bluuebunny/crossref_metadata_2025 huggingface dataset:
Consultation for Thrombocytopenia in Hospitalized Patients
Authors: Boxer Michael
Published: October 2019

Abstract 4659

Thrombocytopenia is a frequent cause for hematologic consultation in an acute care hospital. During a 22 month period one hematologist assessed 93 patients with thrombocytopenia. Thrombocytopenia was the sole reason for the consultation. No previous thrombocytopenia had been recognized previously in any if these patients.

The cause for thrombocytopenia was liver diease in 21 patients, sepsis in 12, immune (ITP) in 12, antimetabolites or antiseizure medications in 7, heparin induced thrombocytopenia (HIT) in 6, platelet clumping in 4, thrombotic thrombocytopenic purpura (TTP) in 3, myelodysplasia (MDS) in 3, acute progranulocytic leukemia (APL) in 3, rhabdomyolysis in 2, viral infections in 2, systemic lupus (SLE) in 2, and intravenous antibiotics in 2.

The following conditions were each diagnosed once as the cause for thrombocytopenia:pregnancy, disseminated intravascular coagulation (DIC), post coronary bypass grafting, brain injury, acute myelogenous leukemia (AML-M5), antiphospholipid syndrome (APLS), and intra aortic ballon pump.

No diagnosis could be made in 13 patients. In 28 patients the initial diagnosis was incorrect. No diagnosis could be made in 13 patients. In 6 patients two causes for thrombocytopenia were present. One patient was admitted twice with two different causes for thrombocytopenia (sepsis and ITP).

Review of the peripheral smear is both necessary and mandatory. All three patients with APL were initially diagnosed with ITP, and the diagnosis of APL was not made until the smear was assessed. Smear review confirmed or made the diagnosis of TTP. Platelet clumping could not be made without reviewing the peripheral smear.

Primary hematologic diagnoses (ITP, HIT, clumping, TTP, MDS, APL, AML, APLS) were present in 33 patients. Secondary causes for thrombocytopenia were present in the majority of patients.

In summary thrmobocytopenia a frequent cause for hematologic consultation, and most patients will not have a primary hematologic etiology for the thrombocytopenia. Advanced liver disease is an underappreciated cause for thrombocytopenia and was never considered to be the cause for the thrombocytopenia by the referring physician.

Despite exhaustive testing the cause for thrombocytopenia could not be made in 13 patients. However in these patients the platelet count rarely was below 100,000.

The importance of reviewing the peripheral smear cannot be overemphasized.

Disclosures:

No relevant conflicts of interest to declare.
Nilotinib Exhibits an in Vitro Antiviral Activity Against Human Cytomegalovirus (HCMV): Potential Clinical Applications
Authors: Wolf Dana , Djian Esther , Beider Katia , Shimoni Avichai , Nagler Arnon
Published: October 2019

Abstract 4666

Chronic myeloid leukemia (CML) is a clonal disorder associated with chromosomal translocation t (9; 22), which produces the Philadelphia chromosome. The fusion gene encodes for the chimeric oncoprotein BCR-ABL, associated with deregulated constitutive tyrosine kinase (TK) activity, leading to leukemogenesis. Imatinib mesylate, the first potent selective inhibitor of BCR-ABL TK, has revolutionized the current management of CML. The second generation TK inhibitor (TKI) Nilotinib (Novartis, East Hanover, NJ) is an aminopyrimidine derivative of imatinib with an increased binding affinity to the chimeric p210 BCR-ABL. Nilotinib is active in imatinib-resistant or -intolerant patients in chronic phase, accelerated phase (AP), and blast crisis (BC) CML.

Allogeneic stem cell transplantation (SCT) is currently reserved for patients(pts) in CP after the failure of second line TKI or for patients in advanced phase disease.SCT remains the treatment of choice in patients with Ph+ALL. TKIs can be used successfully pre -SCT as a bridge to SCT in advanced disease and post-SCT in order to prevent disease recurrence. HCMV is one of the leading causes of morbidity and mortality post SCT in particular in pts transplanted for advance CML and pts with graft vs. host disease (GVHD) While preemptive antiviral therapy has reduced the occurrence of HCMV disease post SCT, the use of all currently available antiviral drugs is often limited by toxicity, low oral bioavailability, and drug resistance.

