bluuebunny/crossref_metadata_2025_split huggingface dataset:
Zanolimumab, a Fully Human Monoclonal Antibody: Preliminary Results of an Ongoing Clinical Trial in CD4+ Peripheral T-Cell Lymphomas (PTCL).
Authors: d’Amore Francesco , Relander Thomas , Jerkeman Mats , Hagberg Hans , Osterborg Anders , Johnson Peter , Radford John A. , Hancock Barry W. , Gramatzki Martin , Schmitz Norbert , Tilly Herve , Morschhauser Franck , Lisby STeen , Baadsgaard Ole 
Published: October 2019

Abstract
Zanolimumab (previously referred to as HuMax-CD4) is a fully human monoclonal IgG1k antibody. It is specific for the CD4 antigen expressed on a subset of T cells and has been shown to possess cytotoxic and anti-proliferative effects. We report initial clinical efficacy and safety results following treatment with 980 mg of zanolimumab in 15 patients diagnosed with biopsy-proven, treatment-refractory or relapsed CD4+ systemic PTCL. The following PTCL subtypes were included : enteropathy-type T-cell lymphoma (n=1), anaplastic large T-cell lymphoma (ALCL, n=3, 2 alk-negative, 1 alk-status unknown), angioimmunoblastic T-cell lymphoma (AIL, n=8) and PTCL-unspecified (PTCLu, n=3). The primary endpoint of the study was objective tumor response. Responses were based on CT scan and clinical examination and classified according to the Cheson criteria. Out of 15 patients enrolled, 4 responses have been seen; 2 PR (one patient later achieved a CRu) and 2 CRu verified by CT scan. Further, clinical improvements (marked decrease in the size of peripheral lymph nodes) were recorded in 2 additional patients. So far, a total of 21 SAEs has been reported. Of these, 4 were considered related to the study treatment: febrile neutropenia (n=1), thrombocytopenia (n=1), infusion related reaction (n=1) and hyperthermia (38.8° C) with hypotension (n=1). Eight unrelated deaths have been reported so far, all secondary to disease progression. In conclusion, in this heavily pretreated patient population, zanolimumab demonstrated a good tolerability profile and the present results indicate efficacy in the treatment of CD4-positive PTCL.
European Collaborative Paediatric Database on Cerebral Venous Thrombosis: Risk Factors for Recurrent Venous Thromboembolism.
Authors: Kenet Gili , Kirkham Fenella , Niederstadt Thomas , Kurnik Karin , Schobess Rosemarie , Knofler Ralf , Zieger Barbara , Nowak-Gottl Ulrike 
Published: October 2019

Abstract
Background: Risk factors for cerebral venous thrombosis (CVT) in children include local and systemic underlying conditions, drug toxicity, and hereditary prothrombotic risk factors. Their relevance to the risk of a second venous thrombosis (VT), compared with other clinical, neuroimaging and treatment variables, is unknown.
Methods: 330 of 407 consecutively enrolled CVT-patients aged newborn to <18 years (median 5.3 years; male 55%) were prospectively followed for a median (range) of 33 (12–84) months. In accordance to international treatment guidelines, 259 children (78.5%) received acute antithrombotic therapy (AT) with UFH or LMWH, followed by long-term AT with LMWH or warfarin in 218 cases (66.0%).
Findings: Recurrent VT was diagnosed in 19 of 330 children (5.8%) at a median (range) age of 6 (0.5–84) months following CVT, with no difference observed between the study centres (p=0.84). Multivariate Cox regression analysis revealed that the risk of recurrence was significantly higher in those with older age at first CVT onset (p=0.02), those in whom AT was not administered prior to recurrence (p=0.0003), those who did not have complete patency rates on repeat venography at 3–6 months (p=0.01), and those subjects carrying the factor II G20210A mutation (p=0.03).
