bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mesenchymal Stem Cells Induce a Reversible Suppression of Alloimmune Reactions.
Authors: Tschiedel Sabine , Bartsch Kristina , Reinhardt Annette , Chiara Gentilini , Dietger Niederwieser 
Published: April 2020

Abstract
Objective Bone marrow contains pluripotent mesenchymal stem cells (MSC) that form bone, cartilage, adipose tissue and muscle. These cells are not immunogenic and escape recognition by alloreactive T cells and natural killer cells. MSC can be readily isolated from bone marrow and expanded ex vivo without modification in phenotype or loss of function. They are capable of differentiating along multiple mesenchymal lineages, play a crucial role in the bone marrow microenvironment and have profound immunosuppressive properties. In the present investigation we analysed the antiproliferative capacity of MSC using primary and secondary immune responses in vitro.
Methods MSC were isolated from heparinized bone marrow of healthy donors (n=5). Bone marrow aspirates were layered onto a Percoll cushion (density 1,073g/ml) and the MSC-enriched mononuclear fraction was collected, washed and resuspended in Dulbecco modified Eagle medium with low glucose and 10% fetal bovine serum. The cells were plated at 2x105/cm2 and maintained at 37°C in a humidified atmosphere and subcultured prior to confluency.
MSC stained positive for specific markers as SH2, SH3 and SH4 but were negative for CD14, CD34 and CD45. Immunsuppressive effects were assessed by adding third party MSC to primary mixed lymphocyte reactions (MLR) in decreasing concentrations (10%, 1%, 0,1%) on days 0–5. Cell proliferation was measured on day 6 by means of an 18-hour pulse with 3H-thymidine (3H-TdR). Secondary MLRs were performed by restimulating MSC co-cultured MLRs with the primary PBMC on day 10. In addition secondary MLRs were also tested in presence of MSC and cultured for 1, 2, 3 and 6 days in 96-well plates and proliferation was detected by 3H-TdR uptake.
Results Primary MLRs show significant inhibition of proliferation with 10% MSC added on day 0 (85% inhibition), 1 (69% inhibition) or 2 (70% inhibition) (p&lt;0,01). Similar effects were observed by adding 1% MSC on day 0 (42% inhibition) and 1 (10% inhibition). Co-cultivation with MSC over 10 days lead to delayed proliferation in secondary MLRs with peak on day 3. Secondary MLRs were also inhibited by the addition of 1% (47% inhibition) and 10% (56% inhibition) MSC independent from co-cultivation with MSC in the primary culture (11% and 44% inhibition respectively). Restimulation with third party PBMC on day 10 lead to a primary response with a proliferation peak after 6 days.
Conclusions Our findings emphasize the immunosuppressive effects of MSC. We observed that MSC-induced inhibition of primary MLRs was reversible in secondary MLRs. This clearly demonstrates that MSC do not induce tolerance and could be used in a clinical setting due to their immunomodulatory properties.
Flow Cytometric Detection and Phenotype of Circulating CD34+ Cells in Patients Affected by Chronic Lymphocytic Leukemia.
