bluuebunny/crossref_metadata_2025_split huggingface dataset:
Allogeneic Hematopoietic Cell Transplantation (AHCT) for Primary Cutaneous T Cell Lymphoma (CTCL): a Center for International Blood and Marrow Transplant Research (CIBMTR) Review
Authors: Lechowicz Mary Jo , Agovi Manza , Carreras Jeanette , Lazarus Hillard M. , Laport Ginna G. , Montoto Silvia , Maloney David G , Hari Parameswaran 
Published: October 2019

Abstract
Abstract 364
Patients (pts) with advanced stage CTCL have a poor prognosis with standard therapy whereas AHCT possibly is curative. Published experience with AHCT for CTCL is modest. We report the outcomes of a large cohort of CTCL pts receiving AHCT (N=133) reported to the CIBMTR from 2000–2009. The majority of pts 129 (97%) were diagnosed with Mycosis Fungoides or Sezary Syndrome. Only 8 (6%) received AHCT in a complete remission (CR) while 46 pts (35%) never achieved CR pre-AHCT. Most pts (83%) were transplanted beyond 12 months from diagnosis. Median (range) time from diagnosis to transplant is 29 (4-206) months. Peripheral blood was the main graft source (83% vs. 14% marrow and 3% cord blood). Donor types included: 66 HLA-identical siblings (50%), 56 unrelated donors (42%), 11 from other related donors (8%). Reduced intensity conditioning (RIC) was used in 64%. Recipients of RIC were on average older (median age 51 vs. 44 yr for myeloablative) and transplanted in later study years. Among the myeloablative regimens, cyclophosphamide with total body irradiation was the most common (38%) whereas among RIC, fludarabine in combination with busulfan, cyclophosphamide or melphalan was used in 50%. GVHD prophylaxis regimens varied. Overall mortality at 30 and 100 days was 7% and 16%, respectively. Overall survival at 6 mo and 2 yr was 75% (95% CI 67–82%) and 44% (95% CI 34–53%), respectively. Conditioning intensity was not different statistically for the overall survival of these pts (figure 1). Treatment related mortality (TRM) at 6 mo and 2 yr was 10% (95% CI 5–17%). Progression/relapse of CTCL was 44% (95% CI 34–54%) at 6 mo and 58% (95% CI 47–68%) at 2 yr. Progression free survival at 6 mo was 46% (95% CI 36–57%) and at 2 yr 32% (95% CI 23–42%). The incidence of grade II-IV acute GVHD was 36% (95% CI 22–50%) and of chronic GVHD 31% (95% CI 19–45%) in a subset of reported pts. Infection as a cause of death does not seem to be increased in CTCL transplanted pts. Conditioning intensity did not impact TRM or the risk of progression. This very large series of allogeneic HCT demonstrates feasibility in pts with advanced CTCL with a low TRM and long term progression free survival in approximately one third of pts. Progressive disease was the primary cause of treatment failure in this cohort of pts with advanced disease.
Figure 1. Figure 1.

Disclosures:
No relevant conflicts of interest to declare.
Cytokine-Induced Killer Cells Facilitate Immune Reconstitution After Allogeneic BMT In Mice.
Authors: Mase Shintaro , Nishimura Ryosei , Kuroda Rie , Maeba Hideaki , Naka Kazuhito , Araki Raita , Ikawa Yasuhiro , Koizumi Shoichi , Yachie Akihiro 
Published: October 2019

