bluuebunny/crossref_metadata_2025_split huggingface dataset:
Preemptive Donor Lymphocyte Infusion Induces Polyspecific T-Cell Responses in a Patient with AML with NPM1 Mutation
Authors: Hofmann Susanne , Götz Marlies , Herbst Cornelia , Schneider Vanessa , Zhang Lu , Bunjes Donald , Döhner Hartmut , Wiesneth Markus , Greiner Jochen 
Published: October 2019

Abstract
Abstract 4311
Delayed donor lymphocyte infusion (DLI) already plays a key role in supporting the graft-versus-tumor effect after allogeneic hematopoietic stem cell transplantation (HSCT) although the potency of DLI differs for each disease entitiy. The graft-versus-leukemia (GvL)-effect observed after DLI is based on the CTL-mediated immunity which is reactive against minor histocompatibility antigens (mHAg). To date, the role of leukemia-associated antigens (LAAs) in GvL is not clear.
We therefore analysed peripheral blood of a 57-year old woman with AML with NPM1 mutation (NPM1mut) who developed molecular relapse without detection of myeloid blasts in the peripheral blood and bone marrow at d+171 after allo-HSCT and received preemptive DLI at d+ 260. In parallel to the molecular relapse, chimerism continuously decreased to 60 % in the polymorphonuclear and 75 % in the mononuclear cell fraction. Blood samples were taken before and after DLI and investigated for specific T-cell responses against several LAAs for which cytotoxic T-cell responses were already described. In addition, the immune status of the patient was determined by measuring the counts of CD8+ and CD4+ T-cells as well as B-cells in the peripheral blood at several time points. To test specific CD8+ T-cell responses, ELISpot analysis for interferon gamma and granzyme B were perfomed. Two epitopes derived from the NPM1mut peptide (NPM1-P3 and –P9) and PRAME (P300 and P435) were tested as well as one epitope obtained from Survivin, RHAMM, Proteinase 3 and WT-1.
CD8+ T-cell responses against NPM1-P3, -P9, P300, Proteinase 3, Survivin and WT-1 were detected in blood samples after preemptive DLI but not in samples before DLI. In parallel, former detectable NPM1mut transcripts were no longer traceable – the patient achieved molecular complete remission - and former mixed chimerism became complete after DLI.
Here, we could demonstrate for the first time polyspecific cytotoxic CD8+ T-cell responses against several known LAAs in a patient with AML with NPM1mut after preemptive DLI in molecular relapse. Interestingly, the immune responses against LAAs were associated with MRD negativity. Whether specific cytotoxic T-cell responses against epitopes derived from the NPM1mut peptide or the polyspecificity of the cytotoxic T-cells against several LAAs are decisive in the elimination of the myeloid blasts with NPM1mut remains to be determined in a larger cohort of patients. All in all it is of interest to perform preemptive DLI in patients with molecular relapse systematically. In addition, boostering of T-cells against specific LAAs could possibly be an approach to enhance GvL-effect with reducing GvHD-effect at the same time, but the most suitable LAA or combination of LAAs for each disease entitiy still has to be investigated.

Disclosures:
No relevant conflicts of interest to declare.
Molecular Mimicry in the Adulteration of Heparins. Chemical Basis for the Development of Oversulfated Chondroitin Sulfate and Related Contaminants
Authors: Fareed Jawed , Wahi Rakesh , Thethi Indermohan , Hoppensteadt Debra , Bansal Vinod , Walenga Jeanine M. , Messmore Harry L. 
Published: October 2019

