bluuebunny/crossref_metadata_2025_split huggingface dataset:
Important Role for Absolute Lymphocyte Count in Predicting Risk of Relapse in Pediatric Acute Lymphoblastic Leukemia Post Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Ishaqi Mohammed Kashif , Afzal Samina , Dupuis Annie , Doyle John , Gassas Adam 
Published: October 2019

Abstract
Allogeneic hematopoietic stem cell transplantation (HSCT) is currently an established treatment for children with relapsed or high risk acute lymphoblastic leukemia (ALL). Relapse is a major risk post HSCT and an important cause of treatment failure with abysmal outcome. An important therapeutic mechanism of allogeneic HSCT is the immune-mediated destruction of recipient leukemic cells by donor lymphocytes, the graft-versus-leukemia-effect (GVL). Our hypothesis was to determine if delayed lymphocyte recovery measured by the absolute lymphocyte count (ALC) at two time points post HSCT at day 21 and day 30 correlates with leukemia relapse. We reviewed 136 consecutive paediatric patients with ALL who received allogeneic HSCT between 1994 and 2005 in the Hospital for Sick Children, Toronto, Canada. All patients were in complete morphological remission prior to HSCT and remission status at time of HSCT were as follows; Complete remission 1 (CR1, n=36); Complete remission 2 (CR2, n=79); Complete remission 3 (CR3, n=21). Conditioning regimens included single dose of VP16 (60mg/kg infused over 4 hours) and fractionated total body irradiation (TBI; 1200cGy) in six fractions over 3 days (VP16/TBI) in 49 patients (1994–1998) and cyclophosphamide 50mg/kg infused over 1 hour daily for 4 days followed by the same dose of fractionated TBI (CY/TBI) in 83 patients (1999–2005) and 4 patients received other conditioning regimens. Fifty-six patients received matched sibling donor (MSD), 12 patients received one antigen mismatched related donor (MMRD), 64 received unrelated donors and 4 patients received cord progenitor stem cells. Two groups were identified based on absolute lymphocyte count at day 21 and day 30 post HSCT. Patients with absolute lymphocyte count <0.3×109/L at day 21 (n=104) had more than 5 times risk of relapse compared to those with ALC count >0.3 × 109/L (n=32) (Hazard ratio 0.19; P=0.0004). Patients with ALC <0.3×109/L (n=48) at day 30 were more than twice likely to relapse compared to those with ALC >0.3×109/L (n=88) (Hazard ratio 0.46; P=0.01).
Conclusion: ALC, at day 21 and day 30 post HSCT identifies a patient population of pediatric ALL with delayed lymphocyte recovery and a significant risk of relapse post HSCT. Such patients could be considered for early intervention targeted to prevent relapse such as reduction of immunosuppressive therapy or other means of immunotherapy to enhance the graft versus leukemia effect.
Optimizing Immunotherapy in Multiple Myeloma: Restoring the Function of Patient’s Monocyte-Derived Dendritic Cells by Inhibiting p38 or Activating MEK/ERK MAPK and Neutralizing Interleukin-6 in the Progenitor Cells.
Authors: Yi Qing , Wang Siqing , Hong Sungyoul , Yang Jing , Qian Jianfei , Zhang Xiang , Shpall Elizabeth , Kwak Larry W. 
Published: October 2019

Abstract
Previous studies have demonstrated that circulating dendritic cells (DCs) in myeloma patients are functionally abnormal. However, the phenotypic and functional properties of monocyte-derived DCs (MoDCs) from myeloma patients, which are commonly used for immunotherapy in patients, are still poorly defined. This study was undertaken to examine the quality of MoDCs from myeloma patients compared with cells from healthy donors. We analyzed and compared cells from 12 newly diagnosed, stage II or III myeloma patients and 10 healthy donors. MoDCs were generated by culturing adherent cells or purified CD14+ monocytes in medium containing GM-CSF and IL-4. A cocktail of IL-1β, IL-6, TNF-α, and PGE2 was used to induce DC maturation. We found that patient-derived MoDCs are phenotypically and functionally defective. Compared with their normal counterpart, patient-derived, immature and mature MoDCs expressed significantly lower levels of CD1a, CD40, CD80, and HLA-DR. The mature cells were also poor at activating alloreactive T cells, presenting recall antigen and activating autologous antigen-specific T cells, and inducing myeloma-specific cytotoxic T-lymphocyte response. These abnormalities may be attributed to elevated production of autocrine cytokines such as IL-6, constitutively activated p38 and STAT3, and inhibited MEK/ERK signaling pathways in the progenitor cells. Treatment with neutralizing IL-6-specific antibody and more importantly, p38 inhibitor, or both, could correct these abnormalities. Treating patient-derived cells with these agents not only significantly increased cell yield, but also produced MoDCs that were as functional as their normal counterpart. We further showed that the p38 inhibitor inhibited the activity of p38 and drastically increased MEK1/2 and ERK1/2 phosphorylation and activation, and a combination of IL-6 neutralizing antibody with the p38 inhibitor further upregulated phosphorylation of MEK and ERK. By examining the role of STAT3 in MoDC differentiation, we found that STAT3 activation did not impair differentiation of patient-derived cells. These results indicate that the p38 inhibitor improved the generation and function of the MoDCs via inhibiting p38 and activating Raf/MEK/ERK signaling pathways, independent of the STAT3 pathway. Thus, this study has delineated the mechanistic defects of MoDCs from myeloma patients, and identified ways for restoring the function of the cells to improve the efficacy of DC-based immunotherapy against multiple myeloma.
