bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mobilization of Allogeneic Peripheral Blood Stem Cells in Family Donors with Single-Dose Pegfilgrastim.
Authors: Vucinic Vladan , Basara Nadezda , Stiegler Runa , Bartsch Kristina , Kliem Constanze , Leiblein Sabine , Niederwieser Dietger 
Published: October 2019

Abstract
Abstract 2148
Poster Board II-125

Introduction:
The standard procedure for obtaining peripheral blood stem cells (PBSC) is donor mobilization with G-CSF. Pegfilgrastim is a covalently bound conjugate of filgrastim and monomethoxypolyethylene glycol with longer half-life elimination due to decreased plasma clearance and could represent an alternative approach for PBSC mobilization in healthy donors.

Design and Methods:
From July 2006 till August 2009 28 related healthy donors (50% male, 50% female) were treated with single dose of 12 mg pegfilgrastim for mobilization of allogeneic PBSC. The harvests were performed as large-volume, continuous-flow collections using a Cobe Spectra blood cell separator on day 4 and if necessary on day 5 of the mobilization regimen. In case of inadequate CD34+ counts (less than 4×106/kg body weight of recipient on day 5), stimulation was continued with filgrastim. In addition, the serum level of filgrastim was determined twice daily.

Results:
We present the results of 27 donors (the results of the 28th donor are still pending). In all 27 cases the harvests were successful. In 22 out of 27 donors (82%) only a single apheresis was needed to reach the target. Two of the donors required additional treatment with non-pegylated filgrastim.
The maximal concentration of circulating CD34+ cells was achieved on day 4 (median 74.3/μl; range 24.6-136.6). The median yield of CD34+ cells was 5.9×106/kg of the recipients body weight (range 3-14.5), and the median CD3+ count was 9.1×108/kg of the recipient body weight (range 1.4-6.2). Serum filgrastim level peak was on day 2 of the mobilization regimen with a median level of 226 ng/ml (range 35 to 1123 ng/ml), thus preceding the increase of CD34+ cells in blood. The main adverse events were WHO grade 1 and included headaches, bone pain and transient elevations of alkaline phosphatase and lactate dehydrogenase.

Conclusion:
PBSC mobilization with a single dose of pegfilgrastim is feasible for healthy donors. The graft composition was comparable to that obtained with the conventional regimen of short-term G-CSF. Long-term follow-up of healthy donors treated with pegfilgrastim should be further investigated.

Disclosures:
No relevant conflicts of interest to declare.
The Expression Pattern of TCR Vγ I∼III Subfamilies in GVHD Patients After Allogenic Hematopoietic Stem Cell Transplantation.
Authors: Wu Xiuli , Liu Can , Zhang Yu , Fan Zhiping , Liu Qifa 
Published: October 2019

Abstract
Abstract 2219
Poster Board II-196

Objective
The γΔ+ T cells can act as a vast functional setting of immune defense against pathogenic invaders in chronic inflammatory reactions, as well as in modulating systemic and organ-specific autoimmune diseases. Recently, it was reported that γΔ+ T cells might play an important role in regulation of graft versus host disease (GVHD), mediation of graft versus leukemia effect, anti-virus infection, anti-cancer effect and so on. But the frequency of T cell receptor (TCR) Vγ repertoire and the expression patterns of TCR Vγ subfamilies in GVHD remain unknown. To further study on the characteristics of TCR Vγ subfamilies in GVHD, we investigated the frequency of TCR Vγ repertoire and the expression of TCR Vγ I∼III subfamilies genes in the patients with GVHD after allogenic hematopoietic stem cell transplantation (allo-HSCT).

Methods
The expression and cloanlity analysis of TCR Vγ repertoire was detected in peripheral blood mononuclear cells (PBMNCs) from 25 patients with GVHD by RT-PCR and genescan technique. The TCR Vγ I∼III genes expression levels of PBMNCs from patients with GVHD were detected using the real-time fluorescence quantitative polymerase chain reaction with SYBR Green I technique and relative quantification method. Twelve healthy individuals served as normal controls.

Results
The RT-PCR results showed that in GVHD patients, the expression frequency of TCR Vγ I was 88.0% (22/25), TCR Vγ II was 72.0% (18/25), and TCR Vγ III was 96.0% (24/25). All of the three TCR Vγ subfamilies could be detected in PBMNCs from the normal controls. TCR Vγ repertoires with polyclonal pattern were identified in normal controls. However, the skew expression pattern of TCR Vγ repertoire could be detected in patients with GVHD even more than 4 year after allo-HSCT. Oligoclonal or monoclonal expanded T cells were identified in TCR Vγ I∼III subfamilies in GVHD patients. The expression level of TCR Vγ II gene in the PBMNCs from patients with GVHD was significant lower than the normal controls (P<0.05). The pattern of TCR Vγ gene expression levels in GVHD was TCR Vγ I> TCR Vγ III> TCR Vγ II, however, the expression pattern in normal controls was TCR Vγ II> TCR Vγ I > TCR Vγ III.