The mechanisms facilitating HCMV entry into the host cells are not clearly understood. Blocking of the platelet – derived growth factor receptor-α (PDGFRα) has been shown to inhibit HCMV internalization and gene expression. Recently, Imatinib have been shown to inhibit PDGFRα phosphorylation. As Nilotinib is a more potent PDGFR inhibitor, we assessed the in vitro antiviral activity of Nilotinib against HCMV. Nilotinib exhibited a significant dose-dependent inhibition of the virus upon pre-incubation with the drug. Moreover, Nilotinib demonstrated a ∼4.5-fold higher antiviral activity against HCMV when compared to imatinib, with IC50 values of 0.39 μM and 1.75 μM, respectively. No viral inhibition was found upon addition of the drugs after viral adsorption – compatible with inhibition of an early step of infection, involving viral binding/entry into the cell. These findings identify a promising new target for antiviral therapy, representing an alternative paradigm for treatment with compounds combining anti-cancer and antiviral activity. It remains to be determined if the anti-HCMV activity demonstrated for Nilotinib is of clinical relevance in patients undergoing SCT

Disclosures:

No relevant conflicts of interest to declare.
A Clinical Study of Bone Marrow Stromal Cell Therapy in Patients with Refractory Cgvhd by Down-Regulating the Jagged2 Gene
Authors: Weng Jianyu , Huang Xin , Geng Suxia , Luo Chengwei , Wu Suijing , Ling Wei , Guo Rong , Lu Zesheng , Deng Chengxin , Andy Pengxiang , Du Xin
Published: October 2019

Abstract 4673

Refractory chronic GVHD (cGVHD) is an important complication after allogeneic hematopoietic SCT and is prognostic of poor outcome. Bone Marrow Stromal Cells (MSCs) are involved in tissue repair and modulating immune responses in vitro and in vivo. MSCs as salvage treatment for refractory cGVHD have been reported in our previous study, however, the possible mechanism have yet not to be determined.

Between November 2006 and November 2010, 18 patients were diagnosed with refractory cGVHD, 8 patients were treated with in vitro expanded BM-derived MSCs as a compassionate treatment for refractory cGVHD, 10 patients that did not receive BMSCs treatment were control group. The median MSC dose given was 0.6×106/kg body weight. MSCs were harvested fresh from culture and administered to the patients by intravenous infusions over 30 minutes. The median time of MSC administrations was 3 (range, 2–6). The response was assessed monthly after BMSCs treatment, and the total follow-up period was 6 months. The organ response and the overall response were used to determine the therapeutic efficacies of MSC for refractory cGVHD. The expression of the Jagged2 gene of peripheral blood mononuclear cells in patients at the assessment points were analyzed using the TaqMan real-time polymerase chain reaction, with ABL mRNA expression levels as an internal reference.

After BMSCs treatment, a total of 6 patients (75%) had an overall response (PR n=6), and 2 patients had a minor partial response (mPR n=2). The expression levels of Jagged2 mRNA in these cases at the diagnosis of refractory cGVHD were significantly increased, compared with none cGVHD patients (23.94%±18.68% vs 3.76%±1.50%, P < 0.05), and the copies of Jagged2 mRNA in BMSC treatment responsed patients' peripheral blood were significantly reduced (5.15%±3.25%, P <0.05), while Jagged2 mRNA expression levels of the control group were no significant difference (P> 0.05).

Our pilot study showed that Jagged2 gene reproduction upregulated when the cGVHD is active, so, dynamic monitoring of Jagged2 mRNA expression may have the potential effect on predicting the activity of chronic graft-versus-host disease. Mechanism of Bone marrow stromal cells to treat refractory cGVHD may be related to down-regulation of donor T cells Notch ligand Jagged2 gene expression, which suppression of T cell Notch signaling pathway activation, thus inducing immune tolerance.

