bluuebunny/crossref_metadata_2025_split huggingface dataset:
CFU-GEMM Infused Influences Immunologic Recovery in Children Older Than 7 years After Hematopoietic Stem Cell Transplantation.
Authors: Drabko Katarzyna , Choma Marta , Zaucha-Prazmo Agnieszka , Wojcik Beata , Winnicka Dorota , Kowalczyk Jerzy 
Published: October 2019

Abstract
Abstract 4483

Introduction
Reconstitution of immunity is considered as one of the most important factors predicting outcome after hematopoietic stem cell transplantation (HSCT). Age of the recipient as well as graft quality reflect on this process however this influence is not completely understood yet. In this study we aimed to evaluate the influence of graft quality on recovery of lymphocyte subpopulation.

Patients and Methods
The total number of 63 patients who underwent allogeneic transplantation in one center between May 2002 and November 2008, were included into the study. Median age of the patient was 8,6 (range 0.2-18) yrs. Patients were divided into 2 age groups: group 1 (n=26) children age 0,2-7 and group 2 (n=37) children >7 yrs. In all cases quality of graft were assessed by counting WBC, CD34 cells and CFU-GEMM. 45 patients (21 in group 1 and 24 in group 2) were available for evaluation of immune reconstitution: in those patients the absolute number of lymphocytes CD3/CD19 (B) cells, CD3/CD4 (T helper) cell, CD3/CD8 (T suppressor) cells and CD3/CD16+56 (NK) cells were measured 3 and 6 months after transplantation.

Results
Younger children (group 1) received higher median number of transplanted nucleated cells (4.9×108 vs 2.5×108, p=0.008), CD34 cells (5.6×106 vs 2.1×106, p=0.02) and CFU-GEMM (2.25×104 vs 1.17×104, p=0.01) than older ones (group 2). No differences were found in absolute values of B, T-helper, T-suppressor and NK cells at 3 and 6 months after transplantation (Table 1). In older children (group 2), but not in younger ones, the number of GFU-GEMM transplanted negatively correlated with number of T-suppressor cells 3 months post HSCT (p<0,05; r=-0,7) and positively with number of T-helper cells 6 moths after HSCT (p<0,05; r=0,73).

Conclusions
Number of CFU-GEMM infused influences immunologic recovery in children older than 7 years after HSCT. Graft quality correlates with number of T-suppressor cells and T-helper cells during first 6 months after HSCT and these changes may contribute to post transplant complications.
Supported by grant MNiSW of Poland N407 117 35

Disclosures:
No relevant conflicts of interest to declare.
Idiopathic Pneumonia Syndrome in Mice After Allogeneic Bone Marrow Transplantation: Association Between Idiopathic Pneumonia Syndrome and Acute Graft-Versus-Host Disease.
Authors: Liu Qifa , Ning Juan , Zhang Yu , Luo Xiaodan , Fan Zhiping , Wu Xiuli 
Published: October 2019

Abstract
Abstract 4502

Objective
To explore the association between idiopathic pneumonia syndrome (IPS) and acute graft-versus-host disease (aGVHD) in allogeneic hematopoietic stem cell transplantation (allo-HSCT).

Methods
An aGVHD model was established using a mouse transplant (C57BL/6□BALB/c). Chest computed tomography (CT) scans were dynamically performed and histopathology and cytokine levels in lung tissues were kinetically detected using the Linchip system in three experimental groups: mice receiving simple irradiation, syngeneic transplants and allogeneic transplants.

Results
The incidence of aGVHD was 100% in allogeneic transplant mice. CT revealed bilateral diffuse infiltrates in the lungs of most mice with aGVHD vs normal lung tissue in syngeneic transplant mice. Histopathology confirmed mice with aGVHD had acute pneumonitis. Immunohistochemistry showed that during aGVHD onset the infiltrates were mainly CD4+ T cells whereas the domination of CD4+ T-cell was replaced by CD8+ T-cell during aGVHD progression. TNF-α and IFN-γ levels within lung tissues of the three groups were higher than in normal controls on day +3 and +7 post-transplant. On day +7, TNF-α levels were higher in allogeneic than in syngeneic transplant mice, but there was no statistical difference in IFN-γ levels. On day +12 and +16, TNF-α levels were significantly higher in allogeneic than in syngeneic transplant mice, but IFN-γ levels were lower in allogeneic than in syngeneic transplant mice.

