bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification and Characterization of Hemoangiogenic Progenitors during Cynomolgus Monkey ES Cell Differentiation.
Authors: Umeda Katsutsugu , Heike Toshio , Yoshimoto Momoko , Shiota Mitsutaka , Ma Feng , Suemori Hirofumi , Luo Hong Yuan , Chui David H.K. , Torii Ryuzo , Shibuya Masabumi , Nakatsuji Norio , Nakahata Tatsutoshi 
Published: October 2019

Abstract
Previous reports about the close association of development of hematopoietic (HCs) and endothelial cells (ECs) have suggested the presence of the hemangioblast, a putative common precursor of both lineage cells in mouse. The presence of putative ancestors is suggested by the fact that both HCs and ECs share common markers such as vascular endothelial growth factor receptor (VEGFR)-2, CD34, and CD31. The precise analysis about the relationship of HCs and ECs in human embryogenesis, however, remains unclear because of obstacle including the ethical restrictions on experiments using their embryo. To further understand the mechanisms that regulate the differentiation of HCs and ECs in humans, it is currently necessary to employ primate (human and monkey) ES cells. In this study, we identified the intermediate-stage cells with HC and EC differentiation potentials from nonhuman primate ES cells by coculture with OP9 stromal cells. Sequential FACS analysis demonstrated that during in vitro HC and EC differentiation, the expression of common surface markers such as CD34 and CD31 could first be detected, followed by the expression of lineage-specific markers (CD45, CD41 and VE-cadherin) in order. Sequential FACS and immunostaining analyses showed that both lineage developments concomitantly occurred, which were preceded by emergence of two hemoangiogenic progenitors, VEGFR-2high CD34− and VEGFR-2high CD34+ cells. While both VEGFR-2 high cells could differentiate into primitive and definitive HCs as well as ECs, VEGFR-2high CD34+ cells possessed higher hemoangiogenic potential than VEGFR-2high CD34− cells. In contrast, HC and EC production cluster was rare in the VEGFR-2low or − fractions. During HC and EC differentiation, most of VEGFR-2high cells first expressed CD34, and then diverged into both lineage cells at the VEGFR-2high CD34+ stage. We found that the VEGFR-2high cells can generate both HCs and ECs at the single cell level, which strongly supports the existence of hemangioblasts in primates. To our knowledge, this is the first time that hemangioblasts have been shown to be present during primate ES cell differentiation. Furthermore, differentiation pathway into HC and EC lineages can be defined by surface markers, which facilitates further investigations in vitro and in vivo.
Characterization of Endothelial Progenitor Cells in Healthy Subjects: Effects of Different Factors on Immunophenotype and Clonogenic Potential.
Authors: Giorgetti Alessandra , Ciulla Michele M. , Cortiana Michela , Silvestri Ilaria , Montelatici Elisa , Giordano Rosaria , Paliotti Roberta , Cortelezzi Agostino , Rebulla Paolo , Lazzari Lorenza 
Published: October 2019

Abstract
There are experimental evidences that Endothelial Progenitor Cells (EPCs) is a pool of cells that could proliferate and differentiate into mature endothelial cells and that may contribute to endothelial repair, but no systematic studies exist regarding physiologic variations in EP circulating pool and in CFU-Endo capacity in healthy subjects. In this regard, since the level of EPCs may be a biologic marker for vascular function, we decided to characterize primitive and circulating EPCs and their relation with gender, aging and the capacity to form CFU-Endo in a selected healthy population. Each subject underwent a screening evaluation to exclude any underlying cardiac diseases. A 20mL sample of peripheral blood was obtained (h 11:00–12:00 AM) from 15 male and 17 female age matched subjects (mean age 41.6±15.9yrs). EPCs were assessed by flow cytometric analysis on 100,000 events per sample (FACScan, Becton Dickinson). Primitive EPCs, defined as CD45−/34+/133+ and CD45−/KDR+/144−, were 8±1.8SE/uL. Circulating EPCs, defined as CD45−/146+/31+/FCSlow and CD45−/146+/144+, were 18.72±2.5SE/uL. No correlation was found between these populations and aging, gender and BMI. Instead a significant decrease in CD45−/34+/133+ (2.9±0.6SE/uL) subpopulation was observed with aging (r = 0.45; p = 0.009). CFU-Endo were performed as described by Hill JM et al. (NEJM 2003) and scored by microscopy in blind by two operators. The confirmation of endothelial-cell lineage was performed by immunostaining for endothelial-specific antibodies (CD31+, CD105+, CD144+, CD146low, CD133−, vWf-). CFU-Endo were expressed as the mean number of colonies per well in 8–25 separate determinations for each subjects and only 15 subjects gave colonies (mean 2.1±0.05SE). CFU-Endo do not correlate with aging and gender in healthy subjects confirming that endothelial injuries and cardiovascular risk factors are a crucial endothelium stimulus. In fact, recently it has been reported that higher colony counts were found in subjects with various degrees of cardiovascular risks. In conclusion CD45-/34+/133+ cell population correlates with aging but since this population represents Hemato-Endo Precursors that include a majority of endothelial progenitors and a small number of hematopoietic stem cells, further studies will be needed to identify a subpopulation strongly predictive of CFU-Endo capacity and to assess its endothelial lineage switch.
