bluuebunny/crossref_metadata_2025_split huggingface dataset:
Extrafollicular CD4+ T and B Interaction Induces Chronic Gvhd in the Absence of Germinal Center Formation
Authors: Deng Ruishu , Hurtz Christian , Yue Chanyu , Xiao Gang , Yu Hua , Muschen Markus , Forman Stephen J. , Martin Paul J. , Zeng Defu 
Published: October 2019

Abstract
Chronic graft-versus-host disease (GVHD) in patients with allogeneic hematopoietic cell transplantation (HCT) is an immune-mediated syndrome. Both CD4+ T and B cells are required for disease induction, but details of this interaction have not been fully defined. Other groups have reported that germinal centers are enlarged in mice with chronic GVHD and that GC formation is required for induction of the disease. On the other hand, patients with chronic GVHD have a predominance of IgG1 in the serum with little evidence of IgG somatic hypermutation by one year after HCT, suggesting a lack of GC formation (Suzuki et al: Blood. 1996;87:1873-1880; Glas et al: Blood. 2000; 96:1064-1069). Consistently, we observed that GCs were not detectable in mice with chronic GVHD, including MHC-matched C57BL/6 (H-2b) recipients given LP/J (H-2b) grafts and BALB/c (H-2d) recipients given DBA/2 (H-2d) or B10D2 (H-2d) grafts, and MHC-mismatched BALB/c (H-2d) recipients given C57BL/6 (H-2b) grafts. Furthermore, BALB/c recipients given grafts from BCL6-deficient (BCL6 fl/fl Mb1-Cre) C57BL/6 (H-2b) donors lacking the ability to form GC developed chronic GVHD with similar severity to recipients given control BCL6-sufficient grafts. Consistent with lack of GC formation, recipients of BCL6-deficient grafts had profound reduction of serum IgG2b and IgG2c but moderate reduction of IgG1 as compared to recipients given control BCL6-sufficient grafts. The presence of IgG1 indicates extrafollicular CD4+ T and B interaction, which is Stat-3 dependent. Further experiments showed BALB/c recipients given grafts from Stat3-deficient (Stat-3 fl/fl CD4-Cre) C57BL/6 donors did not develop chronic GVHD. Taken together, these results indicate that GC formation is not required for development of chronic GVHD and that extrafollicular CD4+ T and B interaction can induce chronic GVHD. This work is supported by NIH R01 AI066008 and The Beckman Research Institute Excellence Award (to D. Zeng).

Disclosures
Forman: Mustang: Research Funding; Amgen: Consultancy. Martin:Neovii: Research Funding; RegImmune: Research Funding; Enlivex: Consultancy; Janssen: Consultancy; Pfizer: Consultancy; Pharmacyclics: Consultancy.
Host Tissue PD-L1 Separates GVL Effect from Gvhd after Temporary Depletion of Donor CD4+ T Cells Early after HCT
Authors: Ni Xiong , Song Qingxiao , Deng Ruishu , Jin Hua , Cassady Kaniel M. , Zhang Mingfeng , Forman Stephen J. , Martin Paul J. , Wang Jianmin , Zeng Defu 
Published: October 2019

Abstract
Recipient tissue expression of programmed death-ligand 1 (PD-L1, B7H1) down-regulates graft-versus-host disease (GVHD) after allogeneic hematopoietic cell transplantation (HCT), but this information has not been translated clinically. Here we demonstrate a critical role for recipient PD-L1 expression in preventing both acute and chronic GVHD while preserving graft-versus-leukemia (GVL) effects when donor CD4+ T cells are temporarily depleted in vivo early after HCT. Depletion of donor CD4+ T cells increases serum IFN-γ concentrations and enhances recipient tissue expression of PD-L1 and donor CD8+ T cell expression of the PD-L1 receptors CD80 and PD-1. In GVHD target tissues, depletion of CD4+ T cells increases anergy and apoptosis of infiltrating CD8+ T cells in a manner that depends on recipient PD-L1 expression, thereby preventing damage to intestinal Paneth cells and stem cells, hepatocytes, and thymic medullary epithelial cells, and preventing both acute and chronic GVHD. In lymphoid tissues, depletion of CD4+ T cells augments CD8+ T cell proliferation without increasing CD8+ T cell anergy or apoptosis, resulting in expansion of donor CD8+ T cells and strong GVL effects. Therefore, temporary depletion of donor CD4+ T cells early after HCT represents a novel approach for preventing GVHD while preserving GVL effects. This work was supported by NIH R01 AI066008 and Nesvig Lymphoma Fundation (to D.Z.) and by NSFC 81090413, 81270638 (to J.W.).