Abstract
Abstract 4322
The main contaminant in heparin and low molecular weight heparins reported during the heparin crisis (2007–8) was characterized to be oversulfated chondroitin sulfate (OSCS). However, several other sulfated glycosaminoglycans have also been reported to be present. Dermatan sulfate, heparan sulfate and chondroitin sulfate represent heparin byproducts and are invariably present in different proportions in heparin preparations. Hypersulfated derivatives of these are also found as contaminants. The chondroitin sulfate obtained from mammalian tissues and cartilage can be hypersulfated using simple chemical sulfonation techniques such as the chlorosulfonic acid to produce varying degrees of sulfation. The molecular profile of OSCS can be optimized utilizing various depolymerization methods to mimic the required molecular profile of heparin. Moreover, mixing of the molecular mass optimized OSCS to heparin exhibit the anticoagulant potencies in the pharmacopeial assays that are indistinguishable by the methods used prior to 2007. Initial studies carried out to obtain OSCS from bovine, porcine, squid and shark cartilage resulted in the generation of an almost perfect polymer mixture whose molecular profile was similar to heparin (14–16 Kda). The mixing of some of these OSCS preparation to heparin also showed either additive or synergistic effects in the anticoagulant assays. However, the OSCS preparations were resistant to heparinase digestion and some of the other chemical digestion processes. Several other heparinoids such as the galactans, sulfaminoheparosans and sulfated dextrans can also be used to contaminate heparins. Based on these observations, the development of OSCS contaminant was a result of a careful optimization of the molecular profile of non heparin GAG derived oversulfated derivatives and the knowledge of their biologic effects. Despite the implementation of sophisticated analytical techniques such as NMR and mass spectrometry along with the modified biochemical assays for heparin characterization, the heparin molecule renders it to additional approaches for adulteration utilizing chemically and biologically rational approaches.

Disclosures:
No relevant conflicts of interest to declare.
Development of a Large Scale Production Process for BAX 855, a PEGylated rFVIII Product with Prolonged Half-Life
Authors: Siekmann Jürgen , Martin Purtscher , Zöchling Oliver , Kellerer Robert , Rottensteiner Hanspeter , Mitterer Artur , Bossard Marry , Phillips Gayle , Ehrlich Hartmut J. , Scheiflinger Friedrich , Turecek Peter L 
Published: October 2019

Abstract
Abstract 4362
Baxter and Nektar have developed BAX 855, a PEGylated form of Baxter’s recombinant FVIII (rFVIII) product based on the ADVATE™ manufacturing process. The conjugation process for preparing BAX 855 uses proprietary stable PEGylation from Nektar Therapeutics. Similar Nektar technology has been successfully employed for marked and licensed PEGylated drug products and drugs in clinical use. The manufacturing process for BAX 855 comprises several steps, including chromatographic purification on a MacroCap SP resin, a strong cation exchanger, which allows the fractionation of species with different PEGylation degrees and concentration of the conjugate collected by an ultra- / diafiltration step leading to the pre-formulated bulk drug substance (BDS). Final formulation of the BDS includes a filling and lyophilization step to obtain the final drug product. The process described is suited to manufacture BAX 855 in gram scale and showed a good batch to batch consistency, ensuring an equivalent product quality for each batch. BAX 855 manufactured by this process has a specific activity similar to that of rFVIII in ADVATE™ and PEGylation degrees in the narrow range of 2 to 3 mols PEG / mol rFVIII. SDS-PAGE and Western blot analysis of BAX 855 confirm the stable PEGylation and demonstrate an increase in the molecular weight of the various FVIII domains. PK studies in different species display longer survival of BAX 855 compared to ADVATE™.

Disclosures:
Siekmann: Baxter Innovations GmbH: Employment. Martin:Baxter Innovations GmbH: Employment. Zöchling:Baxter Innovations GmbH: Employment. Kellerer:Baxter Innovations GmbH: Employment. Rottensteiner:Baxter Innovations GmbH: Employment. Mitterer:Baxter Innovations GmbH: Employment. Bossard:Nektar Therapeutics: Employment. Phillips:Nektar Therapeutics: Employment. Ehrlich:Baxter Innovations GmbH: Employment. Scheiflinger:Baxter Innovations GmbH: Employment. Turecek:Baxter Innovations GmbH: Employment.
Serial D-dimer in Pregnant Women Compared to Antiphospholipid-antibodies Level, a History of Fetal Loss Syndrome and Invitro Fertilization
Authors: Atmakusuma Tubagus Djumhana 
Published: October 2019

Abstract 4364

Background
We have reported in abstract P-Th-374 in a 2011 Kyoto ISTH Congress that an increased level of D-dimer in combination with umbilical artery systolic diastolic (UASD) ratio can be used as a timing to start with prophylactic anticoagulants In women with pregnancy failure, regardles of the antibody antiphospholipids (APAs) level.
The question is: Are there any differences of a serial D-dimer level if is compared to the APAs level?.