Modulation of Cord Blood CD8+ T Cell Effector Differentiation by 4-1BB and CD30: Cross-Talk with TGF-β1.
Authors: Kim Young-June , Stringfield Teresa M. , Chen Yan , Broxmeyer Hal E. 
Published: October 2019

Abstract
Transforming growth factor-β1 (TGF-β1), an immunosuppressive cytokine, potently suppressed cytotoxic T lymphocyte (CTL) differentiation of human cord blood naïve CD8+ T cells as determined by reduced induction of characteristic phenotypes of effector cells and cytotoxic activity. In contrast, 4-1BB (CD137), a costimulatory molecule in the TNF receptor family, promoted effector and memory CTL differentiation. We present evidence that the suppressive TGF-β1 effects can be effectively reversed by anti-4-1BB costimulnation during in vitro CTL differentiation driven by IL-15 from exclusively naïve cord blood CD8+ T cells. 4-1BB may counteract TGF-β1 effects at the level of TGF-β1 signaling by inhibiting Smad2 phosphorylation induced by TGF-β1. In contrast to 4-1BB, recent studies show that CD30, another member of the TNF receptor family, may play an important role in down-modulating effector or memory CTL responses. Thus, we hypothesized that CD30 may interact with 4-1BB in responding to the TGF-β1 suppression. TGF-β1 inhibits 4-1BB expression on CTLs. Conversely, TGF-β1 greatly up-regulates CD30 expression. IL-4 augments the TGF-β1-mediated up-regulation of CD30 expression resulting in further increase of CD30 expression. In contrast, IL-12 abrogates TGF-β1-induced CD30 expression. Such IL-4 and IL-12 effects on CD30 expression appear to be exactly opposite for 4-1BB expression: down- and up-regulating 4-1BB expression, respectively. Co-engagement of CD30 abrogates 4-1BB expression as well as its activity for reversing TGF-β1-mediated suppression of CTL proliferation particularly in the presence of IL-4. Expression of NKG2D, an important stimulatory receptor for NK and CTL cytotoxic activity is inhibited by TGF-β1. 4-1BB stimulation restores the NKG2D expression inhibited by TGF-β1. Again, co-engagement of CD30 significantly inhibits the 4-1BB activity for restoration of NKG2D expression. Taken together, our data suggest that 4-1BB promotes effector CTL differentiation and CD30 acts as a negative feed-back molecule, promoting TGF-β1-mediated CTL suppression and counteracting 4-1BB activity. In conclusion, we have delineated a novel link between CD30 expression and TGF-β1 which is pivotal to the down-regulation of 4-1BB-induced CTL effector differentiation.
Interleukin-7 Over-Expression Regulates T Cell Versus B Cell Lineage Development in the Thymus.
Authors: El Kassar Nahed , Lucas Philip J. , Flomerfelt Frank , Merchant Melinda , Bare Catherine V. , Mahmud Semhar , Mackall Crystal L. , Bhandoola Avinash , Gress Ronald E. 