Interpretation: Age at CVT onset, administration of AT, poor patency rates, and the factor II G20210A variant were of relevance to recurrent VT in children with CVT. Until evidence-based data derived from randomized treatment trials are available the message of this follow-up study is to administer any AT prophylaxis on an individual patient basis in paediatric CVT cases in newly identified VT risk situations.
Evaluation of Minimal Residual Disease in Blood and Bone Marrow for CLL as Assessed by 6 Color Flow Cytometry.
Authors: Maloum Karim , Sutton Laurent , Garff-Tavernier Magali Le , Fiocconi Caroline , Bonnemye Patrick , Brissard Martine , Merle-Beral Helene 
Published: October 2019

Abstract
MRD evaluation is becoming critical in CLL patients since the new therapeutic approaches lead to higher and higher biological response. Multicolor flow cytometry is demonstrated to be one of the best technique for assessing MRD. In a recent work, Moreno et al (Blood, 2006) suggested that peripheral blood could be sufficient for MRD evaluation. We here report our experience which compares peripheral blood and bone marrow aspirate for MRD assessment using a 6 color flow cytometry (FC) method. Two months after the end of the treatment, blood and bone marrow were harvested at the same time for 64 CLL patients in complete remission according to the NCI criteria. Patients were treated by chemotherapy followed or not by autologous stem cell transplantation (ASCT). None of them received immunotherapy. Until now 30 out of the 64 couples of blood and marrow samples have been studied. Immunophenotyping was performed using the FacsCanto Device (Becton Dickinson) and the following monoclonal antibody (moAb) combinaisons: k-FITC / l-PE / CD19-PerCP-Cy5.5 / CD5-PE-Cy7 / CD79b-APC / CD20-APC-Cy7; FMC7-FITC / CD38-PE / CD19PerCP-Cy5.5 / CD5-PE-Cy7 / CD20-APC-Cy7 for peripheral blood assessment and k-FITC / l-PE / CD45-PerCP-Cy5.5 / CD5-PE-Cy7 / CD19-APC / CD20-APC-Cy7; FMC7-FITC / CD19-PE / CD45PerCP-Cy5.5 / CD5-PE-Cy7 / CD79b-APC / CD20APC-Cy7 ; CD38-FITC / CD23-PE / CD45-PerCP-Cy5.5 / CD5-PE-Cy7 / CD19-APC. In the bone marrow, we have discriminated hematogons from CLL cells by using moAb combinations containing CD45 and CD38. Between 4500 and 5500 lymphocytes were analyzed in each test. By using a method of serial dilutions, we demonstrated a sensitivity of 10−4 for our 6-color FC technique. CLL cells were defined as CD19+CD5+CD79b low CD20low lymphocytes with low expression of monotypic Ig light chain. Results were expressed as a percentage of leukemic cells among total B lymphocytes. We showed a complete correlation between blood and bone marrow for MRD evaluation using this technique. A complete phenotypic remission defined by the absence of leukemic cells in the blood and in the bone marrow, was observed in 17 out of 30 cases. In the other 13 patients, persistent leukemic cells were detected in the marrow as well as in the peripheral blood with a median rate of 1.56 % [0.15 – 41] and 0.56% [range 0.06 – 36] respectively. Our results showed that, by using 6 color FC, blood assessment for detecting MRD in CLL after chemotherapy or ASCT could provide a simple surrogate to bone marrow assessment. Analysis of the remaining couples of samples for the last 34 patients is ongoing.
CD38 Expression in B-CLL Is Dynamic and Changes Due to Contact with Activating Stimuli Found within the Leukaemic Microenvironment.
Authors: Patten Piers E.M. , Buggins Andrea G.S. , Richards Julie , Wotherspoon Andrew , Hamblin Terry John , Mufti Ghulam , Devereux Stephen 
Published: October 2019

Abstract
High levels of CD38 expression in B-cell chronic lymphocytic leukaemia (B-CLL) confer a poor prognosis. Although its role in B-CLL is unknown, signalling through CD38 has been implicated in cell survival, trafficking and proliferation. Since proliferation in B-CLL is thought to take place within both bone marrow (BM) and secondary lymphoid tissue, we investigated whether CD38 expression might vary in response to stimuli that occur in these tissue compartments.
