bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Novel Case of Co-Inheritance of Rare Hemoglobin Variant; Hb Rancho Mirage [βCd 143 (CAC→GAC) His→Asp] with Hb D-Punjab
Authors: Zaabi Eiman A. Al , Mahmoud Amira , Baysal Erol , Spence David G. 
Published: October 2019

Abstract
Abstract 4759

Introduction
Co-inheritance of Hb Rancho Mirage and Hb D-Punjab produces a clinically significant condition through increasing oxygen affinity that causes a compensatory erythrocytosis. We report a first case of co-inheritance of Rancho Mirage with Hb D-Punjab which differs in its clinical presentation from that described in the literature.

Case presentation
A 59-year-old Pakistani male was admitted to hospital because of chest tightness. The imaging studies showed no marked organomegaly or masses. Laboratory investigations revealed erythrocytosis with high hematocrit and borderline MCV and MCH. Arterial blood gas testing (ABG) showed low pO2 and JAK2 V617F mutation was not detected. Erythropoietin level was normal. Hemoglobin analysis through high performance liquid chromatography (HPLC) revealed two hemoglobin variants; Hb D-Punjab (32.5%) and the remaining Hb comprised of a fast moving variant. The variant was faster than Hb F expressing at a considerably high level. There was no detectable Hb A. (figure 1, HPLC shows both variants)
At the molecular level, the β globin gene was amplified and sequenced using specific primers. DNA analysis demonstrated the presence of two mutations in Exon 3. The first one was a rare mutation in βCd 143 (CAC→GAC) causing a substitution of His→Asp that was identified as Hb-Rancho Mirage. It is speculated that the replacement of the ‘His‘ residue with that of ‘Asp’ would unequivocally alter the oxygen binding characteristics of the heme pocket. The other mutation was the common variant defined as Hb D-Punjab βCd 121 (GAA→CAA). (figure 2, DNA sequencing shows both variants). The patient had undergone regular venesection to alleviate erythrocytosis.

Discussion
The co-inheritance of Hb Rancho Mirage with Hb D-Punjab has never been reported previously. Both mutations exist in the β globin gene giving rise to concomitant beta chain variants. Carriers of Hb Rancho Mirage are anticipated with mild anemia. However, coexistence of both abnormal hemoglobins showed functional defects manifested as polycythemia, likely due to the disruption of the 2,3 DPG binding site which is actively involved in the allosteric equilibrium during oxygen transport.
The current case depicts the first occurrence of a double heterozygosity for two beta chain variants affecting exon 3 of the β globin gene with a relatively mild to moderate phenotype. Family studies are recommended to understand further the structure and function relationship of this rare hemoglobin variant in association with a more common Hb D- Punjab.

Disclosures:
No relevant conflicts of interest to declare.
The Impact of Epstein Bar Virus On Hodgkin Lymphoma's Histopathological Patterns in Morocco
Authors: Anoun Soumaya , Marouane Sofia , Kabbali Meryem , Zamiati Soumia , Benchekroun Said , Howard Scott C , Quessar Asmaa 
Published: October 2019

Abstract
Abstract 4781

Background:
In Morocco, improvement of living conditions and access to healthcare has increased life expectancy from 58 years in 1980 to 72 years in 2010. There has been a corresponding decrease in early childhood infections, including Epstein Barr virus (EBV) infection, which is associated with Hodgkin lymphoma (HL). Mixed cellularity (MC) was the most common histopathological subset of HL in Morocco. Prior studies have shown the shift to nodular sclerosis (NS) subset since 1998. Currently, the incidence of NS subtype is still predominant in our country.
Our aim is to study the incidence of EBV infection in HL Moroccan population and correlate its decrease to explain the shift of HL histopathological pattern.

Patients and methods:
We compared children and adults with HL diagnosed during 3 periods: 1980–1995, 1998–2005 and 2010–2011 in the same university medical centre. All histological samples were prepared for examination by paraffin method, than stained by Hematoxylin staining protocol. Immunuhistochemical features of Hodgkin Reed Sternberg cells were performed to assess the expression of antibodies markers (CD30, CD15). The EBV profiling was identified by immunohistochemical stains for EBV latent membrane protein 1 (LMP-1) for the last periods. Statistical comparison of all the features was performed by Epi Info software.

