bluuebunny/crossref_metadata_2025_split huggingface dataset:
Successful Treatment with Intrathecal Tocilizumab(IL-6 antibody) for 2 Cases of Graft-Versus Host Disease of the Central Nervous System after a Second Allogeneic HSCT from Haploidentical Donor
Authors: Gui Ruirui , Zhao Juanjuan , Zu Yingling , Han Lu , Zhang Yanli , Yu Fengkuan , Li Zhen , Zhao Huifang , Fang Baijun , Lin Quande , Gao Quanli , Song Yongping , Zhou Jian , Liu Delong 
Published: October 2019

Abstract
Graft-versus-host disease (GVHD) of central nervous system (CNS) is a rare complication after allogeneic hematopoietic cell transplantation (allo-HSCT), which is easily misdiagnosed and has limited treatment and poor efficacy. In this study, we reported two children(1 case of severe aplastic anemia and 1 case of acute myeloid leukemia) who were subjected to the secondary haploid hematopoietic stem cell transplantation of another door after the first implantation failure. Two patients developed epileptic seizures during hematopoietic recovery. Peripheral blood and cerebrospinal fluid cytokines were detected and diagnosed as acute GVHD in the central nervous system. The cytokine elevation is mainly IL-6(IL-6 level in peripheral blood was 2276.25 pg/ml, and in cerebrospinal fluid was 2353.89pg/ml of case 1). We administered multiply consecutive plasmapheresis, intravenous infusion of tocilizumab combined with intrathecal injection of tocilizumab and dexamethasone. The epileptic symptoms of 2 cases are effectively controlled after the first dose. The IL-6 levels in peripheral blood and cerebrospinal fluid of both patients decreased significantly after 4(IL-6 level in peripheral blood was 6.74 pg/ml, and in cerebrospinal fluid was 195.72pg/ml of case 1) and 5 doses, respectively. Central nervous system aGVHD is prone to hematopoietic recovery after secondary transplantation; intrathecal injection of tocilizumab is safe and effective in the treatment of central nervous system aGVHD with elevated IL-6 mainly, which is worth further exploring.
Figure. Figure.

Disclosures
No relevant conflicts of interest to declare.
B-Cell Mediated Humoral Responses Are Impaired in Sepsis Survivors
Authors: Rana Minakshi , Sherry Barbara , Diamond Betty 
Published: October 2019

Abstract
Sepsis survivors exhibit a sustained compromise in innate immunity, and high mortality rates due to increased vulnerability to infections. Previously we demonstrated in a cecal ligation and puncture (CLP) model, that sepsis survivors had a reduced TNFα response to LPS-challenge in vivo, which is accompanied by an increase in CD4+ChAT+ T cells. These data together with clinical observations of low endogenous levels of IgG in sepsis survivors led us to hypothesize that T-cell dependent (T-D) B cell responses might also be altered after CLP. In order to assess alterations in the adaptive humoral response in sepsis survivors, we immunized C57BL/6J sham surgery and CLP mice with a single intraperitoneal (I.P.) injection of NP (29)-KLH (T-D antigen) in alum 4 weeks post-CLP (or sham surgery). We observed sepsis survivors showed reduced serum NP-specific IgG and IgG1 antibodies compared to sham controls. Furthermore, to confirm whether this reduced immune response is mediated through a T-D or T cell-independent (T-I) type II mechanism, we used other carrier molecules CGG (T-D) and Ficoll (T-I), respectively. To evaluate time dependent kinetics, we immunized mice with single I.P. injection of either NP (20)-CGG or NP (55)-Ficoll. We analyzed both low-affinity and high-affinity anti-NP antibodies through NP (23)- and NP (7)- BSA ELISAs, respectively. We found a significant reduction in anti-NP IgG and IgG1 antibodies in CLP mice post-NP-CGG immunization (p<0.01 and p<0.001, respectively) while the response was not diminished after NP-Ficoll immunization at day 7; suggesting that compromised antibody response is mediated through a T-cell dependent pathway. Flow cytometry data also revealed that the splenic frequency of NP-specific CD138+ intracellular IgG1 plasma cells was decreased in CLP mice immunized with NP-CGG as compared to sham mice at day 7, 14 and 21 (p<0.001, p<0.05 and p<0.05, respectively). Since follicular dendritic cells (FDCs) play a key role in B-cell activation and affinity maturation of antibodies, we performed immunofluorescence staining of FDCs in spleens from sham surgery and CLP mice. Interestingly, we found a decreased number of FDC clusters in CLP mice compared to sham controls. In summary, our study is clinically relevant as it may explain the hypogammaglobulinemia seen in sepsis surviving patients.

