bluuebunny/crossref_metadata_2025 huggingface dataset:
CD11b Expression Is An Independent Adverse Prognostic Factor in Pediatric Acute Myeloid Leukemia Treated with Allogeneic Stem Cell Transplantation,
Authors: Albitar Maher , Alseraihy Amal , Ayas Mouhab , Al-Jefri Abdullah , Al-Ahmari Ali , Belgaumi Asim F , Walter Claudia Ulrike , Nounou Randa , Khalil Salem , Bakshi Nasir , Owaidah Tarek , El-Solh Hassan
Published: October 2019

Abstract 4092

Background:

The adhesion molecule CD11b, which associates with the beta2-integrin to form the Mac-1 complex, is expressed in monocytic leukemias as well as other myeloid leukemias. Its expression on the lekemic cells has been reported to correlate with more aggressive course in adult patients with AML. The clinical relevance of CD11b expression in pediatric AML is not known. More importantly, the clinical relevance of CD11b expression on pediatric AML when these patients are treated with allogeneic hematopoietic stem cell transplant (HSCT). We investigated whether HSCT can modify the expected adverse effects of CD11b expression in pediatric AML patients.

Methods:

Immunophenotype data of 62 pediatric patients with AML, all treated with allogeneic HSCT, was reviewed. The expression of the CD11b on the blast population as determined by flow cytometry was correlated with clinical data and outcome. The median age of patients was 8 years (range: 8 months–14 years) and included 47 (76%) patients transplanted in their first complete remission (CR1). Fourty six (74%) of all patients had intermediate-risk cytogenetics and the remaining (26%) had adverse-risk cytogenetic abnormalities.

Results:

Twenty five (40%) of all studied patients expressed CD11b on the surface of the blasts at diagnosis. Patients with CD11b expression had significantly shorter overall survival (P=0.038) with median survival of approximately 11 months while the median survival in the CD11b negative patients has not been reached. Similarly, CD11 positive patients had significantly shorter event free survival (EFS) (P=0.03). When we considered only patients treated with HSCT in CR1, the OS and EFS for the CD11b positive patients were both significantly shorter (P=0.04 and P=0.05, respectively). Multivariate analysis including CD11b expression and cytogenetic risk identified CD11b expression as an independent prognostic factor for survival and EFS, while cytogenetic-risk stratification became no longer a predictor.

Conclusion:

Our data suggests that the expression of CD11b in pediatric AML is a significant independent poor prognostic factor and HSCT does not change this trend. Most of the CD11b positive patients die in their first year after HSCT and the management of these patients should be modified to address the biological basis of this subset of AML that expresses CD11b adhesion molecule.

Disclosures:

No relevant conflicts of interest to declare.
Impact of High Dose Cyclophosphamide on the Outcome of Autologous Stem Cell Transplant in Patients with Newly Diagnosed Multiple Myeloma,
Authors: Bacon Wendi A. , Long Gwynn D. , Rizzieri David A. , Horwitz Mitchell E. , Chute John P. , Sullivan Keith M. , Yopp Amanda , Johns Angela , Chao Nelson J. , Gasparetto Cristina
Published: October 2019

Abstract 4127

In the age of novel targeted agents, autologous stem cell transplant (ASCT) remains the standard of care for younger patients with newly diagnosed multiple myeloma (MM), offering similar treatment responses and overall survivals as standard chemotherapeutic agents but with the added benefit of a prolonged treatment-free period. Nevertheless, a standard of care for stem cell mobilization for ASCT has yet to be determined. Even in the era of new mobilization agents such as Plerixafor, Cyclophosphamide (Cy) and G-CSF combination remains the preferred mobilizing approach for patients with MM. Several studies have shown that Cy improves the stem cell yield at the expense of increased toxicity, but whether the administration of this chemotherapeutic agent pre-transplant has any impact on the long-term event-free and/or overall survival of myeloma patients remains controversial.