Results:
The features of 1200 HL patients were enrolled. The average number of HL cases per year was 46 patients for the first period versus 74 patients for the last period. The mean age was respectively 24 years (range 2–64 years) for the first period, 32 years (range 2–60 years) for the second period and 34 years (range 3–76 years) for the third one. Sex-Ratio showed male predominance. The comparison of HL incidence through the age over the past 30 years is further characterized by a significant higher incidence in elderly patients (2% versus 10%), and lower incidence in young children (32% versus 7%). The expression of LMP1 in HRS is significantly reduced between the two last periods (P=0.001). LMP1 positivity is higher in patients under fifteen years than in young adults. Currently, NS is the most prevalent histological subtype with a significant increasing frequency through the three periods (p< 0.001) instead of MC subtype. (Table1).

Conclusion:
HL pattern in Morocco tends to be like in developed countries. The shift of HL histological subtype from CM to NS could be explained by the decrease of EBV infection in Hodgkin Reed Sternberg cells. More patients need to be enrolled to confirm this hypothesis

Disclosures:
No relevant conflicts of interest to declare.
CD8 Suppressor Cells Can Be Induced by Pre-B Acute Lymphoblastic Leukemia As a Potential Mechanism of Immune Escape
Authors: Shand Jessica C , Kanu Alieu , Fry Terry J 
Published: October 2019

Abstract
Abstract 4797
Suppressor CD8 T-cells have been associated with poor antiviral immunity and, more recently, poor prognosis in hematologic malignancy. We have previously observed that nonprofessional antigen presenting cells, including B-lymphocytes, can induce IL10-secreting CD8 suppressors in the setting of lymphopenia. In this study, we hypothesized that pre-B cell acute lymphoblastic leukemia could also induce CD8 suppressors as a potential mechanism of immune escape.
To test this hypothesis, naïve murine (B6) CD8 T-cells (CD45.2) specific for the immunodominant HY peptide (HY-CD8), were cultured with a B6 HY-expressing murine pre-B cell acute lymphoblastic leukemia (CD45.2) carrying the human E2A-PBX transgene (E2APBX-ALL). Male B6 CD45.1+ dendritic cells and B-cells served as controls, as did female (HY-) E2APBX-ALL. Production of IFNγ and IL10 was determined by enzyme-linked immunoadsorbent assay (ELISA) at 48–72 hours, and specific identification of IFNγ, IL10 and FoxP3 expression by HY-CD8 was measured by intracellular flow cytometry by gating on live, CD8a+CD45.2+ cells. Surface expression of B7 family costimulatory molecules such as PD1 was also measured.
As expected, coculture of activated male dendritic cells (DC) with HY-CD8 generated high levels of IFNγ (1036 pg/ml +/− 40), with 67.2 +/− 9.4% CD8a+CD45.2+IFNγ+ cells at 72 hours. HY-CD8 did not produce detectable IL10 or express FoxP3 by either immunoassay or intracellular stain by 72 hours of culture in the presence of male DC. In contrast, HY-expressing E2APBX ALL induced IL10 production in HY-CD8, significantly less interferon gamma production by immunoassay (576 pg/ml +/− 150, p<0.0034) and intracellular flow cytometry (26.8% +/− 9.4), with 72.8% + 4.7 HY-CD8 expressing FoxP3 and nearly 100% expressing PD1. Interestingly, although female E2APBX-ALL did not stimulate IFNγ or IL10 production, suggesting antigen-specific effects, a similar induction of PD-1 on HY-specific T cells was produced by exposure to either male or female E2APBX-ALL.
In conclusion, our data indicates that pre-B acute lymphoblastic leukemia is capable of inducing an IL10-producing, FoxP3+ suppressive phenotype in CD8 T-cells. In this in vitro system, the suppressive phenotype is only induced when ALL bears an antigen recognized by the CD8 T-cell receptor, suggesting antigen-specific regulatory blockade as a potential target. Further studies that address the in vivo function and molecular signature of ALL-induced CD8 suppressors in a preclinical model are ongoing.

