bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hematopoietic Progenitor Cell Response to SDF-1 alpha and SDF-1 alpha Production Are Impaired in Chronic Phase Chronic Myeloid Leukemia at Diagnosis.
Authors: Prebet Thomas , Chabane Kaddour , Tigaud Isabelle , Wattel Eric , Michallet Mauricette , Nicolini Franck E. 
Published: April 2020

Abstract
SDF-1 alpha and its cognate receptor CXCR-4 are involved in normal Hematopoietic Progenitor Cells (HPC) regulation of adhesion, migration and survival. In Chronic Phase (CP) Chronic Myeloid leukaemia (CML), HPC exhibit a deregulation of all of those phenomena. Here, we investigated by different ways, the involvement of SDF-1 alpha in the pathogenesis of CP CML. Samples of bone marrow (BM) and peripheral blood (PB) from normal donors (ND) (n=10), CP CML (100% Ph1+ metaphases, all selected for Ph1+ LTC-IC) patients at diagnosis (n=35) and CP CML patients in Complete Cytogenetic Response (CCR) (0% Ph1+) (n=10) were explored. First, SDF-1 alpha concentrations were determined by ELISA assays in PB and BM samples, and showed higher yields for CP CML PB (2060±700 pg/ml) compared to CCR PB [1570±300 pg/ml (p=0.002)] and lower concentrations for CP CML BM (3080±1015 pg/ml) compared to CML in CCR BM [3990±680 pg/ml (p=0.01)] and ND BM [4060±1020 pg/ml (p=0.03)]. CCR BM and PB, and ND BM and PB SDF-1 alpha concentrations were not different. Further, we analyzed CD34+Lin− HPC migration in a 4h-Transwell™ assay, in SFM conditions ± SDF-1 alpha or ± one of its specific inhibitors SDF-1(G2) vs control. After migration, median percentages of migrating cells were consistently higher for CML vs ND BM (32±9 vs 20±1, p=0.002) in control arm, but equivalent for CML samples with SDF-1 alpha (39±10) vs control (32±9) indicating a relative insensitivity of CML HPC to this chemokine. In addition, SDF-1(G2) significantly inhibited the migration of CML HPCs (28±7) vs SDF-1 alpha (p=0.01). The proportion of CFC and LTC-IC in the migrating fraction was equivalent in the 3 experimental arms (SDF-1 alpha vs SDF-1(G2) vs control). As assessed by FACS, CD34+Lin− cells CXCR-4 surface expression was equivalent for CP CML (31±2.3%) and ND BM (43±2%) before migration, and there was no modification of the CXCR-4 expression after migration for CP CML. As SDF-1 alpha is involved in the regulation of the proliferation of normal HPC, we assessed CP CML CD34+Lin− cells in 3H-Thymidine cycling assays after 14 days culture on M210B4 murine feeder cell line (producing residual concentrations of SDF-1 alpha). CP CML HPC (CFC and LTC-IC) cycling status was not modified either after SDF-1 alpha or SDF-1(G2) exposure whereas ND BM cycling was paradoxically up-regulated by SDF-1(G2) vs control, suggesting a specific functional defect of CP CML HPCs in response to SDF-1 alpha. Taken together, all these results suggest that SDF-1 alpha production by stromal cells is impaired in CP CML, and that migratory and proliferative responses of CP CML HPCs to SDF-1 alpha are likely to be perturbed. This results might illustrate some pathologic interactions between BCR-ABL and signal transduction pathways activated through CXCR-4 ligation to its sole ligand SDF-1 alpha.
Cell Adhesion and Migration Regulated by EphB6 Expressed on Human Hematopoietic Progenitors.