Disclosures:

No relevant conflicts of interest to declare.
Donor Lymphocyte Infusion for the Treatment and Prevention of Recurrence After Hematopoietic Stem Cell Transplantation
Authors: Liao Jianyun , Long Lina , Li Chunfu , Zhou Xiaohui , Liu Qifa
Published: October 2019

Abstract 4674

Background—Treatment and prevention of relapse and recurrence after HSCT is challenging. DLI from the same donor is an effective therapeutic clinical option for patients. Method—we retrospectively analyzed the data of 14 patients with diagnoses included ALL (n=3), AML(n=3), thalassemia (n=7), aplastic anemia(n=1), who received DLI after HSCT. We analyzed the relationship between the during of DLI to the occurrence of aGVHD. Results—A total of 25 infusions were given to 14 patients with a median infusion time of 116(44–620) days post-HSCT. acute graft-versus-host disease (GVHD) grade I occurred in 9 patients at a median day of 33.4(14–70) after DLI. GVHD prophylaxis (using CsA) reduced the incidence of aGVHD. Our data showed that the median time to DLI in patients with aGVHD was 206 (44–521) days, and aGVHD developed in about 50% of the patients receiving DLI less than 180 day post-transplantation, while GVHD occurred in 33.3% of the patients who received DLI over 180 day (p=). the risk of developing GVHD was also dependent on the dose of DLI, with 7 patients developing GVHD when the dose of infused cells is 1¡Á107 lymphocyte cells/kg however, ? patients. Conclusion—the response to DLI is dependent on the time post-transplant. It seems that it is more easy to develop GVHD to DLI less than 180 days post-transplantation and to receive a dose of 1¡Á107 lymphocyte cells/kg. GVHD would possibly occur in 4 weeks after DLI.

Disclosures:

No relevant conflicts of interest to declare.
Pancytopenia a Presentation of disseminated Histoplasmosis
Authors: Hussain Sakeer
Published: October 2019

Abstract 4687

A 70 year old female with diabetes mellitus, rheumatoid arthritis with worsening fatigue, intermittent shaking chills and fever for 2 weeks. She had been on oral prednisone 5 mg daily and methotrexate 15 mg every week for her rheumatoid arthritis. Vital signs were stable and physical examination was significant for 3+ lower extremity edema and pallor. The CBC showed pancytopenia, white cell count of 2.5×109/L (4.3–10.8×109/L), hemoglobin was 9.2g/dl (11.0–15.0g/dl), and platelet counts were 93 × 109/L (130–400 × 109/L). SHe had mildly elevated total bilirubin 1.7mg/dl (0.0–1.2mg/dl), elevated alkaline phosphate 1212U/L (38–126 U/L) liver enzymes, SGPT 83U/L (0–40U/L), SGOT 126U/L (0–35U/L), and elevated blood sugar of 260mg/dl. Blood cultures remain negative for bacterial growth for multiple days. Her SPEP, ANA, HIV, CMV, and EBV serology were negative. CT scan of the chest, abdomen and pelvis were consistent with hilar, mediastinal lymphadenopathy and upper abdominal lymphadenopathy with splenomegaly. A bone marrow aspiration and biopsy was consistant with histoplasma capsulatum infection. Patient was treated with IV amphotericin B for 4 weeks. Histoplasma capsulatum was identified from fungal culture from the bone marrow aspirate. Retrospectively urine analysis for histoplasma antigen was positive by ELISA. Subsequently patient recieved itraconazole for 6 months. Cytopenia and symptoms improved and lymphadenopathy regressed. Most patoents who develop disseminated histoplasmosis are immunosuppressed It often mimic the presentation of malignancy like lymphoma or infection like tuberculosis and inflammatory process like sarcoidosis. Misdiagnosis can lead to inappropriate and harmful therapy with glucocorticoides and cytotoxic chemotherapy. Diagnosis requires high index of suspicion and familiarity with clinical presentation and appropriate diagnostic test.

Disclosures:

No relevant conflicts of interest to declare.
Evaluating Conflicts of Interest in Academic Research On Erythropoietin Receptors in the Cancer Setting
Authors: Bennett Charles , vanSlooten Eleasa , Bialkowski Matthew , Trenery Alyssa , Byrne Ashlyn , Murday Alanna , Yarnold Paul , Qureshi Zaina Parvez , Sartor A. Oliver
Published: October 2019

Abstract 4695

Background:

We previously reported that among preclinical cancer articles published prior to 2008, academic researchers without financial conflicts of interest were significantly more likely than academic researchers with financial conflicts or researchers employed by erythropoiesis stimulating agent (ESA) manufacturers to identify erythropoietin (Epo)-induced cancer-related signaling events and Epo-induced changes in functions of cancer cells.(Arch Int Med 2010) In 2007, the National Cancer Institute convened a workshop on Epo-receptors for 14 academic researchers and six researchers employed by ESA manufacturers. We update our analysis following this workshop.