Conclusions
The aGVHD is the underlying cause of IPS. T cells and TNF-α may play a role in the pathogenesis of aGVHD-induced IPS. IPS progression may be associated with decreasing levels of IFN-γ within the lung tissues.

Disclosures:
No relevant conflicts of interest to declare.
Defining Outcomes in Hispanic Aggressive Non-Hodgkin Lymphoma Patients.
Authors: Paraskos Lara M , Blum Mariela A. , Escalon Maricer , Ferrell Annapoorna , Kobetz Erin , Rosenblatt Joseph 
Published: October 2019

Abstract
Abstract 4542

Background
Multiple studies evaluating outcomes for oncology patients have demonstrated inferior overall survival rates among African-Americans. Hispanics comprise the largest ethnic minority in the United States. Surprisingly outcomes for Hispanic aggressive non-Hodgkin lymphoma (NHL) patients have not been evaluated and are therefore largely unknown. We aim to identify differences in epidemiology, treatment modalities and outcomes of Hispanic patients treated for aggressive NHL at our institution between the years 2000-2004.

Methods
We reviewed the medical records of 82 patients with aggressive type NHL who were identified using the tumor registry. Exclusion criteria include indolent and highly aggressive lymphomas per WHO classification as well as lymphomas related to AIDS or organ transplantation. Standard statistical analyses, including Kaplan-Meier survival estimates and Chi-squared analysis for group comparisons were used. P value of <0.05 was considered statistically significant.

Results
Of the 82 patients, 46 were self-reported Hispanic in origin; 36 were non-Hispanic. There were no significant differences with respect to age at diagnosis, LDH level, stage, or complete remission rate. Kaplan-Meier estimate of median overall survival (OS) of the cohort was 3.8 years with no statistically significant differences observed between the Hispanic and non-Hispanic ethnic groups. (P = 0.816).

Conclusion
Ethnic patterns of disease occurrences have been reported, but responses to treatment and outcome for Hispanic patients have not been established. Interestingly from our preliminary analysis, there appears to be no statistically significant difference in overall survival between Hispanic and non-Hispanic patients. Further evaluation is warranted.

Disclosures:
No relevant conflicts of interest to declare.
Implementation of a Hematological Malignancy-Based Survivorship Clinic.
Authors: Steinberg Amir , Asher Arash 
Published: October 2019

Abstract
Abstract 4550

BACKGROUND
There are approximately twelve million cancer survivors in the United States. Cancer survivors are at increased risk for health complications such as heart disease, secondary malignancies, and psychological distress. Reflecting the significant needs of cancer survivors as part of our health care system, the Institute of Medicine issued a seminal report, From Cancer Patient to Cancer Survivor: Lost in Transition, recognizing cancer survivorship as a “distinct phase of cancer care.” A dedicated survivorship clinic with creation of personal care plans and treatment summaries can serve the needs of this growing cohort of patients.

OBJECTIVE
To determine the viability of a hematological malignancy-based survivorship clinic with treatment summary and care plan for survivors. The malignancies include all lymphomas, leukemias, plasma cell dyscrasias, myeloproliferative disorders, and hematopoietic stem cell transplant patients.

METHODS
A literature search was initiated examining recommendations and guidelines regarding follow-up of cancer patients. We incorporated and consolidated guidelines from ASCO, NMDP, NCCN, and COG, pertinent articles from the MEDLINE/PubMed database, and the growing literature from cancer survivorship textbooks and lectures. Awareness of the program was to be initiated with letters to affiliated physicians, lecture series, patient-oriented educational handouts, and articles in the hospital literature.

DATA SYNTHESIS
A treatment care plan was devised for all survivorship clinic patients. This care plan summarized patients' prior care with recommendations for follow-up – such as cardiac and cancer screening, osteoporosis, thyroid disease, and vaccination schedules–all based on our consolidative review of available evidence-based or expert guidelines. A referral base was developed with specialists focused on lymphedema, psychiatric issues (needs determined by screening tools), social work, art therapy, exercise, and nutrition.

RESULTS
Patients from affiliated hematologists/oncologists were seen in the survivorship clinic. Unique care plans were devised for them. A satisfaction survey was to be implemented. Preliminary satisfaction from both physicians and the patients themselves was positive.

CONCLUSION
A hematological-based survivorship clinic is feasible and fills an important need in our large, urban, hospital-based cancer center. Further investigation will focus on developing a computer-based treatment summary and care plan, increasing patient referrals, and developing a lecture series devoted to survivorship issues.