Authors: Matsui Toshimitsu , Matsuoka Hiroshi , Tamekane Akira , Inoue Ryuichi , Shimoyama Manabu , Okamura Atsushi , Obama Hiroya , Kelly Meghan L. , Nakamoto Masaru 
Published: April 2020

Abstract
Normal human hematopoietic progenitors as well as leukemia/lymphoma cells express kinase-defective EphB6 receptors. The only unique high affinity ligand for EphB6 among eight known mammalian ephrins, ephrins-B2 is expressed not only on hematopoietic malignancies, but also on mesenchymal stem cells. However, the biological functions of the receptor and its ligand in hematopoietic cells are largely unknown.
In the present study, we showed that the interaction between EphB6 and ephrinB2 could initiate forward as well as reverse signaling in vitro. Both pre-clustered and unclustered ligands could trigger the signal transduction, but pre-clustered ones more rapidly down-regulated the signaling. We also examined the EphB6/ephrinB2 function in cell adhesion and migration.
Figure Figure
HEK-EphB6 cells placed in the upper chamber of a Transwell apparatus, in which the lower side of filter was coated with different concentrations of ephrin-B2-Fc or Fc, were allowed to migrate to the lower side at 37°C overnight. Vector-transfected cells were used as controls. The cells that had migrated to the lower side of filter were stained, photographed. A BSA-coated filter is shown as a control. EphB6 exerted biphasic effects in response to different concentrations of the ephrin-B2. EphB6 promoted cell adhesion and migration when stimulated with low concentrations of ephrin-B2, whereas it induced repulsion and inhibited migration upon stimulation with high concentrations of ephrin-B2. A truncated EphB6 receptor lacking the cytoplasmic domain showed monophasic positive effects on cell adhesion and migration, indicating that the cytoplasmic domain is essential for the negative effects. We further explored the signal transduction of the biphasic effects.
Figure Figure
The Src family kinase, Fyn was co-immunoprecipitated with anti-EphB6 antibody in the absence or presence of ephrin-B2 stimulation. High concentrations of ephrin-B2 induced tyrosine phosphorylation of EphB6 through a Src family kinase activity. These results indicate that EphB6 can both positively and negatively regulate cell adhesion and migration, and suggest that tyrosine phosphorylation of the kinase-defective EphB6 receptor by a Src family kinase acts as the molecular switch for the functional transition.
Thus, EphB6 expressed on hematopoietic cells may play an important role in the regulation of cell homing to hematopoietic tissues as well as leukemia cell infiltration.
Abnormal Bone Marrow-Derived Mesenchymal Stem Cell: Another Clone in Hematological Malignancy?.
Authors: Yeh Su-Peng , Chiu Chang-Fang , Liao Yu-Min , Lin Chiao-Lin , Lo Wen-Jyi , Lee Cheng-Chun 
Published: April 2020

Abstract
The growth, differentiation, and survival of normal hematopoietic stem cells as well as bone marrow (BM) cancer cells are regulated closely by the BM microenvironment. In case of lymphoma, recent study has demonstrated that the endothelial cells (not lymphoma cells) in the lymphoma microenvironment have the same chromosomal abnormality to that of lymphoma cells themselves. Besides, in our prior study, the mesenchymal stem cells (MSCs) isolated from BM of patients with various hematological malignancies may have different surface antigens expression as comparing to normal control (Leukemia, 2005, doi:10.1038/sj.leu.2403795, published online 19 May 2005). We therefore hypothesize that the MSCs isolated from BM of patients with hematological malignancies are “different” to those of normal and may probably has the same chromosomal abnormality as to that of the hematological cancer cells. In this study, cytogenetic studies were performed on both BM-blood cells and BM-derived MSCs at the same time by using method of conventional G-banding. At submission of this abstract, we have just examined 4 specimens obtained from patients of various diseases as well as normal adult (see table). Surprisingly, we identified a new chromosomal translocation, t(3;9)(q25;q31), on the BM-derived MSCs of a patient (Case 1 of the table, Figure 1) with acute promyelocytic leukemia harboring t(15;17) on the BM blood cells. Besides, a complex chromosomal abnormalities was found in a male patient of AML (Case 2 of the table, Figure 2). Though he has received a successful allotransplant 2 years ago and the BM blood cells had completely converted to donor type (normal 46 XX), the BM-derived MSCs were still recipient in origin and having a chromosomal abnormality that was different to the BM blood cells (45, -Y) at initial diagnosis of AML. BM-derived MSCs from a patient of CML and normal adult were normal. It is still too early to making a solid conclusion at present and we are now analyzing more specimens to look into the phenomenon we found. However, this is a new field in leukemic research that deserved further investigation and our novel finding indicates that abnormal cells (another clone?) may present in the BM stroma of patients with hematological malignancies.