Widening the Applicability of Human Neutrophil Elastase and Proteinase 3 Peptide Vaccines by Elucidating Immunogenic Non-HLA-A2 MHC Class I Restricted Epitopes.
Authors: Hewitt Rachel E. , Melenhorst J. Joseph , Price David A. , Gostick Emma , Hensel Nancy F. , McCoy J.P. , Barrett A. John 
Published: October 2019

Abstract
Human Neutrophil Elastase (HNE) and proteinase 3 (Pr3) belong to a group of myeloid tissue restricted serine proteases, aberrantly expressed by myeloid leukemia cells of interest as a source of myeloid-restricted antigens applicable to immunization strategies. HNE shares with Pr3 the antigenic PR1 peptide sequence inducing HLA-A*0201 restricted cytotoxic T cells (CTL) against leukemic cells. Our previous studies with full length HNE protein have demonstrated its potential for inducing both CD4+ and CD8+ T cell responses in healthy individuals including HLA-A2 negative individuals, suggesting an immunogenic potential of HNE beyond the PR1 peptide (Fujiwara H et al Blood 2004). To find epitopes other than PR1 in HNE and Pr3, peripheral blood mononuclear cells (PBMC) from 15 AML patients were screened at diagnosis or relapse with a peptide matrix consisting of 15mers overlapping by 11 amino acids spanning HNE and Pr3.The corresponding Pr3 and HNE 15mers were also used to generate and test short term T cell lines derived from patient samples. Responses to the peptide matrix pools were measured by a CFSE flow cytometry-based proliferation assay where the proliferating fraction in response to peptide pools was compared to the background co-stimulatory antibody only control for each assay. A positive response was defined as a proliferating fraction minimum 0.1% of the total CD8/CD3 population after subtraction of background proliferation and at least one and a half times background for each assay. These steps helped to control for variability between samples. Using this strategy we identified PR1 and 6 new candidate epitopes contained within HNE and Pr3. Corresponding immunogenic Pr3 sequences in 2 epitopes differed by only 3 amino acids from the immunogenic HNE sequence. CD8 short term T cell lines generated to these epitopes were restricted by HLA-A2, B35 and A68. Optimal peptide length was predicted using RANKPEP (http://www.mifoundation.org/Tools/rankpep.html) and confirmed with flow cytometric assays measuring responses to candidate epitopes, by interferon-gamma, tumor necrosis factor-alpha, IL-2 production and degranulation in response to HLA-matched or mismatched EBV-transformed B cell antigen-presenting cells pulsed with relevant and irrelevant peptides. This study confirms the presence of multiple immunogenic epitopes in HNE and Pr3 and represents the first step in developing a multi-peptide leukemia vaccine broadly applicable to individuals of diverse HLA type.
Rapid Regression of Bone Marrow Fibrosis after Dose-Reduced Allogeneic Stem Cell Transplantation in Patients with Myelofibrosis.