Abstract
Abstract 3719
Cytokine-induced killer (CIK) cells are ex vivo–expanded T lymphocytes expressing both natural killer (NK)– and T-cell markers. We have reported that adoptive transfer of allogeneic CIK cells in a murine model caused minimal graft versus host disease (GVHD) with retention of antitumor activity mediated by NKG2D, which is an activating receptor expressed on NK cells. With the purpose of potential application of CIK cells in a clinical hematopoietic stem cell transplantation, the problem we have to consider next is whether CIK cells could promote an engraftment and facilitate an immune reconstitution. To this end, lethally irradiated BALB/c mice were injected with minimal number of MHC incompatible C57BL/6 bone marrow (BM) cells alone, in which almost half of mice died because of graft failure, or with CIK concurrently. The mice receiving BM plus CIK cells survived with 93% without GVHD, demonstrating that CIK cells significantly promote an engraftment (P<0.05). In particular, recovery of CD3+ T-cells was significantly faster (p<0.05) in the mice receiving BM plus CIK cells than those receiving BM cells alone. Next we further evaluated whether CIK cells also promote an engraftment in the mice receiving non-myeloablative conditioning using low dose total body irradiation. As expected, CIK cells promoted an engraftment and favored immunoreconstitution, especially from the early time point after BMT. In conclusion, our study clearly demonstrated that CIK cells promote an engraftment and facilitate an immure reconstitution without GVHD. As recent studies demonstrated that sufficient number of CIK cells could be expanded even from washouts of cord blood units bags, infusion of CIK cells would be a potent strategy for preventing a graft-failure in clinical settings, especially after cord blood transplantation.

Disclosures:
No relevant conflicts of interest to declare.
Augmented Regulatory T Cell Response After Photochemical Treatment Alleviates Acute Graft-Versus-Host Disease and Improves Survival.
Authors: Kanathezhath Bindu , Larkin Sandra K , Kaplan Julika , Walters Mark C. , Kuypers Frans 
Published: October 2019

Abstract
Abstract 3740
Graft-versus-Host disease (GVHD) causes significant morbidity and mortality after allogeneic transplantation. Regulatory T cells of both donor and host origin can limit GVHD. Acute GVHD early post transplantation is heralded by a cytokine storm induced by a dominant T helper type 1(Th1) response, which damages host tissues like skin, gut, liver and lungs. We hypothesized that co-transplantation of photochemically (S-59) treated T cells modulates T cell effecter subsets and their cytokine milieu, and thereby reduces acute GVHD. To this end, we transplanted whole bone marrow cells in a major histocompatibility complex (MHC) antigen mismatched murine model using marrow with T cells from C57BL/6J AKR (acute GVHD model), with and without S-59 treatment. We observed equivalent elevation of CD4+Th1 (IL-2 and IFN-g), cytotoxic CD8+ (FAS, IFN-g and TNF-a) and CD4+Th2 (IL-4, IL-5, IL-6, IL-13) cytokines in recipients of both groups during the first week after transplantation. While the Th1 cytokines persisted as long as 10 days after transplantation, there was a shift to a regulatory T cell (Treg) cytokine profile (Transforming growth factor b [TGF-b] and IL-10) in the group that received the S-59 treated T cells. TGF-b and IL-10 levels were higher in the peripheral blood and bone marrow of the study group compared to controls (table). This was accompanied by the appearance of FoxP3High CD4+ CD25+ Tregs in the spleen and CD4+ Th17 cytokine (IL-17) elevation in the thymic compartment of recipients that received S-59 treated T cells (mean-28.44pg/ml versus 1.45pg/ml, p=0.0059). In-vivo tracking of S-59 treated T cells demonstrated the disappearance of these cells in the peripheral blood, spleen, bone marrow and thymus by 48 hours after transplantation. Nonetheless, we noted that recipients of S-59 treated T cells had significantly less acute GVHD and better overall survival (p=0.0001). In summary, our experiments indicate that there is an initial dominance of inflammatory and CD4 Th1 cytokines immediately post transplantation. Co-transplantation of S-59 treated T cells shifts the effecter CD4 T cell profile to resemble a Treg phenotype. Despite the absence of circulating photochemically treated T cells, significant alterations in the recipient cytokine milieu persisted after transplantation. Thus, S-59 treated T cells appear to exert an important immunomodulatory effect to ameliorate GVHD and improve survival after MHC-mismatched allogeneic transplantation.
Blood (pg/ml) Bone marrow (pg/ml) Thymus (pg/ml) TGF-b [S59] 8482.7 * 7062.1 2926.9 TGF-b [control] 1.1 301.55 46.5 IL-10 [S59] 419.0 ** 174.0 30.4 IL-10 [control] <1 82.1 33.44 ρ value- * 0.0001 ** 0.0006