Methods:

Articles identified in MEDLINE and EMBASE databases (1989–2012) investigating preclinical ESA findings were reviewed. Outcomes included Epo-receptors, Epo-induced signaling events, Epo-induced cellular function events, and qualitative conclusions among reports published since the 2007 meeting. Recent publications before versus after the NCI workshop according to source of research funding were analyzed. Outcomes were reported according to academic investigators without financial conflicts of interest (26 publications post-2008; 64 publications pre-2008), academic investigators with financial conflicts (5 versus 7 publications) and investigators employed by ESA manufacturers (2 versus 3 publications).

Results:

Since 2008, academic investigators without financial conflicts of interest and academic investigators with financial conflicts differed from researchers employed by ESA-manufacturers with respect to identifying the following EPO-induced events: signaling associated with tumor promotion (86%, 100%, versus 0%; p=0.06), cellular function changes associated with tumor growth/metastases (86%, 67%, and 0%; p=0.06). We qualitatively concluded that preclinical findings raised concerns that ESAs could be harmful clinically (77%, 40%, and 0%, p=0.03). Prior to 2008, academic investigators without financial conflicts of interest differed from academic investigators with financial conflicts and researchers employed by ESA manufacturers for these same outcomes (signaling events, 94%, 0%, and 0%, p=0.001; cellular changes (57%, 0%, and 0%, p=0.007), and qualitative conclusions regarding potential harm (57%, 0%, and 0%, p=0.007), respectively.

Conclusions:

The potential for conflicts of interest to affect preclinical research findings dissipated, following an NCI-convened workshop, for academic researchers with financial conflicts of interest, but not for researchers employed by ESA manufacturers.

Disclosures:

Bialkowski: Eli Lilly: family member employment Other. Sartor:Amgen: Consultancy.
Improving the Quality of Our Clinical Work for Coumadin Anticoagulation Therapy with Electronic Medical Record (EMR) Based System Redesign
Authors: Thomas-Hemak Linda , Thomas Lisa , Shi Qi , Sheth Jignesh , Praveen Etikala , Choudhary Gunjan , Abdelmalek Cherif , Muppidi Vijayadershan , Eikins Tiffany
Published: October 2019

Abstract 4714

Introduction:

Coumadin anticoagulation therapy is indicated in a variety of thromboembolic conditions. Close INR monitoring is required to maintain therapeutic range in order to decrease complications, emergency room visits, and medical costs associated with Coumadin therapy. To achieve this, tight quality control in primary health care systems is undeniably important. The purpose of this study is to assess the effects of a redesigned system for coumadin anticoagulation therapy in an outpatient setting. Our project includes: (1) Meaningful use of a new electronic medical record (EMR) based INR flow sheet; (2) Improved physician and staff efficiency using new EMR based protocol for INR tracking and clinical decision support, including drug interaction information, dose adjustment, per-procedure anticoagulation bridging, and an education/training program; (3) Improved patient education, including enhanced Coumadin educational materials. Vitamin K prescriptions will be given to the patients in the event of complications with coumadin.

Method and Materials:

INR data were collected from 55 patients receiving Coumadin therapy from 10/01/2009 to 10/31/2011 at an outpatient setting. Total blood tests for INR were 1679 samples. We summarized results from 55 patients twelve months before and after using our redesigned system.

Result:

Our data shows the new system helped to: (1) Increased the percentage of INRs (average) in the therapeutic range from 51.35% to 54.05% and the percentage of INRs (average) in extended therapeutic range (+/−0.2) from 63.78% to 70.43%; (2) Decreased the highest value from 15.4 to 10.2; (3)Decreased annual total number of INR checkups for all patients from 881 to 798; and (4)Decreased average number of INR checkups per month from 1.8 to 1.3 (average).