Disclosures
Forman: Amgen: Consultancy; Mustang: Research Funding. Martin:Neovii: Research Funding; RegImmune: Research Funding; Enlivex: Consultancy; Janssen: Consultancy; Pfizer: Consultancy; Pharmacyclics: Consultancy.
Effect of Transfusion of the Third Party Umbilical Cord Blood on Haplo-Identical Hematopoietic Stem Cell Transplantation
Authors: Wang Jing Bo , Fei Xin Hong , Cheng Hao Yu , Yin Yu Ming , Zhang Wei Jie , Zhang Shu Qin , Li Qian , Xue Song , Gu Jiang Ying , He Jun Bao , Yang Fan , Liu Meng Qi , Zhao Jie , Liu Dai-Hong 
Published: October 2019

Abstract
Objective To retrospectively evaluate the effect of the third party umbilical cord blood on haplo-identical hematopoietic stem cell transplantation.
Methods From June 2012 to May 2015, 125 leukemia patients were enrolled, including 41 cases of ALL, 62 cases of AML, 12 cases of MDS, 7 cases of CML-BP, 2 cases of acute mixed leukemia and 1 case of Mother cell dendritic cell tumor. Inclusion criteria: 1) AL patients; 2) halpo-identical HSCT; 3) 3/6 matched cord blood was available. Patients were divided into two groups, ie. group A (HSCT, n=65) and group B (HSCT plus umbilical cord blood transfusion group, n=60). Myeloablative conditioning regimens consisted of BuCy, TBI/FLAG, TBI/Cy, and FLAG that followed by reduced-intensified BUCY. The median dose of mononuclear cells in group A and B were 8.58×108/kg and 9.01×108/kg, respectively. The median dose of CD34+ cells for transfusion in each group were 3.67×106/kg and 2.94×106/kg, respectively. The dose of grafted UCB MNCs and CD34+ cells for group B were 3.5×107/kg and 2×105/kg, respectively. All patients received cyclosporineA, MMF and methotrexate for GVHD prophylaxis. The endpoints of this study were hematological engraftment, incidence of acute and chronic GVHD, incidence of relapse, transplant-related mortality(TRM), non-relapse mortality(NRM), Overall survival(OS) and Disease-free survival(DFS) in each group.
Results The median follow-up time was 17(3-29) months in group A and 18(3-35) months in group B. Patients in group A reached a sustained ANC of more than 0.5*109/L at a median of 11 days, whereas 14 days in group B. Platelet more than 20*109/L occurred at a median of 19 days in group A, whereas 17 days in group B (P= .4). The rate of aGVHD was not significantly different in the two groups, 56.9% in group A and 48.3% in group B (P= .21). The accumulative incidence of II-IV grade aGVHD was 35.4% in group A and 30% in group B (P= .42). The incidence of chronic GVHD was 79.2% in group A and 71% in group B (P= .47). The incidence of extensive type was lower in group B, 69.2% Vs 35%, P=0.09. The incidence of CMV was lower in group B, 80% Vs 60% (P= .01). The accumulative incidence of EBV was lower in group B, 35.4% Vs 3.3% (P<0.01). At two years, the accumulative incidence of relapse was 24.4% in group A and 17.2% in group B, P=0.13. The two-year accumulative incidence of OS was 72.9% in group A and 84.8% in group B, P=0.07. The one-year accumulative incidence of DFS in each group were 65.7% and 79.4%, respectively, P=0.03. Conclusion Our clinical results have shown that HSCT with transfusion of the third party umbilical cord blood is a promising modality for induction of immunity reconstitution. Better survival results may benefit from lower incidence of GVHD and virus infection.