Firstly, we compared the percentage CD38 expression of CD5/19 cells on 35 paired PB and BM aspirate B-CLL samples. The mean CD38% was significantly higher in BM than PB in all samples (27% vs 19%, p=0.009) including samples with a PB CD38 of 7% or more (33% vs 42%, p=0.047), indicating that factors present in the BM up regulate CD38 expression. Next, CD38 expression and cell division of B-CLL peripheral blood mononuclear cells (PBMCs) were examined in an in vitro system aimed at mimicking the proliferation centre microenvironment where leukaemic cells are situated in close proximity to activated T lymphocytes. Positively selected T cells from 15 B-CLL patients were activated overnight with CD3/28 beads and subsequently cultured with autologous B-CLL PBMCs. Both the percentage of CD19+ CD38+ cells (29.9% vs 59.9%, p=0.003) and CD38 mean fluorescence intensity (75.1 vs 830.8, p=0.005) increased over the 6 day culture period. B-CLL cell division was assessed using the dye carboxyfluorescein diacetate succinimidyl ester (CFSE) in the same co-culture system. This showed that co-culture with autologous activated T-cells can result in B-CLL cell division, and is preceded by CD38 up regulation. In addition, significantly more B-CLL cells underwent at least one division from patients with an initial CD38 level of 7% or more, as compared to under 7% (24.6% vs 10.9%, p=0.031). To further investigate the relationship between B-CLL cell proliferation, CD38 expression and the role of T-cells we examined tissue sections known to contain paraimmunoblasts and other proliferating B-CLL cells. Four colour confocal microscopy using CD3, Ki67, CD38 and CD23 to label frozen B-CLL lymph nodes was employed. Large Ki67+ CD23+ cells were present in close proximity to CD3+ T-cells and these large B-CLL cells had higher CD38 expression than the surrounding small B-CLL lymphocytes.
These results support the proposal that CD38 expression in B-CLL is dynamic and may reflect exposure to T-cell derived stimuli which contribute to proliferation in the BM or LN microenvironment. A possible explanation for the poorer prognosis of patients with higher CD38 expression may be that their disease has more proliferative potential.
Expression of ZAP-70 in Chronic Lymphocytic Leukemia Cells Enhances CD79b Phosphorylation Following Surface IgM Ligation.
Authors: Chen Liguang , Apgar John , Tang Li , Kipps Thomas J. 
Published: October 2019

Abstract
CD79b is B-cell surface molecule that non-covalently associates with CD79a and surface immunoglobulin (sIg), which together serve as the B-cell receptor complex (BCR). Both CD79a and CD79b have cytosolic immunoreceptor tyrosine-based activation motifs (ITAMs) that can become phosphorylated following sIg ligation, thereby allowing for recruitment to the BCR complex of cytosolic kinases, such as p72Syk , which then can initiate downstream intracellular signaling events. Compared to normal B cells, chronic lymphocytic leukemia (CLL) B cells typically expresses low levels of CD79b, which is speculated to contribute to the relatively poor capacity of CLL cells to initiate intracellular signaling following BCR ligation despite having apparently adequate levels of p72Syk. BCR signaling in CLL cells can be enhanced by expression of the zeta-associated protein of 70 kD (ZAP-70), a tyrosine kinase that initially was identified in T cells, where it plays a critical role in the phosphorylation of ITAMs of the accessory molecules of the T-cell receptor (TCR) complex for antigen following TCR ligation. We investigated for phosphorylation of CD79b following BCR