bluuebunny/crossref_metadata_2025_split huggingface dataset:
In Vivo Effects of Cyclosporine A, Mycophenolate Mofetile and Rapamycin on Adoptively Transferred CD4+CD25+ Regulatory T Cells in Prevention of Experimental Acute Graft-Versus-Host Disease.
Authors: Zeiser Robert , Nguyen Vu , Beilhack Andreas , Baker Jeanette , Schulz Stephan , Contag Christopher H. , Negrin Robert S. 
Published: October 2019

Abstract
CD4+CD25+ regulatory T (Treg) cells reduce GVHD without abrogating GVT effects in murine models and hold promise for clinical application. However, in contrast to most clinical situations, Treg have been investigated in models without immunosuppressive drugs. Therefore, we aimed to assess the impact of Cyclosporine A (CSA), Mycophenolate mofetile (MMF) and Rapamycin (RAPA) on Treg function both in vitro and in a lethal acute GVHD model. Gfp+ Treg reisolated from mixed leucocyte reactions (MLR) containing CSA but not RAPA showed significantly reduced ability to suppress CD4+CD25− T-cell proliferation in secondary MLRs (p=0.025) in a CFSE based assay. The CSA effect could be reversed to 85±3.4% by the addition of IL-2 (50 IU/ml) to the primary culture. In vivo bioluminescence imaging after major mismatch bone marrow transplantation demonstrated reduced early proliferation of donor derived luciferase-labeled conventional T-cells (Tcluc+) in animals treated with Treg. The addition of RAPA and MMF did not interfere with this suppressive effect. Conversely, combining Treg with CSA led to a significantly increased Tcluc+ proliferation as measured in photons/second/mouse (p=0.002). Treatment with Treg/CSA was associated with increased GVHD risk and lowest survival of the Treg/drug combinations. Immunofluorescence microscopy of secondary lymphoid organs revealed less donor derived Treg (CD4+FOXP3+H-2q+) per high power field in animals receiving CSA as compared to RAPA or MMF. Our data indicate that CSA, but not RAPA or MMF interferes with Treg function possibly through an IL-2 dependent mechanism. In vivo, the combination of Treg with RAPA or MMF allowed for adequate GVHD suppression and may therefore provide guidance for clinical trials.
Discrepancies in p21ras, p53, and Mdm2 Protein Expression Predict Worse Prognosis in Acute Myeloid Leukemia.
Authors: Schulz Elizabeth , Lorand-Metze Irene , Koch Rainer , Costa Solange Campelo , Costa Fernando Ferreira , Saad Sara Terezinha Olalla 
Published: October 2019