Disclosures
No relevant conflicts of interest to declare.
Personalized Prophylaxis in Hemophilia a: A Pediatric Center Experience in Mexico
Authors: Aquino Efrain 
Published: October 2019

Abstract
Summary. Prophylaxis is the current standard of care for Hemophilia patients. However, the regimen should be individualized as much as possible based on age, bleeding phenotype and physical activity. PK based regimens have been proposed as an effective way to achieve more successful outcomes. According to 2016 WFH data, Mexico has 1.425 IU/capita mean use, which barely overcomes the limit of 1IU/ minimum target to achieve survival of the hemophilia population (WFH Report on the Annual Global Survey). Objectives. To examine a Mexican Center experience obtaining limited sampling PK profiles in pediatric patients. To determine the difference in frequency dosification in standard prophylaxis vs PK guided. Methods. Limited sampling was done in 14 pediatric patients with Hemophilia A without inhibitors, moderate to severe, in prophylaxis aged 2 to 14 years old using population PK tool myPKFiT. PK profiles based on the same data points plus patient characteristics were generated by a Bayesian algorithm with myPKFiT. Results. The mean age was 5.5 years (range 1-14 years) with a mean standard dose of 35 IU/k 2 times a week. The PK based mean dose was 27IU/kg 4 times a week. The mean half-life was 9.5 hours (range 6.8-11.6 hours. Conclusions. Shorter half life should be expected in pediatric patients, and be taken into account with other variables to achieve a personalized prophylaxis. Tailored prophylaxis facilitate patient education and adherence. Pediatric PK suggest shorter intervals or higher dosing are needed to achieve a desired target level. Use of myPKFiT and clinical, radiological and adherence scores to improve outcomes, in particular in the pediatric population. Integrating PK data into routine hemophilia care could provide promising results in developing countries.

Disclosures
No relevant conflicts of interest to declare.
Knockout Tumor Microenvironment HO-1 Neutralizes Myeloid-Derived Suppressor Cells and Enhances the Antitumor Effect of PD-1 Inhibition in Murine Models of Acute Myeloid Leukemia
Authors: Zhou Zhen , Fang Qin , Ma Dan , Wei Danna , Hu Xiuying , Liao Yudan , Wang Jishi 
Published: October 2019

Abstract
Background/Aims: Myeloid-derived suppressor cells (MDSCs) has been shown to be involved in tumor immune escape mechanisms and B cell-specific immunity in Acute myeloid leukemia (AML) patients. In our previous study shown that HO-1 protein have immunomodulatory activity including targeting of immune suppressor cells in the tumor microenvironment. Thus, we decided to assess whether HO-1 could enhance anti-PD-1 treatment and investigate those alterations in the immunosuppressive tumor microenvironment that contribute to the combined antitumor activity.
Method: We utilized C57/BL6 (HO-1 knockout) mouse were radiation with 6.5GY once in a row to repress residual immunity. Malignant tumor HL-60 cells respectively (1×107 cells) per animal were injected subcutaneously into the right abdomen. The xenograft mouse models of AML were euthanised on the 14th day after treatment with a PD-1 inhibitor once a day (20 mg/kg). The tumor volumes were measured and calculated by ruler. Survival curve of individual groups was evaluated from the first day of treatment until death using Kaplan-Meier curves.
Results: HO-1 gene knockout enhanced the antitumor effect of PD-1 inhibition in xenograft mouse models of AML by reducing tumor growth and increasing survival. HO-1 gene knockout inhibited the immunosuppressive function of both polymorphonuclear (PMN)- and monocytic-myeloid derived suppressor cell (M-MDSC) populations. Analysis of MDSC response to HO-1 gene knockout revealed significantly reduced arginase-1, iNOS, and COX-2 levels, suggesting potential mechanisms for the altered function. We also observed significant alterations in cytokine/chemokine release in vivo with a shift toward a tumor-suppressive microenvironment.
Conclusions: Our results demonstrate that HO-1 gene knockout enhances the antitumor effect of PD-1 targeting through functional inhibition of MDSCs and a transition away from an immune-suppressive tumor microenvironment.
[Key words] HO-1 gene knockout; PD-1 inhibitor; tumor microenvironment; MDSCs; AML