Published: October 2019

Abstract
We have shown that high IL-7 transgene (Tg) over-expression (39-fold at day 1 in thymic tissue) under the T cell specific, proximal lck promoter had a dose effect on TCRαβ that was accompanied by active B cell development in the thymus. To further characterize these affects in the thymus of IL-7 transgenic mice, we analyzed thymi from day 18 embryos and newborn Tg mice, as well as fetal thymic organ culture (FTOC) derived from using day 16 embryos. We show that arrested T-cell and increased B-cell thymic development is initiated during fetal development. Using mixed bone marrow chimeras and anti-IL-7 monoclonal antibody injection, we further demonstrate that abnormalities in thymic T and B cell development are non-cell autonomous and are due to IL-7 over-expression. Recently, it was shown that only the early thymocyte progenitor (ETP, c-kit+IL-7R−/lo) fraction within the DN1 subpopulation had a T-cell proliferative potential in contrast to the c-kit−IL-7R+ DN1 subset. Here we show that in Tg mice the ETPs were decreased, while the c-kit−IL-7R+ cells are increased in both percentage and absolute count when compared to normal controls. In order to explore the T vs. B ETP potential, we seeded re-aggregate thymic organ cultures with sorted lin−CD44+CD25−c-kit+IL-7R+ cells. While ETPs derived from normal controls were able to proliferate and produce 83% of DP thymocytes, ETPs sorted from Tg mice developed poorly (10-fold less) into DP cells (30%) and produced 14% of B220+ cells vs. 6% in controls. Moreover, sorted Pro/Pre B derived thymic B cells from Tg mice, but not BM-derived Pro/Pre B cells had the TCRβD-J rearrangement, suggesting a T-specific origin. Since the B-cell differentiation pathway in normal mice is selectively inhibited by thymic presentation of Notch ligands, we hypothesized that IL-7 down-regulates Notch signaling. To test this hypothesis, we analyzed thymocyte progenitors (DN1-DN4) in normal and Tg mice for the intra-cytoplasmic part of Notch, that is cleaved upon Notch/Notch-ligand activation. Notch staining was decreased in the lin−CD44+CD25inter representing the only DN2 population present in these Tg mice. These data favour a decrease of Notch signalling in mice with high IL-7 Tg over-expression, inducing a block in TCRαβ development, and skewing of thymic B cell development by T vs. B lineage subversion. These conclusions may have implications for IL-7 in the clinical setting.
The Peripheral Cannabinoid Receptor CB2 and CD40 Are Novel Biological Markers That Predict Outcome in Diffuse Large B-Cell Lymphoma of Elderly Patients.
Authors: Rayman Nazik , Lam King H. , Mulder Dries A.H , van der Holt Ronnie B. , Doorduijn Jeanette , Lowenberg Bob , Sonneveld Pieter , Delwel Ruud H.R 
Published: October 2019

Abstract
Since diffuse large B-cell lymphomas (DLBCL) are biologically and clinically heterogeneous, various attempts have been made to distinguish subtypes of DLBCL. Using gene expression profiling two main subtypes have previously been identified, i.e germinal center B-cell like (GCB) with a favorable prognosis and non-GCB-cell like DLBCL with an unfavorable outcome (Alizadeh et al. Nature403, 503–511, 2000). Applying immunohistochemical analysis, using antibodies directed against CD10, BCL-6 and MUM1, the same investigators were capable of identifying the same subclasses of favorable (GCB) and non-favorable (Non-GCB) DLBCL phenotypes (Hans et al. Blood103, 275–282, 2004). To investigate the applicability of these three markers in de novo DLBCL patients at an age of 65 years or older with advanced stage disease, a cohort of 138 cases was studied. All patients were included in a multicenter clinical trial setting and received uniform treatment. Using the markers CD10, BCL-6 and MUM1, we assigned 40 cases (29%) into the GCB (CD10+ and/or BCL6+, MUM1−) and 98 cases (71%) into the non-GCB-like (CD10−, BCL6+, Mum1+ or CD10−, BCL6−, MUM1+) DLBCL group. In contrast to previous reports, no clinical significance was observed between the two subtypes (overall survival (OS), event-free survival (EFS), progression-free survival (PFS) and disease-free survival (DFS) P-values >0.2), indicating that the prognostic significance of this classification was not evident in this cohort of elderly patients. We previously identified a novel combination of markers that could discriminate between GCB and non-GCB in normal lymphoid tissue, i.e. CB2 and the activation protein CD40. B-cells present in the germinal center (GC) of healthy individuals appeared highly positive for both CD40 and CB2, a previously identified proto-oncogene that encodes for the peripheral cannabinoid receptor. Non-GCB-lymphocytes were mainly CB2 and CD40 negative or weakly positive. We used these two markers for immunohistochemical analyses in the same cohort of novo DLBCL. Single expression of CB2 (PFS (P<0.04) and DFS (P=0.02)) and CD40 (P<0.04) was associated with a favorable clinical outcome. A combined immunophenotypic profile of N-CB2 and CD40 resulted in improved outcome prediction (PFS, P=0.02). Moreover, our data show that CB2 and CD40 are novel markers that recognize DLBCL with a particular GC-like phenotype and have impact on the clinical outcome in a cohort of elderly DLBCL patients. We suggest that CB2 and CD40 may serve as novel markers in DLBCL outcome prediction.