Chromosomes of BM-blood cells and MSCs Case No. Diseases Chromosomes of BM-blood cells Chromosomes of BM-MSCs Case 1 APL, at diagnosis 46 XX, t(15;17)(q21;q11) 46 XX, t(3;9)(q25;q31). (see figure 1) Case 2 AML (45, -Y at diagnosis), after allotransplant Normal 46 XX (male patient, female donor) 46 XY, t(7;22)(p22;q11); del(13)(q12q22); del(15)(q15?). (see figure 2) Case 3 CML, at diagnosis 46 XY, t(9;22)(q34;q11) Normal 46 XY Case 4 Normal adult Normal 46 XY Normal 46 XY
Figure Figure
Molecular Characterization of Unique Junctional Complexes as Communication Pathways among Mesenchymal Stem Cells.
Authors: Wuchter Patrick , Boda-Heggemann Judit , Straub Beate , Krause Ulf , Seckinger Anja , Kuhn Caecilia , Grund Christine , Gottschling Sandra , Franke Werner W. , Ho Anthony D. 
Published: April 2020

Abstract
The interaction between stem cells and their niche is essential for the balance between self-renewal and differentiation. Mesenchymal stem cells (MSC) from bone marrow are able to differentiate into various tissues, such as bone, cartilage, muscle and fat. We have characterized the intercellular contacts among MSC and previously demonstrated the occurrence of a novel kind of adhering junction, consisting of villiform-to-vermiform cell projections (processus adhaerentes). The molecular compositions of these junctions have now been elucidated. Using a panel of antibodies specific for various components of tight, gap and adhering junctions (the latter comprising desmosomes and adhering junctions), we systematically analyzed the junctional complexes of MSC obtained from bone marrow aspirates from healthy voluntary donors and compared the data to bone marrow tissue in situ. Light and electron microscopy was used and several biochemical analyses were applied, including immunoprecipitation and RT-PCR. We could show that intercellular connections in MSC are realized in vitro by occasional gap junctions and frequent adhering junctions. Intercellular projections are noted, often deeply intruding into the neighbouring cell. These findings have been confirmed in situ in bone marrow biopsy material. Furthermore, we analyzed the molecular composition of these junctions and cell-projections. In most of them, we found the transmembrane glycoproteins cadherin 11 and N-cadherin, together with the cytoplasmic plaque proteins alpha- and beta-catenin and protein p120ctn. Interestingly, similar junctions had been already noted in primary mesenchymal stem cells of day 7 to 8.5 mouse embryos. Our hypothesis is that this junction type might be a prerequisite for early mesenchymal cells. Our findings indicate that intercellular communication in MSC in human marrow is realized through a unique type of junctional complexes not described before. These structures are crucial for the long-term fate and differentiation of the cells and might have significant impact on the interaction with hematopoietic stem cells.
Two Cellular Components of Bone Marrow Origin Contribute to Pulmonary Irradiation Fibrosis.
Authors: Greenberger Joel S. , Niu Yunyun , Zhang Xichen , Cao Shaonan , Carlos Timothy , Epperly Michael W. 