Authors: Stagno Fabio , Parrinello Nunziata Laura , Fargione Giovannella , Triolo Anna , Privitera Antonella , Seria Elisa , Mannino Francesca , Di Raimondo Francesco , Guglielmo Patrizia , Giustolisi Rosario 
Published: April 2020

Abstract
Flow cytometric determination of peripheral blood CD34+ cells provides reliable measurements of circulating hemopoietic progenitors. Since the detection of the absolute number of circulating CD34+ cells has been found of clinical utility in the setting of chronic myeloproliferative disorders, we investigated whether peripheral CD34+ cells could play any role in the clinical work-up of B-cell chronic lymphocytic leukemia (B-CLL). In this view, we determined by flow cytometry the absolute number of circulating CD34+ cells in the peripheral blood of 28 patients (16 males and 12 females, median age 67 years) affected by typical B-CLL (Matutes score 5,4,3) and in different Rai stages of the disease (19 early stage: Rai 0, I, II; 9 advanced stage: Rai III, IV). Conventional and multiparameter flow cytometric analysis was performed utilizing a FACSCalibur cytometer (Becton Dickinson). Our data showed a significant increase in the number of circulating CD34+ cells in the peripheral blood of patients with B-CLL (median CD34+ cells:7.8mL) as compared to controls (median CD34+ cells 0.1mL) (p=0.008). No statistical difference between B-CLL patients in early versus advanced stage (p=0.5) and between untreated versus treated (p=0.7) was found, as well as there was no correlation with some of the clinical characteristics of B-CLL (WBC-count, LDH levels, Beta-2M). In 10 out of 28 B-CLL affected patients, circulating CD34+ cells were correlated with ZAP-70 and CD38 antigen but no correlation was found. In addition, we detected in the peripheral blood of 22 out of 28 patients small numbers of circulating CD34+ cells displaying the CD19+/CD5+ phenotype (median CD34+/CD19+/CD5+ cells:5.7mL) whereas these cells were absent in normal controls. This unexpected finding, whose significance remains to be clarified and still restricted to a small number of cases, could be directly correlated to the underlying lymphproliferative disease and might represent a pool of leukemic stem cells. However, further studies are warranted.
Deletion of p53 as Detected by Fluorescence In Situ Hybridization Is Associated with Significant Shorter Event-Free Survival in Patients with Multiple Myeloma Who Are Receiving Allogeneic Blood Stem Cell Transplantation.
Authors: Schilling Georgia , Hansen Timon , Perez-Simon Jose Antonio , Schwerdtfeger Rainer , Bornhaeuser Martin , Otterstetter Svenja , Seeger Doris , Dierlamm Judith , Ayuk Francis , Kurt Hossfeld Dieter , Carsten Bokemeyer , Axel Zander R. , Nicolaus Kroeger 
Published: April 2020

Abstract
Deletion of chromosome band 17p13 (P53) is known to be an adverse prognostic factor for time to progression (TTP) and overall survival (OS) in myeloma patients receiving conventional or high-dose chemotherapy. In this retrospective multicenter study, we investigated the impact of P53 deletion as detected by fluorescence in situ hybridization combined with immunofluorescent cytoplasm immunoglobulin staining (cIg-FISH) on outcome after allogeneic blood stem cell transplantation (SCT) in 50 patients (pts) with advanced or relapsed multiple myeloma (MM). Median age was 51 years (34 – 67 ys.), 32 patients were male, 18 female. Thirty pts received a stem cell graft from an unrelated donor, and 20 pts from a HLA-identical sibling. P53 gene deletion was found in 5 out of 49 pts (10,2 %). Deletion 13 detected by cIg-FISH was found in 20 out of 47 pts (42%). There was a strong correlation between P53 deletion and deletion 13: four out of the five patients with P53 deletion also showed deletion in chromosome band 13q14. The estimated event-free (EFS) and overall survival (OS) at three years for the entire study population was 43% and 65%, respectively. For patients with del 13 only a trend for a worse 3 year EFS was seen (38% vs 42%, p=0.2), while pts with P53 deletion had a significant reduced EFS (0% vs 46%, p=0.0001). Three out of five pts with P53 deletion relapsed very early after allogeneic SCT (day 81, 108 and 287 respectively). These data suggest that P53 is a risk factor for patients with MM treated with allogeneic SCT which can not be overcome by this treatment strategy.
Gene Expression Profiling of MLL-PTD in AML Identifies Differentially Regulated Members of the HOX Gene Family.
Authors: Austin Stephen J. , Gilkes Mandy F. , Mills Ken I. , Burnett Alan K. 