Conclusions
The expression patterns of TCR Vγ I∼III subfamilies were changed in GVHD. The low expression of TCR Vγ II subfamily might be related with the pathogenesis of GVHD.

Disclosures:
No relevant conflicts of interest to declare.
Low Sensitivity of the (1-3)-Beta-D-Glucan Assay for Diagnosis of Invasive Aspergillosis in Hematologic Malignancy Patients.
Authors: Yonezumi Masakatsu , Kurosawa Mitsutoshi , Fukuhara Takashi 
Published: October 2019

Abstract
Abstract 2242
Poster Board II-219
Invasive aspergillosis (IA) is a common opportunistic infection in hematologic malignancy patients. The diagnosis of IA is difficult and the outcome is poor despite recent progress in treatment. The detection of galactomannan antigen is useful in diagnosis of IA and is defined as one of EORTC/MSG diagnostic criteria. The (1-3)-beta-D-glucan assay is also used in diagnosis of invasive fungal disease, but the sensitivity and specificity of these tests for IA are variable in recent reports. To evaluate the correlation of them, we analyzed retrospectively the results of these tests in IA cases of hematologic malignancy patients.
From January 2006 to December 2008, we reviewed 2914 hematologic malignancy patients (AML=492, ALL=198, MDS=284, NHL=1373, HL=92, MM=375) retrospectively who were recieved chemotherapy or stem cell transplantation(SCT). 607 cases received SCT are included in this study. We diagnosed IA according to the EORTC/MSG criteria, and extracted the data of galactomannan antigen and (1-3)-beta-D-glucan assay examined at diagnosis of each case.
43 cases(1.5%) are diagnosed as invasive fungal disease (Aspergillus=24, Candida=9, Mucor=6, Trichosporon=2, Cryptococcus=1, Geotrichum=1). 23 IA cases are analyzed (proven: 3 cases, probable: 20 cases). All cases are positive for galactomannan antigen test(range, 0.5-5.0< ODI, cut-off value:0.5), but only 9 cases for (1-3)-beta-D-glucan assay (range, 24.5-313pg/ml, cut-off value:20). In the three cases of proven IA, only one case are positive for this assay. There is no difference in patient characteristic and prognosis between the positive and negative group.
We conclude that the (1-3)-beta-D-glucan assay is not useful tool for diagnosis of invasive aspergillosis.

Disclosures:
No relevant conflicts of interest to declare.
Prospective Evaluation of Proteomic Screening with An Agvhd-Specific Proteomic Pattern MS-17.
Authors: Mischak-Weissinger Eva M. , Metzger Jochen , Krons Annika , Kontsendorn Julia , Krauter Jürgen , Stadler Michael , Mischak Harald , Ganser Arnold 
Published: October 2019

Abstract
Abstract 2246
Poster Board II-223
Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the only curative treatment for many hematologic malignancies or hematopoietic dysfunction syndromes in adults, but the application is still limited due to major complications, such as severe graft versus host disease (GvHD) and infectious complications. Diagnosis GvHD is based on clinical features and biopsies, a non-invasive, unbiased laboratory test using proteomics was published previously and evaluated in a multicenter setting (Weissinger et al., 2007). In order to maintain good quality data of the proteomic pattern data and due to the fact that the electrospray ionization time of flight mass spectrometer (ESI-TOF) from ABI (mariner) is no longer produced, we applied a more sensitive mass spectrometer (MS; ESI-TOF/Bruker). In order to validate the quality of the data produced with the Bruker, ESI-TOF was used since December 2007 in parallel to the mariner. Our first and previously published “aGvHD-specific pattern” (Weissinger et al., 2007) was used to evaluate and improve the pattern with the more sensitive decreased (6) or increased (11) signal intensity in patients with GvHD grade II to IV. Patients post HSCT and patients with GvHD grade I have the same pattern. In addition, the MS-17-pattern yielded at least comparable if not superior results to the previous one yielding sensitivities around 95% and specificity about 80%.
To date the proteomic pattern specific for aGvHD MS17 was evaluated blindly on 400 samples collected from about 90 patients undergoing allo-HSCT at MHH since Dec. 2007.
The peptide pattern MS-17 consists of 17 peptides - either absent/decreased (6) or present /increased (11) in aGvHD grade II to IV when compared to patients with clinical aGvHD grade I or without GvHD. Prospective and blinded evaluation of the patients included in this parallel diagnostic study revealed the correct classification of patients developing aGvHD about 7 days prior to the development of clinical signs and symptoms for aGVHD with a sensitivity and specificity of about 85%. Sequencing 3 of the 17 markers yielded the following results:
Peptide No 5: Albumine fragment (N-terminus); Peptide No 7: ψ2 microglobulin; Peptide No 10: fragment of CD99 (MIC2; a single-chain type-1 glycoprotein). CD99 and β-2 microglobulin are particulary interesting because of their involvement in immune responses, both being expressed on leukocytes and CD 99 especially on thymocytes with involvement in apoptosis of double positive T-cells and binding cyclosporin A. Data on the pattern, signal intensity (3 dimension, mass and charge are shown in Figure 1)