An IELSG International Survey of Primary Effusion Lymphoma (PEL).
Authors: Conconi Annarita , Spina Michele , Ascoli Valeria , Lopez-Guillermo Armando , Cortelazzo Sergio , Alessandro Re , Ichinohasama Ryo , Sata Tetsutaro , Luppi Mario , Vallisa Daniele , Bergonzi Cesare , Provencio Mariano , Rossi Davide , Levine Alexandra , Raphael Martine , Klepfish Abraham , Gloghini Annunziata , Prevot Sophie , Gaidano Gianluca , Carbone Antonino 
Published: October 2019

Abstract
PEL is a rare B-cell neoplasm characterized by a preferential involvement of fluid-filled body spaces, consistent infection of the tumor clone by human herpesvirus type-8 (HHV-8) and a close relationship with underlying immunodeficiency status of the host. The International Extranodal Lymphoma Study Group (IELSG) coordinated a retrospective survey involving 15 international institutions to determine the clinico-pathological features and patterns of outcome of PEL. Fourty-three patients (38 males and 5 females) were registered. Median age at diagnosis was 59 years (range 27–102). In 23 (53%) patients an associated human immunodeficiency virus (HIV) infection was reported, in one case the diagnosis of PEL was made after a solid organ transplantation, in two patients other immunodeficiency conditions were present. The tumor HHV-8 infection was demonstrated in 35 out of the 39 tested cases, Epstein-Barr virus infection in 13 of 30 cases. CD4 count was lower than 200/μl in 18 of the 25 cases in whom the data was available. An ECOG perfomance status score ≥ 2 was observed in 29 patients and the presence of B-symptoms in 20 patients. Serum LDH was elevated in 21 of the 39 tested patients. In 4 patients nodal involvement at diagnosis was reported, in 4 cases at least one extranodal site of localization other than serous cavities was present. A low/low-intermediate risk score according to International Prognostic Index was reported in 10 cases, an intermediate-high/high risk score in 29 cases. Twenty-one patients received systemic chemotherapy, in 17 cases an anthracycline-based regimen. Intrapleural cidofovir was administered in 3 patients. Twelve HIV+ patients received highly active anti-retroviral therapy (HAART), four of them as single therapy. Among the 39 patients for whom adequate follow-up data were available, median overall survival was 6.7 months, median cause-specific survival was 13.6 months and median progression-free survival was 8.3 months. Interestingly, cases of tumor complete regression after implementation of the sole HAART, after intrapleural administration of cidofovir or without any treatment were reported. Our data confirm the poor prognosis of PEL but suggest a possible heterogeneity of this entity with respect to its biological and clinical features. A review of the pathological, phenotypical and virological features is forthcoming to validate these preliminary results.
HLA-DR Protein Status Predicts Survival in Patients with Diffuse Large B Cell Lymphoma (DLBCL) Treated with the MACOP-B Chemotherapy Regimen.
Authors: Rimsza Lisa M. , Farinha Pedro , Fuchs Deborah A. , Masoudi Hamid , Connors Joseph M. , Gascoyne Randy D. 
Published: October 2019

Abstract
Background: HLA-DR is one of the MHC Class II antigen presenting molecules. Loss of HLA-DR expression on DLBCL tumor cells has been associated with poor survival in patients on Southwest Oncology Group (SWOG) studies as well as the recent Leukemia and Lymphoma Molecular Profiling project (Rosenwald et al, NEJM 2002). This study was conceived to examine the effect of HLA-DR status on a completely different cohort of patients from a single institution treated uniformly with a single chemotherapy regimen, MACOP-B, in an attempt to validate HLA-DR as an important biomarker in DLBCL.
Methods: HLA-DR immunostaining was performed using a tissue microarray block containing two 0.6mm cores of paraffin embedded tissues from 97 patients treated on the MACOP-B regimen (1981–86) at the BC Cancer Agency in Vancouver, Canada (Semin Hematol 1988, suppl 2: 41–6). Only cases of B cell lineage were used. We used the HLA-DR antibody Class II DR, (IgG2b) at a 1:50 dilution (Novocastra, UK) with epitope recovery, on an automated immunostainer (Ventana Benchmark System) using a biotin-avidin-diaminobenzidine-based detection system. Two pathologists (LR and DF) scored all slides for positive or negative results.