Methods.
A retrospective Cohort study was conducted on a total sampling subjects of pregnancy women who visited two hospitals in Jakarta, Indonesia, at a periode of January 2009 – December 2010. Serial D-dimer using chromogenic assay method (cut off normal level < 500 ng/ml) was tested during antenatal and post partum period. The data were analysed to compare a mean D-dimer with the level of APAs (ACA IgG/IgM, AntiB2GP1 IgG/IgM) tested using ELISA method

Results
Of 83 pregnant women,19 women with high level of APAs (group 1) showed a mean D-dimer level as follows: Trimester I: 892.15 ± 785.15 ng/ml, trimester II: 782.59 ± 440.53 ng/ml, trimester III:1282.62 ± 601.22 ng/ml, post partum 1367.45 ± 581.19 ng/ml; 31 women with normal APAs (group 2) showed: Trimester I: 666.23 ± 396.24 ng/ml, trimester II: 896.66 ± 396.24 ng/ml, trimester II: 896.66 ± 496.32 ng/ml, trimester III: 1313.45 ± 850.20 ng/ml, post partum: 1991.75 ± 1388.70 ng/ml; 33 women with no data of APAs (group 3) showed: Trimester I: 568.47 ± 482.70 ng/ml, trimester 2: 797.95 ± 934.59 ng/ml, trimester III: 966.89 ± 862.10 ng/ml, post partum: 1078.50 ± 836.29 ng/ml

Conclusion
Either in group 1, 2, and 3 mean D-dimer tend to increase from trimester I to trimester II and from trimester II to trimester III, except in group 1 from trimester I to trimester II of pregnancy. Meanwhile, all groups showed a higher mean D-dimer post partum compared to antenatal period.

Disclosures:
No relevant conflicts of interest to declare.
Study on the Pathways to Damage Hematopoiesis by CD8+ Effector T Cells of the Patients with Severe Aplastic Anemia
Authors: Shao Zonghong , Feng Le , Fu Rong , Wang Jun , Liu Chunyan , Li Lijuan , Liu Hui , Zhang Tian , Wang Honglei , Wang Huaquan 
Published: October 2019

Abstract
Abstract 4371

Objective
To investigate the quantity and their pathways to damage hematopoietic cells of CD8+CD25+ and CD8+HLA-DR+ effector T cells in peripheral blood (PB) of the patients with severe aplastic anemia(SAA) and explore the heterogeneous immunopathogenesis of SAA further.

Methods
The quantity of CD8+CD25+and CD8+HLA-DR+ cells in PB and the expressions of perforin, granzyme B, tumor necrosis factor-β(TNF -β) and FasL of 29 SAA (14 untreated and 15 recovered) patients and 12 normal controls were analyzed by flow cytometry.

Results
The ratio of CD8+CD25+T cells in CD8+ T cells was (3.67±2.58)% in untreated SAA patients, (5.19±4.29)% in recovered patients and (4.84±2.31)% in normal controls, and the ratios of CD8+CD25+T cells in CD3+ cells in three groups were (2.25±1.35)%, (2.98±1.35)% and (2.11±1.88)% respectively. There was no statistic difference among 3 groups(P>0.05). The ratio of CD8+HLA-DR+T cells in CD8+T cells was (39.30±8.13)% in untreated patients, which was significantly higher than that of recovered patients[(20.65±5.38%)] and controls [(18.34±6.68%)](P<0.001). There was no statistic difference between recovered patients and controls(P>0.05). CD8+HLA-DR+T cells in CD3+ cells was (27.81±7.10)% in untreated group, higher than that of recovered patients (12.02±3.03)% and controls(8.50±2.33)%(P<0.01). And the ratio in recovered group was higher than in control group(P<0.05). The expressions of perforin, granzyme B, TNF-β and FasL of CD8+HLA-DR+ T cells of untreated SAA patients were 8.51% A96.08% A72.11% and 94.25% respectively, higher than those of recovered patients(1.78% A85.20% A34.38% A51.20%)and controls(1.86% A82.09% A17.92% A32.91%). There was no statistic difference between recovered patients and controls(P>0.05).