Published: April 2020

Abstract
Partial tandem duplications (PTD) of the mixed lineage leukaemia (MLL) gene occur in approximately 7% of acute myeloid leukaemia (AML) patients. MLL-PTD has a frequency of approximately 10% in patients with normal karyotype and is reported to be associated with an adverse prognosis. Recent papers have reported that MLL-PTD may be useful as a marker for minimal residual disease (MRD) in AML patients with no other markers available due to normal cytogenetics. Diagnostic RNA from 133 AML patients was analysed for the presence of the MLL-PTD, twelve samples were positive and three of the twelve were also positive for FLT3 ITD. The MLL-PTD gene dosage was verified using quantitative polymerase chain reaction (QPCR). In order to identify the molecular disruption caused by MLL PTD mutations, fifty-four of the 133 patients were profiled using the Affymetrix U133A chip containing 22,283 probe sets. Four of the 54 samples analysed were MLL-PTD positive and three of these contained FLT3 mutations. Groups of genes were identified that had significance to MLL and showed more than 2-fold change in regulation in the MLL-PTD compared with wild type. Up-regulated genes found common to the two groups include HOXA9, HOX2B, HOX2C, HOX1 and CREBBP/EP 300. Whilst significant genes down-regulated more than 2-fold in MLL-PTD include MONDOA, a transcriptional activator. To identify whether MLL-PTD and FLT3 mutations used a common pathway, a comparison was made between genes with significance to either MLL-PTD or FLT3 mutations and no common genes were found. Hierarchical cluster analysis on the most significant genes, using Genespring software, created 4 main groups and distinguishes non-leukaemic samples, AML patients with MLL-PTD and those with a wild type MLL gene. This gene signature showed that MLL- PTD is a unique entity and could be identified amongst other AML subtypes. Moreover, over expression of Hox genes plays a role in MLL PTD positive AMLs and may provide an insight into the mechanism of the MLL-PTD with potential clinical implications.
Successful Use of Rituximab To Treat Central Nervous System Lymphoma in a Patient with B-Cell Post-Transplant Lymphoproliferative Disorder (PTLD).
Authors: Riggs Shirley A. , Van Buren Charles , Kahan Barry 
Published: April 2020

Abstract
A 27 year old male with end-stage renal failure secondary to IgA nephropathy received a living related renal transplant in 10/2002, complicated by severe cellular and humoral rejection. Immunosuppression included steroids, OKT3, cyclosporine A, rapamycin, thymoglobulin, antithymocyte globulin, and plasmapheresis. He developed multiple complications including severe pancytopenia, pleural effusions, Pseudomonas sepsis, and Vancomycin resistant enterococcus cystitis. Transplant nephrectomy was required 2/2003 for intractable rejection with blood clots in the urine, revealing large nodules of monomorphous B-cell PTLD throughout the renal parenchyma, along with type 2 acute cellular rejection. The B-cells were CD20 positive, and in situ hybridization for EBER was positive in the majority of the cells. PTLD staging was negative in other sites including brain and bone marrow. All immunosuppression was discontinued shortly following the nephrectomy, and he was placed on acyclovir for 3 months until he developed progressive cytopenias. In 8/2003 he underwent open brain biopsy following 2 grand mal seizures and an MRI showing multiple bilateral nonenhancing cortical and subcortical lesions up to several centimeters, with enhancing right temporal and parietal lesions with surrounding vasogenic edema and regional mass effect. Right temporal lobe biospy showed extensive high grade monomorphous B-cell PTLD with necrosis and vascular involvement. Lymphoma staging including lumbar puncture was otherwise negative and LDH was normal. He received a brief course of steroids and valacyclovir in the perioperative period. His performance status remained extremely poor, on hemodialysis, and thus he was not considered a good candidate for systemic chemotherapy, brain radiotherapy, or intrathecal chemotherapy. We elected to give a total of 8 courses of rituximab at 375 mg/m2 intravenously weekly. Although systemic rituximab does not generally penetrate the central nervous system very well, it was presumed that the blood brain barrier in this patient was broken down due to extensive lymphomatous cerebral disease. By 9/2003 there was improvement in the brain parenchymal lesions, and since 11/2003 there has been complete stabilization of residual involuted nonenhancing lesions. He remains in complete clinical remission, off rituximab for 22 months. This excellent response suggests that systemic rituximab can be an effective treatment in selected cases of PTLD in which extensive brain disease is present.