Disclosures
No relevant conflicts of interest to declare.
Trends in Financial Toxicity for Patients of a Community Cancer Clinic
Authors: Santhosh-Kumar Cherupolil R. , Struve Stephanie , Freier Sue 
Published: October 2019

Abstract
Background: Rising health care costs - especially those of new medications for cancer treatment - have caused a severe financial strain on patients, leading to significant alteration of lifestyles. Published information regarding financial distress is scant with data only from large tertiary care institutions that may not be relevant to smaller communities. We sought to analyze the extent of financial toxicity over time in a community cancer clinic staffed by two medical oncologists.
Methods: Since 2010, a team of two oncology social workers prospectively collected data on financial assistance for patients treated at our community cancer clinic. Data regarding medication assistance is available for 2012-2014.
Results: Approximately 500 new patients and 1600 unique follow-up patients are evaluated and treated at the clinic annually. From January 2010 through December 2014, 1,012 requests for assistance were processed. The number of requests increased from 134 in 2010 to 320 in 2014. Most common requests were for medications and financial aid. Trends are shown in the table. Though the number of patients applying for medication assistance increased in 2014, the amount requested was less than previous years because of the influx of specialty pharmacies in the market with their own assistance programs.
Conclusions: Financial toxicity for cancer patients is a real issue in the community clinic setting.Requests for financial assistance have increased over time. The number of FDA approvals for new cancer medications seem to correlate with the increase. Studies are in progress to further clarify the financial impact of cancer on patients.Table.Trends in requests for assistance and aid and FDA approval for new medications.Requests for20102011201220132014Financial aid6411686101143Service311112733Equipment1311111823Transport312357Medications51534449114Patient cost of medications for which aid requested$731,891$1,554,684$1,253,827FDA approvals for new Oncology medications61181513

Disclosures
No relevant conflicts of interest to declare.
Changes in Gene Expression during G-CSF-Induced Emergency Granulopoiesis in Humans
Authors: Pedersen Corinna Cavan , Borup Rehannah , Fischer-Nielsen Anne , Mora-Jensen Helena , Fossum Anna , Cowland Jack B. , Borregaard Niels 
Published: October 2019

Abstract
Emergency granulopoiesis refers to the increased production of neutrophils in bone marrow and their release into circulation induced by severe infection. Several studies point to a critical role for granulocyte colony-stimulating factor (G-CSF) as the main mediator of emergency granulopoiesis. However, the consequences of G-CSF stimulation on the transcriptome of neutrophils and their precursors have not yet been elucidated in humans. Here, we investigate the changes in mRNA and miRNA expression in successive stages of neutrophil development following in vivo administration of G-CSF in humans, mimicking emergency granulopoiesis. Blood samples were collected from healthy individuals after five days of G-CSF administration. Neutrophil precursors were sorted into discrete stages of maturation by flow cytometry and extracted RNA was subjected to microarray analysis. mRNA levels were compared to previously published expression levels in corresponding populations of neutrophil precursors isolated from bone marrow of untreated, healthy individuals. miRNA expression was investigated in the most mature cell population to determine G-CSF-induced changes in circulating neutrophils. G-CSF substantially affected mRNA and miRNA expression patterns, demonstrating significant impact on neutrophil development and function. 1110 mRNAs were differentially expressed more than 2-fold with G-CSF while the treatment induced changes in the levels of 73 miRNAs in the mature population. In addition, G-CSF treatment reduced the levels of four out of five measured granule proteins in mature neutrophils including hCAP-18, which was completely deficient in neutrophils from G-CSF-treated donors. Cell cycle analysis pointed towards an induced proliferative capacity of myelocytes. These results indicate that multiple biological processes are altered in order to satisfy the increased demand for neutrophils during G-CSF-induced emergency granulopoiesis.

Disclosures
No relevant conflicts of interest to declare.
Transcutaneous Immunization with a Solid Nanoscopic Imiquimod Suspension Enhances Tumor Rejection
Authors: Stein Pamela , Gogoll Karsten , Peters Tanja , Denny Mark , Tenzer Stefan , Stevanovic Stefan , Schild Hansjoerg , Theobald Matthias , Langguth Peter , Radsak Markus 
Published: October 2019