In this study, we present a retrospective analysis of 186 patients with newly diagnosed MM who underwent ASCT with high-dose melphalan 200 mg/m2 (HDM) between December of 2000 and 2008 at our Institution. Eighty-three patients were mobilized with single agent G-CSF and 103 patients received high dose Cy (4 gm/m2) and G-CSF combination. Patient characteristics were similar between the treatment groups, including: age, gender, disease stage, and disease status prior to transplant. However, toxicity post-mobilization with Cy/G-CSF was significantly higher compared with G-SCF alone, including: febrile neutropenia (23%), hemorrhagic cystitis (8%), GI toxicity (57%), re-hospitalization due to complications and transplant delay (14%).

The overall post-transplant toxicity was similar in the 2 groups, though the treatment related mortality was slightly higher in the Cy/G-CSF arm (4% versus 2%). Post transplant responses were not significantly different in the 2 groups, with 60% of patients achieving a VGPR or better after ASCT in the G-CSF group and 49% in the Cy/G-CSF group (p = 0.33). The median event-free survivals (EFS) for the Cy/G-CSF and G-CSF cohorts were 21.6 and 22.6 months, respectively, (p = 0.62) yielding no significant difference (Figure 1). Similarly, with a median follow up for surviving patients of 34.3 and 32.7 months, the median overall survivals were 68.2 and 62.3 months (p = 0.23) for the Cy/G-CSF and G-CSF cohorts, respectively (Figure 2). This retrospective analysis confirms that the addition of high dose Cy as part of the mobilizing regimen offers no improvement on the transplant outcome for patients with newly diagnosed myeloma and should therefore only be used in cases of difficult stem cell mobilization.

Disclosures:

No relevant conflicts of interest to declare.
Hematopoietic Cell Transplantation for FA Patients with Acute Leukemia or Advanced MDS,
Authors: MacMillan Margaret L. , Defor Todd E. , Wagner John E.
Published: October 2019

Abstract 4146

Fanconi anemia (FA) patients with acute leukemia or advanced myelodysplastic syndrome (MDS with ≥ 5% blasts) have a poor prognosis and hematopoietic cell transplantation (HCT) experience is limited. We report on the outcomes of 21 patients with FA and acute myeloid leukemia (AML, n=16), acute lymphocytic leukemia (ALL, n=2) or advanced MDS (n=3) who underwent allogeneic HCT at the University of Minnesota from 1988–2009. Five patients had biallelic BRCA2 mutations. Only 2 of 7 patients who received chemotherapy before HCT achieved remission both of whom had biallelic BRCA-2 mutations and T-cell ALL. Median age was 15.5 (range 1.1–48.5) years. Median age for BRCA2 patients was substantially lower at 1.9 (range 1.4–6.6) years. Patients received cyclophosphamide, antithymocyte globulin and either single fraction total body irradiation (n=15) or busulfan (n=6), with (n=11) or without (n=10) fludarabine (FLU) followed by HLA-matched sibling (n=3) or alternative (n=18) donor stem cells consisting of bone marrow (BM, n=14), single (n=4) or double (n=3) umbilical cord blood (UCB). Graft-versus-host disease (GVHD) prophylaxis consisted of cyclosporine A and methylprednisolone. Probability of neutrophil engraftment was 75% without FLU and 100% with FLU based regimens. Probability of acute GVHD was 19%. For the entire cohort, probability of survival at 1 year was 43%. A higher rate of survival was observed in the 11 patients who received a FLU based regimens (50%; figure) and in the 5 patients with BRCA2 mutations (60%). Probability of relapse was 21% in these 11 patients.

These data suggest that HCT with a FLU based preparative regimen offers potential curative therapy for FA patients with acute leukemia or advanced MDS. Except perhaps for BRCA-2 patients with T cell ALL, there is no clear role for chemotherapy prior to HCT in FA patients with AML or MDS.

