bluuebunny/crossref_metadata_2025_split huggingface dataset:
EBV-Associated Follicular Dendritic Cell Tumor of the Spleen.
Authors: Fekrazad M. Houman , Rosenbaum Lizabeth , Vasef Mohammad A. , Rabinowitz Ian 
Published: October 2019

Abstract
Epstein-Barr virus (EBV) expressing follicular dendritic cell proliferations in the spleen is an extremely rare but distinct variant of follicular dendritic cell tumor (FDCT). This neoplasm typically presents as an intra-abdominal mass with a predilection to involve spleen or liver with histologic features reminiscent of inflammatory pseudotumor (IP). However, in contrast to IP, this distinct FDCT consistently is associated with EBV. A subset of splenic tumors previously reported as examples of IP or inflammatory myofibroblastic tumor represent EBV-associated FDCT retrospectively. Despite morphologic similarity with IP, at least focal proliferation of follicular dendritic cells can often be demonstrated using follicular dendritic cell markers. Based on the limited published cases (Cheuk W, et al: Am J Surg Pathol, 2001;25:721–731), these proliferations can recur and/or metastasize but in contrast to classical intra-abdominal FDCT, their clinical behavior remains indolent. We report a case of 37-year-old gentleman who presented with a four month history of left-sided abdominal and flank pain, 8 lbs weight loss over a month, and on and off night sweats during the past two years. Initial blood tests including a CBC, LDH, electrolytes, renal and liver function were all within normal limits. CT scan of abdomen and pelvis revealed a 5.5 cm in diameter solitary splenic mass with cystic and solid components (Figure 1). There was no intra-abdominal or retroperitoneal lymphadenopathy. Hepatitis panel, EBV DNA PCR and also HIV serology were all negative. The patient underwent splenectomy due to clinical impression of a lymphoproliferative disorder. Pathology: The spleen weighed 237 grams and contained a solitary, 6 cm in greatest dimension, well circumscribed tumor. Histologically, the tumor was composed of a mixed population of small lymphocytes, histiocytes, plasma cells, and variable numbers of large oval to spindle cells. The spindle cells in some foci exhibited a storiform growth pattern. Neither overt cytologic atypia nor significant increased mitotic activity was present. Flow cytometric analysis revealed no monoclonal B-cell population or abnormal T-cell proliferation. Cytogenetic analysis revealed normal 46,XY karyotype in 20 metaphases. By paraffin immunohistochemistry, the spindled cells were positive for follicular dendritic cell markers CD21, CD35 and CD23. In situ hybridization using EBV RNA (EBER) probes revealed abundant EBV RNA transcripts in the nuclei of spindled and large oval cells (Figure 2). The spindle cells were negative for S-100 and ALK protein. The systemic symptoms resolved following splenectomy with no additional treatment. Six months after splenectomy, the patient remains asymptomatic and disease free at last clinical follow up.
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CLL Transcript and Serum Profiling Identifies Distinct Oncogenic Pathways in Rai Stage Progression.
Authors: Mahadevan Daruka , Klinkhammer Thomas , Cooke Laurence , Riley Christopher , Choi James , Sinari Shripad , Miller Thomas P. , Spier Catherine 
Published: October 2019

Abstract
Background: Chronic lymphocytic leukemia (CLL) is a common leukemia that is incurable. Survival curves for Rai stage 0/I and III/IV are distinct and a specific gene expression signature may be associated with stage progression (SP). However, distinct oncogenic pathways that differentiate stage progression are not well defined.
Methods: Mononuclear cells from Rai stage 0/I (N=8) and III/IV (N=8) CLL patients were obtained through an IRB approved protocol and analyzed utilizing the HG-U133A 2.0 Affymetrix array after isolating and purifying total RNA (Qiagen, RNAeasy). Control RNA samples are from 4 normal volunteers’ peripheral blood (PB) B-cells (AllCell, CA) and a normal reactive lymph node. Statistical significance of individual genes and pathways was analyzed using ‘BioConductor’. Differential gene expression was analyzed using ‘Limma’. Pathways and cytogenetics regions were analyzed for enrichment using ‘Category’. Real time RT-PCR on specific genes was performed to validate the GEP based on stage. A serum cytokine profile (120 cytokines, RayBiotech) on 8 CLL patients (N=4 low risk; N=4 high risk) and 4 normal volunteers was evaluated to identify a signature for diagnosis and prognosis.