Results: All patients had advanced-stage disease and were treated uniformly with the MACOP-B chemotherapy regimen. The median follow-up of living patients is 17 years. The IPI was predictive of overall survival (OS) in the study group (p = 0.023). Of the 97 B cell cases, 82 had interpretable staining. The other cases were eliminated for lack of tumor (2), necrotic tissue (1), or lack of internal positive control cells in the tissue core (12). Of the remaining 82 cases, 52 were positive for HLA-DR protein with a median OS of 16.2 years while 30 were negative with a median OS of 4.2 years, (p=0.037, figure below). A Cox multivariate model established both IPI (p = 0.031) and HLA-DR status (p = 0.04) as independent predictors of OS.
Discussion: The importance of loss of antigen presenting molecules on cells of DLBCL was a key finding of the previous LLMPP work, and is now further confirmed as impacting OS in a separate set of patients treated uniformly with a different chemotherapy regimen. These findings lend credence to the hypothesis that loss of immunogenicity is of key importance in patient response to treatment and OS and suggest that specific therapies focused on this pathway may benefit patients with DLBCL regardless of their treatment regimen.
Figure Figure
The Novel De-Immunized Anti-CD20-IL-2 Immunocytokine DI-Leu16-IL-2 Is Effective in SCID Mice Bearing Rituximab-Resistant Human Lymphoma Xenografts.
Authors: Staak Jan O. , Colcher David , Lauder Scott , Wang Jianyi , Gillies Stephen D. , Raubitschek Andrew A. 
Published: October 2019

Abstract
Objectives:
Tumor-targeted MAb-cytokine fusion proteins, such as IL-2 based immunocytokines, have demonstrated potent anti-tumor activity in several pre-clinical models as well as early stage clinical trials. For the engineering of anti-CD20-IL-2, the parental murine anti-CD20 MAb Leu16 was deimmunized by removal of potential helper T cell epitopes and then humanized by combination of recombinant V regions with human L chain and H chain constant regions. Fusion of the H chain to human IL-2 resulted in the de-immunized immunocytokine DI-Leu16-IL-2 which retained full CD20 binding activity and showed enhanced ADCC effector function relative to a control MAb (rituximab). This novel molecule has been demonstrated to be effective in vivo in a Daudi Burkitt lymphoma model. We investigated the therapeutic potential in a rituximab resistant CD20+ human lymphoma xenograft mouse model.
Materials & Methods:
The mature plasmocytic B-cell line ARH77 was stably transfected with the firefly luciferase-gene using electroporation. 2x106 luciferase-transfected ARH77 cells (ARH77:luc) were inoculated s.c. into the right flank of 6–8 week old SCID mice. Groups of 8 ARH77:luc tumor bearing mice were injected i.v. with 3 doses of DI-Leu16-IL2 (25μg), Rituximab (20μg) or PBS on day 0, 2, and 4. Tumor burden was monitored by both physical measurements and bioluminescence imaging. Mice were to be sacrificed at a tumor volume > 2000mm3.
Results:
In this regimen DI-Leu16-IL2 treated mice demonstrated significant reduction in tumor growth whereas rituximab or the PBS control had no effect on ARH77:luc tumor progression. An increase of survival from 28 to 44 days, relative to rituximab treated mice and PBS-control, was seen. Re-growth of tumors in the DI-Leu16-IL2 group was observed after an average delay of 16 days. There were no signs of toxicity. Multiple regression analyses between tumor volume and light emission were performed which revealed a correlation between tumor mass and bioluminescence for the DI-Leu16-IL-2 group (r2=0.64), rituximab group (r2=0.61), PBS-group (r2=0.82), and the combined data sets (r2=0.7).
Conclusions:
DI-Leu16-IL2 demonstrated significant anti-tumor activity against rituxan-resistant human lymphoma xenografts using both physical measures and bioluminescence imaging. Despite a shorter half-life it was more effective than rituximab. Previous studies have shown the anti-tumor effects of DI-Leu16-IL2 were not entirely dependent on FcR binding suggesting that both ADCC and IL-2 targeting contribute to the superior anti-tumor activity of the immunocytokine. Ongoing studies include the use of repeat dosing cycles of this novel immunocytokine for best response using the SCID mouse/ARH77:luc tumor model. DI-Leu16-IL2 may offer promising therapeutic potential with presumed low immunogenicity for patients with CD20+ lymphoma.