Disclosures:
Metzger: mosaiques-diagnostics GmbH: Employment. Krons:mosaiques-diagnostics GmbH: Employment. Mischak:mosaiques-diagnostics GmbH: Patents & Royalties, coowner and founder of mosaiques-diagnostics.
Novel Anti-CD19/Idiotype Bispecific Diabody Vaccine for B-Cell Lymphoma.
Authors: Ng Patrick P. , Jia Ming , Virrueta Alejandro , Patel Kedar , Swartz James R. , Levy Shoshana , Levy Ronald 
Published: October 2019

Abstract
Abstract 2712
Poster Board II-688
The unique immunoglobulin idiotype (Id) on B-cell lymphoma is an excellent target for a “custom-made” vaccine therapy for individual patients. Previous studies have shown that vaccine-induced anti-Id humoral responses are effective in protecting mice from the aggressive 38C13 lymphoma. Therefore, we explore a novel strategy of direct activation of Id-specific B cells using a bispecific diabody containing the Id variable fragments (Fv) of 38C13, and those of an anti-CD19 monoclonal antibody. This molecule is designed to cross-link the Id-specific B cell receptor to the CD19/CD21/CD81 co-stimulatory complex, an interaction that has been shown to lower the activation threshold of B cells and result in a dramatic increase in antibody production. The αCD19/Id diabody is a 52 kDa heterodimer of two single chain Fv's (scFv), αCD19VH-38C13VL and 38C13VH -αCD19VL, held together by non-covalent interactions between the cognate VH and VL domains. It was produced in an E. coli extract-based Cell-Free Protein Synthesis (CFPS) System developed by our group, which has been used successfully to rapidly produce cytokine-Id fusion vaccines in previous studies. Within 2 hours, the two diabody scFv's were co-expressed and self-assembled in vitro. The heterodimer product was purified by metal affinity chromatography. Correct folding was confirmed by a flow cytometry assay in which the diabody bound simultaneously to the CD19 on the surface of B cells and also to an anti-38C13 Id antibody labeled with FITC. Most importantly, as a vaccine the αCD19/Id diabody generated a robust anti-Id IgG response comparable to that induced by 38C13 IgM conjugated to the carrier protein keyhole limpet hemocyanin (KLH). Diabody vaccinated mice also showed improved survival compared to control groups when both were challenged with a lethal dose of tumor. Further optimization of the therapeutic efficacy and the development of improved methods for the rapid and economical production of αCD19/Id diabody by the CFPS technology may make it possible to use individualized diabody-based therapy as a frontline treatment, before the use of toxic chemotherapy and radiation therapy.

Disclosures:
No relevant conflicts of interest to declare.
C-KIT with D816Y/V Mutations Cooperate with CBFB-MYH11 to Accelerate Leukemogenesis in Mice.
Authors: Zhao Ling , Melenhorst J. Joseph , Alemu Lemlem , Kirby Martha , Anderson Stacie M , Hoogstraten-Miller Shelley , Kamikubo Yasuhiko , Gilliland D. Gary , Liu Pu Paul 
Published: October 2019