Disclosures:

No relevant conflicts of interest to declare.
Unrelated Donor Hematopoietic Stem Cell Transplantation for the Treatment of Advanced Multiple Myeloma: The John Theurer Cancer Center Experience,
Authors: Donato Michele L. , Vesole David H. , Siegel David S. , Adler-Brecher Barbara , Chichili Eiswarya , Pecora Andrew L , McKiernan Phyllis , Baker Melissa , Nyirenda Themba , Bilotti Elizabeth , Goy Andre , Rowley Scott D
Published: October 2019

Abstract 4162

Allogeneic hematopoietic stem cell transplantation has been associated with a graft-versus-myeloma effect and may be the only potentially curative option for patients with advanced multiple myeloma. Unfortunately, the majority of patients (pts) do not have an HLA-identical sibling. Unrelated donor transplantation is an alternative option for these patients. We conducted a retrospective analysis of patients with advanced multiple myeloma who received an unrelated donor (URD) hematopoietic stem cell transplant at our institution between 11/1997 and 11/2010. Analyses were done using Kaplan Meier survival and Cox regression on the SAS version 9.2 software.

Between 11/1997 and 11/2010, 27 pts with multiple myeloma received an URD transplant. One patient was lost to follow-up and 26 pts are evaluable. Patient characteristics included: median age 52 years (range 37–64), male 58%, all patients had prior autologous transplant (23% of pts had 2 or more prior transplants), and 43% of pts had refractory or progressive disease at the time of transplantation. Transplant regimens were ablative (defined as melphalan dose > 140mg/m2) in 73% of pts. GVHD prophylaxis was tacrolimus-based in all. Hematopoietic cell source was peripheral blood for 92% of pts, 46% of pts received a 10/10 HLA matched URD and 54% had a least one allele mismatch.

Results:

With a median follow up of 3.4 years, 54% of patients are alive and 47% of patients are progression-free. The treatment-related mortality at day 100 was 15%. The incidence of acute graft-versus-host disease (GVHD) was 44% and chronic GVHD 25%. Univariate analysis of risk of progression or relapse and overall survival failed to show any significance for disease status at the time of transplantation, degree of donor match, transplant regimen, refractoriness to bortezomib or lenalidomide, acute or chronic GVHD.

Conclusion:

In this analysis, URD transplantation has an acceptable treatment related mortality, and leads to durable remissions even in patients with relapsed and/or refractory disease. The 3.4 year PFS is similar to the BMT CTN 0102 sibling data and compares favorably to related donor transplantation for relapsed disease. In comparison to the BMT CTN study, the preparative regimen in this current analysis was overall more intense and regimen intensity has been previously shown to be significant. Allogeneic transplantation is an effective treatment strategy in multiple myeloma. Unrelated donor transplantation should be offered to patients with advanced multiple myeloma who do not have an HLA-identical sibling.

Disclosures:

No relevant conflicts of interest to declare.
Gene Transfer to Hematopoietic Stem/Progenitor Cells As a Novel Approach for Immunotherapy Against B-Lineage Malignancies: In Vivo Xenograft Model,
Authors: De Oliveira Satiro N. , Giannoni Francesca , Hardee Cinnamon , Sahaghian Arineh , Cooper Laurence J N , Kohn Donald B.
Published: October 2019

Abstract 4168

Chimeric Antigen Receptors (CAR) against CD19 have been shown to direct T cells to specifically target B-lineage malignant cells in animal models and clinical trials, with efficient tumor cell lysis. But, there has been insufficient persistence of effector cells, limiting the clinical efficacy. We propose gene transfer to hematopoietic stem/progenitor cells (HSPC) as a novel approach to ensure persistent production of effector cells targeting B-lineage malignant cells, exponentially increasing the number of effectors that may be generated against tumor cells. Experiments were performed using NOD-SCID-IL2 receptor gamma chain null (NSG) mice engrafted with human CD34+ HSPCs transduced with lentiviral vectors carrying first and second generations of CD19-specific CAR. There was efficient and stable transduction with 1–2 copies of CAR/cell as determined by qPCR. Differentiation of modified HSPC in vivo was not impaired by gene transfer, as observed in vitro. Results of in vivo studies showed that CAR-transduced human HSPC successfully differentiated into all lineages, with CAR-expressing T, NK and myeloid cells populating bone marrow, spleen and peripheral blood. The human CD19+ B cell populations normally formed in the xenografted NSG mice were significantly reduced when the transplanted HSPC were transduced with the anti-CD19 CAR, demonstrating in vivo biological activity. Cells harvested from bone marrow and spleen of mice engrafted with modified HSPC lysed CD19-positive cell targets ex vivo. Leukemic challenges of engrafted mice are in progress. Our results provide evidence for the feasibility and efficacy of the modification of HSPC with CAR as a protocol for generation of effector cells for immunotherapy against B-lineage malignancies.