Results: Data are presented as “robust” increases or decreases of relative gene expression common to 8 patients in the low or high risk stages. The Wnt/Ror-1 non-canonical, and TGFb pathways are over-expressed in Rai stage 0/I, implicating a program of cell survival and dysregulated migration. The Hedgehog, Proteasome, TNF/FAS/NF-kB, and MAPK pathways are over-expressed in Rai stage III/IV, suggestive of a tumor microenvironment driven program in SP. Genes located on chromosome 3q27.1 are also dysregulated in Rai stage III/IV. A distinct serum cytokine profile also differentiates low from high stage CLL.
Conclusions: GEP of CLL with RT-PCR validation has identified distinct oncogenic pathways that distinguish Rai stage 0/I from III/IV. A serum cytokine profile also differentiates between low and high risk CLL. These data together provide a ‘molecular signature’ for SP in CLL.
Phenotypic Analysis of Plasma Cells in Monoclonal Gammopathy and Multiple Myeloma Subjects.
Authors: Kovarova Lucie , Buresova Ivana , Pour Ludek , Suska Renata , Adam Zdenek , Krejci Marta , Vorlicek Jiri , Penka Miroslav , Hajek Roman 
Published: October 2019

Abstract
Introduction: Multiple myeloma (MM) is a B-cell neoplasia caused by the proliferation of clonal plasma cells (PCs). MM and benign monoclonal gammopathy of unknown significance (MGUS) are routinely distinguished on the basis of paraprotein concentration, level of PCs infiltration and the presence or absence of other clinical features. Flowcytometric detection of PCs according to the expression of CD38 and CD138 has limitation in discrimination between normal and abnormal PCs, but this is possible in multicolor phenotypic analysis. The aim of this study is to compare the numbers of normal and abnormal PCs in MGUS and MM subjects and to find some parameter useful for evaluation PCs distribution.
Materials and methods: 51 newly diagnosed untreated MM patients (63,9±10,0 years old) and 31 non-treated MGUS subjects (64,2±13,8 years old) were analysed. Lysed whole bone marrows were analysed by flowcytometric immunophenotyping and PCs were indentified by expression of CD38, CD138, CD45 and also CD56 and CD19.
Results: Discrimination between normal CD19+ PCs and abnormal CD56+ PCs was done on CD38+CD138+ population. Ratio of normal PCs count and abnormal PCs counts (normal/abnormal=N/A) was used to describe a distribution of PCs. Subjects with MGUS had 0,97±1,65% (average±SD) of CD38+CD138+ cells (median 0,45; range 0,03–7,47%) with average N/A ratio 2,91±3,94 (median 1,36; range 0,01–18,44). Newly diagnosed MM patients had 4,96±9,94% of CD38+CD138+ cells (median 0,85; range 0,02–41,80%) with average N/A ratio 1,70±3,03 (median 0,13; range 0–11,5). In 9,7% MGUS subjects evaluation of distibution of PCs showed transformation of MGUS into MM. In some newly diagnosed MM patients (31,4%) CD38+CD138+ plasma cells were positive for CD19 although they were aberrant and these PCs were mostly CD45+. In some patients (9,7% of MGUS; 17,6% of MM) PC did express neither CD19 nor CD56 and this fact may complicate further evaluation of PCs.
Conclusions: Results confirmed that in MGUS subjects we can find lower numbers of PCs which are mostly CD45+CD19+. A majority of plasma cells in newly diagnosed MM patients are abnormal CD56+ PCs and these plasma cells are usually CD45−. Further follow-up of patients can confirm the N/A ratio as a predictive factor for transformation of MGUS into MM and its value for evidence of relapse or progression to active myeloma.
