bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mid-Treatment Evaluation of 18-FDG-Positron Emission Tomography/Computed Tomography (PET) as Predictive Value of Response Assessment in Aggressive Non Hodgkin Lymphoma (NHL).
Authors: Pregno Patrizia , Chiappella Annalisa , Penna Daniele , Bellò Marilena , Benevolo Giulia , Botto Barbara , Freilone Roberto , Orsucci Lorella , Scaglia Sabrina , Bisi Giovanni , Vitolo Umberto , Gallo Eugenio 
Published: April 2020

Abstract
Introduction: The PET is mostly used in staging of NHL patients at diagnosis and as response assessment at the end of treatment. The evaluation by PET after few cycles of chemotherapy may be useful to predict chemosensitivity and then response, progression-free survival and overall survival in this subset of patients. In our study we introduced the PET in the mid-treatment evaluation (PET-2) of aggressive NHL disease.
Patients and methods: From June 2004 to December 2006, 25 consecutive aggressive NHL patients were evaluated for this study: 17 males and 8 females respectively with median age of 49 years (range 21–67). We included: 18 Diffuse Large B Cell, 1 anaplastic, 2 mantle cell, and 4 follicular grade III NHL. Patients characteristics were: 11/25 bulky disease; 20/25 intermediate or high IPI risk; 3/25 stage II, 3/25 stage III and 19/25 stage IV. All patients were staged according to standard imaging procedures completed by PET at the diagnosis and at the end of treatment. After 2 or 4 cycles of Rituximab-CHOP-like chemotherapy PET-2 was repeated in all of them.
Results: PET-2 demonstrated: 16/25 patients negative and 9/25 patients positive. The conventional and PET restaging performed at the end of treatment were 21/25 negative and 4/25 positive. Among the 16 patients PET-2 negative, 14/16 remained negative at the final PET evaluation and achieved CR, 2/16 became positive with demonstration of progression disease. Among the 9 patients PET-2 positive, 7/9 became negative with achievement of CR. The other 2/9 patients remained positive at the end of therapy: one of them was false positive because of parotid gland carcinoma without NHL involvement. Among 21/25 patients PET negative at the final evaluation, 20/25 patients remained in CR with a median follow-up of 18 months and only one patient negative at the PET-2 and at the final PET demonstrated disease progression. Finally, with a median follow-up of 18 months, FFS was 84% in all pts, 81% in PET-2 negative patients and 89% in PET-2 positive patients respectively.
Conclusions: The PET is an important imaging technique for staging and end-treatment evaluation in lymphoma disease, because it can better define CR patients. In Hodgkin disease several studies demonstrated that the early evaluation by PET is a crucial prognostic factor to test chemosensitivity and then to predict favourable outcome. In our study the mid-evaluation of response by this procedure had not so clear predictive value of response assessment, because patients, even if positive at PET-2, can achieve CR at the end of treatment. More large studies are needed to determine the real impact of on-course PET as response assessment in aggressive NHL patients.
EBV-Specific Cytotoxic T Lymphocytes by LMP2a- and EBNA3a-Specific T Cell Receptor Gene Transfer for Adoptive Therapy in EBV-Associated Hodgkin’s Disease and Post-Transplant Lymphoma.