Disclosures:

No relevant conflicts of interest to declare.
ATP-Hydrolysing Treg in Patients with Acute Myeloid Leukemia
Authors: Schuler Patrick J , Fenton Moon , Hong Chang-Sook , Jackson Edwin K , Whiteside Theresa , Boyiadzis Michael
Published: October 2019

Abstract 4255

We have previously shown that the frequency and suppressor function of regulatory T cells (Treg) is increased in newly-diagnosed patients with acute myeloid leukemia (AML). Here, we show that CD4+CD39+ ATP-hydrolyzing T cells are involved in the production of immunosuppressive adenosine and that this mechanism of suppression characterizes Treg present in the blood and bone marrow of AML patients.

Peripheral blood and bone marrow samples were obtained from AML patients prior to any treatment (n=20) and healthy controls (NC, n=20). The frequency and phenotype as well as cytokine profiles of CD4+CD39+ T cells were determined using multicolor flow cytometry, real-time PCR and western blots. Adenosine production was measured by mass spectrometry. Co-cultures of CD4+CD39+ Treg with conventional T cells were tested for suppression of proliferation.

In NC and patients with AML, CD4+CD39+ Treg contained two subsets of ATP-hydrolyzing T cells in equal proportions (Fig.1): FOXP3+CD25+ and FOXP3negCD25neg.Both subsets were increased in frequency (p < 0.04) in AML relative to NC (Fig.2). In the presence of other immune cells or exosomes positive for CD73 (hydrolyzes AMP to adenosine) both subsets produced immunosuppressive adenosine. In co-cultures, CD4+CD39+FOXP3negCD25neg T cells converted to FOXP3+CD25+ T cells.

Our data suggest that Treg with the ability to produce adenosine are a major subset of suppressor cells in the blood and bone marrow of AML patients. Co-culture experiments indicate that the ability of these cells to suppress immune responses is regulated at the level of FOXP3 and CD25 expression by factors present in their microenvironment.

Disclosures:

No relevant conflicts of interest to declare.
The Natural Product Borrelidin Induces UPR and Apoptosis in AML Cell Lines
Authors: Jackson Leah , Bechler Shelby , Miller Justin , Brownell Amy , Garshott Danielle , Callaghan Michael , Fribley Andrew M
Published: October 2019

Abstract 4260

Acute Myelogenous Leukemia (AML) is the most common form of leukemia. Current therapies are intense and even those fortunate enough to achieve remission often relapse extending extremely poor prognoses to these patient. The most commonly used therapeutics, namely cytarabine aribinoside, the anthracyclines and etoposide, are decades old and target ubiquitous cellular processes. We have previously reported that small molecules and natural products that activate and exacerbate the unfolded protein response (UPR) can effectively and selectively induce cell death in a wide variety of solid tumor cells. We hypothesized that the UPR might be a viable new therapeutic target in AML and sought to determine whether or not the novel UPR-inducing natural product borrelidin might be used as such an agent. A luminescent proliferation assay performed with panel of four AML cell lines treated with the ER stress-inducing antibiotic tunicamycin (Tm) revealed that three of the cell lines displayed IC50 values between 0.47–2.5μ M, doses of Tm which are known to induce a low to moderate level of ER stress. We then repeated the experiment with the more general UPR-inducing natural product borrelidin, which has been shown to have potent anti-inflammatory properties in several murine assays in vivo. All four cell lines were sensitive to borrelidin, displaying IC50 values between 0.032–0.29 μ M. Time course assays performed with borrelidin revealed 4–20 fold increases in active caspase 3 and 7 indicating borrelidin-induced AML decreases in cell proliferation might be the result of apoptosis. Quantitative reverse-transcription real time PCR performed with mRNA isolated from two AML cell lines revealed an increase in the UPR-related transcripts CHOP, ATF4, and GADD34 and the cell death genes Noxa, Puma, DR5 and Bim confirming that borrelidin could induce the UPR and apoptosis in AML cells. Studies currently underway in our laboratory will determine the ability of borrelidin and other UPR-inducing agents to reduce leukemic burden in an in vivo xenograft model.