Conclusion:

Our clinical data shows that Our EMR based system redesign not only helps to increase efficiency and safety of Coumadin anticoagulation therapy, but also decreases the frequency of INR checks, thereby reducing medical expenditures.

Disclosures:

No relevant conflicts of interest to declare.
Interleukin-1 Beta Promotes the Expression of Prox-1 and Vascular Endothelial Growth Factor Receptor Type-3 in Adult Human Dermal Fibroblasts
Authors: Takuji Matsuo , Shirasaki Ryosuke , Tashiro Haruko , Oka Yoko , Yamamoto Tadashi , Akiyama Nobu , Kawasugi Kazuo , Shirafuji Naoki
Published: October 2019

Abstract 4738

Background and Aims:

We recently reported that human interleukin 1 beta (IL-1 beta) stimulates bone-marrow stromal myofibroblasts to express a hematopoietic stem cell marker, CD34 (16th EHA, and 53rdASH meeting). To characterize precisely the effects of IL-1 beta to the stromal cells, adult human dermal fibroblasts (HDF) were cultured with IL-1 beta, and the morphological changes and molecular expressions were observed.

Materials and Methods:

HDF were purchased, and cultured in knockout DMEM medium with 15% KSR with or without recombinant human IL-1 beta for 3 weeks. The morphological changes and the expression of specific hematopoiesis-related genes were analyzed time-dependently. The concentration of hematopoietic growth factors in the culturing supernatants was also measured.

Results:

When HDF were cultured with human IL-1 beta for 3 weeks, the cellular morphology changed to the filamentous appearance. RT-PCR analyses revealed that cells expressed Prox-1, a master molecule for lymphatic duct neogenesis, and also vascular endothelial growth factor receptor (VEGFR) type-3. Smad7 and PAI1 were also expressed; however, LYVE1, a marker for lymphatic vascular endothelial cell, was not induced. VEGF-C production was also demonstrated at RNA and protein levels, and VEGF-C and VEGFR type-3 system played an important role in morphological changes and cell-proliferation.

Discussion:

We previously reported that when HDF were cultured in DMEM/F12 medium with IL-1 beta and EPO, hematopoietic markers were induced. When HDF were cultured in knockout DMEM, a condition for immature cell-growth at ES cell levels, and IL-1 beta, lymphatic duct neogenesis-related genes were activated, and HDF converted their morphology without an introduction of any genes externally. We are now making clear the precise action of IL-1 beta to HDF using microarray analysis.

Disclosures:

No relevant conflicts of interest to declare.
Derivation of Hematopoietic Progenitor Cells From Vector-Free Human Induced Pluripotent Stem Cells
Authors: Schuening Friedrich , Zaboikin Michail , Zaboikina Tatiana , Srinivasakumar Narasimhachar
Published: October 2019

Abstract 4746

Induced pluripotent stem cells (iPSCs), due to their self-renewal and differentiation capability, have tremendous potential in regenerative medicine. Differentiation of IPSCs in vitro to obtain sufficient number of hematopoietic stem cells (HSCs) and their progenitors (HPCs) from iPSCs for therapeutic purposes is a holy grail of cellular therapy. To this end, we are comparing different in vitro differentiation approaches for generation of HSCs/HPCs from IPSCs.

We have generated iPSCs from human adult dermal fibroblasts using two different reprogramming methods: 1) Transduction with retroviral vectors encoding human Klf4, Oct3/4, Sox2 and cMyc or 2) Electroporation with Epstein–Barr virus (EBV) based episomal plasmid vectors encoding Klf4, Oct3/4, Sox2, L-Myc and p53 targeting shRNA. The transduced/electroporated cells were reprogrammed on SNL5 mouse feeder cells. Putative iPSC-like colonies were cloned and adapted to grow under feeder-free conditions on Matrigel (BD) in mTeSR1 (Stem Cell Technologies) medium. From over 30 individual clones isolated, six were further characterized for: 1) expression of pluripotency markers (Tra-1–60, SSEA-3, SSEA-4, Nanog and Oct3/4) by immunofluorescence; 2) endogenous and total mRNA expression by quantitative real-time reverse-transcriptase PCR (RT-qPCR) for Klf4, Oct3/4, Sox2 and cMyc to distinguish between cellular and vector derived expression of reprogramming factors; 3) RT-qPCR to determine expression of other markers of pluripotency such as Nanog and DNA methyl transferease; 4) karyotype analysis to determine chromosomal anomalies. The vector-free IPSC clones were also tested for residual integrated EBV plasmid DNA by qPCR. Trilineage differentiation ability of the clones was determined through embryoid body formation in suspension cultures, and subsequent staining of resulting embryoid bodies after adherence to gelatin coated dishes for makers of ectoderm, mesoderm and endoderm.