Disclosures
No relevant conflicts of interest to declare.
Does Neutrophil to Lymphocyte Ratio (NLR) Predict the Prognosis of Diffuse Large B-Cell Lymphoma?
Authors: Nakaya Aya , Fujita Shinya , Satake Atsushi , Nakanishi Takahisa , Azuma Yoshiko , Tsubokura Yukie , Saito Ryo , Konishi Akiko , Hotta Masaaki , Yoshimura Hideaki , Ishii Kazuyoshi , Ito Tomoki , Nomura Shosaku 
Published: October 2019

Abstract
Background: The international prognostic index(IPI) was widely used to predict aggressive lymphoma patients' outcome and to choose the best therapeutic treatment. However, this scale is unsatisfactory in identifying patients who would receive best benefit from rituximab containing regimens. Recently, NLR has been recognized as a poor prognostic indicator in various solid tumors. Here we quantify the prognostic impact of NLR in de novo DLBCL patients. However, various studies of the usefulness of the NLR have used different cut-off values, and the methods of selecting these NLR cut-offs were unclear. Therefore, we verify the adaptive cut-off value.
Methods: We retrospectively analyzed 543 patients with de novo DLBCL who diagnosed at Kansai Medical University Hospital and Kansai Medical University Medical Center from January 2003 to December 2017. The prognostic value of NLR at diagnosis was assessed. We put the cut-off of NLR; 3, 4, 5, 6, and evaluate which the most predictive cut-off value is.
Results: The median age was 69(20-95) years old, and male was 59%. The Ann Arbor stage III and IV was 60%. The proportion of patients with IPI, Low, Low-intermediate, High-intermediate, High was 36%, 19%, 21%, 24%, respectively. The optimal cutoff for NLR was 6. NLR(6) was associated with overall survival(OS)(HR 1.76, 95%CI: 1.23-2.51, p= 0.002) and progression free survival(PFS) (HR 2.66, 95%CI: 1.65-4.28, p<0.001), either. Multivariate analysis identified NLR(6) remained as a significant factor affecting PFS (hazard ratio: 1.53, 95%CI: 1.02-2.29, p=0.041).
Conclusion: Our result revealed that the optimal cut-off for NLR was 6. NLR predicted both OS and PFS of the patients with de novo DLBCL. NLR is a simple and useful scale and it might be a useful marker for prediction of patients' survival or disease progression in patients with de novo DLBCL.

Disclosures
Ito: Mundipharma: Honoraria; Celgene: Honoraria; Bristol-Myers Squibb: Honoraria, Research Funding; Novartis Pharma: Honoraria; Takeda: Honoraria; Pfizer: Honoraria.
Sumoylation of Pyruvate Kinase M2 Inhibits Myeloid Differentiation in Hematopoietic Cells
Authors: Xia Li , Wang Xin-Ran , Wei Ran , Yan Jin-Song , Chen Guo-Qiang , Lu Ying 
Published: October 2019

