bluuebunny/crossref_metadata_2025_split huggingface dataset:
Generation of CD4+ Regulatory T Cells by Retroviral Transduction of CD4+CD25− T Cells Either with the Full-Length or with a Common Exon-2 Negative Variant of Human Foxp3.
Authors: Mutis Tuna , Verdonck Leo F. , Aarts-Riemens Tineke , Emmelot Maarten 
Published: April 2020

Abstract
The forkhead/winged helix transcription factor, Foxp3 is a key element for the differentiation of CD4+CD25+ regulatory T (Treg) cells. While murine Foxp3 gene is expressed as a single full-length transcript, we observed that transcription of human Foxp3 gene usually reveals two different mRNA products with ~ 100 base pair difference. Sequencing of these products revealed that one transcript represents the full-length Foxp3, while the other transcript appears to be an alternative splicing product that lacks the exon-2. Using a specific primer set that amplifies the region between exon-1 and exon-3 we found that both exon-2pos and exon-2neg variants are preferentially expressed in CD4+CD25+ cells with high levels of expression in CD4+CD25hi cells. Analysis of individual Treg clones generated by limiting dilution of CD4+CD25hi cells revealed that both Foxp3 variants are simultaneously expressed in Treg clones. However, quantitative-real time PCR analyses, performed using a primer set that amplifies only the full-length Foxp3 and another set that amplifies both exon-2neg and exon-2pos variants, indicated that expression levels of the full-length Foxp3 gene do not correlate with expression levels of total Foxp3 in individual Treg clones, in PBMC of healthy donors and in leukemia patients after donor lymphocyte infusions. Remarkably, some donors expressed little or no full-length Foxp3 while expressing considerable amounts of total Foxp3, suggesting a predominant expression of the exon-2neg variant in CD4+CD25+ T cells of these donors. To determine the role of individual Foxp3 isoforms in Treg cell-differentiation, the full-length and the exon-2neg variants of the Foxp3 gene were cloned into retroviral vectors and transduced separately into highly purified CD4+CD25− cells. After a brief in vitro culture, T cells transduced with the Foxp3 isoforms were sorted to high purity by flow cytometry using the GFP marker gene and expanded to high quantities by nonspecific stimulation with allogeneic feeder cells, PHA and IL-2. Phenotypical and functional analyses of expanded cells revealed that T cells transduced with both Foxp3 variants expressed high levels of CD25, intacellular CTLA-4 and CD62L; were anergic to stimulation via CD3 and suppressed the CD3/CD28 triggered proliferation of autologous and allogeneic CD4+CD25− cells. These results reveal that both isoforms of human Foxp3 are functionally intact and can convert CD4+CD25− cells into Treg cells. Current efforts are focused on testing the in vivo capacity of in vitro expanded, Foxp3 transduced T cells to prevent Graft versus Host Disease (GvHD) in a xenogeneic model where GvHD is induced by administration of human T cells in Rag2, gamma chain double knockout mice.
Treatment of Childhood T Cell Lymphoblastic Lymphoma (TLL), Results of the SFOP LMT96.