Disclosures:

No relevant conflicts of interest to declare.
MNPs-Fe3o4 Induce Leukemia Cell Apoptosis by Oxidative Stress
Authors: Wang Ying , Chen Bao-An , Xia Guo-hua , Jian Ziying , Li Zhi , Lin Yun
Published: October 2019

Abstract 4304

The present study evaluated on the safe concentration for normal monocyte cells whether the magnetic nanoparticles of Fe3O4 (MNPs-Fe3O4) could induce caspase3-dependent apoptosis in SKM1 cells and U937 cells by oxidative damage. The average sizes of the particles were carefully determined by transmission electron microscopy. The viability of the cells after exposed to different concentration MNPs-Fe3O4 for 12h, 24h, 48h and 72h were detected by trypan blue staining. Cell cycle after exposure for 72h was analysed by propidium iodide (PI) staining. Apoptosis of MNPs-Fe3O4 group were detected and determined by Annexin V-FITC/PI double staining, 5,5’,6,6’-tetrachloro-1, 1’,3,3’-tetraethylbenzimidazolylcarbocyanine iodide (JC-1) probe and Wright - Giemsa staining. ROS in SKM1 cells and U937 cells after exposure were determined by flow cytometry after dichlorofluoroescein diacetate (DCFH-DA) staining. The changes of caspase3, survivin and bcl-rambo in the cells treatment with MNPs-Fe3O4 with or wtihout trolox for 48 hours were detected by western blot analysis. The significant decreased of cell viability caused by 100 μ M MNPS-Fe3O4 exposure were observed in SKM1 cells and u937 cells, not in normal monocytes cells (p < 0.05), the exposure also induced the SKM1 cells and u937 cells G0/G1 arrest (p < 0.05). Annexin V / PI staining assay show that 100 μ M MNPs-Fe3O4 group appeared more apoptosic rate in the U937 and SKM1 cells than the control group (p < 0.05), and on the same exposed concentration the apoptotic bodies could be frequently found in the U937 and SKM1 cells, while not in normal monocyte cells by Wright - Giemsa staining. The mitochondrial membrane potential in the two kinds of cells significantly decreased after exposed 100 μ M MNPs-Fe3O4 for 24h (p < 0.05), while pretreated with antioxidants trolox, the change was allevated (p < 0.05). DCFH-DA assay show that reactive oxygen species (ROS) generation increased in 6h of exposure in SKM1 cells and U937 cells (p < 0.05). Our results also showed the particle induced caspase3-dependent apoptosis in the SKM1 cells and U937 cells, while did not in normal monocyte cells, and increasing expression of bcl-rambo and decreasing expression of survivin involve in the process. These results suggest that on the safe concentration for normal monocyte cells MNPs-Fe3O4 could induce caspase3-dependent apoptosis in SKM1 cells and U937 cells by oxidative damage, moreover, increasing expression of bcl-rambo and decreasing expression of survivin involve in the process.

Disclosures:

No relevant conflicts of interest to declare.
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 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 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 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 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 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 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 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 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.
Autologous (Auto) Peripheral Blood Stem Cell (SCT) As a Consolidation Therapy for Patients with Acute Myeloid Leukemia (AML) in 1st Complete Remission (CR): A Single Institution Experience
Authors: Buttar Amanpreet , Jagadeesh Deepa , Cerny Jan , Ramanathan Muthalagu , Zhou Zheng , Petrillo-Deluca Laura , Tarrasky Natasha , Bednarik Jayde , Desai Zankar , Walsh William V , Nath Rajneesh
Published: October 2019

Abstract 4505

Introduction:

Patients with AML in 1st CR are frequently consolidated with an auto SCT but the role of this modality in the management of AML is still not fully defined.