Disclosures:
No relevant conflicts of interest to declare.
Utility of SNP Microarray in the Diagnosis of MDS
Authors: Papenhausen Peter , Schwartz Stuart , Dong Henry Y. , List Alan F. , Thomason Ronald 
Published: October 2019

Abstract
Abstract 4806
A series of 206 bone marrow aspirates/blood submitted for diagnostic testing for possible MDS/MPN was studied by high resolution SNP microarray to determine the efficacy of this technology for detecting clonal copy number alterations and copy neutral loss of heterozygosity. The latter, acquired uniparental disomy (aUPD), is associated with oncogene mutations that through mitotic recombination have converted to the homozygous state, offering additional selective advantage to daughter cells. Of the 206 patient samples classified as possible MDS, 76 were abnormal by the array. Thirty one of these were either copy neutral or demonstrated copy number alterations below the resolution of cytogenetics. Twenty-three cases demonstrated aUPD and in 14 of these it was the only abnormality detected. Two of these 14 had multiple aUPD (9q/14q and 1p/4q/14q). When the aUPD clones were accompanied by copy number alterations, they could be seen as either the early primary alteration or as a later evolutionary event. The most common segmental UPD regions in MDS were 4q(9), 7q(4),11q(4) and 14q(4) while 9p(4) was the most common a(UPD in the patients with MPN which was likely to be associated with a homozygous JAK2 mutation (confirmed in two cases). The small MPN group (12 cases) demonstrated eight clones with aUPD, only three of which had copy number alterations. All the aUPDs found in this study involved terminal chromosome arm exchange and almost all involved a percentage of the DNA tested, consistent with an acquired change. Whole chromosome aUPD was not seen in these patients. There were three instances of possible interstitial LOH which were not included in this report until normal patient DNA can be obtained to confirm this uncommon observation. Two of these three showed mosaic LOH consistent with an acquired change. The most common deletions below the resolution of cytogenetics involved the RUNX1 and TET2 genes. In summary, the SNP microarray increased the abnormal clone detection from 45 by cytogenetics to 76 in cases of MDS and from 4 to 8 in the small MPN cohort, underscoring the utility of the testing while laying the groundwork for the discovery of new driver genes in the aUPD regions in the pathogenesis of MDS.

Disclosures:
Papenhausen: LabCorp: Employment. Schwartz:LabCorp: Employment. Thomason:LabCorp: Employment.
A Retrospective Analysis of Gastric Diffuse Large B-Cell Lymphoma with or without MALT Component
Authors: Li Xiaowu , Xia Bing , Guo Shanqi , Sotomayor Eduardo M. , Yu Yong , Yang Hongliang , Zhao Haifeng , Zhao Zhigang , Wang Xiaofang , Wang Yafei , Zhang Yizhuo 
Published: October 2019

Abstract
Abstract 4856

Background:
The aim of this study was to evaluate the clinical characteristics, prognostic factors and survival outcomes of patients with gastric diffuse large B-cell lymphoma (DLBCL).

Patients and methods:
162 patients with gastric DLBCL were evaluated retrospectively. Comparisons were made between patients of gastric DLBCL with component of mucosa-associated lymphoid tissue lymphoma (DLBCL/MALT) and patients of gastric DLBCL without detectable MALT component (de novo DLBCL).

Results:
Results according to the distribution of sex, age, stage, performance status, and other clinical characteristics were similar between de novo DLBCL group and DLBCL/MALT group (p>0.05). The ratio of patients with the germinal center B-cell-like (GCB) subtype to non-GCB subtype did not differ significantly between the two groups (1:1.1 versus 1:1.6, p=0.319). However, the proportion of patients with the stage-modified international prognostic index (m-IPI) ≥2 was higher in DLBCL/MALT groups (18%) than taht in de novo DLBCL groups (34%) (p=0.026). In addition, the H. pylori infection rate was 75% in DLBCL/MALT versus 38% in de novo DLBCL (p<0.001). Patients with de novo DLBCL have better 5-year PFS and OS estimates than those DLBCL/MALT patients (p=0.037 and 0.019 for the 5-year PFS and OS estimates, respectively). Surgical treatment did not offer survival benefit when compared with chemotherapy (p=0.405 and 0.065 for the 5-year PFS and OS estimates, respectively). Multivariate analysis revealed that non-GCB classification and m-IPI≥2 were independently associated with shorter OS and advanced stage was independently associated with shorter PFS.

Conclusion:
Gastric DLBCL is a heterogeneous disease that included de novo DLBCL and DLBCL/MALT lymphoma. Compared with the former, the latter has a higher H. pylori infection rate. And what's more, the proportion of patients with m-IPI≥2 is higher in DLBCL/MALT groups. De novo DLBCL was associated with higher 5-year PFS and OS estimates. Non-surgical treatment should be a primary consideration for gastric DLBCL. Immunophenotype classification and m-IPI were the most reliable factors for OS, and advanced stage was for PFS.