Authors: Bergeron Christophe , Perol David , Pacquement Helene , Perel Yves , Coze Carole , Gandemer Virginie , Vannier Jean P. , Mechinaud Francois , Schmitt Claudine , Leverger Guy , Baruchel Andre , Patte Catherine 
Published: April 2020

Abstract
After LMT81 and SFOP LMT89 protocols (both modified LSA2L2), the SFOP LMT96 was designed according to BFM 86 back bone (Reiter and al, J Clin Oncol, 13:359.1995). The aim of LMT96 protocol was to increase chemotherapy intensity early (day 8) and late (after the intensification phase). From February 1997 to December 2003, 83 unselected patients (pts) with newly diagnosed TLL were included in LMT96 protocol: classic corticosteroid prephase (d1 to d7); induction phase with Cyclophosphamide(CPM: 1g/m2) and high dose methotrexate (HDMTX: 3g/m2 infusion over 3h) at d8, followed by Vincristine, Daunorubicin (40mg/m2) d15, 22, 29; 8 injections of Asparaginase, 2 intrathecal injection MTX at d8 and d15 and prednisone for 4 weeks; two consolidation phases with HDMTX, Cyclophosphamide (CPM), Cytarabine and Asparaginase; one interphase with HDMTX and; an intensification phase similar to a phase II BFM study, a maintenance phase with Mercaptopurine/MTX including initially six intensified monthly pulse phases alternating either HDMTX/Asparaginase, or Cytarabine/prednisone. The total duration of treatment was 18 months for stage I, II and III and 24 months for stage IV. Cranial radiotherapy (18Gy) was only performed in pts with CNS disease. 66% were boys, median age was 10.5 y [1.9 – 17.2] and 88% had a mediastinal involvement. According to St Jude classification, 5 had a stage I or II (6%); 47 had a stage III (57%); 24 had a stage IV [ (63%), 18 bone marrow (BM) involvement, 4 CSF+ and 2 both]. 7 pts received corticosteroid before diagnosis and were classified as stage X and treated as stage IV. After induction phase and consolidation (d 65), the complete remission rate was 95%. 1 pt had a progressive disease and died and 2 pts in partial remission are alive after salvage treatment with local radiotherapy or intensification. Up to now, 8 pts relapsed between 3.7 months and 18 months (median=10.5 months) and 1 pt who developed a glioblastoma before relapsing at 69.2 months. All relapses were mediastinal, 3 were combined with BM and 1 with BM and CSF. The median of FU is 4 years. Five-year EFS is 87% [CI95%: 79–94] and OS is 89% [CI95%: 82–96]. In conclusion these results are as high as those of BFM experience. The intensified monthly pulse courses for 6 months did not seem decrease the relapses on therapy. Early intensification at d8 (with CPM and HDMTX) is feasible with a BFM regimen. Omission of prophylactic cranial irradiation while using a short duration (3 hours) of HDMTX infusion did not jeopardize the outcome. We suggest that our early intensification might be combined with a classic BFM regimen.
Primary CNS Lymphoma Treated with HD-Methotrexate, HD-Busulfan/Thiotepa, Autologous Stem Cell Transplantation and Response-Adapted Whole-Brain Radiotherapy: Results of the Multicenter OSHO-53 Phase II - Study.
Authors: Montemurro Michael , Kiefer Thomas , Schüler Frank , Ali Haifa-Katrin Al , Wolf Hans-Heinrich , Herbst Regina , Haas Antje , Helke Kirsten , Theilig Anja , Lotze Christian , Hirt Carsten , Niederwieser Dietger , Schwenke Matthias , Krüger William , Dolken Gottfried 
Published: April 2020

Abstract
Purpose:
This multicenter study investigated the efficacy and safety of high-dose methotrexate (HD-MTX) induction followed by high-dose busulfan/thiotepa with autologous stem-cell transplantation (HD-BuTT) and response-adapted whole-brain radiotherapy (WBRT) in patients with newly diagnosed primary CNS lymphoma.
Patients and methods:
23 patients (median age 55 years) were treated in five centres. Patients received HD-MTX (4h-infusion; 8g/m2; &gt;60y: 6g/m2) on d1 and d10 followed by leucapheresis. Then patients were stratified according to their results on neuroimaging: In case of at least a partial response, HD-BuTT consisting of 16mg busulfan / 10mg thiotepa per kg body weight followed by peripheral stem cell transplantation was given. Patients without response to induction or without complete response after high-dose therapy received WBRT (45Gy) as further treatment.