Abstract
The pyruvate kinase (PK) is a rate-limiting glycolytic enzyme catalyzing the dephosphorylation of phosphoenolpyruvate to pyruvate. M2 form of PK (PKM2) is expressed during embryogenesis and is the predominant form in tumors of different types. In contrast to the essential role of PKM2 in solid tumors, much less is known about the effects of PKM2 in hematopoietic cells and the development of leukemia. Here we found that PKM2 is modified by small ubiquitin-like modifier 1(SUMO1), which can be reduced by a SUMO1-specific protease SENP1 in hematopoietic cells. SUMOylation induced nuclear localization and conformation change from tetramer to dimer of PKM2. Importantly, SUMOylation of PKM2 is prevalent in a variety of leukemic cell lines as well as primary samples from patients with hematologic malignancies. In consistency, predominant nuclear localization and dimeric forms of PKM2 in leukemic cells were observed. Using in vitro SUMOylation reaction-coupled liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS), we identified K270 lysine residue of PKM2 as the SUMOylation target. Replacement of endogenous PKM2 with mutant PKM2K270 showed a significant shift of PKM2 from tetramer to dimer. To investigate the potential leukemogenic effect of PKM2 SUMOylation, murine hematopoietic progenitor 32D clone 3 (32Dcl3) transfectants expressing wild type(WT) or mutant PKM2K270 were generated and G-CSF-induced differentiation was evaluated by morphology appearance and expression of myeloid associated surface markers CD11b and Gr-1. The results showed that expression of WT PKM2 but not mutant PKM2K270 significantly blocked myeloid differentiation. Further investigations revealed that SUMO1 modification of PKM2 at K270 is essential in mediating the interaction between PKM2 and Runt-related transcription factor 1(RUNX1), a master transcriptional factor implicated in the differentiation of hematopoietic cells. This interaction led to a downregulation of RUNX1 during G-CSF-induced myeloid differentiation of 32D cells, which could be abrogated by expression of mutant PKM2K270. Collectively, these data indicated that SUMOylated PKM2 blocks myeloid differentiation through suppressing RUNX1. These findings reveal a novel nonmetabolic function of PKM2 in modulating myeloid differentiation and highlight the critical role of SUMOylation in leukemogenesis.

Disclosures
No relevant conflicts of interest to declare.
Minit-Livin Conjugated to Targeted Nanoparticles Inhibits Tumorigenicity
Authors: Abd Elrahman Ihab , Nassar Taher , Khairi Noha , Perlman Riki , Benita Simon , Ben-Yehuda Dina 
Published: October 2019

Abstract
We found that Livin, an Inhibitor of Apoptosis Protein, is specifically cleaved to produce a truncated Livin protein (tLivin) and demonstrated the paradoxical pro-apoptotic activity of tLivin. mini-tLivin (mtLivin) is a 70 aa derivative of tLivin that remarkably, exhibits a pro-apoptotic activity as potent as tLivin. Our findings regarding tLivin and mtLivin suggest unique therapeutic avenue against cancer. To improve the delivery and efficacy of mtLivin for treatment we conjugated mtLivin-to nanoparticles (NPs) of poly (lactide-co-glycolide) (PLGA) and targeted the nano-delivery system to diffuse large B-cell lymphoma (DLBCL) using CD40L. Delivery of mtLivin with nanoparticles should enhance stability, provide prolonged bioavailability, intracellular uptake and efficacy.
We generated bifunctional mtLivin-CD40L-NPs to target mtLivin-NPs to DLBCL cells. Various cell lines were used to evaluate the biological activity of free and conjugated (targeted or not) mtLivin including 293T, 721.221 B cells, L428 Hodgkin's lymphoma, OCI-LY19 DLBCL cells and OCI-LY19- GLuc cells that constitutively express a gaussia luciferase reporter.
Targeted, bifunctional mtLivin-CD40L-NPs elicited significant cell death of DLBCL cells while monofunctional mtLivin-NPs had a lower effect. mtLivin-NPs may induce cell death of some cell lines without targeting (e.g. 293T) yet targeting is required to induce marked cell death of DLBCL cells.
To best mimic lymphoma disease in humans, a disseminated lymphoma model was used to evaluate the antitumor effect of targeted mtLivin treatment. NOD/SCID mice were injected with OCI-LY19- GLuc cells or into the tail vein. Gaussia luciferase reporter allows IVIS imaging to monitor tumor size and location in mice. Mice were treated approximately once a week with various mtLivin-NPs formulations. Luciferase signal was observed as early as day 2 after tumor cells injection.
Immunohistochemistry for anti-CD20, a human B-cell marker, showed infiltration of OCI-ly19-Gluc cells in spleen and brain close to tumor cells injection (Day 2). With time, lymphoma infiltrations were detected in brain, bone marrow, lungs, spleen and liver. All control mice exhibited paralysis and did not survive beyond 28 days. In contrast, 37.5% of animals receiving mtLivin-NPs and 71.4 % of animals receiving mtLivin-CD40L-NPs achieved complete pathological tumor response and survived significantly longer. All treatments in this study were well-tolerated.
In terms of mechanism, we show high caspase-3 activity in tumors from mice that were treated with mtLivin-CD40L-NPs demonstrating the ability of the nanoparticles to target the tumors and to induce apoptotic tumor cell death. The resistance of tumor cells to drug-induced apoptosis is a major cause of cancer treatment failure. We designed mtLitvin targeted nanoparticles constituting novel personalized therapeutic approach for resistant cancer.