Activity of ARRY-614, an Inhibitor of p38 Map Kinase and Angiogenic Targets, in Hematologic Malignancies.
Authors: Winski Shannon L. , Gross Stefan D. , Brown Suzy A. , Anderson Deborah , Garrison Augusta , Rodriguez Mareli , Lee Patrice , Munson Mark , Winkler James 
Published: October 2019

Abstract
Multiple Myeloma (MM) is a malignancy characterized by the clonal expansion of plasma cells within the bone marrow microenvironment. Within this compartment, the proliferative capacity of MM cells is enhanced by pro-inflammatory cytokines, and there is growing recognition that bone marrow stromal cells play an important role in the support of MM growth and chemo-resistance. We have identified a series of compounds, of which ARRY-614 is representative, that are potent inhibitors (EC50<10 nM) of cytokine synthesis through the inhibition of p38 MAPK. This compound is also a potent inhibitor of Abl tyrosine kinases and Tie2/Tek receptor tyrosine kinase. ARRY-614 is active against all of these targets on both the isolated enzymes and in cells. To confirm these activities, ARRY-614 was evaluated in relevant in vitro and in vivo models. ARRY-614 inhibited p38α in ex vivo stimulated human whole blood (EC50=2 nM) and the release of IL-6 and TNFα from SEA- or LPS-challenged mice (ED50<10 mg/kg). Further, ARRY-614 administered as a single agent was efficacious at inhibiting bFGF-driven angiogenesis in an in vivo matrigel invasion assay as well as inhibiting tumor growth in subcutaneous K562 (BCR-Abl dependent) and RPMI 8226 (multiple myeloma) tumor xenografts in mice, at doses ranging from 30 to 100 mg/kg qd, PO. In regulated safety studies, this compound was well-tolerated at doses up to 100 mg/kg qd and 30 mg/kg qd in rats and cynomolgus, respectively. Together, these data support the advancement of this agent into clinical trials for hematologic malignancies.
Prospective Evaluation of MRD-Kinetics in 274 Children with High-Risk ALL Treated in Trial ALL-BFM 2000: Insights into Development of Resistance and Impact on Further Refinement of Treatment Stratification Strategies.
Authors: Schrauder André , Stanulla Martin , Flohr Thomas , Cario Gunnar , Köhler Rolf , Panzer-Grümayer Renate , Schäfer Beat , Zimmermann Martin , Peters Christina , Bartram Claus R. , Schrappe Martin 
Published: October 2019

Abstract
In trial ALL-BFM 2000, high-risk (HR) acute lymphoblastic leukemia (ALL) is defined by inadequate initial response to induction treatment [poor prednisone response on treatment day eight, non remission on treatment day 33, and/or a high load of minimal residual disease (MRD, ≥10E-3) after 12 weeks of treatment (TP2)] and/or by cytogenetics [t(4;11 or t(9;22)]. Between August 1999 and November 2006, 494 (15%) out of 3255 study patients were stratified into the HR branch of trial ALL-BFM 2000. 431 (87%) of these HR patients underwent successful MRD monitoring at TP2 with 274 (56%) patients having received additional extensive prospective MRD monitoring subsequent to TP2. Patients with an indication for stem cell transplantation (SCT) and a suitable donor were scheduled for SCT within six weeks after the third HR block of the intensive consolidation phase. The estimated 4-years event-free-survival (4y-pEFS) for the entire HR group was 68%+/−3%, estimated survival was 74%+/−3%. Patients with MRD load of ≤10E-4 at TP2 (n=231) had a 4y-pEFS of 82%+/−3%, patients with MRD levels of 10E-3 at TP2 (n=84) had a 4y-pEFS of 74%+/−6%, and patients with MRD of >10E-3 at TP2 (n=116) had a 4y-pEFS of 35%+/−5%. MRD-kinetics subsequent to TP2 revealed, that 85% of all patients with an MRD level of 10E-3 at TP2 continued to decrease their load below 10E-3 during the pulsatile intensive consolidation phase, whereas this was observed in only 35% of patients with an MRD level of >10E-3 at TP2. The 4y-pEFS of patients with an MRD load persisting at 10E-2 after application of three intensive HR blocks after TP2 was 0% if no SCT was performed, and 33%+/−11% after SCT in CR1. Our data reflect that extensive MRD measurements in HR-ALL patients allow a dynamic insight into the development of resistance, and serve as valuable tool for further clinical treatment adjustment. “MRD non-response” after three BFM HR blocks identifies a patient group in urgent need of alternative treatment elements, closely monitored by MRD, before going into SCT in CR1.