Disclosures:
No relevant conflicts of interest to declare.
Effectiveness of a Novel Humanized VB22B Minibody against Human TPO Receptor Compared with Recombinant TPO and Eltrombopag Using Primary Human CD34+ Cells and Platelets.
Authors: Matsuki Eri , Yamane Akiko , Okamoto Shinichiro , Miyakawa Yoshitaka 
Published: October 2019

Abstract
Abstract 4559
Thrombopoietin (TPO) is a cytokine produced primarily by the liver and kidney that regulates platelet production by stimulating proliferation and differentiation of hematopoietic stem cells, megakaryocytic progenitor cells and megakaryocytes via activation of its receptor, c-Mpl. Recently, TPO receptor agonists such as eltrombopag and romiplostim have been approved for chronic ITP. huVB22B was created as a novel humanized form of murine sc(Fv) 2VB22B minibody (BLOOD, 2005) which activates human c-Mpl by CDR grafting. The advent of these various TPO receptor agonists prompted us to consider the differences in their mechanisms of action, efficacy or potency. However, to date, there has been no in vivo or in vitro study directly comparing the effects of different TPO receptor agonists. In this study, we compared the efficacy of huVB22B on CFU-GM, CFU-E, CFU-Megakaryocyte (CFU-MK), megakaryocyte maturation (DNA ploidy and proplatelet formation) with those of recombinant human TPO (rhTPO) and eltrombopag. Primary human CD34+ bone marrow cells were cultured with various concentrations of rhTPO, huVB22B and eltrombopag using methylcellulose based media. In serum-free condition, 0.286 nM rhTPO, 0.182 nM huVB22B and 17.7 mcM eltrombopag demonstrated almost equivalent efficacy of megakaryocyte colony formation. At these concentrations, all agents demonstrated similar in vitro efficacy for colony formation of CFU-GM and CFU-E, proplatelet formation and nuclear maturation of megakaryocytes. In preliminary results, huVB22B induced maturation of CFU-MK earlier than rhTPO and eltrombopag, suggesting that huVB22B might have some potential to increase human platelets faster than other agents in vivo. This is compatible with the observation that huVB22B induced tyrosine phosphorylation of STAT3, STAT5 and JAK2 faster and stronger than rhTPO and eltrombopag in human primary platelets. Both rhTPO and huVB22B enhanced low-dose ADP and collagen-induced human platelet aggregation in vitro. In contrast, eltrombopag did not enhance ADP or collagen-induced platelet aggregation, although it induced activation of JAK-STAT pathway in human platelets. Contrary to the fact that huVB22B induces phosphorylation of intracellular signaling molecules faster and stronger than rhTPO in human platelets, the priming effect by huVB22B on platelet aggregation was much weaker than rhTPO. In conclusion, we confirmed that newly created huVB22B minibody induced colony formation of CFU-MK, CFU-E, CFU-GM and maturation of megakaryocytes from human bone marrow-derived CD34+ cells in vitro. The differences among TPO receptor agonists observed in our study would lead to further understanding of the basic biology of megakaryopoiesis and the action of TPO receptor agonists.

Disclosures:
Okamoto: Alexion: Research Funding. Miyakawa:GlaxoSmithKline: Consultancy.
CXCR4 Blockade as a New Targeted Therapy for Acute Myeloide Leukemia Characterised by High Cell Surface Density of CXCR4.
Authors: Zhang Yanyan , Jouni Dima , Wittner Monika , Bouamar Hakim , Dombret Herve , Coppo Paul , Opolon Paule , Fujii Nobutaka , Ito Mamoru , Vainchenker William , de Botton Stephane , Louache Fawzia 
Published: October 2019