Disclosures
No relevant conflicts of interest to declare.
Donor Lymphocyte Infusion Prevented Late Post-Transplant Rejection in 30 Patients with Thalassemia Major
Authors: Liao Jianyun , Liu Xiaoting , He Yuelin , Peng Zhiyong , Liu Huaying , Li Chunfu 
Published: October 2019

Abstract
OBJECTIVE:To investigate the role of donor lymphocyte infusions (DLI) in preventing post-transplant graft rejection (GR) in hematopoietic stem cell transplantation (HSCT) for patients with β-thalassemia major (TM);
METHODS: A total of 50 patients with TM who had mixed chimerism (MC) after HSCT from June 2011 to June 2016 was divided into 4 groups. The group 1 included the patients with grade I MC ratio of 90%-95% (n=20) but not DLI; Group 2 patients had same grade I MC ratio as Group 1 and received DLI (n=11); Group 3 had grade II MC ratio of 75%-90% (n=13); Group 4 had grade III MC of <75% (n=6). Group 1 received gradually reduced immunosuppressive agents; Groups 2 to 4 received DLIs of gradually increasing dose after reduction of immunosuppressive agents;
RESULTS: (1) Full donor chimerism (FDC) conversion rates in the groups 1 to 4 were 100%, 100%, 85%, and 66.7%, respectively; The incidence of graft versus host disease (GVHD) in groups 1 to 4 was 15%, 27.3%, 46.2%, and 33.3%, respectively; (2) There was no significant differeces as comparing incidence of GVHD in group 1 and in group 2 (15% vs. 27.3%, P = 0.638); (3) A significant difference was found in comparing patients receiving initial DLI at ≤180 and > 180 days and the incidence of GVHD was 50% vs 10% (P=0.049); (4) Comparing the accumulated doses of DLI >1×107/kg with ≤1×107/kg, the incidence of GVHD was 50% vs. 27.3% (P=0.142);
CONCLUSION: 1. The optimal choice for post-transplant MC grade 1 patient is only observation and decreasing the immunosuppressive drugs; 2. For grade 2-3 patients with high risk of rejection, the fractional increasing dose DLI can effectively reverse the unstable chimera to FDC; 3. Post-DLI GVHD was associated with DLI timing, and the incidence of GVHD was higher in patients who received DLI at ≤180 days after transplantation.