Authors: Nguyen Tuan D. , Gelfort Haike , Sebelin-Wulf Kathrin , Schmetzer Oliver , Uckert Wolfgang , Pezzutto Antonio , Subklewe Marion 
Published: April 2020

Abstract
Adoptive transfer of polyclonal EBV-specific T cell lines has been used as prophylaxis and therapy in patients with EBV-associated malignancies. However, this strategy is time-consuming and demands difficult generation of lymphoblastoid cell lines (LCLs) and corresponding T cells for each individual patient. We applied an alternative strategy to confer T cell immunity against EBV-antigens by isolating EBV antigen-specific T cell receptors (TCRs) for transduction of primary human T cells for adoptive therapy. Previously, we have demonstrated the feasibility of using peptide-pulsed dendritic cells (DC) for generating high-affinity EBV antigen-specific T cell lines and T cell clones. Based on this strategy, T cell clones directed against LMP2a and EBNA3a were generated and functionally analyzed. Monospecificity was demonstrated by homogeneous double staining with CD8 and appropriate tetramers. High avidity of T cell clones (&lt; 0.01 μM) was shown by peptide titration in an ELISPOT assay for IFN-γ secretion. In addition, the cytokine secretion profiles of some of the T cell clones were tested by cytokine bead array assay. High secretion levels of IFN-γ, IL-2 as well as TNF-α after stimulation with the EBNA3a- or LMP2a-peptide were shown for the corresponding T cell clones. Potent TCRs from one LMP2a-specific, HLA-A2-restricted and one EBNA3a-specific, HLA-B8-restricted T cell clone were isolated and cloned into the retroviral vector MP71. Transduction efficiency of TCR-deficient T cell lines was &gt; 40% (TCR-LMP2a) and &gt; 30% (TCR-EBNA3a) as measured by tetramer staining. Both TCR-LMP2a- and TCR-EBNA3a-redirected T cell lines were functional as indicated by NFAT-mediated luciferase expression upon TCR-MHC-peptide ligation. Primary human T cells were successfully transduced with TCR-LMP2a (∼ 12% tetramer-positive) and TCR-EBNA3a (∼ 3% tetramer-positive). Importantly, both TCRs conferred similar cytolytic activity against EBV-transformed B cell lines. Our data support the development of TCR-transduced T cells for adoptive transfer in EBV-associated malignancies, including Hodgkin′ s disease and nasopharyngeal carcinoma in which only subdominant EBV antigens are expressed. The feasibility and the therapeutic potential of TCR-transduced T cells for adoptive transfer have already been shown in a clinical phase I trial in patients with metastatic melanoma. We believe that redirecting human PBLs is a rapid and efficient tool toward adoptive transfer in EBV-associated malignancies.
Statins Impair Antitumor Effects of CD20 mAb by Inducing Conformational Changes of CD20.
Authors: Golab Jakub , Winiarska Magdalena , Bil Jacek , Wilczek Ewa , Wilczynski Grzegorz , Lekka Malgorzata , Engelberts Patrick J. , Mackus Wendy J.M. , Stoklosa Tomasz , Nowis Dominika , Makowski Marcin , Glodkowska Eliza , Issat Tadeusz , Mrowka Piotr , Lasek Witold , Gorska Elzbieta , Wasik Maria , Warzocha Krzysztof , Sinski Maciej , Gaciong Zbigniew , Dabrowska-Iwanicka Anna , Jakobisiak Marek , Parren Paul W.H.I. 
Published: April 2020

Abstract
A number of monoclonal antibodies (mAb) are presently in development or approved for CD20-directed immunotherapy. Ofatumumab, a human IgG1 anti-CD20 mAb, is currently being evaluated in phase III clinical trials for B-CLL and FL, and in phase II clinical trials for rheumatoid arthritis (RA). Rituximab, a chimeric IgG1 anti-CD20 mAb, has been approved for 1st line treatment of CD20-positive B cell lymphomas alone or in combination with chemotherapy, and in RA. Virtually all rituximab-treated patients relapse after single-agent treatment. The clinical efficacy of rituximab might be further improved by combinations with other drugs such as statins that inhibit cholesterol synthesis and show promising anti-lymphoma effects. We studied the influence of statins on ofatumumab- and rituximab-mediated cytotoxicity. Surprisingly, B cells incubated with statins showed decreased rather than increased CD20 mAb-mediated complement-dependent cytotoxicity (CDC) in cell viability assays. However, cell lysis of statin-treated B cells remained higher when using ofatumumab in comparison to rituximab. Statins decreased CD20 immunostaining in flow cytometry but did not affect total cellular CD20 levels when compared to non-treated cells. Incubation of B cells with other cholesterol depleting agents (methyl-β-cyclodextrin (MβCD) and berberine) established that the presence of plasma membrane cholesterol and not lipid rafts may be required for CD20 mAb-mediated CDC. Cholesterol restitution reversed ofatumumab- and rituximab-mediated CDC and CD20 mAb staining. Statin incubation resulted in conformational changes of CD20 and impaired binding of CD20 mAb to the CD20 molecule as observed by atomic force microscopy. Freshly isolated cells of 4 B cell lymphoma patients were treated with the cholesterol-depleting agent MβCD, and CD20 mAb (B9E9) binding and rituximab-mediated CDC were significantly decreased (P&lt;0.05 t-test, versus control) In addition, 5 hypercholesterolemia patients were treated with atorvastatin. Ofatumumab binding to freshly isolated B cells was found decreased in all patients upon statin treatment (P&lt;0.01, paired t-test). Based on these data, statins may interfere with CD20 detection and anti-lymphoma activity of CD20 mAb and may have significant clinical implications as impaired mAb binding to CD20 conformational epitopes elicited by statins may delay diagnosis, postpone treatment or impair anti-lymphoma activity of CD20 mAb. MAb which are less sensitive to such changes may minimize possible effects of statins on mAb-mediated immunotherapy in these patients.