Disclosures:
No relevant conflicts of interest to declare.
Gpr171, a Putative P2Y-Like Receptor, Counteracts Monocyte Lineage Specification When Over-Expressed In Murine Hematopoietic Progenitors.
Authors: Rossi Lara , Lemoli Roberto M. , Goodell Margaret A. 
Published: October 2019

Abstract
Abstract 3758
Despite intense investigation, the molecular mechanisms underlying the multi-step process that leads from committed progenitors to terminally differentiated cells are still poorly understood. Due to the capability of responding to a variety of chemically different compounds, G protein-coupled receptors (GPRs) have been shown to play a pivotal role in regulating the interaction of hematopoietic cells with the surrounding microenvironment. A novel GPR gene, Gpr171, encodes an orphan receptor putatively related to the family of purinergic receptors (P2YRs) for extracellular nucleotides, a group of mediators previously shown to be involved in the regulation of HSCs. Gpr171 has been indicated as a putative, novel member of the family of P2Y receptors, but no information is currently available on the ligand responsible for its activation. Likewise, its functional role in biological systems remains completely unknown.
By sequence analysis, we reconstructed Gpr171 phylogenesis in mice. We explored the functional role of Gpr171 in the hematopoietic system by transducing Sca-1+ hematopoietic progenitor cells (HPCs) with MSCV bicistronic retroviral vectors (thus driving the enforced expression of Gpr171 along with GFP). Transduced HPCs were analyzed both in vitro and in vivo.
By phylogenetic analysis, we confirmed that Gpr171 is evolutionally related to members of a P2Y gene-cluster localized on mouse chromosome 3. As a first step towards unveiling a role for Gpr171, we investigated its expression profile and function in the murine hematopoietic system. As opposed to other P2YRs, Gpr171 emerged to be significantly repressed in cells undergoing myeloid differentiation. We therefore hypothesized that Gpr171 might be involved in repressing monocyte lineage-specification. Consistent with our assumption, we observed a decreased expression of myeloid markers upon induction of Gpr171 in hematopoietic progenitors (HPCs), as well as a dramatic reduction in the capability of generating myeloid colonies in vitro. Furthermore, mice transplanted with HPCs over-expressing Gpr171 display a significant reduction in the percentage of Mac-1+Gr-1− monocytes.
Our findings establish, for the first time, Gpr171 biological function and unveil its unique role in repressing monocyte differentiation. Gpr171 enforced expression induced a significant decrease in the proliferation rate of myeloid cells, as well as a reduction in the expression of monocyte-specific markers, both in vitro and in vivo. Our findings contribute to the delineation a peculiar evolutionary history for Gpr171 as compared to other members of the P2Y family, suggesting a rather unique role in counteracting myeloid differentiation.

Disclosures:
No relevant conflicts of interest to declare.
Olfactomedin 4 Down-Regulates Neutrophil Killing of Gram-Positive and Gram-Negative Bacteria.
Authors: Liu Wenli , Yan Ming , Liu Yueqin , Coleman William G , Rodgers Griffin P. 
Published: October 2019

Abstract
Abstract 3777

Introduction:
Olfactomedin 4 (OLFM4) is a member of olfactomedin-related glycoprotein family, which is specifically expressed in neutrophils and gastrointestinal tract. OLFM4 expression is upregulated in gastrointestinal cancer and inflammatory diseases such as chronic inflammatory bowel disease and Helicobacter pylori infection. It has been shown that OLFM4 is a target gene of NF-kB and Notch pathways and that OLFM4 down-regulates innate immunity to H. pylori infection. However, its potential biological functions in neutrophils still remain to be defined. The goal of this study is to determine whether OLFM4 is involved in the bactericidal activity of neutrophils using an OLFM4 deficient mouse model.