Disclosures
No relevant conflicts of interest to declare.
Sf3b1 Regulation of Jak/Stat Signaling Is Essential for Hematopoietic Stem Cell Formation
Authors: Bowman Teresa V. , Cameron Rosannah C. , Potts Kathryn S , McKinstry Mia , Gupta Varun , Bai Xiaoying 
Published: October 2019

Abstract
Hematopoietic stem cells (HSCs) maintain the hematopoietic system throughout the lifetime of an organism. During embryonic development, HSCs emerge through an endothelial-to-hematopoietic transition (EHT) from specialized hemogenic endothelial (HE) cells in the dorsal aorta. HSC fate specification depends on gene expression, which is the culmination of coordinated transcription, RNA splicing, and translation. Although transcriptional regulation of HSC fate choice is well studied, the regulatory role of RNA splicing in this process is poorly understood. Using zebrafish loss-of-function mutants for the spliceosomal component splicing factor 3b, subunit 1 (sf3b1), we identified that impaired splicing hindered HSC production. Surprisingly, we found that this constitutive splicing factor selectively regulates the fate of hemogenic endothelium while leaving the identity of closely-related non-hemogenic endothelium unperturbed. To identify Sf3b1-regulated transcripts important in EHT, we performed RNA-sequencing on purified kdrl:gfp+ endothelial cells from sf3b1 mutant and wild-type siblings at 24 hpf. Approximately 900 genes were mis-spliced, 144 of which were differentially expressed. Ingenuity Pathway Analysis identified Janus Kinase (Jak)/Signaling Transducer and Activator of Transcription (Stat) signaling, in particular Stat3, as one of the top perturbed pathways in the mis-spliced gene set. Stat3 is a transcription factor activated in response to several cytokine and inflammatory signals. To determine if altered splicing of stat3 was critical for HSC formation, we injected antisense splice-blocking morpholinos (MO) targeting the Sf3b1-sensitive stat3 exon19 into wild-type and sf3b1 heterozygous embryos, which normally generate equivalent levels of HSCs. We observed an impairment of HSC production in stat3 morpholino-injected sf3b1 heterozygotes, but not wild-type siblings, indicating a synthetic lethal interaction between sf3b1 and stat3. We also found that overexpression of a constitutively active form of Stat3 significantly suppressed the HSC defects in sf3b1 homozygous mutants. Together, these data indicate that Sf3b1-mediated splicing regulation of the Jak/Stat pathway is critical for HSC emergence.

Disclosures
No relevant conflicts of interest to declare.
The Molecular Genetic Analysis of Coagulation Factor XI (FXI) Deficiency in Two Chinese Patients
Authors: Zhou Rong-Fu , Li Qian , Zhou Min 
Published: October 2019

Abstract
Objective: To investigate the molecular pathogenesis of two coagulation factor XI (FXI) deficiency patients.
Methods: The diagnosis was validated by coagulant assays: APTT and correct test, PT, INR and coagulation factors activities. Coagulation factor activity were tested with clotting assay. The patients' DNA were extracted and all exons and flanking sequences of FXI gene were amplified using PCR. After purified, the products were sent for sequencing directly, the mutations were detected by comparing with wild sequences and analyzed using some bioinformatics software.
Results: The two patients were diagnosed as coagulation factor XI deficiency due to prolonged APTT and low activities of coagulation factor FXI. The results of APTT, FXI:C was 88.1s, 1.1% and 107.1s, 3.8% , respectively. Genetic analysis found that compound heterozygous mutations g.1251-1G > A and g.1271delT in the first patient and the sequencing results of TA plasmid clones showed that the two mutations were located on different single strands of chromosomes. Double heterozygous mutations g.1070A >G and g.1446C > G were detected in the second patient, resulting in Lys357Arg and Cys482Stop. Software analysis indicated the mutations probably brought amino acid sequence changed, protein features affected and splice site changed.
Conclusion: Compound heterozygous mutations g.1251-1G > A, g.1271delT and g.1070A > G , g.1446C > G had been identified in two coagulation factor XI deficiency patients, which might be the cause of their prolonged APTT and low FXI:C. To the best of our knowledge, the four mutations are reported for the first time in the literature.
Figure. Figure.

Disclosures
No relevant conflicts of interest to declare.