Abstract
Tumor progress is a multistep process in which clones of cells become abnormal by accumulating growth alterations until they are transformed and/or immortalized. In order to verify a potential role for p53, mdm2, and Ras genes in acute leukemogenesis, we investigated the expression and mutational status of these genes in a cohort of 35 adult patients with acute myeloid leukaemia (AML) at diagnosis with a minimum follow up of 4,5 years. The expression of p21ras, p53, and Mdm2 proteins was studied in bone marrow cytospins stained by APAAP (alkaline phosphatase anti-alkaline phosphatase procedure) using monoclonal antibodies Y13-219, PAb 1801 and 2A10, respectively. N-, K-, H-ras and p53 gene mutations were assessed by PCR-SSCP (polymerase chain reaction/single strand conformation polymorphism) and DNA sequencing to exclude the presence of mutations. We found 2 N-Ras gene and 2 p53 gene mutations in 4 patients. Stratification of AML patients by different cytoplasmic expression patterns (p21ras/p53, p53/mdm2, or p21ras/mdm2, and p21ras/mdm2/p53) revealed that patients with phenotypes p53+/mdm2+ (n=2) had significant best prognosis, patients p53—/mdm2— (n=16) an intermediate prognosis and patients with discrepant phenotypes p53—/mdm2+ or p53+/mdm2— (n=6) a worse prognosis (p=0.003 by Log Rank and p=0.008 by Wilcoxon). Patients with phenotypes p21ras+/mdm2+ (n=2) showed significant best prognosis, patients p53—/mdm2— (n=16) an intermediate prognosis and patients with discrepant phenotypes p53—/mdm2+ or p53+/mdm2— (n=8) a worse prognosis (p=0.07 by Log Rank and p=0.02 by Wilcoxon). Patients with phenotypes p21ras+/p53+/mdm2+(n=2) had a tendency to exhibit best prognosis, patients p21ras—/p53—/mdm2— (n=13) a tendency to intermediate prognosis and patients with discrepant phenotypes p21ras/p53/mdm2 (n=8) had a tendency to worse prognosis (p=0.06 by Log Rank and p=0.06 by Wilcoxon). However, stratification of AML patients by different expression values 0(—), 1(+) or 2(++) for each isolated variable p21ras, p53 and mdm2 using the same survival analysis demonstrated that there were no statistically significant difference among these groups. We found that discrepancies in cytoplasmic expression of p53/mdm2, p21ras/mdm2 and p21ras/p53/mdm2 confer worse long term prognosis to these categories of AML patients. These findings suggest that the signalization pathways p53/mdm2, ras/mdm2 and ras/p53/mdm2 may be active in AML and that an intact p53/Mdm2 axis is important in the prognosis of these patients.
Avascular Necrosis of Femoral Head in Childhood Acute Lymphoblastic Leukemia.
Authors: Tang JingYan 
Published: October 2019

Purpose This study was to determine the frequency of avascular necrosis of femoral head(AVNFH), clinical manifestation, following up results and risk factors in children with acute lymphoblastic leukemia (ALL) and non-Hodgkin lymphoma (NHL). Method Acute non-lymphoblastic leukemia and solid tumor as the background, to assess AVNFH in ALL and NHL between October 1998 and June 2003. Glucocorticoid was included in ALL and T-NHL therapy, but not in B-NHL and other diseases. The criteria of AVNFH is clinical hip pain and/or limp with avascular necrosis sign at MR imaging of the femoral head, but no sign of cancer relapse in clinical and laboratory checking. Results We treated ALL 164 cases, T-NHL 28, B-NHL 20, and solid tumor 126 between October 1998 and June 2003. Forty-three patients with ALL and T-NHL are older than 10 years, including 29 boys and 14 girls. Five cases were diagnosed as AVNFH through MR imaging, including 3 with ALL and 2 with T-NHL. No AVNFH happened in B-NHL and other groups. All of AVNFH are older than 14, average 15 years, and presented with hip pain and/or limping after 1 year glucocorticorid contained chemotherapy. It is 2.6%(5/192) in all ALL and T-NHL cases, but 11.6%(5/43) in the group of older than 10 years. Of those 5 patients, 4 girls and 1 boy. So in the group of older than 10 years girls with ALL and T-NHL, AVNFH happened as high as 28.5%(4/14). After medical interfering, 3 recovered, 2 remained slightly limping but no surgical replacement. Conclusion AVNFH incidence in childhood ALL and T-NHL under our therapy protocol is at least 2.6%. Girls, old than 10, with glucocorticoid contained chemotherapy, are the risk factors of AVNFH. Early detection and interfering may make most of them recover. So, regular MR imaging of the hips for girls who are older than 10 years with glucocorticoid contained chemotherapy longer than 1 year, is reasonable if financially tolerated.
Clinical data of AVNFH in childhood ALL and T-NHL No sex disease chemo-time AVNFH predinision* status of cancer status of AVNFH * or equile to predinision 45mg/m2.d 1 F LR-ALL 42 months left 150 days CR stable, limp 2 M HR-ALL 12 months right 65 days CR stable, limp 3 F T-NHL 17 months right 90 days CR recover 4 F T-NHL 22 months both sides 105 days CR recover 5 F HR-ALL 42 months both sides 150 days CR recover
Detection of 24 TCR Vβ-Dβ1 sjTRECs in T Cells from Cord Blood, Peripheral Blood of Normal Individuals and Patients with AML-M2.
Authors: Li Yangqiu , Yin Qingsong , Chen Shaohua , Yang Lijian 
Published: October 2019