Results:
1. OLFM4 expression in neutrophils is upregulated in response to Staphylocococus aureus (Gram positive) and Escherichia coli (Gram negative) bacteria. 2. We have shown that neutrophils from OLFM4 deficient mice have increased intracellular killing of S. aureus and E. coli bacteria in vitro. 3. The OLFM4 deficient mice displayed enhanced bacterial clearance in vivo when the mice were challenged with intra-peritoneal injection of S. aureus and E. coli. 4. To elucidate the molecular mechanisms that mediate these effects, we performed a yeast 2-hybrid screen and found that OLFM4 interacts with cathepsin C (dipeptidyl peptidase I or DPPI), a lysosomal cysteine protease that has a degraditive function as an exopeptidase and is essential for activation of neutrophil granule-associated serine proteases including neutrophil elastase, cathepsin G and proteinase 3. The direct association of OLFM4 with cathepsin C was confirmed in human primary neutrophils. 4. We have demonstrated that OLFM4 is a direct substrate of DPPI and inhibits DPPI activity in transfected 293T cells. 5. The cathepsin C activity in neutrophils from OLFM4 deficient mice was significantly higher than that in neutrophils from wild-type littermate mice in the absence or presence of bacterial infection, suggesting that OLFM4 is an endogenous inhibitor of cathepsin C in neutrophils. 6. We have also demonstrated increased activities of neutrophil elastase, cathepsin G and proteinase 3 (whose processing and maturity require cathepsin C activity) in OLFM4 deficient neutrophils compared with wild-type neutrophils. 7. Activation of NADPH oxidase, myeloperoxidase (MPO) activity, and neutrophil phagocytosis were not altered in OLFM4 deficient neutrophils compared with wild-type neutrophils.

Conclusion:
These results suggest that OLFM4 is an important regulator of neutrophil bacterial killing activity via negative regulation of cathepsin C activity and its down stream granule-associated serine proteases.

Disclosures:
No relevant conflicts of interest to declare.
Adult Langerhans Cells Derive Directly From Embryonic Macrophages.
Authors: Bertrand Julien , Wittamer Valérie , Traver David 
Published: October 2019

Abstract
Abstract 3788
Dendritic cells (DCs) represent the most potent antigen-presenting cells in vertebrates. Upon immune challenge, DCs can uptake antigens and migrate to lymphoid organs to activate naïve T cells. Therefore, understanding their biology is important for designing new treatments. Langerhans cells (LCs) are a subset of DCs residing in the adult epidermis. In adult teleosts, many evidences point to the existence of DCs, but no tools have been generated to enable in vivo imaging, prospective isolation and functional analyses of this subset.
By combining two new transgenic lines, cd45:DsRED and mhc2dab:eGFP, we could isolate mononuclear phagocytes (macrophages/DCs) in the CD45+DAB+ fraction from adult tissues. Zebrafish DC-like cells displayed cytological features that resembled those of mammalian DCs. In the skin, the majority of CD45+DAB+ cells exhibited a typical morphology of LCs. Thus, we can isolate a pure population of LCs from the adult skin.
Most DC subtypes are constantly replenished from hematopoietic stem cell derived progenitors. However, recent studies indicate that LCs self-renew independently from the bone marrow in steady-state conditions, suggesting the existence of a local progenitor in the skin. Recent reports showed that mouse LCs derive from embryonic myeloid precursors that seed the skin before undergoing proliferation. By combining imaging and gene expression analyses, we could show that mhc2dab is first expressed in myeloid cells (CD45+) at 7 days post fertilization (dpf). Accordingly, we can first observe CD45+DAB+ cells at 8 dpf, in the skin of developing embryos, suggesting that LCs derive from early CD45+ cells. In order to unravel their origin, we traced CD45+ cells by using the tamoxifen-inducible cd45:CRE-ERt2 transgenic line, combined to a switch reporter that allows us to permanently label cells. This switch-reporter consists of the b-actin promoter driving the expression of DsRED after a STOP cassette is excised by the CRE recombinase. By exposing double-transgenic embryos to tamoxifen between 20 and 26 hpf, we could induce CRE expression in primitive macrophages only. Several weeks after tamoxifen treatment, we could identify the progeny of primitive macrophages in the skin, showing LC morphology. This strongly suggests that primitive macrophages can persist to adulthood and contribute to the LC network. Additional experiments will be required to prove if they are the only source of LCs in steady-state conditions.