Abstract
Abstract 274
CBF leukemia is a group of acute myeloid leukemia (AML) caused by the fusion genes involving core binding factors (CBFs), which include RUNX1 and CBFB. C-KIT mutations are common in CBF leukemias. For example, 10–45% of AML cases with inv(16), which generates a fusion gene between CBFB and MYH11, carry activating mutations in c-KIT. In general, the presence of c-KIT mutations in inv(16) AML suggests poor clinical prognosis. Therefore, it is important to verify that c-KIT mutations cooperate with CBFB-MYH11, and to investigate the underlying mechanism during leukemogenesis. Here, we transduced bone marrow (BM) cells from conditional Cbfb-MYH11 knockin mice with retroviral vectors carrying c-KIT mutations,D816V and D816Y (located in the kinase domain of c-KIT and can cause constitutive activation of c-KIT), which are common in inv(16) AML. Wild type c-KIT and empty retroviral vectors were used as controls. The transduced BM cells were then transplanted to sub-lethally irradiated recipient mice. In 2–5 months, 60% of mice carrying D816Y and 50% of mice carrying D816V c-KIT mutations developed leukemia, while none of the mice transduced with the control vectors developed leukemia up to one year after transplantation. In addition, BM cells from wildtype mice transduced with the c-KIT mutant vectors did not induce leukemia >5 months after transplantation. Analysis of signaling pathways revealed that Stat3-serine phosphorylation and P44/42 MAPK pathways, but not AKT and Stat5 pathways, were activated (phosphorylated) in the leukemia cells from the Cbfb-MYH11 knockin mice expressing mutated c-KIT. In vitro colony forming assays indicated that Cbfb-MYH11 BM cells expressing c-KIT D816V/Y mutants produced similar number of colonies as Cbfb-MYH11 BM cells transduced with wild type c-KIT control vector. However, they had increased CFU-G and CFU-GEMM and decreased CFU-E colonies, suggesting that the c-KIT mutants drove Cbfb-MYH11 BM cells towards an earlier progenitor state. Secondary transplantation with 1×106 leukemia cells from donors with c-KIT mutations showed an average life span of three weeks compared to six weeks from donors without c-KIT mutation. Further limiting dilution transplantation with 100 leukemia cells from one donor with c-KIT mutation led to lethal leukemia in 4 out of 5 mice 2.5 months after transplantation, indicating a very high frequency of leukemia initiating cells in this sample. These data suggest a strong cooperation between the mutant c-KIT and Cbfb-MYH11 for leukemogenesis in mice. This mouse model can also serve as a clinically relevant model for small chemical screening and the development of novel therapeutic approaches.

Disclosures:
No relevant conflicts of interest to declare.
DNA Methylation Changes Following 5-Azacitidine Treatment in Patients with Myelodysplastic Syndrome.
Authors: Tran Huong Thi Thanh , Kim Hee Nam , Kim Yeo-Kyeoung , Ahn Jae-Sook , Lee Il-Kwon , Yang Deok-Hwan , Lee Je-Jung , Kook Hoon , Hwang Tai Ju , Kim Hyeoung-Joon 
Published: October 2019

Abstract
Abstract 2791
Poster Board II-767
Gene silencing by promoter methylation is as potent as functional inactivating of tumor suppressor genes by mutations. DNA methyltransferase inhibitor, 5-azacytidine (AC) and 5-aza-2 -deoxycitidine (DAC), which is proved to be effective in myelodysplastic syndromes (MDS) can induce re-expression in cancer; however their mechanism remains controversial. 25 tumor suppressor genes by MS-MLPA (methylation-specific multiplex ligation-dependent probe amplification) were analyzed in 44 MDS patients treated Vidaza® (5-azacitidine, AC). Hypermethylation of at least one gene was detected in 9/44 patients (20.5%), including four genes CDKN2B, FHIT, ESR1 and IGSF4. Interestingly, of 9 hypermethylated patients, 8 patients showed demethylation in concordance with their clinical responses after three to five cycles AC treatment. Lack or decrease methylation was observed in four patients with hematological improvements. Persistence methylation was observed in four others who became AML transformation or no response after treatment, especially reinforcing methylated gene in a case progressed to leukemia later. Our study also founds out IGSF4 gene hypermethylation in MDS as a first report. Additionally, mRNA expression of CDKN2B, IGSF4, and ESR1 in MDS were significantly lower than those in the control group (p < 0.05).
Our results suggest that the methylation changes of specific genes contributes to disease pathogenesis and might present a molecular marker that can be used to monitor the efficacy of AC treatment in MDS.

Disclosures:
No relevant conflicts of interest to declare.
Heparanase Promotes Osteolysis by Upregulating RANKL– A Novel Role for Heparanase in Multiple Myeloma.
Authors: Yang Yang , Nan Li , Suva Larry J , Sanderson Ralph D. 
Published: October 2019