Authors: Kroeger Nicolaus , Thiele Juergen , Zander Axel , Kvasnicka Michael 
Published: October 2019

Abstract
Myelofibrosis with myeloid metaplasia [MMM] is a myeloproliferative disease of which bone marrow histology is characterized by substantial collagen fibrosis and osteosclerosis. Bone marrow histology changes in MMM are reactive and cytokine mediated. Studies suggested a pathogenic role of transforming growth factor β [TGF-β], platelet derived growth factor, and basic fibroblast growth factor. In the current study we investigated the dynamic and kinetic of fibrosis regression in 18 patients with myelofibrosis who underwent allogeneic stem cell transplantation after a dose-reduced conditioning. The conditioning regimen consisted of Busulfan 10 mg/kg, Fludarabin 180 mg/m2, and Antithymocyte globulin 30 – 60 mg/kg followed by peripheral stem cell transplantation (SCT). 18 patients with a median age of 45 [range 35 to 64] were included and bone marrow histology was investigated prior SCT, between day+30 and +40 after engraftment, on day+100, and 1 year after stem cell transplantation. The grading of myelofibrosis [MF] was performed according to the recently published European Consensus on Grading Bone Marrow Fibrosis [Thiele et al., Haematologica2005;90:1125–1132]. This consensus distinguishes between:
MF - 0 Scattered linear reticulin with no intersections [cross-overs] corresponding to normal bone marrow MF - 1 Loose network of reticulin with many intersections, especially in perivascular areas MF - 2 Diffuse and dense increase in reticulin with extensive intersections, occasionally with only focal bundles of collagen and/or focal osteosclerosis MF - 3 Diffuse and dense increase in reticulin with extensive intersections with coarse bundles of collagen, often associated with significant osteosclerosis
At time of transplantation all patients suffered from advanced myelofibrosis 2° [39 %] or 3° [61 %], resulting in a median MF grade of 3 [range 2–3]. After engraftment on day+30 a regression of MF grade to a median of 2 [range 0–3] was observed. Grade 2 or less was observed in 80 % and grade 1 or 0 in 40 % of the patients. On day+100 the median grade of MF was 1 [range 0–3] and 93 % of the patients had an MF-grade of 2 or less, while 73 % of the patients had MF grade-1 or 0. One year after transplantation the median MF-grade was 0 [range 0–2] and 100 % of the patients had a MF-grade of 2 or less and 92 % of the patients had MF grade 1 or 0. We conclude that allogeneic stem cell transplantation after a dose-reduced conditioning induces a rapid and nearly complete regression of bone marrow fibrosis within the first year after transplantation.
Efficacy and Safety of Once Weekly Epoetin Beta 30 000 IU in 691 Patients with Solid or Lymphoid Malignancies Receiving Chemotherapy: Results from the NAUTICA Study.
Authors: Spaeth Dominique , Salles Gilles , Facon Thierry , Garidi Reda , Pichon Francois , Damaj Ghandi , Bergougnoux Loic 
Published: October 2019

Abstract
Anemia is the most frequent hematologic complication of cancer, affecting 68% of patients at some time (Ludwig et al, Eur J Cancer 2004). Anemia causes debilitating fatigue and has a major impact on quality of life. Epoetin beta* (NeoRecormon®) is an effective treatment for anemia that has traditionally been administered three times weekly; however, a once weekly (QW) schedule would be more convenient and may encourage patient compliance. This open-label, single-arm study evaluated the efficacy and safety of epoetin beta 30 000 IU QW in patients with solid or lymphoid malignancies. Adult patients with anemia (hemoglobin [Hb] levels 8–12 g/dL), a WHO performance status 0–2 and who were scheduled to receive chemotherapy were enrolled from 87 centers in France between 31 Dec 2003 and 30 Aug 2005. Patients received SC epoetin beta 30 000 IU QW for up to 16 weeks. Primary efficacy parameter was change in Hb level. Hb response was defined according to patients’ baseline Hb level: in patients with Hb levels of 11–12 g/dL, response was defined as achievement of Hb level of ≥ 13 g/dL; in patients with Hb levels of <11 g/dL, response was defined as Hb increase of ≥ 2 g/dL or Hb level of ≥ 12 g/dL. Of the 691 patients included in the intent-to-treat population, 540 had baseline Hb levels <11 g/dL and 151 had baseline Hb levels of 11–12 g/dL. Fifty-three percent had solid tumors and 47% had lymphoid malignancies. The median duration of treatment was 14 weeks. Mean (±SD) baseline Hb level was 10.1±1.1 g/dL. At endpoint, mean Hb level had increased to 12.0±2.2 g/dL. Hb levels increased rapidly in the subgroup of patients with baseline Hb <11 g/dL, which corresponds to the intervention level recommended by the EORTC (figure). Hb response was seen equally in patients with lymphoid malignancies (60%) or solid tumors (61%). Median time to response was 49 (range 10–130) days. Hb response was seen with all types of chemotherapy. Epoetin beta was well tolerated. Seven percent of patients presented with thromboembolic events, a rate consistent with information provided in the current label for epoetin beta. Epoetin beta 30 000 IU QW is effective in anemic patients with solid tumors or lymphoid malignancies. This regimen provides a convenient and well tolerated treatment which may encourage patient compliance.