Conclusion
There were elevated quantity of CD8+HLA-DR+ T cells and high expressions of perforin, granzyme B, TNF-β and FasL in SAA, which might contribute to the bone marrow failure of SAA.

Disclosures:
No relevant conflicts of interest to declare.
Expression Feature of CD3 and FcεRIγ Genes in Patients with Aplastics Anemia
Authors: Li Bo , Niu Yu Zhe , Liu Si Chu , Chen Shao Hua , Jian Yang Li , Li Yang Qiu 
Published: October 2019

Abstract
Abstract 4372
Aplastic anemia (AA) is characterized by pancytopenia and bone marrow hypoplasia, resulting from immune-mediated suppression of hematopoiesis. Understanding of the immune pathophysiology, especially T-cell immune of the disease over the past 3 decades have led to improvement the treatment of AA. However, primary and secondary failures after immunosuppressive therapy remain frequent. Thereby, more knowledge of the immune mechanisms leading to the aplastic anemia is crucial to more understanding the mechanism of the disease. In order to elucidate the feature of TCR signal transduction in AA, the expression levels of CD3γ, δ, ε and ζ chain and FcεRIγ genes which involed in TCR signal transduction and negative correlation of the expression levels between CD3ζ and FcεRIγ genes in actived T-cell in peripheral blood mononuclear cells (PBMCs) were analyzed. Real-time RT-PCR with SYBR Green technique was used for detecting the gene expression level in PBMCs from 27 patients with AA and 9 healthy individuals. The β2-microglobulin gene (β2M) was used as an endogenous reference. The expression levels of CD3γ, CD3ε and CD3ζ gene in AA patients (19.96±21.46, 17.50±17.53 and 5.50±3.53, respectively) were significant increased compared with healthy group (4.00±1.70, 4.48±2.37 and 1.95±1.77, respectively) (P< 0.01), whereas the expression level of FcεRIγ gene was significant decreased in AA patients (17.58±10.23) in comparison with healthy group (40.20±25.46) (P< 0.01), while no significant difference of the CD3δ expression level was detected between AA and healthy groups. Moreover, there was lost the negative correlation of the expression levels between CD3ζ and FcεRIγ genes (r=0.19,p=0.33). In conclusions, this is, to our knowledge, the first description of the feature of CD3γ, CD3δ, CD3ε, CD3ζ and FcεRIγ gene expression in AA patients. The abnormality expression of CD3γ, CD3ε, CD3ζand FcεRIγ gene might related to the T-cell dysfunction in AA. Further research is needed to investigate the different expression pattern of these genes in different subset of T cells.

Disclosures:
Li: China Postdoctoral Science Foundation funded project (No. 20070410840): Research Funding; Natural Science Foundation of Guangdong province (No. 07301046): Research Funding; The Fundamental Research Funds for the Central Universities (No. 21610408): Research Funding; Medical Science Technology Program of Guangzhou (2009-ZDI-18): Research Funding; National Natural Science Foundation of China (No. 30972455): Research Funding. Li:Medical Science Technology Program of Guangzhou (2009-ZDI-18): Research Funding; The Fundamental Research Funds for the Central Universities (No. 21610408): Research Funding; Natural Science Foundation of Guangdong province (No. 07301046): Research Funding; China Postdoctoral Science Foundation funded project (No. 20070410840): Research Funding; National Natural Science Foundation of China (No. 30972455): Research Funding.
Efficacy and Safety of Biosimilar G-CSF and Originator G-CSF for Haematopoietic Stem Cell Mobilisation: A Randomised Comparison
Authors: Dmoszynska Anna , Manko Joanna , Walter Croneck Adam , Grzasko Norbert , Klimek Piotr , Nowaczynska Aleksandra , Gorska-Kosicka Magdalena 
Published: October 2019