Abstract
Thymic function is characterized its importance of thymus to T-cell diversity in the periphery of both children and adults during both health and disease. The generation of TCR diversity occurs in the thymus through recombination of gene segments encoding the variable parts of the TCRα and β chains. During these processes, by-products of the rearrangements are generated in the form of signal joint T-cell receptor excision circles (sjTRECs). As sjTRECs are stable extrachromosomal DNA fragments, are not replicated during mitosis and thus diluted with each round of cell division, and are therefore most frequent in naïve T cells that have recently left the thymus, their quantification is actually considered as a very valuable tool to estimate thymic function. Quantitative of δRec-ψJα sjTRECs can direct evaluate the recent thymic output function, but it is unable to analyze the particular thymic output function of different TCR Vβ subfamily naive T cells. The complexity of TCR Vβ repertoire is an important factor for immune reconstitution, it should be established the quantitative analysis of series TCR Vβ-Dβ sjTRECs to evaluate the levels of different Vβ subfamily naive T cells. In the present study, analysis of 24 TCR Vβ-Dβ sjTRECs was established by semi-nested PCR using 24 Vβ subfamily antisense primers and 2 Dβ1 sense primers. TCR Vβ-Dβ sjTRECs were amplified in genomic DNA from mononuclear cells of 10 cord blood samples, 10 cases of peripheral blood from normol individuals and 11 cases with AML-M2. Different amounts of DNA (corresponding to 2*105, 5*104, 1*104 and 1*103 cells respectively) from all samples were amplified to estimate the frequency of TCR Vβ-Dβ sjTRECs. The results showed that most Vβ subfamily sjTRECs could be detected in all samples from cord blood and peripheral blood at 2*105 or 5*104 cells level, some of Vβ subfamily sjTRECs could be detected in 1*103 cells level. Whereas the frequency of Vβ subfamily sjTRECs were lower in peripheral blood T cells from patients with AML-M2 than in normal individuals (p<0.05). Vβ19, Vβ23 and Vβ24 subfamily sjTRECs could not be detected in all samples at 5*104 cells level. The results indicated that Vβ subfamily naive T cells could be detected with different frequency in peripheral blood of normaol individuals as well as in some patients with AML-M2. Lower frequency of Vβ subfamily naive T cells was found in most AML-M2 patients.
The delta Assay - A Suitable Tool To Detect the Cellular Immune Response Against Acute Myeloid Leukemia Blasts.
Authors: Kaiser Sabine E. , Mautner Josef , Schmid Christoph , Schmetzer Helga , Gaeta Antonia , Kolb Hans-Jochem 
Published: October 2019

Abstract
Adoptive immunotherapy with donor T-cells has been used successfully for the treatment of both relapsed chronic and acute myeloid leukemia after hematopoietic stem cell transplantation. Here we describe a method for the detection of T cell immunity by the inhibition of cytokine driven growth of blasts. According to the heterogeneous growth pattern of various cases of acute myeloid leukemia (AML) the culture requirements had to be determined individually. The basic cytokine cocktail consisted of SCF (50 ng/mL), GM-CSF (100 ng/mL), IL3 (50 ng/mL), G-CSF (100 ng/mL) and EPO (2 U/mL); this cocktail succeeded to induce blast-proliferation in 15 out of 16 AML samples tested. In a first step we determined optimal growth conditions regarding cell density and the day of linear growth. The peak proliferation was measured by the incorporation of tritium labeled thymidine at various time points. It ranged between days 2 and 5. In a second step, donor cells were stimulated for 10 days with irradiated AML-cell lines (MonoMac6, THP1) or patient derived AML blasts. After harvesting, the donor cells were irradiated (15 Gy) and co-cultured with the AML-cell lines, or the patients cytokine stimulated blast suspensions. Inhibition of growth was measured by the incorporation of tritium labeled thymidine at the previously determined day of linear proliferation of the AML-blasts. A significant inhibition of blast proliferation was observed in the co-culture of normal T cells with both AML-cell lines. In HLA-identical combinations T cells were primed by dendritic cells derived from AML blasts and re-stimulated on days 6 and 10. T cells primed by this method inhibited the growth of AML blasts in higher effector target ratios, in lower ratios no inhibition or even stimulation was observed. In summary, the delta-assay is a suitable tool to monitor specific anti-leukemic immune responses of donor lymphocytes even against very heterogeneous blast populations, if individualized culture conditions are considered.
Familial Acute Myeloblastic Leukemia with a Dominant Negative Mutation in the CEBPA Gene.
Authors: Nanri Tomoko , Kawakita Toshiro , Hoshino Koyu , Mitsuya Hiroaki , Asou Norio , Uike Naokuni 
Published: October 2019