Loss of FOXP3 Expression and Emergence of Cytokine-Producing Cells after In Vitro Expansion of Human CD4+CD25+CD127 - Regulatory T Cells.
Authors: Hoffmann Petra , Eder Ruediger , Boeld Tina J. , Huehn Jochen , Floess Stefan , Wieczorek Georg , Baron Udo , Olek Sven , Andreesen Reinhard , Edinger Matthias 
Published: October 2019

Abstract
We and others previously showed that the adoptive transfer of donor-type CD4+CD25+ regulatory T (Treg) cells protects from graft-versus-host disease (GVHD) after allogeneic stem cell transplantation (SCT) in animal models. Exploring this strategy in human SCT, we currently perform a first Phase I clinical trial using freshly isolated Treg cells. For future trials requiring large Treg cell numbers for repetitive treatments, we described in vitro culture conditions that permit a more than 3-log expansion of polyclonal Treg cells (Blood 104:895; 2004). A highly enriched starting population proved to be crucial for the generation of pure Treg cell products and we could show that the naive, CD45RA+ subpopulation of CD4+CD25high T cells fulfills this criterion (Blood108:4260; 2006). A recently proposed alternative approach for the isolation of pure Treg cells relies on the exclusion of CD127+ cells, as activated (i.e. CD25+) conventional T cells express high levels of CD127 while CD4+CD25+ Treg cells show no or only weak expression levels (Seddiki et al. and Liu et al., JEM203:1693; 2006). To directly compare these two approaches, we isolated CD4+CD25+CD127low/neg T cells (CD127-Treg) and CD45RA+CD4+CD25high T cells (RA+ Treg) from the same leukapheresis products and analyzed the cells after 2 and 3 weeks of expansion. Whereas both populations were > 94% FOXP3+ upon isolation, only RA+ Treg maintained this high level of FOXP3+ cells throughout the expansion period (93% (range: 78 to 97%; n=11) FOXP3+ after 2 and 87% (range: 71 to 97%; n=9) FOXP3+ after 3 weeks). In contrast, the proportion of FOXP3+ cells in CD127-Treg cultures was already reduced after 2 weeks (82% (range: 56 to 96%; n=11)) and highly variable and significantly lower than that of RA+ Treg cultures after 3 weeks (57% (range: 18 to 93%; n=9; p=0.006)). When further subdivided into CD45RA+ and CD45RA- subpopulations before expansion, cultures of CD45RA-CD127-Treg cells lost FOXP3 expression and comprised substantial numbers of FOXP3- cytokine producing cells (on average 54% and 38% IL-2 and IFN-γ producers, respectively) after 3 weeks, whereas CD45RA+ CD127- Treg behaved similar to RA+ Treg cells, as they maintained FOXP3 expression over time and contained only low numbers of cytokine producers. Furthermore, when we analyzed the DNA methylation status of Treg cells, we found the Treg-specific CpG demethylation pattern within the FOXP3 gene (EJI 37, 2007) in RA+ Treg cell lines, while CD127- Treg cell cultures showed increased methylation over time and even more so RA- Treg cell cultures. Based on these findings, we suggest that isolation and expansion of CD45RA+CD4+CD25high T cells at present represents the best strategy for adoptive cell therapies requiring in vitro expanded Treg cells.