Abstract
Abstract 4392

OBJECTIVES:
Recombinant granulocyte colony-stimulating factor (G-CSF) is widely used to mobilise haematopoietic stem cells. Biosimilar filgrastim is now available in Europe. No differences were observed between biosimilar filgrastim (n=40) and a retrospective cohort of patients receiving originator filgrastim for stem cell mobilisation in a previous comparison, although no safety findings were reported (Lefrère et al. Adv Ther 2011;28:304−10). We compared the efficacy and safety of a biosimilar filgrastim (EP-2006, Sandoz Biopharmaceuticals) with originator filgrastim (Neupogen®, Amgen) in patients with haematological malignancies.

METHODS:
A total of 108 patients were included in this study, 59 of whom were female (49 male), with an overall median age of 51 years (range 19–69). Patients had multiple myeloma (n=46), Hodgkin’s lymphoma (n=26), non-Hodgkin’s lymphoma (n=28) or other diagnosis (n=8). Median time from diagnosis to mobilisation was 10 months (range 3−122). After administration of mobilising regimens (primarily high-dose etoposide, high-dose cyclophosphamide, intermediate-dose Ara-C or ESHAP), patients were randomised to a standard daily 10 μg/kg dose of EP-2006 (n=54) or originator filgrastim (n=54).

RESULTS:
Median duration of G-CSF administration was 8 days with both EP-2006 (range 4−17) and originator filgrastim (range 4−14). Both groups had a median of one apheresis with a median time until first apheresis of 11 days. There were no statistically significant differences between groups in the median (range) number of mobilised CD34+ cells/μL in peripheral blood (EP-2006, 62.0 [2−394]; originator filgrastim, 47.5 [2−370]) or the number of CD34+ cells/ kg body weight (EP-2006, 9.1 [0−23]; originator filgrastim, 9.4 [6−48]). Five patients (9%) in each group did not mobilise sufficient CD34+ cells. The adverse event profile was comparable between the EP-2006 and originator filgrastim groups, with similar occurrence of neutropenic fever (9 vs 11 patients) and bone pain (8 vs 6 patients).

CONCLUSION:
EP-2006 demonstrated similar efficacy and safety as the reference filgrastim in haematopoietic stem cell mobilisation in patients with haematological malignancies.

Disclosures:
Dmoszynska: Mundipharma: Advisory Board; Roche: Honoraria.
Argentine Registry of Hematologic Disease (RAEH) Chronics Myeloid Leukemia
Authors: Enrico Alicia Inés , Moiraghi Beatriz , Klein Graciela , Cabrejo Maria del Rosario , Crisp Renee , Bendek Georgina , Gil Mariana , Garcia Juan , Alzueta Abel , Cicchetti Alejandro , Rojas Francisca , Perez Mariel , Burgos Ruben , Bengió Raquel 
Published: October 2019

Abstract
Abstract 4455

Introduction:
The CML Registry was developed by the Argentine Society of Hematology as a part of the centralized RAEH. Results from epidemiologic and clinical data related to CML, will enable to determine the geographic distribution of the target population, to establish associated environmental causes, and mainly to rationalize resources supply.

Objective:
a) To analyze characteristics of CML patients registered in the RAEH; b) To evaluate the CML Registry performance through its first year.

Materials and Methods:
Patients with CML registered in the RAEH from January 1st, 2011 up to July 31st, 2011. The protocol allowed to enroll de novo patients as well as patients diagnosed from 2000 on.