Rituximab and Yttrium-90 Ibritumomab Tiuxetan Radioimmunotherapy May Be Feasible Options in Patients with Low-Grade Lymphoma of the Orbit.
Authors: Esmaeli Bita , Pro Barbara , Saville M. Wayne , McLaughlin Peter 
Published: April 2020

Abstract
Objective: To report a cohort of patients with lymphoid tumors of the orbit, including 1 with benign lymphoid hyperplasia of the orbit whose disease responded to immunotherapy with rituximab alone or rituximab followed by Zevalin (yttrium-90 ibritumomab tiuxetan) radioimmunotherapy.
Methods: Between October 2002 and January 2005, 8 patients with low-grade non-Hodgkin’s lymphoma and 1 with benign lymphoid hyperplasia of the orbit were treated with monoclonal antibodies against CD20. Five patients with low-grade lymphoma of the orbit (3 with follicular B cell or 2 with MALT) received rituximab followed by yttrium-90 ibritumomab tiuxetan, 2 patients with low-grade orbital lymphoma (1 with follicular B-cell and the other with small lymphocytic lymphoma) and 1 patient with benign lymphoid hyperplasia received rituximab alone. Three out of the 5 patients who received yttrium-90 ibritumomab tiuxetan were part of a prospective trial evaluating the efficacy of yttrium-90 ibritumomab tiuxetan for low-grade B-cell lymphoma of the orbit; the other 2 had been treated in other open label trials at M. D. Anderson Cancer Center. For each patient, clinical records and imaging studies were reviewed to establish the diagnosis and document response.
Results: Three men and 5 women in this cohort were between the age of 23 and 83 years old (median age, 63 years). Of the 7 patients with orbital lymphoma, 4 had stage IE and the other 3 had stage IVE, and 6 had previously untreated disease. All 8 patients experienced resolution of the orbital tumor in response to treatment with monoclonal antibodies against CD20. Follow-up time ranged from 6 months to 32 months (mean, 12 months) after completion of immunotherapy. There were no serious systemic or ocular side effects during the study period. The most commonly experienced side effect was mild fatigue. All 5 patients treated with yttrium-90 ibritumomab tiuxetan had transient pancytopenia, which normalized as expected within 3 months after treatment. One patient who received yttrium-90 ibritumomab tiuxetan developed shingles 6 weeks after treatment. There were no sequelae from this infection, which resolved with the usual course of oral antiviral therapy.
Conclusions: Rituximab and yttrium-90 ibritumomab tiuxetan may be considered as alternative treatment modalities to external beam radiation therapy for low-grade B-cell follicular non-Hodgkin’s lymphoma or MALT of the orbit. Systemic targeted immunotherapy may potentially have the advantage of lower rate of distant (out-of-field) relapse and less ocular toxicity compared with external beam radiotherapy; these potential advantages would have to be verified in long-term studies and in larger numbers of patients.
Promising Activity of Bortezomib, Rituximab, and Dexamethason (BORID) in Patients with Relapsed Mantle Cell Lymphoma.