Abstract
Transcription factors involved in myeloid cell differentiation are frequent targets of chromosomal translocations or point mutations in patients with acute myeloblastic leukemia (AML). Familial AML harboring a mutation of a transcription factor should provide an association with clinical features and functions of the transcription factor. Recently, two pedigrees of AML carried a germ-line mutation in the CEBPA, the gene encoding transcription factor C/EBPα. have been reported. We here present clinical and molecular features of a Japanese family in whom two members affected by AML had an identical CEBPA mutation. Father had received the diagnosis of AML M2 in 1988 at the age of 39 years. His blast cells contained Auer rods and aberrantly expressed CD7 antigen. Following a relapse, he received autologous stem cell transplantation, after which he has been in a lasting complete remission (CR). His son was diagnosed to have M2Eo in 2004 at the age of 26 years. His marrow showed blasts including Auer rods and 6.8% eosinophils. He has achieved a continuous CR. Bone marrow cells at the time of diagnosis in both patients showed a 4-base pair insertion in the CEBPA (350_351insCTAC). The corresponding protein is predicted to terminate prematurely at codon 107 (I68fsX107). Therefore, this heterozygous mutation causes truncation of the 42-kD C/EBPα protein and overproduction of a 30-kD isoform, which functions in a dominant negative fashion, causing a decrease in C/EBPα activities. Peripheral blood cells obtained during CR in both patients also had the same mutation. Interestingly, the germ-line mutation identified in the affected individuals we present is almost identical to those in the familial AMLs carrying a C/EBPα mutation, 212delC or 217insC. N-terminal C/EBPα mutations in sporadic AML patients are associated with M1/M2, presence of Auer rods, CD7 expression, normal karyotype, and favorable prognosis. Familial AMLs with a C/EBPα mutation demonstrate links of these unique features to a dominant negative C/EBPα mutation. Although the mechanism underlying the development of AML is yet unclear, carriers of the CEBPA mutation appear to have a large population of undifferentiated myeloid cells associated with an increased risk of a second genetic hit, leading to AML over a long latency.
The Role of Leukemia Derived B7-H1 in Tumor-T-Cell Interactions in Humans.
Authors: Krusch Matthias , Wintterle Sabine , Chen Lieping , Kanz Lothar , Wiendl Heinz , Salih Helmut R. 
Published: October 2019