Non-Classical Anti-Factor VIII C2 Domain Inhibitors Are Present in the Plasmas of Most Factor VIII Inhibitor Patients.
Authors: Meeks Shannon L. , Healey John F. , Barrow Rachel T. , Parker Ernest T. , Lollar Pete 
Published: October 2019

Abstract
Approximately 30% of patients with severe hemophilia A will develop inhibitory antibodies to factor VIII (fVIII inhibitors). The immune response to fVIII currently is the most significant complication in the management of patients with hemophilia A. In addition, autoimmune antibodies to fVIII can develop in non-hemophiliacs, producing acquired hemophilia A, which frequently produces life- or limb-threatening bleeding. These inhibitors primarily are directed against the A2 or C2 domains of fVIII. The human response to the C2 domain of fVIII classically has been thought to inhibit fVIII activity by blocking its binding to phospholipid. We recently characterized the antibody response to the C2 domain of human fVIII in a murine hemophilia model and described 5 structural groups of antibodies. Groups A, AB, and B are classical anti-C2 antibodies. Groups BC and C consist of non-classical anti-C2 antibodies that inhibit the proteolytic activation of fVIII but do not block the binding of fVIII to phospholipid. Most non-classical antibodies have inhibitor titers greater than 10,000 Bethesda units/mg IgG. To determine if non-classical antibodies are present in fVIII inhibitor patients, patient plasmas were tested in an ELISA for their ability to block the binding of representative antibodies from the different anti-human fVIII C2 antibody groups. Classical and non-classical monoclonal antibodies (MAbs) were biotinylated and serially diluted into either fVIII deficient plasma or patient inhibitor plasma and then added to microtiter wells coated with fVIII. The ability of patient plasma to block the binding of the murine MAbs to fVIII was determined. A total of 16 patient plasmas were assessed: 4 from patients with a C2 predominant response, 2 with a non-C2 predominant response, and 10 with unknown specificities. Three of the 4 patients with C2 predominant responses had non-classical anti-C2 antibodies, while the 2 with non-C2 predominant responses did not. In the unknown plasmas, 6 of 10 had evidence of non-classical antibodies. Figure 1 shows representative results of the effect of 3 patient plasmas on the binding of a biotinylated non-classical MAb to fVIII. Patient plasmas 1 and 2 blocked MAb binding while patient plasma 3 did not. This study indicates that the majority of patients with fVIII inhibitors have non-classical anti-C2 antibodies in their response to fVIII.
Figure Figure
RhoH Functions as an Adaptor Molecule for ZAP-70 in TCR Signaling Pathway.
Authors: Chae Hee-Don , Siefring Jamie E. , Hildeman David A. , Gu Yi , Williams David A. 
Published: October 2019

Abstract
RhoH is a hematopoietic-specific and GTPase-deficient Rho GTPase of the RhoE family. In a previous report, we showed that RhoH plays a critical role in T cell receptor (TCR) signaling via recruitment of ZAP-70 in T cell development (Gu et al., Nat. Immunol., 2006). RhoH-deficient T cells show impaired CD3ζ phosphorylation which is critical for the recruitment of ZAP-70 to the TCR complex. In this study, we investigated whether RhoH acts as an adaptor molecule for ZAP-70 membrane targeting or a regulator of the Lck activity. Although Lck activity was not significantly decreased in RhoH-deficient T cells as determined by the level of Lck phosphorylation and in vitro kinase assay, Lck protein did not translocate to the immunological synapse in these cells and the association between Lck and CD3ζ was impaired. ZAP-70 has been reported to mediate the recruitment of Lck into TCR complex with subsequent CD3ζ phosphorylation. In RhoH-deficient T cells, ZAP-70 did not translocate to TCR complex in the plasma membrane, especially the detergent-insoluble (cytoskeletal) fraction. To further determine if RhoH mediated membrane localization of ZAP-70 and if this localization was responsible for normal T cell differentiation, we generated a myristoylation sequence-fused ZAP-70 (Myr-ZAP-70). Myr-ZAP-70 localized to the membrane fraction and resulted in the partial rescue of RhoH−/ − T cell phenotype as measured by T cell reconstitution in Rag2−/ − recipient mice (Table 1). These results suggest that RhoH plays a key role in TCR signaling as an adaptor for ZAP-70 plasma membrane localization.