Published: April 2020

Abstract
Following lung irradiation of C57BL/6NHsd mice, migration of marrow origin macrophages into the lungs is detectable at 100 days followed by migration of marrow origin fibroblasts into areas of developing organizing alveolitis/fibrosis. To demonstrate the importance of each cellular migration, mice were irradiated to 20 Gy to the pulmonary cavity. Beginning at day 80, the mice were injected (daily for 15 days I.P.) with antibodies to either mouse macrophages or to a pan-T-cell antigen (M1/70 or TIB-207, respectively). Groups of mice were injected at day 80 with DS-red labeled Smad3+/+ clonal bone marrow stromal cell line cells or a GFP+ Smad3−/− bone marrow stromal cell line. The mice were then followed for survival. Mice injected with the M1/70 antibody had increased survival compared to those receiving TIB-207 or control irradiated mice (percent survival at 180 days 90% and 60% compared to 40%, p=0.0225 and 0.8263). There was a decrease in the number of macrophages in the lungs of mice injected with M1/70 compared to TIB-207 injected or control irradiated mice. Mice injected with bone marrow stromal cells from the Smad3−/− line, which have abrogation of the TGF-β signaling pathway, showed decreased numbers of labeled cells in areas of organizing alveolitis/fibrosis compared to mice that were injected with Smad3+/+ cells at 120 days (5.4 ± 1.6% vs. 21.0 ± 3.3% cells per 100x high powered field in areas of organizing alveolitis/fibrosis, respectively, p = 0.0003). These results demonstrate a critical importance of both marrow hematopoietic (macrophage) and stromal fibroblasts to the development of irradiation-induced pulmonary fibrosis.
A ComParison of Retroviral Integration Sites in Donor T-Lymphocytes In Vivo and In Vitro.
Authors: Giordano Frank A. , Laufs Stephanie , Fehse Boris , Nagy K. Zsuzsanna , Hotz-Wagenblatt Agnes , Lauterborn Daniel , Fellenberg Kurt , Kuehlcke Klaus , Naundorf Sonja , Zander Axel R. , Zeller W. Jens , Fruehauf Stefan 
Published: April 2020

Abstract
Genetically modified T-Lymphocytes (TLCs) have been used for adoptive immunotherapy in the context of allogeneic hematopoietic stem cell transplantation (SCT). Infused TLCs have been shown to be susceptible to elimination through exposure to ganciclovir in the event of graft-versus-host disease (GvHD).
Yet, reports on insertional mutagenesis in a mouse gene marking study and a clinical gene therapy trial for X-chromosomal severe combined immunodeficiency (X-SCID) reminded us the actual risk of insertional oncogene activation and subsequent leukemia development.
We investigated retroviral integration sites in vitro and in vivo. Therefore, donor TLCs transduced with the MoLV-based TK/neoR vector Mo3TIN for a clinical HSV-Tk study were examined. TLCs of four different donors as well as whole blood samples of two patients transplanted with donor TLCs were analyzed either using highly sensitive and specific ligation-mediated PCR (LM-PCR).
A total of 114 retroviral integration sites were detected in vitro. 41.2% of all integrations appeared near the transcription start regions (+/−10kb) of genes. Further analysis showed that 57 (50%) of all integrations targeted RefSeq genes. 24 of those appeared in intron 1 (42% of all integrations into genes) while 18% (10/57) of all integrations into genes landed in exon sequences whereas 6 hit the first exon. 18 of the targeted genes (15.8% of all integrations) could be at last assigned to signal transduction pathways, whereas the transcription factor family was afflicted 13 times (11.4% of all integrations). Among the targeted genes we found integrations into the CD8, CD100, CD44, CX3CR1, HLA-DMP and IL10-receptor genes. Within at a range of 5kb up- and 5kb downstream of vector integrations 15 genes were located that were not hit. 5 are known as transcription factors, whereas two of those are involved in leukemia, namely the homo sapiens myeloid/lymphoid or mixed-lineage leukemia 5 gene (MLL5) and the homo sapiens ALL1 fused gene from 5q31 (AF5Q31).