Abstract
Abstract 4570
Stromal cell-derived factor 1 (SDF-1)/CXCR4 axis plays key roles in hematopoiesis regulating the interactions between hematopoietic cells and their stromal microenvironment within the bone marrow, their trafficking from the bone marrow to blood, their proliferation and survival. SDF-1/CXCR4 interactions also participate in the development of leukemic blasts in acute myeloid leukaemia (AML) influencing their trafficking, survival and differentiation. We used the xenotransplantation model of nonobese diabetes/severe combined immunodeficiency γcnull (NOG) mice reconstituted with human primary cells from AML patients and 2 small molecule competitive antagonists of CXCR4, AMD3100 and TN140 to better define the role of CXCR4/SDF-1 on the leukemic blast burden within the bone marrow and in extramedullary sites. In a first set of experiments, we investigated whether there was a correlation between CXCR4 expression or function on leukemic blasts and their ability to engraft the bone marrow of NOG mice in 34 patients. Using flow cytometric analyses, we observed that CXCR4 membrane expression was highly variable between patients. This expression did not correlate with engraftment. In addition, SDF-1 responsiveness evaluated by transwell migration only marginally correlated with engraftment. However, in these initial analyses, we observed that the differences between engrafters and nonengrafters were significant if the cut-point migration is set to 20%. Patients with higher chemotactic response to SDF-1 had a significantly increased NOG repopulating ability. In further studies, NOG mice reconstituted with AML cells from 5 different patients were treated with optimal concentration of either AMD3100 or TN140 for 1 week. We observed that CXCR4 inhibition by TN140 (used as a single therapy) had profound inhibitory effects on the proliferation and the development of extramedullary dissemination of the disease in this xenotransplantation. This patient had FAB M1 leukemia and initially exhibits high CXCR4 level on blast. Our study demonstrated that CXCR4 inhibition in selected patients might be a potent therapy against leukemic development.

Disclosures:
Bouamar: Association pour la Recherche sur le Cancer: Employment; Cancéropôle IDF: Employment.
Human Umbilical Cord Wharton's Jelly-Derived Mesenchymal Stem Cells Differentiate Into Insulin-Producing Cells.
Authors: Ma Lian , Wang Hongwu , He Hongyan , Lin Limin , Li Weizhong , Huang Tianhua , Ma Guixia , Wang Aihong 
Published: October 2019

Abstract
Abstract 4578

Introduction
Islet transplantation is an effective way of reversing type 1 diabetes. However, islet transplantation has been hampered by problems, such as immune rejection, and the scarcity of donor islets. Human Umbilical Cord Wharton's Jelly-derived Mesenchymal Stem Cells (huMSCs), which can be differentiated into insulin-producing cells could provide a source of cells for transplant.

Methods
Vitro Research We isolated and cultured huMSCs, and induced huMSCs differentiated into insulin-producing cells in the condition of islet cells grows. The morphology of huMSCs after induction were monitored by under inversion phase contrast microscope?GImmunocytochemical methods were used to detect the insulin and glucagon protein, and reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect Human insulin gene and PDX-1 gene. Dithizon-stained was used to detect zinc hydronium and radio-immunity was used to detect insulin level of culture supernatant.Vivo Research huMSCs were transplanted into the body of diabetic rats through vena caudalis, and then we observed the change of blood glucose?Abody weight ?Aserum insulin levels and survival ratio in STZ-induced diabetic rats. We detected human insulin by immunohistochemistry and RT-PCR. HE stain was used to detect the morphological changes of rat's pancreatic island.

Results
Vitro Research The morphology of huMSCs under medicine induction gradually changed from fibroblast to round and some of then had the tend of forming clusters.?GThe result of immunocytochemical showed that the expression of human insulin and glucagon was positive after treatment with medicine?GhuMSCs induced by medicine can express insulin and PDX-1 gene by RT-PCR?GDithizon stain show that the cytoplasm of huMSCs after induction were stained in Brownish red color?Gthe results of radio-immunity manifested that the insulin quantity secreted by medicine induction were significant differences compared with control group(t??6.183,P<0.05). Vivo Research When transplanted into Streptozotocin(STZ)-treated diabetics rats, huMSCs can decreased blood glucose, increased body weight and survival ratio in diabetic rats?GAfter being transplanted for one month, we discovered that it can be planted into rat's pancreas and liver by Hoechst33258?Gimmunohistochemistry and RT-PCR show that the pancreas of rat can express human insulin?Gthe morphology of rats' pancreatic island was repaired obviously if compared with diabetic rats before the transplantation through HE-stain.

Conclusion
huMSCs can be differentiated into insulin-producing cells in vitro or in vivo. Therefore, huMSCs have the potential to become an excellent candidate in β cell replacement therapy of diabetes.