Abstract
Introduction:
Transcutaneous immunization (TCI) is a novel vaccination strategy to induce strong therapeutic cytotoxic T-lymphocyte (CTL) responses by directly targeting skin-resident professional antigen-presenting cells (APC). This vaccination approach is very promising to overcome current limitations of standard vaccination approaches that are mostly effective in prophylaxis, but not in the treatment of diseases. In this context, we have developed a TCI method based on a synthetic TLR7 agonist imiquimod that partial tumor protection in experimental rodent models. In our present work, we describe a novel optimized formulation of imiquimod in a solid suspension of crystalline imiquimod (IMI-Sol) that induces superior CTL responses and enhanced tumor protection.
Methods:
We determined imiquimod-penetration into the skin as well as systemic distribution by HPLC in C57BL/6 mice (at 6-12 weeks). For TCI, C57BL/6, Tlr7 (TLR7-/-) or Myd88 (MyD88-/-) gene deficient mice IMI-sol or imiquimod (in the commercially available formulation Aldara®) together with the CTL epitope SIINFEKL (derived from chicken ovalbumin) was applied on two consecutive days on the dorsum of the animals after hair removal by electric clippers. Induction of CTL responses was characterized after 7 days by the analysis of peptide-specific CTLs (ELISpot, tetrameric MHC I complexes) and in vivo cytolytic activity by flow cytometry. Tumor protection was assessed using the B16OVA tumor model.
Results:
Despite equal imiquimod penetration into the skin, we found superior vaccination capacity of IMI-Sol compared to Aldara® leading to enhanced tumor-protection. Using TLR7-/- or MyD88-/- mice, we confirmed that the induction of CTL responses with IMI-Sol was completely dependent on the TLR/MyD88 pathway. The enhanced immune response are due to more efficiently activated skin-derived DCs upon IMI-Sol treatment found in skin-draining lymph nodes.
Conclusions:
We developed of a superior imiquimod formulation for transcutaneous vaccination that boosts vaccination efficacy by the enhanced activation of skin-resident APCs leading to the induction of therapeutic CTL responses underscoring the importance of pharmaceutical aspects affecting vaccination potency beyond understanding of the underlying basic mechanisms. Combined efforts are necessary to establish TCI as a promising novel next generation vaccination platform that can be used for the treatment of persistent infections and cancer.

Disclosures
No relevant conflicts of interest to declare.
Identification of the Protein S Binding Site in the Factor IXa Serine Protease Domain Presents Therapeutic Possibilities
Authors: Plautz William E. , Sekhar Satish , Majumder Rinku , Sheehan John P. 
Published: October 2019

Abstract
Protein S (PS) has been shown to inhibit Factor IXa (FIXa); however, its binding sites on FIXa have not been fully elucidated. By structural analysis of FIXa in conjunction with previous studies of FIXa's ability to bind heparin, an FIXa inhibitor, we identified that multiple residues that make up the heparin binding exosite, including Lys126, Asn129, Lys132, Arg150, Arg165, and Arg170 (chymotrypsin numbering), might contribute to binding PS; these residues were substituted with either Ala or Leu in separate FIXa mutants. Factor Xa (FXa) generation assays along with fluorescence analyses provided both the catalytic efficiencies and dissociation constants for the FIXa mutants under conditions of varied PS. Furthermore, anisotropic measurements of the fluorescent DEGR-FIXa's binding interaction with PS, when titrated against each of the FIXa mutants, supported the data found in the previous assays. These studies suggested that multiple residues, which are critical for the binding of heparin, such as Lys126, Lys132, Arg165, and Arg170, are also required in binding PS. To additionally verify this data, further ex vivo studies, including co-immunoprecipitation and flow cytometry of PS and FIXa are underway. Although previous studies have shown that the first EGF-like domain of FIXa is required to bind PS, these novel results have not only localized the main inhibitory site of PS on FIXa, but also provided enough information to propose a binding mechanism, which may rely on the reciprocal association between the head groups of PS and FIXa with the other's EGF-like domains, forming a bridging structure upon the cell membrane. Further studies are required to verify this hypothesis, but when clarified, it could provide immense benefits in the production of therapeutics for thrombosis.

Disclosures
No relevant conflicts of interest to declare.
Super Cross-Presentation of Tumor Antigens to Elicit Anti-Lymphoma Immunity By Synthetic Design of an Anti-Phosphatidylserine Bridge Protein
Authors: Corey Daniel , Weissman Irving L 
Published: October 2019