Disclosures:
No relevant conflicts of interest to declare.
Myelomatous Plasma Cells Display An Intermediate Gene Expression Pattern Between a Normal Plasma Cell and a Memory B Cell
Authors: Baez Alicia , Piruat Jose Ignacio , Caballero-Velazquez Teresa , Barbado María Victoria , Alvarez-Laderas Isabel , Millan-Ucles Africa , Medrano Mayte , Garcia-Guerrero Estefania , Sanchez-Abarca Luis Ignacio , Espigado Ildefonso , Perez-Simón Jose Antonio 
Published: October 2019

Abstract

Introduction
Memory B cells (MBCs) remain viable in a non-proliferative state for years. These cells express genes involved in cell survival and anti-apoptotic factors, while repress the expression of cell cycle regulatory genes. During their differentiation into plasma cells (PCs), these cells develop an opposite gene expression pattern, with a higher expression of genes implicated in cell proliferation and activation, and a lower expression of survival genes. In multiple myeloma (MM) the PCs accumulate into the bone marrow due, at least in part, to failure of pro-apoptotic mechanisms normally expressed in PCs. In the present study we analyzed the gene expression patterns of MBCs and PCs from healthy donors and patients with MM, in order to determine whether or not myelomatous PCs share characteristics of MBCs and/or normal PCs, and to identify possible genes related to the pathophysiology of the disease.

Methods
MBCs were obtained by immunomagnetic separation from buffy coats of 5 aged healthy donors. Likewise, PCs were isolated from bone marrow of 6 healthy donors and of 5 patients with MM. Using microarray techniques we analyzed the expression of 45000 genes in all samples. We performed unsupervised hierarchical cluster of gene expression data using the average linkage and the Euclidean distance. To identify differentially expressed genes among experimental groups we applied non-parametric Kruskal Wallis test. The differences in expression with a p value < 0.05 were considered significant. All analyses were performed using the Multi-experiment Viewer 4.7.1 software.

Results
From the hierarchical cluster obtained we clearly identified two groups, one which included normal PCs samples and the other one containing myelomatous PCs and MBCs. Interestingly, myelomatous PCs displayed intermediate features between MBCs and normal PCs (Figure 1). We found 5159 genes differentially expressed between normal and myelomatous PCs. Among these, we identified 3455 genes which displayed a similar expression pattern between MBCs and myelomatous PCs, including caspases inhibitors, MAP kinases, ubiquitins, transcription and translation factors.

Conclusion
Myelomatous PCs display an intermediate gene expression pattern between normal PCs and MBCs. These cells display high expression of genes involved in cell survival that should be normally inactivated in the transit of MBC to a normal PC, so that its expression pattern is closer to a MBC than a normal PC.

Disclosures:
No relevant conflicts of interest to declare.
Development of Donor Cell Derived Leukemia After Allogeneic Stem Cell Transplant
Authors: Ahmed Sairah , Di Stasi Antonio , Shpall Elizabeth J. , Khouri Issa F. , Alousi Amin M. , Ciurea Stefan O. , Bashir Qaiser , Qazilbash Muzaffar , Jones Roy B. , Nieto Yago , Parmar Simrit , Kebriaei Partow , Shah Nina , Rezvani Katy , Hosing Chitra M , Oran Betul , Andreeff Michael , Anderlini Paolo , Andersson Borje S , Champlin Richard E , Popat Uday 
Published: October 2019

Abstract

Background
Donor cell derived leukemia (DCL) is a rare event after allogeneic stem cell transplantation. First described in 1971, several cases of acute myeloid leukemia (AML) or myelodysplasia (MDS) arising from donor cells have been reported. The true incidence of DCL is difficult to ascertain, however the European Group for Blood and Marrow Transplantation survey reported in 2005 an estimated rate of 124 cases per 100,000 transplants (Hernstein et al., Hematologica 2005;90:969-975). The aim of our study was to determine the incidence and natural history of DCL after allogeneic stem cell transplantation (allo-HCT).