Methods:

Retrospective analysis was performed on all patients who underwent an auto SCT for AML in 1st CR at UMass Memorial Medical Center from January 2000 to December 2010. Data was analyzed as of August 2011. The study was approved by UMass Memorial Medical Center IRB.

Results:

25 Patients were identified from the database. There were 13 males and 12 females. The median age was 46 years range (19–72). Cytogenetics was good risk in 6 patients, intermediate risk in 14, poor risk in 3 and unknown in 2 patients. Induction chemotherapy was standard dose Ara-C based in 13 patients and high dose Ara-C based regimen in 12 patients. Median time from diagnosis to transplant was 133 days range (107–362). Stem cell mobilization regimen consisted of Ara-C/Etoposide (E) in 21 (80%), Cyclophosphamide (Cy) in 3 (12%) and Mitoxantrone/Ara-C/E in 1 (4%). Preparative regimen included Busulfan (Bu) 4/ E in 17 (68%), Bu4/Cy in 5 (20%) and high dose Melphalan in 3 (12%). A median of 4.56 ×106 range (1.05 × 106 – 9.67 × 106 ) CD34 cells/kg were infused. 3 patients failed to engraft. Median time to neutrophil recovery was 11 days range (9 to 16) and platelet recovery was 20 days range (9 to 116). 100 day mortality was 12% (3/25). Median follow of the entire cohort is 6.5 years range (0.6 – 10.7). Total of 9 patients died. The cause of death included disease in 7, sepsis in 1 and unknown in 1 patient. Kaplan Meier estimate for progression free survival (PFS) and overall survival (OS) was 72% and 80% at 6 months and 62% and 61% at 3 years.

Conclusion:

ASCT is an effective and safe intensive consolidation therapy for patients with AML in 1st CR.

Disclosures:

No relevant conflicts of interest to declare.
Granulocytes Infiltrate The Intestine In a Zebrafish Model Of Inflammatory Bowel Disease
Authors: Geiger Brenda , Zhen Aileen W. , Kokkotou Efi , Fraenkel Paula G.
Published: October 2019

Inflammatory bowel disease (IBD) is a condition in which lymphocytes and neutrophils infiltrate the intestine, stimulate the production of various inflammatory cytokines, and injure the intestinal mucosa. Because of the conserved aspects of hematopoiesis in the zebrafish, we hypothesized that a zebrafish model of IBD would reproduce human features of the disease, while retaining desirable aspects of zebrafish disease models including small size, low cost, and ease of chemical and genetic screening. We used adult transgenic zebrafish expressing eGFP under control of a myeloperoxidase promoter (Tg(mpx:eGFP) to trace the activity of granulocytes following intrarectal injection with trinitrobenzene sulfonic acid (TNBS), 1.6 mmole/gram body weight, or an equivalent volume of vehicle alone (30% ethanol). The zebrafish were sacrificed 6 hours’ post-treatment for assessment of changes in intestinal architecture, intestinal infiltration by myeloperoxidase-positive leukocytes, and cytokine transcript levels using histology, flow cytometry, and quantitative realtime RT-PCR, respectively. At 6 hours post-treatment, we detected a mononuclear infiltrate in the intestinal mucosa and a 10-fold increase in the number of myeloperoxidase-positive leukocytes in the intestines of TNBS-treated fish vs controls (0.614%±0.202% vs. 0.052%±0.014%, p=0.0269; n=6-7 fish per group). We also observed a significant increase in the transcript levels of interleukin-1beta (6,935±2,454 AU vs 100±28 AU, p=0.0179), interleukin-8 (246 ± 63 AU vs 100 ± 21 AU, p=0.05) and interleukin-10 (385 ± 84 AU vs 100± 8 AU p=0.0055) in the intestines of fish injected with TNBS vs vehicle alone (n=9-10 fish per group). These findings resemble those seen in patients and mammalian chemical injury models of IBD. They will provide the basis for further study of the mechanisms controlling intestinal inflammation in IBD.

Disclosures:

No relevant conflicts of interest to declare.