Abstract
Cell loss by apoptosis is a common feature in tumors. Dying tumor cells induce immune tolerance within the tumor microenvironment largely through highly conserved homeostatic clearance programs that restore tissue immune homeostasis and contribute to the formation of an immunosuppressive niche. The translocation of phosphatidylserine (PS) on cellular membranes, during the initial phases of apoptosis, functions as a recognition and removal signal that limits the immunogenicity of cell death. We examined whether altering clearance of dying cancer cells to elicit inflammatory turnover potentiates immune responses against lymphoma cells. To remove inhibitory signals in the homeostatic clearance pathway we utilized a molecular bridge scaffold to engineer a modified phosphatidylserine bridge protein (FA58C2-hIgG1 or C2-hIgG1) that works as a bridge between apoptotic cells expressing aminophospholipids and phagocytes bearing Fc receptors. In vitro administration of C2-hIgG1 to murine bone marrow derived macrophages promotes engulfment of apoptotic murine lymphoma cells (38C13 cell line) and ablates the secretion of the anti-inflammatory cytokine interleukin-10 (IL-10) and suppression of pro-inflammatory cytokines tumor necrosis factor (TNF-α) and IL-12p40 to the presence of apoptotic cells. Similarly, uptake of C2-hIgG1 treated lymphoma cells triggers upregulation of the costimulatory markers CD80, CD86, and MHC class II on macrophages and promotes secretion of Th1-recruiting lymphocyte chemokines CXCL9, CXCL10, and CCL5. Accordingly, in vivo administration of C2-hIgG1 partially restores immune responses to dead lymphoma cells in antigen cross presentation assays and promotes recruitment and retention of tumor antigen specific CD8+ T cells, dendritic cells, and natural killer cells into tumors. These effects combine to elicit anti-lymphoma immunity, improve responses to immune checkpoint inhibitors, and enhance the effectiveness of adoptive T cell transfers using engineered T Cell Receptors (TCRs) but not CD19-directed chimeric antigen receptor engineered (CAR-T) T cells.

Disclosures
No relevant conflicts of interest to declare.
NEDD4-1 Modulates the Resistance of Multiple Myeloma to Bortezomib
Authors: Huang Xi , Zhang Enfan , Guo Xing , Chen Jing , Lin Xuanru , Chen Qingxiao , Yan Haimeng , Yang Li , Li Yi , He Jingsong , Huang He , Cai Zhen 
Published: October 2019

Abstract
Background: Multiple myeloma (MM) is among the most common hematologic malignancies. Proteasome inhibitor bortezomib (Bor) is one of the most effective drugs for treatment of MM. However, during long-term Bor treatment, MM cells may eventually develop acquired-resistance to Bor which results in recurrence and a poor prognosis of MM. Several researches show that E3 ubiquitin ligases (E3s) primarily determine the substrate specificity of ubiquitin proteasome system and play an essential role in Bor resistance of MM. NEDD4-1 E3s, a founding member of the Neural precursor cell-Expressed Developmentally Downregulated gene 4 (NEDD4) family, was proved to involve in the proliferation, migration, invasion of cancer cells and the sensitivity of anticancer therapies. Our current study aims to explore the role and underlying mechanism of NEDD4-1 in acquired resistance of Bor in MM.
Methods: The mRNA and protein levels of NEDD4-1 and its substrates in MM cell lines (H929, LP-1, RPMI8226, OPM-2 and ARP-1) and MM patients were detected by Quantitative Realtime PCR and Western Blotting. Lentiviral plasmids containing shRNA against NEDD4-1 were transfected into MM cells. Cell viability, proliferation and apoptosis of MM cells were measured by Cell Counting kit8 (CCK8) and flow cytometry. Gene array was used to compare the gene expression profiles of a panel of Bor treated MM cells vs vehicle-treated MM cells.
Results: Gene array showed NEDD4-1 was significantly increased in MM cells treated with Bor. MM cells (CD138+ plasma cells of the bone marrow) from refractory/recurrence patients expressed lower NEDD4-1 than primary patient myeloma cells. Also, MM cell lines H929, ARP-1, LP-1 highly expressed NEDD4-1 at mRNA and protein levels. RPMI8226 and OPM-2 were relatively low expressed. Cell growth assay displayed no significant difference in proliferation between the NEDD4-1 knockdown (KD) and the control group (P>0.05). After suppression of NEDD4-1 using shRNAs, the killing effect of Bor in MM was significantly weaker than the control group (P<0.05). We also found that PTEN was decreased in the NEDD4-1 KD H929 cell line. Otherwise, phospho-STAT3 (ser727) and oncoprotein c-Myc and Bcl-2 were upregulated.
Conclusion: Collectively, our study reveals that inhibition of NEDD4-1 can reduce MM sensitivity to Bor via regulating PTEN, c-Myc and Bcl-2, may be related to JAK/STAT signaling pathway, which suggests that NEDD4-1 probably acts as a novel drug target and therapeutic paradigm in the battle against multiple myeloma.

Disclosures
No relevant conflicts of interest to declare.