Abstract
Objective: Expression of the B7-homologue B7-H1 (PD1-Ligand) has been proposed to enable tumor cells to evade immune surveillance. Recently, B7-H1 on murine leukemia cells was reported to mediate resistance to cytolytic T-cell destruction. In this study we investigated the expression and functional role of the B7-homologue B7-H1 in human leukemia.
Patients and Methods: Leukemia cells from 20 patients and 9 human leukemia cell lines were investigated for B7-H1 expression by flow cytometry. Functional relevance of B7-H1 for tumor-immune interactions was assessed by coculture experiments using purified, alloreactive CD4 and CD8 T-cells in the presence of a neutralizing anti-B7-H1 antibody.
Results: Significant B7-H1 expression levels on leukemia cells were detected in 13 of 20 patients and in 8 of 9 cell lines. In contrast to various other tumor entities and the data reported from a murine leukemia system we did not observe any significant inhibitory effect of leukemia-derived B7-H1 on CD4 and CD8 cytokine production (IFN-g, IL-2) or expression of T-cell activation markers (ICOS, CD69). In the presence of a neutralizing B7-H1 antibody (mAb 5H1) no significant changes in T cell IFN-g or IL-2 production were observed.
Conclusions: Our data demonstrate that leukemia-derived B7-H1 seems to have no direct influence on T-cell activation and cytokine production in humans. Further experiments are warranted to delineate factors and characterize yet unidentified B7-H1 receptor(s) that determine inhibitory and stimulatory functions of B7-H1 in human leukemia.
Early Intervention in Minimal Residual Disease after Allogeneic Transplant for Philadelphia Chromosome-Positive Acute Lymphoblastic Leukemia: Prolonged Survival by Imatinib.
Authors: Miyamura Koichi , Yoshihisa Kodera , Skamaki Satoshi , Nakao Shinji , Sasaki Satoshi , Yokozawa Toshiya , Kato Chiaki 
Published: October 2019

Abstract
Minimal residual disease (MRD) was examined using quantitative polymerase chain reaction in 34 patients with Philadelphia chromosome- positive acute lymphoblastic leukemia (Ph+ ALL) who received allogeneic hematopoietic stem cell transplantation (HSCT). Patients were divided into two groups according to the time of transplant (Group A; n=17, – 2000 and Group B; n=17, 2001-). Group B had a significantly higher overall survival rate than Group B (74.2% vs 29.4%, P<0.01). In Group A, MRD+ after HSCT indicated imminent hematological relapse with subsequent death. On the contrary, in Group B, 8 patients showed MRD after HSCT. However, only one died of leukemia during the observation. Among the MRD+ patients after HSCT, the probability of hematological relapse was higher in group A than in group B (100% vs 20%, P<0.05). MRD-based clinical interventions such as Imatinib administration, donor lymphocyte infusion, or second HSCT may contribute to longer survival. In particular, imatinib introduce MRD-negativity in most patients, resulting better performance status and time for preparing next strategies. Two patients who became MRD-negative after imatinib treatment developed molecular relapse after imatinib discontinuation. Thus, Imatinib may contribute to the improvement of overall survival, however, an allogeneic power may be required for the cure of leukemia.
CXCR4 Antagonists Mobilize Acute Lymphoblastic Leukemia Cells into the Peripheral Blood and Inhibit Engraftment in Mice.
Authors: Juarez Julius , Hewson John , Cisterne Adam , Baraz Rana , Bradstock Kenneth F. , Bendall Linda J. 
Published: October 2019

Abstract
The role of CXCL12 in the growth of B cell progenitor acute lymphoblastic leukemia (ALL) and the homing of these cells to the bone marrow has been well established. However the effect of modulating CXCL12/CXCR4 interactions on the growth of ALL cells in vivo has not been examined. In this study we used specific peptide and small molecule antagonists of CXCR4 to examine the importance of CXCL12/CXCR4 interactions in the development of leukemia in an in-vivo murine model of ALL. CXCR4 antagonists induced mobilization of human and murine B cell progenitor ALL cells into the peripheral blood, with a 3.8±1.9 and 6.5±3.3 fold increase in leukemic cells/ml one hour after administration of the antagonist respectively, similar to that observed for normal progenitors. Daily administration of AMD3100 commencing the day following the injection of cells and continuing for 21 days resulted in a mean reduction in peripheral blood white cell count of 50±12% and the leukemic cell count of 63±4%. There was also a significant reduction in both the total cells in the spleen of 58±1% and the leukemic cell number in this organ of 75±11%. A significant reduction in leukemic cell numbers in the bone marrow was observed in one (44% reduction) case. There was reduced infiltration of other organs including kidney, liver and skeletal muscle. This study demonstrates that disrupting the CXCL12/CXCR4 axis in B cell progenitor ALL reduces the tumor burden. Whether this is due to direct inhibitory effects on proliferation and survival, or results from disruption of the leukemic cell interactions within the bone marrow remains to be determined.
Expression of CD25+ Cell Subsets after Induction Chemotherapy in Adult Patients with Acute Myeloid Leukemia.
Authors: Min Woo-Sung , Kim Hee-Je , Choi Young , Kim Sung-Yong , Eom Ki-Sung , Kim Yoo-Jin , Lee Seok , Min Chang-Ki , Lee Jong-Wook , Kim Chun-Choo 
Published: October 2019