Improved Responses and Cures with Less Hematologic Toxicity Using Anti-CD20 Bispecific Antibody Pretargeted Radionuclides Compared to Directly Radiolabeled IgG in a NHL Model.
Authors: Goldenberg David M. , Sharkey Robert M. , Karacay Habibe , Chang Chien-Hsing , Rossi Edmund A. , McBride William J. 
Published: April 2020

Abstract
Directly radiolabeled anti-CD20 IgGs approved for the treatment of follicular and transformed non-Hodgkin lymphoma produce higher response rates than rituximab. However, these treatments result in severe and protracted hematologic toxicity, which is directly related to the slow blood clearance of these radiolabeled products. Pretargeting procedures separate radionuclide-targeting from the slow antibody-targeting step, and because the radiolabeled compound is small in size, it clears rapidly and thoroughly from the blood and body in just a few hours, minimizing radiation exposure. We have developed a novel recombinant and humanized Tri-Fab bispecific antibody (bsMAb) for pretargeting using the modular Dock-and-Lock procedure. This bsMAb divalently binds to CD20, with the monovalent Fab binding to a unique peptide hapten (HSG) that carries the therapeutic radionuclide. Nude mice bearing established (0.5 to 0.9 cm3) subcutaneous Ramos human B-cell lymphomas were given a single dose of either a 90Y-humanized anti-CD20 IgG (150 and 175 μCi; 50 μg) or a bsMAb-pretargeted 90Y-DOTA-HSG-peptide (150, 250, 500, and 700 μCi). Tissue counting data derived 24 h after injecting the radiolabeled product showed similar tumor uptake between the IgG and pretargeted peptide, but tumor/blood ratios were &gt;1000:1 for the pretargeted peptide vs. 1:1 for the IgG. At both 90Y-IgG doses, blood counts decreased ∼90% for 2 weeks after treatment, while the pretargeted groups only decreased ∼60% at the 700 μCi dose to as low as a 25% decrease at the 250 μCi dose, with full recoveries occurring within 2 weeks of the nadir. The severe hematologic toxicity also resulted in 2/10 and 4/10 deaths within 2 weeks of the 150- and 175-μCi 90Y-IgG doses, respectively. In the remaining animals, after experiencing an initial response, all tumors progressed to ≥1.5 cm3 within 3 weeks of treatment. Only 1/10 animals given 500 μCi of the pretargeted dose showed tumor progression, with all others in this group and the 700-μCi-group showing no evidence of disease. Earlier studies have shown 40%, 59%, and 90% cure rates after 14 to 18 weeks evaluation with 150, 250, and 500 μCi, respectively, of the bsMAb-pretargeted 90Y-peptide (n = 10 to 22 animals). These studies demonstrate that bsMAb pretargeting significantly improves response rates, with durable cures and without the excessive hematologic toxicity commonly associated with directly radiolabeled antibodies.
(Supported in part from USPHS grant P01-CA103985.)
Development of a New Fully Human Anti-CD20 Monoclonal Antibody for the Treatment of B-Cell Malignancies.
Authors: Bornstein Gadi Gazit , Queva Christophe , Tabrizi Mohammad , VanAbbema Anne , Chavez Carlos , Wang Ping , Laing Naomi , Raja Sandhya , Wen Shenghua , Rong Xianhui , Green Larry L. , Yang Xiaodong , Blakey David 
Published: April 2020

Abstract
In spite of the widespread use of Rituximab, a chimeric monoclonal antibody with demonstrated efficacy in the treatment of non-Hodgkin’s lymphomas, there is a recognized need to develop fully human antibodies with improved efficacy. Towards this end, using XenoMouse™ technology, a fully human IgG1 monoclonal antibody specific to human CD20 was generated. This antibody, denoted mAb 1.5.3, evoked enhanced pro-apoptotic activity in vitro, as compared to Rituximab, in the Ramos human lymphoma cell line. In addition, mAb 1.5.3 was active in mediating complement dependent cytotoxicity (CDC) and elicited improved antibody-dependent cellular cytotoxicity (ADCC) relative to Rituximab in Ramos, Raji, and Daudi human B-lymphoma lines. To recapitulate various aspects of acquired resistance to Rituximab, as observed in a subpopulation of patients, Rituximab-resistant clones were established from lymphoma lines. Interestingly, mAb 1.5.3 demonstrated superior cytolytic activity against engineered Rituximab-refractory lymphoma clones, as well as across multiple human B-lymphoma and chronic B-cell leukemia lines in an in vitro whole blood assay. Furthermore, mAb 1.5.3 exhibited enhanced anti-tumor activity in Rituximab-sensitive cell lines and -refractory engineered lymphoma clones in vivo. Lastly, mAb 1.5.3 produced a superior B-cell depletion profile in lymph node organs and bone marrow as compared to Rituximab in a primate PD model. In contrast to Rituximab, mAb 1.5.3 is a fully human antibody and is thus anticipated to exhibit a longer serum half-life with minimal immunogenicity following repeated administration. In sum, these results demonstrate the superior anti-tumor activity of mAb 1.5.3 relative to Rituximab and its potential for improved clinical activity in the treatment of B-cell malignancies.