HSCs/HPCs were obtained from IPSCs by 1) coculture with OP9 stromal cells, or 2) step-wise differentiation in feeder-free conditions on Matrigel under defined conditions in the presence of appropriate growth factors [Niwa A et al. PLoS One. (2011); 6(7):e22261.]. The resultant HSCs/HPCs were subjected to colony forming assays in semi-solid medium containing hematopoietic cytokines. Both erythroid and myelomonocytic colonies could be readily identified. The influence of ambient oxygen concentration on the HSC/HPC derivation procedure is being investigated. The results of these studies will be presented.

Disclosures:

No relevant conflicts of interest to declare.
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 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 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 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 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 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

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

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.
Allogeneic Hematopoietic Stem Cell Transplant Reverses The Phenotype Of DOCK8 Deficiency
Authors: Cuellar-Rodriguez Jennifer , Freeman Alexandra F , Gea-Banacloche Juan , Su Helen , Grossman Jennifer K. , Fry Terry J , Holland Steven M. , Hickstein Dennis D.
Published: October 2019

Background

DOCK8 deficiency— a combined immunodeficiency characterized by recurrent sinopulmonary infections and severe cutaneous DNA viral infections, eczema, food and drug allergies, and increased risk of viral driven malignancies— results from homozygous or compound heterozygous mutations in the dedicator of cytokinesis 8 protein. Allogeneic hematopoietic stem cell transplantation (HSCT) represents a potential curative therapy for DOCK8 deficiency.

Methods

We treated 4 patients with DOCK8 deficiency with allogeneic HSCT using a reduced toxicity, myeloablative-conditioning regimen consisting of busulfan 3.2 mg/kg/day IV for 4 days and fludarabine 40 mg/m2/day for 4 days. Two patients received matched related donor allogeneic HSCT, and 2 received 10/10 HLA matched unrelated donor allogeneic HSCT. All patients received tacrolimus, and short methotrexate on days 1,3, 6, and 11, for graft-versus-host-disease (GVHD) prophylaxis. Indication for transplant was severe recurrent infections with end organ damage in three patients, and a refractory EBV-virus driven lymphoma at the time of unrelated donor PBSC transplant in the fourth patient.

Results

The ages of the patients at the time of transplant were 27, 25, 18, and 16. The median follow-up for patients was 7 months (range 5-18 months). All 4 patients engrafted at a median of 12 days, and all 4 had complete reconstitution of the CD3/CD4 and CD3/CD8 compartments with donor cells. Clinical correction of the DOCK8 clinical phenotype occurred within 3-6 months of engraftment and correlated with lymphocyte reconstitution. Mucositis was the major side effect of the transplant-conditioning regimen. All had transient worsening of their pre-transplant infections 1-3 months post-transplant. In the patient with an EBV-virus driven lymphoma that had been refractory to chemotherapy, there was complete resolution of the disease on PET scanning at 100 days post-transplant. One patient, who received unrelated donor bone marrow cells, had skin GVHD that responded to a short course of steroids. There was no chronic GVHD.

Conclusions

Allogeneic HSCT in DOCK8 deficiency results in reconstitution of the deficient lymphocyte compartments that are present pre-transplant and complete reversal of the infection susceptibility phenotype. There was minimal regimen-related toxicity, and the incidence of GVHD was low despite complete donor chimerism. With genetic testing for DOCK8 deficiency now more widely available, we anticipate that earlier diagnosis will enable patients to be transplanted earlier in their clinical course, before significant organ damage or the development of viral-driven malignancies

Disclosures:

No relevant conflicts of interest to declare.