Disclosures:
No relevant conflicts of interest to declare.
First Report and Results From VSCO.NET, the Virtual Society of Clinical Oncology, An Independent Internet-Based Haematology/Oncology Platform.
Authors: Weihrauch Martin R , Brinker Florian , Breuer Jonas 
Published: October 2019

Abstract
Abstract 3832

Introduction.
VSCO.NET, the Virtual Society of Clinical Oncology was founded as an independent internet platform for healthcare professionals working in the field of hematology and oncology. The aim is to bundle all accessible knowledge and experience of the oncology community in one place and to grant an open, free-of-charge source of information.

Methods.
The concept was developed by hematologists/oncologists and was realized together with a team of web programmers. The software was developed in HTML, PHP and JavaScript and runs on a Linux web-server. To enable an internationally accepted platform, VSCO.NET offers its pages in English, Spanish, French and German.

Results.
VSCO.NET went public in December 2009 in a beta-stage with the following functions: 1. a comprehensive chemotherapy order system with over 100 included chemotherapy protocols, 2. staging tools for Hodgkin's and Non-Hodgkin's lymphoma, multiple myeloma, CLL, and CML, 3. texts and tools for oncological emergencies, 4. a knowledge database/file repository for information texts, presentations and pictures, 5. a news channel with oncological abstracts, and 6. a virtual tumor board (discussion forum). Currently, over 400 physicians, mostly hematologists/oncologists, have registered with VSCO.NET. The most frequented tool is ChemoBOS, the chemotherapy order system, which quickly generates chemotherapy orders for patients. Due to its open and free protocol template generator, many physicians have created their own adapted chemotherapy protocols, which can be used by the entire VSCO.NET community. Also, the staging tools were generally well accepted and have a fairly high usage. However, the virtual tumor board as a way to communicate about difficult cases has not been used so far, which could show a reluctance of physicians to discuss their patient cases in an internet-based forum. It should be noted that many physicians registered from developing countries, who cannot afford expensive chemotherapy software and now have the opportunity to improve patient care at their centers.

Conclusions:
As the first internet-based society for hematology/oncology with a wide variety of interactive functions, VSCO.NET shows great potential to internationally connect physicians, expand the accessibility of oncological knowledge and to facilitate the daily clinical routine.

Disclosures:
No relevant conflicts of interest to declare.
Exportin 7 (RanBP16) Plays An Essential Role In Terminal Erythroid Differentiation
Authors: Hattangadi Shilpa , Burke Karly , Lodish Harvey 
Published: October 2019