Figure. Increase in Hb levels during epoetin beta therapy: patients with baseline Hb levels <11g/dl (n=540) Figure. Increase in Hb levels during epoetin beta therapy: patients with baseline Hb levels <11g/dl (n=540)
Alemtuzumab-Cyclosporin A for Bone Marrow Failure Syndrome.
Authors: Kim Hawk , Shin Soojin , Park Jeo-Hoo , Min Young Joo 
Published: October 2019

Abstract
Patients with bone marrow failure syndrome are suffered from cytopenia and resulting requirement of transfusion. Allogeneic hematopoietic stem cell transplantation (alloHSCT) is the standard therapy for severe aplastic anemia (AA) with suitable sibling donor. However, it is true in many cases that alloHSCT can not be performed even in very severe AA due to no suitable donor. In that regards, immunosuppressive therapy (IST) is another option for those patients. Alemtuzumab (ALM) targets not only B-cell but also T-cell, which is known to play an important role in pathogenesis of bone marrow failure syndrome (BMFS). Although there were several reports of ALM for immune cytopenia and pure red cell aplasia, no report were published regarding ALM for BMFS. We studied the feasibility of ALM and cyclosporine A (CsA) for patients with BMFS. Inclusion criteria were transfusion-dependent BMFS including AA, MDS, PNH. Prior IST or other drugs for BMFS except for ALM were not excluded. ALM was infused for 3 days (10mg on day 1, 20mg on day 2, and 30mg on day 3) along with CsA for at least 6 months. Prophylactic ciprofloxacin, acyclovir, and bactrim were given for first 2 months. Without partial response (PR) or more within 6 months, subsequent alloHSCT or antithymoglobulin therapy was allowed. Total 12 patients were enrolled. Female were 9 (75%). Median age was 43 (16–74) years. Disease were 9 severe AA, 2 moderate AA and 1 hypoplastic MDS. All patients were transfusion-dependent. Median amount of transfused red cell, platelet concentrate and platelet apheresis prior to ALM-CsA were 2, 2 and 7 units, respectively. Prior therapy were none in 9 (75%), anti-thymoglobulin in 2 (16.7%), and danazol in 1 (8.3%). Only 1 patient could not receive full scheduled CsA because the patient underwent alloHSCT. Median follow-up was 326 (140–466) days. Response to ALM-CsA were no response in 7 (58.3%), PR in 4 (33.3%), and CR in 1 (8.3%). Therefore, overall response rate was 5/12 (41.7%). All responsive patients showed their responses within 3 months. Median time to response was 47 (24–71) days [Figure 1]. PR was converted to CR in 1 patient at 12 months after treatment. Neither fever nor skin rash during treatment did not affect the possibility of response to ALM-CsA (p=0.523, p=0.523, respectively). Toxicity during ALM was fever in 3 (25%), skin rash in 3 (25%), G1 ALT elevation in 7, G1 AST elevation in 4, and hyperbilirubinemia in 3 (G1 in 2, G3 in 1). There were no anaphylaxis, serum sickness or CMV reactivation.
In spite of small number, AML-CsA showed comparable response rate within 3 months after treatment. Therefore in conclusion, ALM-CsA is feasible and tolerable therapy for BMFS.
Figure Figure
Cyclosporine A Effect the CD28 Expression on CD8+T Cells of Patients with Aplastic Anemia through down Regulate FLIP.
Authors: Huang Haiwen , Wu De Pei , Chen Guanghua , Fu Jinxiang 
Published: October 2019

Abstract
Objective The exact reason of aplastic anemia is still unknown. Recently more and more researchers realized that immune factors play an important role in aplastic anemia. In this study, we detected the expression of CD28 on different subgroups of T cells in patients with aplastic anemia and furthermore studied the effect of cyclosporine A to CD28 and FLIP on these cells. Method 21 patients and 15 healthy controls were investigated. The Expression of CD4CD28 and CD8CD28 on patients’ T cells was detected with FCM; patients’ CD8+T cells were purified with Mini MACR system, then the expression of FLIP, caspase-8 in CD8+T cells were analyzed by RT-PCR; we also detected the expression of CD28, FLIP and caspase-8 in patients’ CD8+T cells after incubation with cyclosporine A.