Result:
Data reported by 15 hospitals were included: 224 patients were registered. Mean age was 50 (18–86 y) and gender distribution was female: 102, male: 122. Occupational data showed no a characteristic pattern. 96.5% of patients were diagnosed in chronic phase, while 3.5% were diagnosed in accelerated phase/blast crisis. In 6% of patients cytogenetic tests detected 8 abnormalities besides t(9,22): double Philadelphia chromosome and monosomy 12 were the most frequent findings. FISH tests were recorded for 18% of patients at the time of diagnosis. Bone marrow biopsy was reported as a diagnostic procedure in 51%. Qualitative BCR/ABL was recorded for 31% of patients at the time of diagnosis. Molecular RQ-PCR tests for follow-up of treatment response were reported for 51% of patients.
Of the registered population, 21% received interferon as previous therapy to Imatinib (IM); 89% received IM 400mg daily; 6% required dose increase. Second line treatment with dasatinib or nilotinib was recorded in 34% and 14% of patients, respectively.
At 60 months mean follow-up 8% of the registered population had developed blast crisis and 6% had died.

Conclusions.
The RAEH’s first year of performance in CML was assessed. This only reflects the experience with 15 sites. Data registered will allow us in the following years to learn about disease epidemiology and available resources to improve patient accessibility.

Disclosures:
No relevant conflicts of interest to declare.
Late Mortality After Hematopoietic Stem-Cell Transplantation for a Childhood Malignancy
Authors: Schechter Tal , Pole Jason , Darmawikarta Denise , Gassas Adam , Greenberg Mark L. , Ali Muhammad , Doyle John J , Nathan Paul 
Published: October 2019

Abstract
Abstract 4458
Hematopoietic stem cell transplantation (HSCT) is used increasingly as a curative therapy in children with malignancies. However, survivors of HSCT are at risk for both disease recurrence and life-threatening complications beyond the acute period of transplantation. The risk for late mortality (> 2 years) after autologous and allogeneic HSCT has not been well reported in pediatric patients. The aim of this population based retrospective cohort study was to assess late mortality in children (0–18 years of age) who underwent a HSCT in the province of Ontario, Canada. All but a small minority of the province’s pediatric transplantations are performed at a single center (The Hospital for Sick Children [SickKids], Toronto). Indication for HSCT, type of transplant, donor source and survival were determined via a record linkage between the SickKids’ clinical transplant database and POGONIS, the provincial pediatric cancer registry. Mortality information contained in POGONIS is captured at the time of death in hospital, or by record linkage to the provincial mortality file. Records of all 842 children with a malignancy who underwent HSCT at SickKids between 1985 and 2009 were retrieved. Underlying diagnoses were leukemia (N=464), lymphoma (N=88), central nervous system tumor (N=60), neuroblastoma (N=167), sarcoma (N=44) and other (N=19). Four-hundred and nineteen children underwent allogeneic HSCT and 423 underwent autologous HSCT. Among the allogeneic HSCT group, 252 received stem cells from a related donor and 167 from an unrelated donor. Overall, 537 children (67.8%) survived for at least 2 years after HSCT. The median follow-up of these 537 survivors was 10.2 years, with 134 (25%) of the patients being followed for ≥15 years. Of these 537 survivors, 103 (19.2%) subsequently suffered a late death. A greater proportion of patients who underwent autologous HSCT had a late death (67/279; 24.0%) than patients who underwent allogeneic HSCT (36/258; 14.0%). In a multivariate analysis of patients who underwent autologous HSCT, males had increased risk of death compared to females (HR=2.3, 95% CI: 1.29–4.0), and patients treated for neuroblastoma had a greater risk of death than those treated for leukemia (HR=10.9, 95% CI: 3.0–40.0). Among patients treated with allogeneic HSCT, none of gender, donor source (related vs. unrelated), age at transplant or underlying diagnosis was associated with increased late mortality. In conclusion, children with cancer who are treated with a HSCT continue to be at risk for premature death even if they survive for two or more years after their transplant. Further investigation will focus on examining the contributions of treatment toxicity and disease recurrence to late mortality.

Disclosures:
No relevant conflicts of interest to declare.