Results:
16 patients received the planned treatment with HD-MTX followed by HD-BuTT. CR / PR rates for these patients were 19 % / 69 % after HD-MTX, 69 % / 13 % after HD-BuTT, 81 % / 6 % after HD-BuTT plus WBRT, respectively. Included the patients with early WBRT due to toxicity (n=2) and non-responders to HD-MTX induction (n=4) the overall response rate for all 23 patients was 83 % (intention-to-treat). Outcome was significantly influenced by the response to MTX-induction. There were three treatment-related deaths. Irradiated patients (n=9) had a high incidence of severe neurotoxicity leading to death in 3 patients. At a median follow-up of 15 months the median EFS and OS for all patients were 17 and 20 months, after HD-BuTT 27 months and “not reached”, respectively. Patients older than 60 years and younger patients have achieved similar outcomes.
Conclusion:
This study showed that HD-methotrexate induction followed by HD-BuTT is a feasible treatment option for newly diagnosed primary CNS lymphoma. Patients achieving CR after HD-BuTT show no signs of clinical neurotoxicity with median survival not reached yet. Time on treatment is 2–3 months only, but the induction treatment needs improvement to be more effective. WBRT in this study was associated with a high incidence of severe neurotoxicity and should therefore be avoided.
International Primary Central Nervous System Lymphoma Collaborative Group (IPCG) Study on Low-Grade Primary Central Nervous System Lymphoma in Immunocompetent Patients.
Authors: Korfel Agnieszka , Jahnke Kristoph , O’Neill Brian P. , Blay Jean-Yves , Abrey Lauren , Martus Peter , Poortmans Philip M.P. , Shenkier Tamara N. , Batchelor Tracy T. , Neuwelt Edward A. , Raizer Jeffrey J. , Pels Hendrik , Herrlinger Ulrich , Thiel Eckhard 
Published: April 2020

Abstract
Low-grade primary central nervous system lymphoma is a very rare subtype of primary central nervous system lymphoma (PCNSL), for which almost no data is currently available. The purpose of this retrospective study was to characterize the clinical presentation, course and outcome of patients with low-grade PCNSL.
Forty patients (18 male, 22 female) from 18 cancer centers in five countries were identified with a median age of 58 (range, 19–78) years and a median Eastern Cooperative Oncology Group (ECOG) performance status of 1 (range, 0–4). The mean time to diagnosis was 14.8 months (range, 0.25–84). Thirty-two patients (80%) had a B-cell and eight a T-cell lymphoma. Thirty-seven patients (92.5%) showed involvement of a cerebral hemisphere or deeper brain structures, while two evidenced only leptomeningeal involvement, and one patient had spinal cord disease. Treatment was performed in 39 patients: chemotherapy and radiotherapy in 15 (38%), radiotherapy alone in 12 (30%), chemotherapy alone in 10 (25%), and tumor resection alone in two. The median progression-free survival (PFS), disease-specific survival (DSS) and overall survival (OAS) were 61.5 (range, 0–204), 130 (range, 1–204), and 79 (range, 1–204) months, respectively. An age ≥60 years was associated with a shorter PFS (P = .009), DSS (P = .015) and OAS (P = .001) in multivariate analysis.
Low-grade PCNSL differ from the high-grade subtype in pathological, clinical and radiological features. In this study, the long-term outcome was better as compared to the results obtained in PCNSL in general with age ≥60 years adversely affecting survival.
Phase II Study of Denileukin Diftitox (Ontak) for Relapsed/Refractory T-Cell Non-Hodgkin’s Lymphoma.