Table 1: Engraftment and T cell reconstitution of the transduced RhoH−/ − BM cells in Rag2−/ − recipient mice Donor cells (retroviral vector transduction) CD4+ CD8− cells (x 10e5) CD4− CD8+ cells (x 10e5) Spleen PB (ml) Spleen PB (ml) RhoH−/− (empty vector) 3.33 0.9 1.2 0.36 RhoH−/− (ZAP-70) 16.7 1.2 3.1 0.6 RhoH−/−(Myr-ZAP-70) 35.2 3.3 9.1 2.2 RhoH−/− (HA-RhoH) 65.0 3.9 13.6 1.3
Role of CD37SMIP a Novel Engineered Small Modular Immunopharmaceutical in the Treatment of CLL.
Authors: Lapalombella Rosa , Zhao Xiaobin B. , Baum Peter R. , Ledbetter Jeffrey A. , Muthusamy Natarajan , Byrd John C. 
Published: October 2019

Abstract
CD37 is a lineage-specific B-cell antigen highly expressed on both normal and transformed B-cells. The significant B-cell selective CD37 expression makes it a valuable target for therapy against B cell malignancies including chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), and non-Hodgkins lymphoma (B-NHL). A novel CD37-specific small modular immunopharmaceutical (CD37SMIP) was engineered to contain a single chain variable region (scFv) linked to a human IgG1 hinge, CH2 and CH3 domains. CD37SMIP induces potent apoptosis in the presence of a crosslinker and antibody dependent cellular cytotoxicity (ADCC) against primary CLL cells. Significant in vivo therapeutic efficacy was demonstrated in a SCID mouse xenograft leukemia/lymphoma model that is dependent upon NK cell function. The induced cytotoxicity in primary CLL cells was independent of activation of caspase cascade, and consistent with this, the pan-caspase inhibitor z-VAD-fmk failed to rescue CD37 SMIP drug induced cytotoxicity. In an attempt to define the CD37 mediated early signaling events and their role in cytotoxicity, we investigated protein tyrosine phosphorylation as a potential early activation event responsible for CD37 SMIP drug mediated cytotoxicity. Primary B CLL cells were stimulated with increasing concentration of CD37 SMIP with or without crosslinker. Western blot analysis of cellular lysates with anti-phosphotyrosine antibody revealed several tyrosine phosphorylated proteins including a predominant ∼65KDa protein in response to the cross-linking of CD37 SMIP within 10 minutes. Further, pre-treatment with the tyrosine kinase inhibitor herbimycin prevented tyrosine phosphorylation of the predominant 65KDa protein and inhibited CD37 SMIP induced apoptosis in a dose dependent manner. Attempts to identify the tyrosine phosphorylated proteins, using MALDI-TOF mass spectrometry, and to define the potential role of these target proteins in the apoptotic pathway are ongoing. To further examine the molecular mechanism whereby CD37 SMIP triggers cell death, we investigated the involvement of mitochondrial pathways including changes in mitochondrial potential (Δψm) and generation of reactive oxygen species. Treatment with CD37 SMIP induces a time dependent mitochondrial membrane depolarization and increased production of ROS. Further, primary B CLL cells cultured with CD37SMIP induced cleavage and mitochondrial translocation of Bax and upregulation of Bim in a time dependent manner indicating a potential role for Bim and Bax pro apoptotic proteins in CD37 induced apoptosis. These findings provide strong justification for CD37 as a therapeutic target aimed at modulation of pro apoptotic proteins and introduce small modular immunopharmaceuticals as a novel class of targeted therapies for B-cell malignancies. (SMIP trademark is owned by Trubion Pharmaceuticals).