Abstract
Abstract 2821
Poster Board II-797
Heparanase, an enzyme that cleaves the heparan sulfate chains of proteoglycans, is upregulated in a variety of human tumors including multiple myeloma. We have previously demonstrated that heparanase promotes robust myeloma tumor growth and supports spontaneous metastasis to bone. In addition we showed that expression of heparanase is a major determinant of the osteolytic phenotype in myeloma and that heparanase expression in a distal tumor can promote systemic osteolysis due to enhanced osteoclastogenesis within distal bones prior to metastasis. Because it has been shown that myeloma cells can produce and secrete RANKL, we hypothesized that heparanase upregulates the expression of RANKL thereby leading to osteolysis. To test this idea, CAG myeloma cells transfected with human heparanase cDNA (heparanase-high) or empty vector (heparanase-low) were injected into SCID mice and the levels of human RANKL expression were evaluated by immunohistolochemical staining and western blotting. A significant elevation of RANKL was observed in heparanase-high tumors compared to heparanase-low tumors formed in human bones implanted in SCID mice (SCID-hu), as well as in subcutaneous myeloma tumors. In addition, RANKL levels were significantly elevated in both in vitro cell extracts and conditioned medium of myeloma cells when heparanase expression was enhanced. The elevation of RANKL expression was also found in other myeloma cell lines (MM.1S and U266) when cells were cultured in the presence of recombinant human heparanase (50 ng/ml and 100 ng/ml each cell line, added every 12 hours) for 48 hours. In addition, in vitro osteoclastogenesis is significantly enhanced by the medium conditioned by heparanase-high cells as compared to heparanase-low cells and this enhanced osteoclastogenesis is completely abolished by addition of OPG (a RANKL inhibitor) to the medium. From these data, we conclude that elevated heparanase in myeloma cells increases RANKL expression and secretion, resulting in the increased osteolysis in myeloma.

Disclosures:
No relevant conflicts of interest to declare.
A Prospective Randomized Phase I/II Study of Lenalidomide, Melphalan, Prednisone and Thalidomide (RMPT) for Relapsed/Refractory Multiple Myeloma Patients.
Authors: Cavallo Federica , Larocca Alessandra , Petrucci Maria Teresa , Federico Vincenzo , Falcone Antonietta Pia , Sanpaolo Grazia , Canepa Letizia , Crugnola Monica , Boccadoro Mario , Palumbo Antonio 
Published: October 2019

Abstract
Abstract 2864
Poster Board II-840

Background.
New drug associations may improve patients outcome in relapsed/refractory multiple myeloma (MM). In newly diagnosed MM patients the addition of lenalidomide or thalidomide or bortezomib, to the standard oral melphalan and prednisone combination significantly increased response rate and event-free survival, showing synergistic effects.

Aims.
This is a multicenter, randomized, open label phase I/II trial designed to evaluate the safety/efficacy profile of the salvage treatment, Lenalidomide, Melphalan, Prednisone and Thalidomide (RMPT) in patients with relapsed/refractory myeloma.

Methods.
Oral lenalidomide was administered at 10 mg/day on days 1-21, in combination with oral melphalan at 0.18 mg/kg on days 1–4 and oral prednisone at 2 mg/kg on days 1–4. Thalidomide was administered at 50 mg/day (Arm A) or 100 mg/day (Arm B) on days 1-28. Each course was repeated every 28 days for a total of 6 courses. Maintenance therapy included Lenalidomide alone at 10 mg/day on days 1-21. Aspirin 100 mg/day was given as a prophylaxis for thrombosis.

Results.
Forthy-four patients with relapsed/refractory MM were enrolled in the study between May 2007 and March 2008, including 22 patients in arm A and 22 patients in arm B. After a median of 5 cycles 75% of patients achieved at least a partial response (PR), including 20% very good partial response (VGPR) and 14% near or complete response (nCR or CR). Among patients who received thalidomide 100 mg, the PR rate was 82% (including 23% VGPR and 23% CR/nCR). Higher response rate was observed among patients who received RMPT in first relapse. The 1-year-progression-free survival was 51,5% and the 1-year survival from study entry was 72%. Hematologic toxicity was the most frequent adverse event. Grade 3-4 hematologic adverse events included: neutropenia (68%), thrombocytopenia (36%) and anemia (32%). Grade 3-4 non hematologic adverse events included: infections (22,7%), neurological toxicity (4,5%) and fatigue (6,8%). No grade 3-4 thromboembolic events were reported.

Conclusion.
This is the first trial exploring the association of two immunomodulatory drugs, Thalidomide and Lenalidomide. RMPT is an active regimen in patients with relapsed/refractory MM. Toxicities were manageable and the incidence of neutrotoxcities was low. Updated results will be presented at the time of the meeting.

Disclosures:
Cavallo: CELGENE: Honoraria. Petrucci:CELGENE: Honoraria. Canepa:CELGENE: Honoraria. Boccadoro:CELGENE: CONSULTANCY, ADVISORY COMMITTEES, Research Funding. Palumbo:CELGENE: Honoraria.