ligation with F(ab)2 anti- μ antibody in CLL cell samples that did or did not express ZAP-70. All CLL cell samples expressed similar amounts of surface IgM and p72Syk, as assessed via flow cytometry and immunoblot analysis. Within 10 minutes after treatment with anti-μ the CLL cell samples that expressed ZAP-70 (n = 28) experienced a mean increase in phosphorylation of CD79b of 21.5% (± 14.0% S.D.), which was significantly greater than the 7.5% increase (± 7.9% S.D.) experienced by similarly treated CLL cell samples that did not express ZAP-70 (n = 19) (P< 0.01). Immune precipitation studies demonstrated association of CD79b with p72Syk in CLL B cells. CLL cell samples (n = 5) lacking expression of ZAP-70 were transfected with a control vector or an expression vector encoding ZAP-70, allowing us to examine the effect that engineered-expression of ZAP-70 has on CD79 phosphorylation following treatment with anti-μ. Anti-μ treatment induced significantly higher mean levels of CD79b phosphorylation in CLL samples made to express ZAP-70 (33% ± 16%) than in control mock-transfected CLL cells (4% ± 2%). This also was associated with enhanced anti-μ induced phosphorylation of p72Syk. We conclude that expression of ZAP-70 in CLL B cells enhances phosphorylation of the accessory molecules in the BCR complex following sIg ligation, potentially allowing for improved recruitment of cytosolic kinases and adapter proteins to these accessory molecules for enhanced BCR signaling.
The Vascular Endothelial Growth Factor (VEGF) Autocrine Survival Signaling Pathway in Chronic Lymphocytic Leukemia (CLL) Is Regulated by Lysophosphatidic Acid (LPA).
Authors: Hu Xiaojie , Sun Jinmie , Johnston James B. , Gibson Spencer B. 
Published: October 2019

Abstract
Vascular endothelial growth factor (VEGF) is a survival and angiogenesis factor that has been the target of chemotherapy in a variety of cancers including chronic lymphocytic leukemia (CLL). The VEGF survival response is regulated by an autocrine loop where cells constitutively express VEGF and activate the VEGF receptors leading to up-regulation of anti-apoptotic Bcl-2 members and prevention of apoptosis. Herein, we have demonstrated that VEGF is elevated in the plasma of CLL patients compared to normal individuals. We have previously shown that the growth factor, lysophospatidic acid (LPA), is a survival factor in CLL and its receptor LPA1 is up-regulated in CLL cells (J Biol Chem, 280:9498, 2005). Now, we demonstrate that inhibiting VEGF receptor activation blocks the survival response seen with LPA in CLL. LPA treatment was found to induce VEGF mRNA and protein levels within 1 hour. LPA receptor inhibitor Ki16425 blocked the VEGF production and reversed the protective effect of LPA against apoptosis. Combining LPA and VEGF failed to increase survival responses in these cells. In addition, LPA protection against fludarabine and TRAIL induced apoptosis was eliminated using the VEGF receptor kinase inhibitor, epigallocatechin-3-gallate (EGCG), which is present in green tea, or VEGFR:Fc, that inhibits VEGF receptor activation. Furthermore, LPA induced AKT activation and up-regulation of the expression of the Bcl-2 family member, Mcl-1, were eliminated by the VEGF receptor inhibitor. Thus, the protective effect of LPA against apoptosis in CLL is related to increased production of VEGF. Targeting the LPA pathway similar to the VEGF pathway may thus be a novel approach for the treatment of CLL.
Long Term Outcome of Patients with Steroid Refractory Acute Intestinal Graft Versus Host Disease after Treatment with Pentostatin.