Authors: Dang Nam H. , Pro Barbara , Hagemeister Fredrick , Samaniego Felipe , Jones Dan , Rodriguez Maria , Goy Andre , Romaguera Jorge , Walker Pamela L. , Younes Anas , Neelapu Sattva , Kwak Larry , Fayad Luis 
Published: April 2020

Abstract
Denileukin diftitox (ONTAK) is a fusion protein combining interleukin-2 (IL-2) and the enzymatically active domain of diphtheria toxin that targets tumor cells expressing the IL-2 receptor. We initiated a phase II study at MD Anderson Cancer Center to evaluate its efficacy in relapsed/refractory T-cell lymphoma excluding CTCL. Denileukin diftitox was administered at 18 mcg/kg/day by IV infusion daily for 5 days every three weeks for up to 8 cycles. Corticosteroid was given before every infusion to reduce the incidence and severity of acute hypersensitivity reaction. Twenty-five patients are currently evaluable for response. Median age was 55 years (range 26–80) and mean number of prior treatments was 2 (range 1–6). Tumor CD25 status was determined by flow cytometry and/or immunohistochemistry, with CD25 positivity being defined as 10% or more tumor cells expressing detectable CD25. Of the 13 patients with CD25+ T-cell lymphomas, there were 4 CR (30.8%) and 4 PR (30.8%) for an overall response rate of 61.6%. Of the 10 patients with CD25- tumors, there were 1 CR (10%) and 3 PR (30%) for an overall response rate of 40%. Two patients had a tumor CD25 of undetermined status. The overall response rate was 48% with 5 CR (20%) and 7 PR (28%). The median progression-free survival for responding patients was 6 months (range, 1–24+ months), with 1 patient with CD25+ ALK-1 negative ALCL having an ongoing CR at 24+ months. Of note is the fact that one patient who experienced a CR and another who had a PR went on to receive allogeneic transplantations while in remission induced by denileukin diftitox. Treatment was well tolerated, with the majority of toxicity being grade 1or 2 and transient. Most common toxicities seen were transient transaminase elevation, hypoalbuminemia and edema. Denileukin diftitox has activity in relapsed/refractory T-cell NHL, and is well tolerated at the dosing scheduled tested. While response to denileukin diftitox is seen in both CD25+ and CD25− T-NHL, drug activity in CD25 expressing tumors is particularly striking.
CD4 T-Helper Responses to the ALK Protein in Patients with ALK-Positive ALCL.
Authors: Ait-Tahar Kamel K. , Hatton Chris S.R. , Pulford Karen K. 
Published: April 2020

Abstract
Anaplstic Lymphoma Kinase (ALK)-positive anaplastic large cell lymphoma (ALCL) has a favourable prognostic outlook compared to ALK-negative AlCL, possibly as a result of the immune recognition of the ALK protein. We have previously shown the presence of both a cytotoxic T cell and an antibody response to the ALK protein in patients with ALK-positive ALCL. The aim of our present study was to investigate the presence of a CD4 T-helper (Th) response in patients with ALK-positive ALCL and in control individuals. Using the TEPITOPE web-based predicitive search algorithm, three 24-mer promiscuous peptides were identified from the ALK sequence as being potentially immunogenic in the context of MHC class II. A gamma-interferon (γ-IFN) and IL-4 ELISPOT assay was used to detect a T cell response in the peripheral blood cells from patients with ALK-positive and ALK- negative ALCL, as well as healthy controls after 6–11 days of culture with the three peptides. ALK278–301 and ALK233–256 were shown to be highly immunogenic in the majority of the ALK-positive patients (see Table). ALK411–434 was immunogenic to T cells from only one of the ALK-positive patients (Patient 4). Cells from none of the two ALK-negative ALCL patients or the five healthy donors showed any reactivity to the ALK peptides. No response to the control irrelevant peptide was observed in any of the ALCL patients or healthy donors. With the exception of one ALK-positive ALCL patient (Patient 2), no significant IL-4 response was recorded in any of the patients or controls. All of the ALK-positive patients presented antibodies to the ALK protein at time of diagnosis.These findings further demonstrate the immunogenicity of the ALK protein and are suggestive of a Th1 type of immune response to the protein. Our findings are of potential prognostic value and open up therapeutic options for those ALK-positive patients who do not respond well to chemotherapy.