Current analyses are focusing at the in vivo pattern of retroviral integration in DNA of TLCs obtained from transplanted patient’s TLCs. Therefore we developed a new high sensitive PCR method (HS-PCR), an improved LM-PCR to even detect minimal quantities of transduced DNA.
Impact of Dendritic Cell CD16+ Recovery on Outcome after Reduced-Intensity Conditioning Allogeneic Stem Cell Transplantation.
Authors: Talarn Carme , Urbano-Ispizua Alvaro , Martino Rodrigo , Batlle Montse , Herrera Concha , Perez-Simon Jose Antonio , Torrebadell Montse , Fernandez-Aviles Francesc , Gaya Anna , Granell Miquel , Marin Pedro , Sierra Jordi , Montserrat Emili 
Published: April 2020

Abstract
Dendritic cells (DCs) play a critical role in the regulation of alloimmune responses. Therefore, the number and function of these cells after allogeneic stem cell transplantation (allo-SCT) might influence patients’ outcome, an aspect scarcely investigated, particularly in the setting of reduced-intensity conditioning transplants (allo-RIC). Against this background, we studied DCs recovery in 92 patients (median age 50, range 17–67; male 57%), undergoing allo-RIC from HLA identical donors in five Spanish institutions between October 2002 and December 2004. Median follow up was 250 days (range, 25–708). Conditioning regimen consisted on fludarabine 150 mg/m2 + melphalan 140 mg/m2 for lymphoid malignancies (n=54), and fludarabine 150 mg/m2 + busulphan 10 mg/kg for myeloid malignancies (n=38); 71 (90%) patients had advanced disease. Peripheral blood samples were obtained at 1, 3, 6 and 12 months after transplant. DCs were identified as positive for HLA-DR and negative for lineage markers (CD3, CD14, CD19, and CD56). The expression of CD33, CD123 and CD16 was used to identify DC1, DC2 and DC CD16+ subsets, respectively; the immunophenotypic analysis being centralized at a single institution. The most significant association with clinical outcome was found with DC CD16+ levels at three months after transplantantation. Thus, at that time point, patients with a DC CD16+ count lower than the median (&lt;1.5×104/mL) had a significantly higher incidence of relapse (59% vs. 45%; p=0.04), more transplant related mortality (30% vs. 0%; p=0.01), worse event free survival (EFS) (24% vs. 51%; p=0.002) and overall survival (0% vs. 55%; p=0.002). Only two factors were associated with worse EFS in the multivariate analysis: autologous transplantation before allo-RIC (RR 6.5, 95%CI 1.6–26.2; p=0.008), and DC CD16+ count lower than the median at +3m (RR 13.1; 95% CI 1.7–100.5; p=0.01). In conclusion, this study shows that besides a well-known poor prognostic factor (i.e., prior autologous transplantation) a low DC CD16+ count at three months after allo-RIC is associated with a poorer outcome, this probably reflecting impaired immunosurveillance mechanisms.
An Analysis of Angiotensin Converting Enzyme Gene Polymorphisms and Non-Infectious Pulmonary Dysfunction after Hematopoietic Stem Cell Transplantation.
