanics; however the nodular sclerosis subtype remains the most common histological subtype. Though more Hispanics had an early clinical stage disease at diagnosis, they had a lower complete response rate compared to the other ethnic groups. How this observation will translate into a poorer survival outcome remains to be seen.
Results: Hispanic African American Caucasians N=65 26 27 12 Median age 26 27 38 Nodular Sclerosis subtype 26 27 12 (P=0.08) Mixed Cellularity subtype 6 1 1 Clinical Stage 1–2 16 9 9 (P=0.03) Clinical Stage 3–4 10 18 3 Completed Treatment N=50 23 19 8
An Intact Spleen Is Required for Alloimmunization to Transfused Red Blood Cells Due to Intrasplenic Activation of CD4+ T Cells.
Authors: Hendrickson Jeanne E. , Roback John D. , Hillyer Christopher D. , Zimring James C. 
Published: October 2019

Background: Factors influencing rates of alloimmunization to antigens on transfused red blood cells (RBC alloimmunization) are poorly defined. In particular, the role of the spleen with respect to alloantibody formation is unclear, with conflicting clinical reports in the literature. Moreover, the complexities of multiply mismatched antigens and antigen priming due to transfusions prior to splenectomy make human studies difficult to interpret. To better define the role of the spleen in RBC alloimmunization, we utilized a murine model of transfusion medicine (with the model RBC antigen mHEL (membrane bound hen egg lysozyme)).
Methods: Cohorts of splenectomized and non-splenectomized mice (C57BL/6 × B10.BR) were transfused with the human dose equivalent of 1 unit of leukoreduced mHEL RBCs. RBC alloimmunization was assessed by anti-HEL IgG specific ELISA. The role of antigen-specific CD4+ T cells was studied by the adoptive transfer of 1.5 × 106 HEL-specific CFSE-labeled CD4+ T cells from 3A9 TCR transgenic donors. Adoptively transferred cells were visualized using a congenic marker (Thy1.1); enumeration and division of these cells were monitored by flow cytometry in liver, spleen (if applicable), and lymph node preparations.
Results: Splenectomy dramatically decreased RBC alloimmunization; 14 of 14 splenectomized mice (from 3 experiments) had undetectable to very low levels of anti-HEL IgG following transfusion with mHEL RBCs (average 6.3 fold less than non-splenectomized mice, 95% C.I. 4.6). Moreover, ten of ten splenectomized mice failed to make detectable levels of anti-HEL IgG even following the adoptive transfer of HEL-specific CD4+ T cells. In comparison, elevating the precursor frequency of HEL-specific CD4+ T cells increased RBC alloimmunization by 10,000 fold in non-splenectomized mice. Proliferation and division of CD4+ T cells were detectable in both spleen and liver preparations of non-splenectomized mice by day 3 following transfusion; in contrast, no expansion nor division of CD4+ T cells was seen in liver nor lymphatic preparations of splenectomized animals.
Conclusions: The low level of RBC alloimmunization seen in non-splenectomized mice in this system is limited by existing CD4+ helper T cell responses, as increasing the naive helper T cell precursor frequency dramatically increased RBC alloimmunization. Furthermore, the spleen itself is critical to CD4+ helper T cell function during alloimmunization given that, in splenectomized mice, adoptively transferred HEL-specific CD4+ T cells fail to expand, divide, or stimulate production of detectable alloantibody. Ongoing studies are investigating the phenotype of HEL-specific CD4+ T cells (as effector cells, anergic cells, or regulatory cells) in splenectomized and non-splenectomized mice. These studies have implications for preventing alloimmunization in transfusion-dependent patient populations.