Summary of the CD4 Th responses to ALK in ALCL patients and healthy donors None PHA (10 μ g/ml) ALK233–256 (10 μM)- (IFN- γ/IL-4) ALK278–301(10 μM)- (IFN- γ/IL-4) ALK411–434 (10 μM)- (IFN- γ/IL-4) Irrelevant peptide (10 μM)- (IFN- γ) Antibody titres to ALK (IgG isotype) ND= Not done. Results are of triplicate cultures ALK+ve patients Patient 1 12 188 56/10 44/18 22/6 8 1/2250 Patient 2 20 240 126/48 78/52 40/26 18 1/2250 Patient 3 14 48 38/ND 24/ND 12/ND 16 1/6750 Patient 4 6 108 64/8 72/8 22/6 10 1/60750 Patient 5 10 48 36/13 26/18 12/12 14 1/6750 Patient 6 15 132 74/ND 58/ND 28/ND 18 1/6750 Patient 7 10 180 34/28 56/32 12/9 12 1/750 ALK-ve patients Patient 8 14 122 12/10 10/6 12/14 22 −ve Patient 9 16 82 14/ND 12/ND 10/ND 24 −ve Healthy Donors Normal 1 22 148 18/14 22/24 26/12 10 −ve Normal 2 12 18 2/4 6/8 12/2 4 −ve Normal 3 10 38 12/10 12/16 9/4 18 −ve Normal 4 9 172 9/ND 10/ND 6/ND 12 −ve Normal 5 4 108 8/12 8/12 2/1 10 −ve
Dasatinib (BMS-354825), a Multi-Targeted Kinase Inhibitor, Inhibits the Kinase Activity of Wild-Type, Juxtamembrane and Activation Loop Mutant Kit Isoforms Associated with Human Malignancies.
Authors: Schittenhelm Marcus M. , Shiraga Sharon , Schroeder Arin , Corbin Amie S. , Griffith Diana , Lee Francis Y. , Deininger Michael W. , Druker Brian J. , Heinrich Michael C. 
Published: April 2020

Abstract
Activating mutations of the activation loop (AL) of KIT are associated with certain human neoplasms, including a subset of patients with AML,systemic mast cell disorders (SM), seminoma, and Gastrointestinal Stromal Tumors (GIST). KIT AL mutations such as D816V that are typically found in AML and SM are resistant to imatinib (IM, IC50 &gt; 5–10 μM). Dasatinib (BMS-354825) is a novel, oral, multi-targeted kinase inhibitor that targets BCR-ABL and SRC. Due to its potent inhibition of these kinases, dasatinib is currently being tested in clinical trials of patients with imatinib resistant/intolerant CML/Ph+ ALL. Based on previous observations of the ability of certain SRC/ABL inhibitors to also inhibit KIT kinase, we hypothesized that dasatinib might inhibit the kinase activity of both WT and mutant KIT isoforms. The inhibitory potential of dasatinib against WT KIT, KIT mutant isoforms and KIT-dependent downstream pathways was evaluated by immunoblotting. In addition, we evaluated the effects of dasatinib on cellular proliferation and induction of apoptosis. Dasatinib potently inhibited WT, juxtamembrane- (JM) and AL-mutant KIT autophosphorylation. Based on the ability of dasatinib to bind to BCR-ABL irrespective of the ATP AL conformation (inactive versus active), dasatinib was expected to be insensitive to KIT AL conformation. In contrast, we found that the IC50 for KIT autophosphorylation varied significantly among the various KIT mutant isoforms: WT KIT, D816Y, V560G (JM mutation) [IC50 1–10 nM] &lt;D816F [IC50 100 nM] &lt;D816V [IC50 200–250 nM]. These results indicate that the conformation of the KIT AL does influence dasatinib potency. Inhibition of KIT kinase activity by dasatinib reduced cellular proliferation and induced apoptosis in mast cell/leukemia cell lines expressing mutant KIT isoforms. In these cell lines, KIT activates downstream pathways important for cell viability and cell survival such as RAS/MAPK, JAK/STAT and PI3K/AKT. Dasatinib potently blocked activation of MAPK1/2 and STAT3. Inhibition of KIT by dasatinib abrogated phosphorylation of AKT S473, but not AKT T308. This partial inhibition of AKT activation was insufficient to inhibit phosphorylation of p70S6K, a kinase downstream of AKT and mTOR. Combining dasatinib with rapamycin, a known mTOR inhibitor, had an additive to synergistic anti-proliferative effect on cells expressing KIT D816V. Our studies suggest that dastatinib may have clinical activity against human neoplasms that are associated with gain-of-function KIT mutations such as AML, SM, seminoma, and GIST. Combining dasatinib with mTOR inhibitors may further increase efficacy against KIT-driven malignancies.