Result: The percentage of CD8+CD28−T cells in the patients is significantly higher than that of normal controls (30.45±5.26% versus 19.32±4.72%, p<0.05) and it can be induced to return to normal after effective treatment with Cyclosporine A; the expression of FLIP in patients’ purified CD8+ T cells is higher than that in healthy controls’ CD8+ T cells, but there is no difference in the expression of caspase-8; after being cultured with Cyclosporine A, the expression of FLIP in patient’s CD8+T cells was down regulated and the quantity of CD28 on the surface of patient’s CD8+T cells increased significantly (28.3±5.1% versus 40.6±7.71%40.6±7.71%).
Conclusion: The increased expression of FLIP in patients’ CD8+T cells is related to the increased percentage of CD8+CD28−T cells in patients’ peripheral blood. Cyclosporine A can down regulate the FLIP expression and correct the abnormal percentage of CD8+CD28−T cells in patients with aplastic anemia.
Role of JAK 2 Mutation Detection in Budd-Chiari Syndrome (BCS) and Portal Vein Thrombosis (PVT) Associated to MPD.
Authors: Kiladjian Jean-Jacques , Cervantes Francisco , Leebeek Frank W.G. , Chevret Sylvie , Cazals-Hatem Dominique , Plessier Aurélie , Marzac Christophe , Cassinat Bruno , Raffa Sebastian , Garcia-Pagan Juan Carlos , Murad Sarwa Darwish , Janssen Harry L.A. , Valla Dominique , Casadevall Nicole , Fenaux Pierre 
Published: October 2019

Abstract
Background: MPD is found in up to 50% of BCS and 30% of PVT, but its diagnosis is often difficult, bone marrow biopsy (BMB) and endogenous erythroid colony (EEC) formation being the most reliable tests in our experience (Br J Haematol, 2005;129:553) and that of others. We assessed the diagnostic and prognostic value of V617F JAK2 detection in this situation.
Methods: Underlying prothrombotic factors were studied in 241 pts (104 BCS, 137 PVT) from 3 centers (Paris, Barcelona, Rotterdam). Based on BMB results (blindly reviewed), 31% of patients had MPD, 67% no-MPD, and 2% were uncertain. Based on combined BMB+EEC results, 29% had MPD, 41% no-MPD and 31% had discordant features. V617F JAK2 mutation was studied in all patients by quantitative RT-PCR.
Results: V617FJAK2 was detected in 45% of BCS and 34% of PVT (p=.08). Mean % circulating V617F allele was 37% and 28% in BCS and PVT pts, respectively (p<.04). In pts diagnosed with MPD using conventional criteria, V617F mutation was found in 84% pts with positive BMB, and in 97% pts with concordant BMB+EEC results (table). In pts previously diagnosed as no-MPD, V617F was present in 19% pts with negative BMB and in 7% pts with both negative BMB and EEC. This discrepancy was greater in BCS patients, where V617F was found in 25% pts previously not diagnosed with MPD. V617F was also found in 58% of pts with discordant BMB and EEC results, allowing to increase the rate of definite MPD diagnosis. On the other hand, 16% pts with wild type JAK2 had MPD features on BMB and 20% had positive EEC. Taking into account V617F JAK2 in addition to BMB and EEC, definite MPD incidence increased from 30 to 49% overall; from 32% to 60% in BCS patients; and from 29% to 49% in PVT patients. V617F JAK2 was significantly associated with higher Hb, WBC, and platelet counts, larger spleen size, lower serum EPO, and higher red cell mass. Other prothrombotic states were found in 19/92 V617F patients and 29/144 wtJAK2 patients (NS). Baseline BCS prognostic scores (Child Pugh, Clichy and Rotterdam scores) did not significantly differ between V617F and wtJAK2 patients. BCS and PVT survival was not affected by the presence of V617F JAK2.
Conclusion: Detection of V617FJAK2 (1) is almost as common in PVT as in BCS but the % of mutant allele is higher in BCS; (2) is not fully redundant with EEC or BMB findings and increases the diagnosis of underlying MPD by 28% in BCS and by 20% in PVT; (3) has no discernible impact on medium term outcome of BCS or PVT. (Supported by EU: QLG1-CT-2002-01686).
% of V617F JAK2 positive All patients BCS PVT BMB / EEC + / + 97 100 94 − / − 7 25 0 Discordant 58 53 62 BMB + 84 75 92 − 19 27 12 EEC + 77 84 72 − 25 38 18 inconclusive 33 36 30