Authors: Onizuka Makoto , Ando Kiyoshi , Kishi Kengi , Kawada Hiroshi , Morishima Yasuo , Hotta Tomomitsu , Inoko Hidetoshi 
Published: April 2020

Abstract
Non-infectious pulmonary dysfunction (NIPD) is a common but often fatal complication associated with allogeneic hematopoietic stem cell transplantation (HSCT). An insertion/deletion (I/D) polymorphism in the angiotensin-converting enzyme (ACE) gene has extensively been studied in relation to pulmonary fibrosis, and the D-allele frequency was reported to be significantly higher in patients with pulmonary fibrosis than in the control. We have previously found that increased D-allele frequency is also associated with NIPD after HSCT from HLA haplo-identical sibling donor. In this study, we performed a retrospective analysis to assess the relation between ACE I/D polymorphism and unrelated donor HSCT. 772 patients were enrolled in this study (461 males and 311 females, median age 23; range 0 to 52). The underlying diseases were 229 ALL, 188 ANLL, 168 CML, 66 MDS, 74 aplastic anemia, 24 lymphoma, 12 inherited disease and an other disease. DNA samples and clinical information were obtained from the Japan Marrow Donor Program registry. Determination of the ACE genotype was accomplished by polymerase chain reaction. NIPD occurred in 73 patients (9.5%). The D allele frequency and the D/D genotype frequency were not significantly different between the patients with NIPD and the patients without NIPD. However, the D/D genotype patients, whose donor also had D/D were frequently observed in the NIPD population (23.8% vs 9.1%; p = 0.05). Furthermore, an investigation of the relation between ACE genotype and onset of NIPD revealed that NIPD patients with the D/D genotype tended to be diagnosed faster than patients with the I/D or I/I (mean onset day 48.5, 95.2 and 110.6; p = 0.037). Our results suggest the involvement of ACE gene polymorphism in early onset NIPD, which may be different from late onset NIPD in respect to the pathogenesis.
Haploidentical Stem Cell Transplantation in Children: A Comparison between Positive Selection of Stem Cells and Depletion of T- and B-Cells.
Authors: Lang Peter , Bader Peter , Greil Johann , Klingebiel Thomas , Niethammer Dietrich , Handgretinger Rupert 
Published: April 2020

Abstract
Profound depletion of T- and B-cells is a fundamental prerequisite for haploidentical 1–3 loci mismatched stem cell transplantation. Here, we compare hematopoietic engraftment and occurrence of GvHD after positive selection procedures (with anti-CD34 coated or anti-CD133 coated beads, n=64 patients) and after a direct depletion procedure with anti-CD3/anti-CD19 coated microbeads (n=12) in children with leukemias (ALL, AML, CML), lymphomas and nonmalignant diseases. Median purity of stem cells was comparable after CD34+ selection and CD133+ selection, whereas stem cells were only slightly enriched after CD3+/CD19+ depletion (97.5%, 93.4% and 1.02%). Indirect depletion of T-cells by positive selection methods resulted in 10 000 residual CD3+ cells/kg (7000–30 000). Patients with grafts depleted with anti-CD3/anti-CD19 coated microbeads, received 32 000 (7000–160 000) residual T-cells/kg. Those grafts comprised also remarkable amounts of effector cells such as NK-cells (median number: 86 × 106/kg), dendritic cells and monocytes/granulocytes (median number: 700 × 106/kg) with antileukemic activity in vitro. Primary engraftment occurred in 83% of patients with positive selected stem cells. After reconditioning, sustained engraftment was achieved in 98%. In patients with CD3+/CD19+ depleted grafts, primary engraftment was observed in 92% (11/12) and sustained engraftment (after reconditioning) in 100% (12/12). GvHD grade II occurred in 6% (positive selection) and 25% (CD3/CD19 depletion). Only one patient had severe GvHD grade III-IV (CD34+). T-cell recovery was significantly faster in patients with CD3/CD19 depleted grafts. In the group with positive selected grafts, 5 year EFS for patients with ALL CR1-3, myeloic leukemias and nonmalignant diseases was 45%, 22% and 60%, respectively. CD3+/CD19+ depleted grafts were given to patients with refractory disease (AML/MDS, ALL) or with high risk of graft failure. 4/10 patients with leukemias and 2/2 patients with SAA/PNH are disease free (median follow-up: 8 months). Conclusions: Haploidentical transplantation with standard CD34+ or CD133+ selected grafts provides stable long term survival rates. CD3+/CD19+ depletion resulted in better and more reliable engraftment but produced slightly more GvHD than positive selection methods. Our preliminary results indicate, that CD3/CD19 selected grafts may be advantageous regarding engraftment and immunoreconstitution. Since effector cell with potential antileukemic activity are cotransfused, such grafts may be suited in particular for patients with insufficient remission.