Evaluation of the Specificity and Cytotoxicity of Three Proteasome Inhibitors.
Authors: Crawford Lisa J.A. , Ovaa Huib , Walker Brian , Chauhan Dharminder , Anderson Kenneth C. , Morris Treen C.M. , Irvine Alexandra E. 
Published: April 2020

Abstract
The proteasome is a mutlicatalytic protease with three main catalytic activities - chymotrypsin-like (CT-L), trypsin-like (T-L) and peptidylglutamyl peptide hydrolising (PGPH). Proteasome inhibition is an emerging therapy for many cancers and is a novel treatment for multiple myeloma (MM). The CT-L activity, considered to be the rate-limiting step in protein degradation, is the primary target of many proteasome inhibitors. We have compared the specificity and potency of the novel proteasome inhibitor BzLLLCOCHO to the previously characterised inhibitors PS-341 (Velcade, bortezomib) and MG-132. Specific fluorogenic substrates were used to measure proteasome proteolytic activity in the presence and absence of the inhibitory compounds. An active site directed probe with a dansyl-sulfonamidohexanoyl hapten tag was used in conjuction with immunoblotting to determine the subunit specificity of the proteasome inhibitors (Nature Methods2005;2:357–362). MM cell lines (U266, OPM-2, KMS-11, KMS-18) were incubated with 10 μM BzLLLCOCHO, 5 nM PS-341 or 1 μM MG-132 for 24 hrs and proteasome activity was measured. Addition of BzLLLCOCHO reduced CT-L activity by 83 ± 13 % in the fluorogenic assay, and T-L and PGPH activities were reduced by 93 ± 6 % and 92 ± 2 % respectively. Immunoblot results revealed a similar pattern, the T-L and PGPH subunits were completely inhibited by BzLLLCOCHO and there was only weak labeling of the CT-L subunit with the active site probe. In contrast, treatment with PS-341 completely inhibited the CT-L and PGPH activities and incubation with MG-132 resulted in weak inhibition of the CT-L and PGPH activities, neither inihibitor significantly affected T-L activity. The ability of the different inihibitors to induce apoptosis in MM cell lines was then evaluated. All three inhibitors were demonstrated to act through both the caspase-8 and caspase-9 signalling pathways. Using Mitosensor™ and Hoescht/Propidium Iodide staining we found that MM cells were more sensitive to the induction of apoptosis by PS-341 and MG-132 than BzLLLCOCHO (U266 cells treated for 72 hrs with BzLLLCOCHO 51 % apoptosis, PS-341 79 % apoptosis and MG-132 84 % apoptosis). BzLLLCOCHO is a cell permeable and potent inhibitor of all three proteolytic activities of the proteasome. PS-341 and MG-132 inhibited only two of the three proteasome activities but were more efficient than BzLLLCOCHO at inducing apoptosis in MM cell lines. MG-132 is known to inhibit non proteasomal proteases such as Cathepsin B and Calpain 1 which may contribute to its potency. Further investigation on the effects of these inhibitors on gene and protein expression in the cell may lead to the development of more specific and targeted inhibitors.