Methods
We retrospectively reviewed patients that received an allo-HCT at MD Anderson from 1/1997 to 6/2012 for hematologic malignancy (n=2148) and chose to study two groups: (1) patients who had a sex mismatched allo-HCT for AML /MDS (n=433) and (2) those who received an allo-HCT for non-hodgkins lymphoma (NHL) (n=773) – subsequently these cases were reviewed for a diagnosis of AML/MDS post transplant. Donor origin of leukemic cells was confirmed by presence of donor specific sex chromosomes on conventional cytogenetic analysis or FISH studies and by donor specific microsatellite polymorphic markers on DNA analysis (chimerism studies). Demographic and disease-related data are presented in table 1.

Results
9 cases of DCL were identified out of 1206 allo-HCTs done (0.7%) from 1997-2012. 4 cases were patients who received sex mismatched allo-HCTs for AML/MDS while the other 5 were patients after allo-HCT for NHL. Median age at transplant was 59 years (range 25-66 years) and median time to diagnosis of DCL was 19 months (range 4-103 months). Only 1 patient was actively treated for GVHD with corticosteroids at diagnosis of DCL. Four of 9 had monosomy 7, 1 of 9 had trisomy 8 while the remainder of patients had diploid cytogenetics. The outcome of DCL was poor with a median survival from diagnosis of 5 months (range 1-89 months).

Conclusions
Our analysis suggests that DCL occurs in no less than 0.7% of patients receiving allogeneic stem cell transplants for AML/MDS or NHL. While the mechanism of leukemogenesis is unclear, monosomy 7 was demonstrated in 45% of these cases. Although the number of cases analyzed is low, the overall prognosis for DCL seems poor.
Cytogenetics (cyto) Recipient (R) Donor (D) Follicular lymphoma (FL) Mantle cell lymphoma(MCL) Large cell lymphoma(LCL) Overall survival (OS)

Disclosures:
No relevant conflicts of interest to declare.
Outcomes Of Allogeneic Hematopoietic Stem Cell Transplantation In AML Patients Registered In Prospective Studies: The JALSG Experience
Authors: Ishiyama Ken , Ohtake Shigeki , Sakamaki Hisashi , Miyazaki Yasushi , Miyawaki Shuichi , Ohnishi Kazunori , Naoe Tomoki 
Published: October 2019

Background
The current guidelines for acute myeloid leukemia (AML) issued by the Japan Society for Hematopoietic Cell Transplantation were updated in 2009 to reflect that AML in intermediate-risk or unfavorable-risk patients younger than 55 years of age should be treated with allogeneic stem cell transplantation (allo-SCT) at the first complete remission (1CR). However, most of the eevidence' for this recommendation was based on the results of either prospective studies with randomization according to donor availability or retrospective studies. Therefore, the influence of eselection bias' is inevitable. On the other hands, physicians sometimes have difficulty deciding whether a patient should receive upfront allo-SCT or final treatment with chemotherapy, as the outcomes of patients with 1CR at allo-SCT and 2nd CR (2CR) at allo-SCT are comparable.

Aims
We conducted a subgroup analysis to verify the validity of allo-SCT in AML patients in 1CR using the dataset of patients registered in prospective studies. [Methods] We examined the disease status and outcomes of the AML patients registered in the AML97 and AML201 studies conducted by the Japan Adult Leukemia Study Group (823 patients and 1,064 patients, respectively) who underwent allo-SCT.

Results
A total of 752 patients with AML were registered in the JALSG study and underwent allo-SCT. Their estimated 5-year overall survival (OS) was 44%. The number of patients according to the disease status at allo-SCT was 262 in 1CR, 130 in 1st relapse (1rel), 192 in 2CR and 98 with primary induction failure to CR (PIF). The estimated 5-year OS of the patients with intermediate-risk or unfavorable-risk cytogenetics in 1CR was significantly superior to that of the patients with 1rel or 2CR (57% vs. 41%, p<0.0001). The allo-SCT outcomes of the patients with relapse revealed that the estimated 5-year survival after relapse (SAR) in the patients who achieved 2CR was acceptable at 62%, while that of the patients with 1rel who underwent allo-SCT was significantly superior to that of the patients who did not receive allo-SCT (5-year SAR: 17% vs. 8%, p<0.0001). The outcomes of the patients with relapse and PIF were better among the patients who underwent allo-SCT than among the patients who did not, regardless of the cytogenetic risk category.

Conclusions
The use of upfront allo-SCT in AML patients with 1CR is recommended, as the allo-SCT outcomes of patients with 1CR are superior to those observed after relapse, although the 5-year OS of patients with 2CR is not inferior to that of patients with 1CR.

Disclosures:
No relevant conflicts of interest to declare.