Authors: Drach Johannes , Seidl Sonja , Pichelmayer Oskar , Kaufmann Hannes , Zielinski Christoph , Raderer Markus 
Published: April 2020

Abstract
Bortezomib (B) belongs to a new class of anti-cancer agents, the proteasome inhibitors, and has documented activity in multiple myeloma. Moreover, recent data indicate that B is also effective in other B-cell malignancies, most notably in mantle cell lymphoma (MCL). Phase II studies revealed remission rates between 40 and 55% of patients with relapsed or refractory MCL. Preclinical studies suggest that B has synergistic activity with rituximab (R), which provides a rationale for the exploration of such a treatment combination. We have therefore initiated a phase II study in relapsed/chemotherapy refractory MCL in order to evaluate the activity and safety of B in combination with R and dexamethasone (BORID). A treatment cycle consists of B at 1.3 mg/m2 administered on days 1, 4, 8, and 11, R at 375 mg/m2 administered on day 1, and dexamethasone 40 mg orally on days 1 to 4. Cycles are repeated every 3 weeks for a total of 6 treatment cycles. Patients with progressive MCL after at least one prior line of therapy (including CHOP or a CHOP-like regimen) are eligible. Up to now, we have enrolled 7 patients at a median age of 65 years (range, 52 to 74 years); they had received one to 6 lines of prior therapy including R in 5 of them and high-dose chemotherapy followed by autologous stem cell transplantation in 2 patients. Severe adverse events (&gt; grade II) included 3 infections (herpes zoster, bacterial pneumonia, pneumonia of potential viral origin), grade III peripheral neuropathy in one patient, and nodular skin infiltrates in one patient. All adverse events were managable by standard means of supportive care and prolongation of the treatment interval between cycles. No severe hematological toxicity has yet been encountered. Of 6 patients evaluable for efficacy, 5 have achieved a response (1 CR, 4 PR), and 1 patient experienced stable disease. The patient in CR (previously treated by R-CHOP, autologous transplantation, and R plus thalidomide) was also negative for disease activity by PET scanning. None of the patients has yet progressed at 4 to 9 months after initiation of BORID. Recruitment of patients is ongoing, and updated results will be presented. Data obtained thus far indicate that BORID has promising activitiy and managable toxicity in patients with relapsed MCL, which warrants further investigation not only in MCL but also in other B-cell lymphomas.
Apigenin Induces Apoptosis in Primary Effusion Lymphoma.
Authors: Uddin Shahab , Al-Kuraya Khawla S. 
Published: April 2020

Abstract
The mechanisms that regulate induction of the antiapoptotic state and mitogenic signals in primary effusion lymphoma (PEL) are not well known. In efforts to identify novel approaches to block the proliferation of PEL cells, we found that apigenin (4′,5,7,-trihydroxyflavone), a flavonoid, induces apoptosis in a dose dependent manner in several PEL cell lines. Such effects of apigenin appear to result from suppression of the constitutively active AKT, FOXO transcription factor and GSK3. Our data also demonstrate that apigenin induces loss of mitochondrial membrane potential with subsequent release of cytochrome c and activation of caspase-3, followed by polyadenosin-5′-diphosphate-ribose polymerase (PARP) cleavage. Altogether, our findings suggest a novel function for apigenin, acting as a suppressor of AKT/PKB pathway in PEL cells, leading to the induction of caspase-dependent apoptosis. Therefore, apigenin may have a future therapeutic role in PEL and possibly other malignancies with constitutive activation of AKT/PKB pathway.
Role of Phosphatidylinositol 3- Kinase/AKT Pathway in Diffuse Large B-Cell Lymphoma Survival.
Authors: Uddin Shahab , Hussain Azhar R. , Bavi Prahant , Siraj Abdul K. , Al-Kuraya Khawla S. 