Disclosures:
No relevant conflicts of interest to declare.
Differentiation of Human Male Germ Cells From Human Umbilical Cord-Derived Mesenchymal Stem Cells in Vitro and In Vivo.
Authors: Ma Lian , Wu Xiaoying , Lin Limin , Lin Guangyu , Tang Qiuling , Huang Tianhua , Feng Xueyong , Ma Guixia , Xie Qingdong 
Published: October 2019

Abstract
Abstract 4581

Introduction
Infertility affects 15% of couples, about 50% infertility caused by male and growing evidence suggested an increasing problems in male reproductive. Recent studies have demostrated that adult stem cells have more flexible potentials than expected, and possessed the plasticity and capacity to transdifferentiate into mutilineage cells, including germ cells. Human umbilical cord-derived mesenchymal stem cells (HUCMSCs) possess stem cell properties. In this study, we cultured HUCMSCs, and assessed the possibility of HUCMSCs differentiated into human male germ cells in vivo and in vitro, and find a new source of cells for the transplantation to the male infertility.

Methods
The ethics committee of our institution approved this study. HUCMSCs were isolated from the Wharton's jelly of the umbilical cord, clonally expanded. To investigate the capacity of differentiation in vitro, HUCMSCs were treated with human menopausal gonadotropinn (HMG) and retinoid acid (RA) in vitro. While investigate the capacity of differentiation in vivo, HUCMSCs were transplanted into the seminiferous tubules of busulfan-treated mice testes after labeled with pIRES2-EGFP or bromodeoxyuridine (BrdU). After induction in vitro, the morphologic changes of the differentiated cells were detected by phase contrast microscopy?Aelectron microscopy and transmission electron microscope?Gthe male germ cell markers were detected by immunohistochemistry, Western-blot and PT-PCR. HUCMSCs were also transplanted into the seminiferous tubules of the busulfan-treated mice by microinjection. To assess the fate of HUCMSCs in the testis, the survival?Amigration and germ cell markers of the HUCMSCs in the infertility mice testis were detected by immunohistochemistry?A immunofluorescence.

Results
HUCMSCs can express some some molecular markers of germ cells after induction. Immunohistochemistry revealed that HUCMSCs can survive in the mice testis at least 120 days, and they can migrate from the lumens to the basement membrane then to lumens again. Immunofluorescence showed that HUCMSCs can go further differentiation in the mice favorable testicular environment, and express the germ cell markers.

Conclusions
These suggested that HUCMSCs may differentiate into male germ cell-like cells after induced by HMG and RA in vitro; and it could survive also in a favorable testicular environment, may differentiate into germ cell lineages. This finding may provide a new strategy for the treatment of male infertility.

Disclosures:
No relevant conflicts of interest to declare.
Distinct Mechanisms Regulate the Expression of flt3 Gene in Normal and Leukaemia-Like Stem Cells.
Authors: Volpe Giacomo , Walton David , Dassé Emilie , del Pozzo Walter , O'Neill Laura , Göttgens Berthold , Frampton Jonathan , Dumon Stephanie 
Published: October 2019

Abstract
Abstract 4586
The interaction between the tyrosine kinase receptor Flt3 and its ligand FL leads to crucial signalling during the early stages of the commitment of haematopoietic stem cells (HSCs). Consequent to mutations or over-expression, constitutive activation of the Flt3/FL pathway is also a key factor in the enhanced survival and expansion of leukaemic cells. Flt3 expression is in particular a feature of Acute Myeloid Leukemia (AML) as it is found in the vast majority of cases. As well as a powerful inducer of leukaemogenesis, Flt3 expression could therefore be viewed as a measurable consequence of the leukaemic transcriptional programme. Hence, understanding its transcriptional regulation would improve our knowledge of the molecular events underlying normal stem cell commitment and AML disease. Accounting for 40% of AML cases, normal cytogenetic AML (NC-AML) is a heterogeneous group of diseases that present a number of different alterations in transcriptional regulators of haematopoiesis, including mutations (MLL, C/EBPα, AML1, PU.1, GATA-1) or altered expression brought about by silencing (C/EBPα, PU.1) or activation (MN1, MLL, ERG1, Cdx2, Hox proteins). Here, we show that a substantial number of these transcription factors are implicated in the regulation of the flt3 gene, and by comparing cell line models of normal HSCs and AML, we pinpoint the importance of HoxA9, Meis1, C/EBPα and c-Myb, and remodelling of the epigenetic environment in the regulation of flt3 expression in leukaemic cells. C/EBPa mutations are found in 10% of AML cases and are considered to be associated with a good prognosis in the absence of Flt3 constitutive activation. We present a model linking C/EBPα and c-Myb activity to flt3 gene expression and FLT3 signalling.

Disclosures:
No relevant conflicts of interest to declare.