Activating Mutation of Heme Oxygenase (HO)-1: A Novel Disease Entity Characterized by Microcytic, Hemolytic Anemia, a Defect of Bilirubin Synthesis and Hereditary Hemophagocytic Lymphohistiocytosis.
Authors: Greil Johann , Verga-Falzacappa Mavi , Tsukahara Hirokazu , Behnisch Wolfgang , Heimpel Hermann , Schneider Marion , Janka Gritta , Claus Maren , Muckenthaler Martina , Kulozik Andreas E. 
Published: April 2020

Abstract
Primary hemophagocytic lymphohistiocytosis (HLH) is a severe inflammatory condition that is caused by an abnormal accumulation of activated macrophages. The common NK- and cytotoxic T-cell dysfunction in HLH has been related to genetic defects of vesicle transport and apoptosis. Here, we identify a novel form of hereditary HLH that is related to and possibly caused by a homozygous missense mutation (G139V) of the active centre of heme oxygenase-1 (HO-1). The clinical picture of this male patient is characterized by persistent, severe microcytic hemolytic anemia without hyperbilirubinemia since birth. In the second year of life, the clinical syndrome of HLH developed with persistent fever, marked hepatosplenomegaly, thrombocytopenia, hyperferritinemia, hypertriglyceridemia, and elevated levels of soluble CD25. Liver histology showed marked lympho- and erythrophagocytosis, which confirmed the diagnosis of HLH. The clinical syndrome responded well to immunosuppressive treatment with etoposide, dexamethasone and cyclosporine A. PRF1, UNC13-D and STX11 mutations were excluded. Functional analysis of NK-activity showed only slightly decreased killing activity with normal responses of NK-cells to cytokine stimulation. Persistent endothelial damage was indicated by a pronounced elevation of vWF and D-dimer concentrations. The persistently very low bilirubin levels despite marked hemolysis suggested a defect of hemoglobin degradation. HO-1 is the rate limiting enzyme of this pathway resulting in bilirubin synthesis. Normally, HO-1 is constitutively expressed at a low level but strongly induced by heme, and hypoxia. Functional analysis of HO-1 protein in the patient’s mononuclear blood cells showed a high constitutive expression that did not respond further to heme treatment. Sequence analysis revealed a homozygous GGT (Gly) to GTT (Val) point mutation of HO-1 codon 139. This mutation of the enzyme’s active center is known from in-vitro analyses to change enzyme activity from an oxygenase into a peroxidase. Consistent with this change in activity, quantitative analysis of reactive oxygen species (ROS) metabolites in the urine of this boy showed high amounts of bilirubin oxidative metabolites, 8-hydroxy-2′-deoxyguanosine and acrolein-lysine. In this report we thus identify a novel disease entity which is characterized by an activating mutation of HO-1 which results in a defect of hemoglobin degradation, bilirubin synthesis and in excessive oxidative stress and inflammation.
Intravenous, Intraperitoneal, and Subcutaneous Routes of Administering 64Cu-DOTA-HB22.7 Display Equivalent Tumor Targeting Ability.
Authors: Martin Shiloh M. , O’Donnell Robert T. , McKnight Hayes , Sutcliff Julie L. , Tuscano Joseph M. 
Published: April 2020

Abstract
BACKGROUND: Chemotherapy for Non-Hodgkin’s Lymphoma (NHL) is initially effective, but often limited by toxicity and resistance. New agents for the treatment of NHL are needed. HB22.7 is a monoclonal antibody that binds CD22 and has previously been shown to reduce human lymphoma xenograft volume in nude mice.