Authors: Klein Stefan A. , Mousset Sabine , Bug Gesine , Waßmann Barbara , Hoelzer Dieter , Martin Hans 
Published: October 2019

Abstract
Steroid refractory intestinal acute Graft versus Host Disease (aGvHD) is a mayor complication after allogeneic hematopoietic cell transplantation. Survival rate of severe intestinal aGvHD is low. No therapy has been proven to increase the survival rate in steroid refractory aGvHD so far. A promising strategy is the use of pentostatin, a purine nucleotide analogue and inhibitor of adenosine deaminase, which is known to decrease function and number of lymphocytes. Here we report the six year follow-up of a pilot study on salvage therapy of aGvHD by pentostatin. We treated 18 patients with steroid refractory intestinal aGvHD stage III or IV with Pentostatin (8 female and 10 male patients). The mean age was 45 (range: 25–61 years). The underlying diseases were ALL (4), AML (10), Multiple Myeloma (2), Hodgkin’s Disease (1) and CML (1). Eight patients were allografted with HLA-identical sibling donors, three with related donors with a single HLA class I mismatch, seven with matched unrelated donors. All patients were allografted with peripheral blood stem cells. As GvHD prophylaxis ciclosporine A in combination with mycophenolate mofetil or methotrexate were used. Eleven patients had aGvHD stage III, seven stage IV. All patients had a severe gastrointestinal-tract involvement (grade III or IV). After failure of steroid treatment (prednisolone >2 mg/kg for at least 3 days) pentostatin was applied as a salvage therapy (1 mg/m2 for three consecutive days). Eight patients received between one and three further courses of pentostatin every three to four weeks. Therapy was well tolerated. No severe neutropenia was observed. Except one case of hemolytic uremic syndrome (HUS) no impairment of renal function was observed. Eleven patients achieved a complete, five a partial remission of aGvHD. Time until a clinical improvement could be observed was 12–14 days. Two patients died before day 12 without improvement of GvHD symptoms. Seven patients (39%) are alive (510–2060 days post first cycle of pentostatin), two with extensive chronic GvHD. All one-year-survivors stayed alive. Eleven patients died (two due to relapse of leukemia, nine transplant related [1 x interstitial pneumonitis, 1 x pneumonia, 1 x chronic GvHD, 4 x aGvHD and multiple organ failure (MOF), 1 x parvo B19 infection and MOF, 1 x HUS and MOF]). In conclusion, in this pilot study pentostatin has been demonstrated to be a highly effective and well tolerated drug for salvage therapy of steroid refractory acute GvHD with intestinal involvement. Moreover, compared with earlier studies on steroid refractory aGvHD a high rate of long term survivors could be observed. By a multicenter prospective randomised study we are currently investigating whether pentostatin can improve the outcome in steroid refractory intestinal aGvHD.
Deleterious Roles of the KIR Ligand Mismatch on Acute Graft Versus Host Disease in Unmanipulated HLA-Mismatched/Haploidentical Blood and Marrow Transplantation.
Authors: Huang Xiao Jun , Zhao Xiang Yu , Liu Dai Hong , Liu Kai Yan , Xu Lan Ping , Chen Yu Hong , Han Wei 
Published: October 2019

Abstract
The beneficial effect of KIR ligand mismatch with a very low incidence of rejection, acute graft versus host disease (aGVHD), and leukemia relapse had been demonstrated by Perugia’s group in the Haploidentical mismatched Hematopoitic cell transplantation (HCT) with extensive T cells depletion in vitro. However, many analytical results of the haploidentical and unrelated mismatched transplantation appeared to be inconsistent with Perugia’s work. The disputes in the inconsistent roles of KIR ligand mismatch seem to be caused by the different transplant protocols with different extent of T cells depletion in vitro or in vivo. In recent years, we successfully established a novel protocol—conditioning including antithymocyte globulin followed by un-manipulated HLA-mismatched/haploidentical blood and marrow transplantation, which can achieve comparable outcomes to HLA-identical sibling transplantation. Following the contradictory results about the KIR ligand mismatch in the haploidentical related and mismatched unrelated HCT, we have evaluated the roles of the KIR ligand mismatch in 94 leukemia patients undergoing unmanipulated HLA-mismatched/haploidentical blood and marrow transplantation. Multivariate analysis showed that both KIR ligand mismatch (HR 2.833, CI, 1.286–6.241, p=0.01) and doses of T cells (HR 3.059, CI, 1.292–7.246, p=0.011) were independent risk factors causing the acute graft versus host disease (aGVHD). In addition, compared to patients without KIR ligand mismatch, KIR ligand mismatch worsened the adverse effect of ‘high’ dose T cells (>1.48×108/kg) on aGVHD (100% vs 63.3%, p=0.036), and increased the incidence of aGVHD in patients with HLA-C mismatch (80% vs 57.4, p=0.056). Since multivariate analysis demonstrated that high risk leukemia is the only predictor for TRM, relapse and OS, we further analyzed the effect of KIR ligand mismatch on prognosis in standard and high risk patients. The differences in TRM (50% vs 7.6%, p=0.005) and OS (50% vs 88.4%, p=0.014) between patients with and without KIR ligand mismatch were most striking for standard risk. Therefore we conclude that due to the presence of large dose T cells in the allograft, the alloreactivity of NK cells had been inhibited and KIR ligand mismatch directed alloreactive T cells played crucial roles in our model.