Published: April 2020

Abstract
Phosphatidylinositol 3-kinase (PI3-kinase) is a key player in cell growth signaling in a number of lymphoid malignancies including myeloma and primary effusion lymphoma. However, its role in diffuse large B-cell lymphoma (DLBCL) has not been elucidated. Therefore, we have studied the PI3-kinase pathway and apoptosis in a panel of DLBCL cell lines (SUDHL4, SUDHL8, SUDHL10 and OCI-LY19). Our data show that inhibition of PI3-kinase by a specific inhibitor, LY294002, induced apoptosis as detected by Annexin V/Propidium Iodide dual staining in the majority of DLBCL cell lines. We then dissected the PI3-kinase pathway by analyzing the downstream targets of phosphorylation by Western blot. We found that AKT/PKB was constitutively phosphorylated, and thus activated, in all DLBCL cell lines. The downstream elements of AKT, ForkHead (FKHR) and GSK3 were also constitutively phosphorylated in all DLBCL cell lines. Similarly, treatment with LY294002 prevented this phenomenon in all the cell lines regardless of their final apoptotic endpoint. Inhibition of PI3-kinase activity further downstream induced cleavage of Bid in all DLBCL cells and subsequently loss of mitochondrial membrane potential and release of cytochrome c from mitochondria in all DLBCL cell lines. The release of cytochrome C led to activation of Caspases 9 and 3 and cleavage of PARP. Finally expression of the inhibitor of apoptosis, XIAP, which is also a downstream target of AKT, was compromised in the all cell lines following LY294002 treatment. Our data demonstrate that the PI3-kinase pathway plays a major role in the survival and growth of DLBCL cells. Altogether, these results suggest that blocking the PI3-kinase pathway may be a potential target for therapeutic intervention in diffuse large B-cell lymphoma.
Loss of Expression of the Pro-Apoptotic Bcl-2 Family Proteins Bak and Bax in Rituximab- and Chemotherapy-Resistant Non-Hodgkin’s Lymphoma Cells.
Authors: Olejniczak Scott H. , Hernandez-Ilizaliturri Francisco J. , Clements James L. , Czuczman Myron S. 
Published: April 2020

Abstract
Sustained remissions following standard chemotherapy are achieved in less than half of patients diagnosed with non-Hodgkin’s lymphoma (NHL). A major case of treatment failure is the development of resistance to current therapies. Early studies demonstrated that rituximab was a safe and effective monotherapy for patients with indolent B-cell lymphoma refractory or relapsed following prior chemotherapy. In one study, Davis et al. demonstrated a loss of responsiveness upon rituximab re-treatment in 60% of patients suggesting the development of antibody resistance. To study mechanisms by which cells become resistant to rituximab we induced rituximab resistance in several well characterized B-cell lymphoma cell lines (Raji, RL, SU-DHL-4) by exposing them to increasing concentrations of rituximab or rituximab plus human serum as a source of complement. Individual clones were than generated from rituximab-resistant cell lines (RRCL) by limiting dilution. Characterization of the cell lines and clones generated by repeated exposure to rituximab revealed that in addition to gaining a rituximab-resistant phenotype they also developed concurrent chemotherapy resistance. In rituximab-sensitive parental cell lines, chemotherapy induced apoptosis via the intrinsic pathway. However, apoptotic cell death was completely blocked in resistant cell lines and clones. Upon investigation of potential mediators of both rituximab and chemotherapy resistance, we observed a significant down-regulation of the pro-apoptotic Bcl-2 family proteins Bax and Bak. We therefore hypothesize that resistance to chemotherapy- and rituximab-induced apoptosis is due to the down-regulation of Bax and Bak observed in all rituximab-resistant cell lines and clones characterized to date. We are currently investigating the mechanism(s) underlying the down-regulation of Bax and Bak protein in RRCL. Preliminary data suggests that wild-type Bax and Bak mRNA is expressed at a comparable level in RRCL and parental cells. This strongly suggests that Bax and Bak protein expression may be differentially controlled via post-transcriptional mechanisms in resistant cells. Currently, we are evaluating expression of Bax and Bak in archived lymphoma biopsy specimens and their correlation to treatment response or resistance. If a clinical correlation is identified, therapies aimed at restoring Bak and/or Bax expression may someday prove useful in circumventing clinical resistance to currently used immuno +/− chemotherapy-based regimens.