Abstract
An attempt was made to correlate clinical outcomes with specific patterns of the expression of CD25+ cells after induction chemotherapy (IC) in adult patients with acute myeloid leukemia (AML). Seventy-five newly diagnosed AML patients received the same initial IC and serial bone marrow (BM)- or peripheral blood (PB)-samples were taken. The gated CD45/CD25/CD4 cell populations were used to compare for the intensity of immunophenotypic signals and the different cell subsets, according to the treatment timeline. As one of the best predictive prognostic parameters, patients who responded poorly to IC showed exceptionally higher levels of PB CD45+CD25+ cells on days 7 (P=0.002) and 21 (P=0.05) post-IC. The results of patients in complete remission (CR)(n=61), as well as those of the patients who showed continuous CR, showed relatively lower levels of PB CD45+CD25+ and higher CD4+CD25+ regulatory T cells in the steady PB after the standard IC, which was accurately discernible in every patient and in normal healthy individuals (n=21). We found considerably lower expression levels of BM/PB CD4+CD25+ regulatory T cells in the patients. These results suggest that this tool can be used to predict the outcome of adult AML patients at an early period in treatment.
Retroviral Gene Therapy of RAG-1-/- Mice: Balance between Efficiency and Toxicity.
Authors: Lagresle-Peyrou Chantal , Yates Frank , Malassis-Seris Michele , Hue Christophe , Morillon Estelle , Garrigue Alexandrine , Liu Allen , Hajdari Philippe , Stockholm Daniel , Danos Olivier , Lemercier Brigitte , Gougeon Marie-Lize , Rieux-laucat Frederic , de Villartay Jean-Pierre , Fischer Alain , Cavazzana-Calvo Marina 
Published: October 2019

Abstract
Patients affected by severe combined immunodeficiency (SCID) have a profound defect in T cell development and the only curative treatment is hematopoietic stem cell transplantation (HSCT). When an HLA-identical family related donor exists, the success rate of HSCT is 90% while in all others cases, it ranges between 40 and 70%. In SCID patients also presenting B cell differentiation defects, such as in RAG-1 and RAG-2 deficiencies, the survival rate is significantly reduced. We have thus been interested in the development of a potential alternative treatment by using retroviral gene transfer of a normal copy of RAG-1 cDNA. To this end, murine RAG-1 deficient hematopoietic stem cells were transduced with a retroviral modified Moloney leukemia virus vector containing the RAG-1cDNA and transplanted into RAG-1-deficient mice. B and T cell development was achieved in all the recipient mice and the reconstitution is stable over time, attesting for a selective advantage of transduced progenitors. The T cell compartment was found to be diverse and functional as shown by Va and Vb TCR immunoscope analysis as well as proliferation assays in the presence of mitogens. The mature circulating B lymphocytes were not abundant but functional as the serum immunoglobulin levels and specific antibody response against the T cell dependent KLH antigen were always within normal range. Noteworthy, a high transgene copy number was detected in all lymphoid organs and this was associated with a risk of insertional mutagenesis as observed in one mouse. Altogether, these results demonstrate that RAG-1 gene transfer can correct immunodeficiency associated with RAG-1 defect but that human application would require the use of lentivirus vectors with self inactivating long terminal repeats in order to decrease the risk of lymphoproliferative diseases.