Polymorphism in the Promoter Region of the Hemeoxygenase I Gene of the Donor Influences Overall Survival and Graft Versus Host Disease.
Authors: Gerbitz Armin , Hillemanns Patrick , Schmid Christoph , Wilke Andrea , Jayaraman Rajshri , Kolb Hans-Jochem , Bremm Heike , Eissner Guenther , Holler Ernst 
Published: October 2019

Abstract
Genetic polymorphisms of various donor and host genes have been found to be associated risk factors for graft versus host disease and transplant related mortality. Most of these genes are involved in the regulation of immune response and inflammatory reactions towards pathogens. The heme oxygenase (HO) protein is the rate limiting enzyme in heme degradation to biliverdin, free iron and carbon monoxide. Its inducible form I (HO-I) is constitutively expressed in the spleen due to the abundance of its substrate. It is inducible in a wide range of tissues upon exposure to cellular stress factors such as irradiation, bacterial lipopolysaccharide, proinflammatory cytokines and heat shock. Further, HO-I is involved in regulating inflammatory response and has been described as a “protective gene” in solid organ transplantation. In man the promoter region of HO-I displays length polymorphism due to a variable number of gt repeats within a 500bp region upstream of the TATA box. Individuals exhibiting 25 or less gt repeats express HO-I upon cellular stress at a higher level than individuals with more than 30 gt repeats. We retrospectively analysed length polymorphisms of 92 donor and host pairs undergoing allogeneic stem cell transplantation for various malignancies. We show here that donor promoter length polymorphism leading to low expression of HO-1 (>30 gt repeats) is associated with improved overall survival, relative to donors with ≤ 30 repeats (80.6% versus 54.4%, p=0.023). When subgrouped, having HLA identical siblings as donors had no influence on outcome (see figure below, right panel) whereas having matched unrelated donors strongly influenced overall survival (82.6 vs. 44.2%, p=0.015). Combination of high HO-1 expressing unrelated donors and high expressing hosts displayed a 27.8% survival (left panel, solid line vs.81.3% for individuals with one allele >30 repeats dotted line). Polymorphisms of the recipient HO-1 genes did not influence outcome after transplantation. When matched unrelated donors were used, the incidence of grade III gut GvHD was significantly higher when (p=0.032) and overall GvHD (≥ grade III) displayed a trend towards significance (p=0.098). Transplant related mortality was strongly influenced by having a donor who expresses HO-1 at a high level upon cellular stress (p=0.006). In a multivariate Cox regression analysis of pretransplant factors such as age, disease stage, sibling/unrelated donor etc. polymorphism in the donor HO-1 gene had a significant influence on overall survival (p=0.027). These data suggest a role for HO-I in GvHD pathophysiology and a possible role for HO-1 as a non MHC risk factor for unrelated transplantation.
Figure Figure