Abstract
Abstract 3877
Members of the nuclear transport receptor family of importins and exportins regulate the passage of proteins between the nucleus and cytoplasm. Although evolutionarily conserved across several species, Exportin 7 (Xpo7 or RanBP16) and its cargo are not well understood. In our study, we find that Xpo7 is highly erythroid-specific, as all other exportins are downregulated during terminal erythroid differentiation, a process including the induction of a highly specialized erythroid expression program, a set number of 3–5 terminal cell divisions, and chromatin condensation and eventual enucleation. Xpo7, in contrast, is highly induced during terminal erythropoiesis. Using retroviral shRNA knockdown of Xpo7 in in vitro fetal liver erythroid cell cultures, we demonstrate that exportin 7 is necessary for normal cellular proliferation and terminal erythroid differentiation, specifically for normal enucleation. Through microarray and biocomputational analysis of mRNA isolated from the knockdown cultures, we have found that the promoters of genes that are dysregulated after Xpo7 knockdown are enriched for binding sites for the activating transcription factor 4 (ATF4). Given that the erythroid phenotype of the ATF4 knockout mouse is very similar to the specific erythroid defects we observe in our in vitro knockdown analysis, our data suggests that either ATF4 or its binding protein may be Xpo7's cargo during terminal erythroid differentiation. Ongoing studies aimed at confirming this mechanism, the interaction between ATF4 and Xpo7, and the role and cargo of Xpo7 in terminal erythroid differentiation, are underway.

Disclosures:
No relevant conflicts of interest to declare.
Expression of the c-Met Oncogene Correlates with Favorable Progression Free Survival In Classical Hodgkin Lymphoma
Authors: Xu Chuanhui , Van Den Berg Anke , Plattel Wouter , Ruther Nele , Xin Huang , Wang Miao , Van Imhoff Gustaaf , Viardot Andreas , Möller Peter , Poppema Sibrand , Visser Lydia 
Published: October 2019

Abstract
Abstract 3880

Background
The HGF/c-Met signaling pathway regulates a variety of biological processes, including proliferation, survival and migration. Aberrant c-Met activation has been implicated in the pathogenesis of many human cancers. Furthermore, c-Met is of prognostic significance in several types of cancer such as diffuse large B cell lymphoma, bladder cancer, breast cancer, colorectal cancer and ovarian cancer. Expression of c-Met in Hodgkin lymphoma (HL) patients, as well as increased levels of HGF in the urine of HL patients has been reported, but no prognostic studies or functional data have been reported.

Methods
We studied the prognostic significance of c-Met expression by immunohistochemistry on paraffin sections of classical HL (cHL) patients from two independent patient cohorts. Functional studies were performed on HL cell line L428 that showed high c-Met levels, constitutive phosphorylated c-Met and no HGF expression. The effect of stimulation by HGF and inhibition by c-Met kinase inhibitor SU11274 was investigated by Western blot, cell proliferation and cell cycle progression analysis.

Results
Expression of c-Met was detected in Hodgkin Reed-Sternberg (HRS) cells in 52% (79/152) of primary cHL tissue samples and expression of its ligand, hepatocyte growth factor (HGF), was detected in HRS cells in 8% (10/120) of the cHL patients. Co-expression of HGF and c-Met in HRS cells was observed in only 3% (4/120) of cHL patients. A variable percentage of infiltrating cells stained positive for HGF, supporting a predominant paracrine activation route. In contrast to its adverse prognostic impact in other cancers, high c-Met expression significantly correlated with favorable 5 year progression free survival in both patient cohorts. To explain these unexpected findings we studied the c-Met/HGF signaling pathway in the L428 cHL cell line. The levels of phosphorylated c-Met, Akt, and Erk1/2 were upregulated upon HGF stimulation and this induction could be blocked by inhibiting c-Met activation with SU11274. Activation with HGF did not effect cell growth, while SU11274 alone suppressed cell growth. SU11274, as well as inhibitors of PI3K, MEK1/2 and Erk1/2 (downstream targets of the HGF/c-Met signaling pathway) induced G2/M cycle arrest.

Conclusion
Expression of c-Met in tumor cells was observed in 52% of cHL patients. In contrast to its prognostic value in other cancers, expression of c-Met in HRS cells of cHL patients correlated with favorable prognosis in two independent cohorts. Although triggering of c-Met in L428 cells with HGF induced activation of its downstream factors, no effect was observed on proliferation. Remarkably, the c-Met inhibitor did suppress cell growth of L428 cells.

Disclosures:
No relevant conflicts of interest to declare.