OBJECTIVES: (1) Develop a 64Cu-DOTA-HB22.7 antibody, for in vivo imaging and potential therapy of NHL. (2) Determine if tumor targeting of 64Cu-DOTA-HB22.7 is equivalent between intravenous (IV), intraperitoneal (IP), and subcutaneous (SQ) routes of administration.
METHODS: DOTA-HB22.7’s ability to bind CD22+ cells was assessed by flow cytometry. 64Cu-DOTA-HB22.7 was injected IV, IP, or SQ into xenograft-bearing nude mice and tumor targeting assessed by positron emission tomography (PET) and biodistribution assay. Pharmacokinetics were determined by measuring radioactivity of blood samples. Serum was analyzed by radio-TLC.
RESULTS: DOTA conjugation does not affect HB22.7’s ability to bind CD22. 64Cu-DOTA-HB22.7 demonstrates specific tumor targeting at 24 and 48 hrs. Targeting is equivalent regardless of route of administration. Pharmacokinetics demonstrate that 64Cu-DOTA-HB22.7 can access the bloodstream (and thus, target tumor) in IP or SQ injections. By 48 hrs, blood 64Cu levels are (a) equivalent, regardless of injection route and (b) below peak levels, indicating clearance from the circulation. Serum analysis shows that right-shifted TLC peaks, possibly 64Cu-DOTA-HB22.7 metabolites, are present only in the IV injected group at 48 hrs.
CONCLUSION: These findings establish 64Cu-DOTA-HB22.7 as an NHL-specific imaging agent and indicate its potential for use in radioimmunotherapy. These findings provide evidence that more accessible routes of administration can achieve equivalent targeting results, and may lead to more efficient and accurate administration of antibody-based therapeutics in mice.
Loss of Pak1 Corrects Multiple Gain of Function Phenotypes in Nf1+/− Mast Cells.
Authors: McDaniel Andrew , Clapp D. Wade , Chernoff J. 
Published: April 2020

Abstract
Neurofibromatosis type 1, caused by mutations in the tumor suppressor gene NF1, is the most common genetic disorder in man and is characterized by the development of tumors called neurofibromas. Studies in genetically engineered Nf1 knockout mice have demonstrated that loss of the Nf1 tumor suppressor in Schwann cells, the known tumorigenic cell in neurofibromas, is necessary but not sufficient for tumor formation. By using adoptive transfer, we have now established that haploinsufficiency of Nf1 in the hematopoietic microenvironment (specifically mast cells) is necessary and sufficient for tumor progression in a Nf1 conditional knockout model. It is known that mast cells promote tumor progression via the secretion of growth factors and proteinases. Thus molecules that target mast cell function may be useful therapeutically. We have previously demonstrated that haploinsufficiency of Nf1 results in hyperproliferation and hyperactivation of the mitogen-activated protein kinase (MAPK) pathway in bone marrow-derived mast cells in response to kit-ligand (kit-L). Cross-cascade signaling between phosphoinositide 3-kinase activated Rho-GTPases and the classical MAPK pathway has been shown to be responsible for these abnormal phenotypes. p21-activated kinases (Paks) are downstream mediators of Rho-GTPase proteins that have been implicated in immortalized cells as positive regulators of MAPK pathway members and as modulators of cell growth and cytoskeletal dynamics. Utilizing an intercross of Pak 1−/− mice with Nf1+/− mice, we studied how Pak1 in regulates cellular functions associated with haploinsufficiency at Nf1 in primary mast cells. Loss of Pak1 corrects the gains of function in proliferation and migration found in Nf1+/− mast cells back to wild type levels via differential regulation of MAPKs. Pak1 regulates proliferation via a Pak/Erk pathway, while a Pak1/p38 pathway is critical for migration. The increased release of preformed inflammatory mediators in Nf1 haploinsufficient mast cells was also abrogated by genetic disruption of Pak1. Pak1 was found to contribute to MAPK signaling by modulating both ERK and p38 activation. Loss of Pak1 reduces the hyperactivation of ERK and p38 found in Nf1+/− mast cells to that of wild type controls. Further, using a model that stimulates local proliferation of mast cells in vivo, we confirm that loss of Pak1 corrects the accumulation and degranulation of Nf1+/− mast cells in vivo to wild type levels. Thus, Pak1 is a novel mediator of Ras-dependent mast cell functions, and treatments that target Pak1 may be useful as a targeted therapy for treatment of neurofibromas.