bluuebunny/crossref_metadata_2025 huggingface dataset:
High-Dose Cytarabine Alone Versus Cytarabine Plus Anthracycline for Consolidation Therapy in Non-Promyelocytic Acute Myeloid Leukemia
Authors: Kim Sung-Hyun , Kim So Yeon , Lee Ji Hyun , Lee Suee , Oh Sung Yong , Kim Hyo-Jin , Kim Dae Sik , Kang Ka-Won , Lee Se Ryeon , Park Yong , Sung Hwa Jung , Choi Chul Won , Kim Byung Soo
Published: October 2019

Standard consolidation therapy of acute myeloid leukemia (AML) contains high-dose cytarabine (HiDC). However, optimal dose of cytarabine and benefit of additional chemotherapeutic agents remain unresolved problems. One hundred and forty-two patients among 173 newly diagnosed AML patients, who achieved complete remission following induction chemotherapy and subsequently received different consolidation therapies in 4 independent institutes, were retrospectively analyzed. Patients were divided into 3 groups by consolidation regimens: HiDC (1.5-3g/m2, bid, Days 1,3,5) alone in 44 patients (Group A), HiDC plus anthracycline in 46 patients (Group B), and intermediate-dose cytarabine (1g/m2, bid, Days 1,3,5) plus anthracycline in 52 patients (Group C). Overall survival (OS), relapse-free survival (RFS), disease-free survival (DFS), and event-free survival (EFS), were not significantly different in Group A vs. Group B. However, Group B showed longer RFS, DFS, and EFS, as compared with Group C. Treatment-related mortality was significantly higher in Group B, as compared with Group A (P=0.040) and Group C (P=0.013). Cytogenetic risk was the only significant prognostic factor in OS, RFS, DFS and EFS.

In this study, the addition of anthracycline to HiDC as consolidation chemotherapy in AML patients did not improve treatment outcomes, and had more toxicity. HiDC enhanced RFS, DFS, and EFS, as compared with intermediate-dose cytarabine. Therefore, HiDC single therapy could be considered as standard consolidation therapy for non-promyelocytic AML

Disclosures

Choi: Alexion Pharmaceuticals: Research Funding.
Effect of Early Removal of Central Catheterization in Febrile Neutropenic Patients with Acute Leukemia
Authors: Hwang Su Hyun , Choi Yong Won , Jeong Seong Hyun , Park Joon Seong
Published: October 2019

Background: A guideline of central line-related blood stream infection (CRBSI) is only for immune-competent patients with end-stage renal disease on hemodialysis. We need widely acceptable guideline whether remove or maintain a central catheter in febrile neutropenic patients.

Method: Medical records from 293 cases (95 patients) of febrile neutropenia with Central Catheterization were obtained retrospectively, and clinical characteristics, particularly duration of fever, were analyzed according to the early removal of central catheterization under anti-cancer chemotherapy for acute leukemia.

Results: Forty-eight cases (16.4%) have already baseline fever and 25 cases (8.5%) showed baseline pneumonic infiltration prior to chemotherapy. The insertion sites of catheterization were subclavian (85.3%), internal jugular (12.6%), and others (2.0%). Early removal of central catheter was done at 93 cases (31.7%) and the cause of early removal was fever (63.4%), malfunction (11.8%), and others (24.7%). The duration of fever was significantly shortened by early removal of central catheterization (5.4 vs 7.5 days, p=0.003, Figure 1A). In addition, the duration of fever was related with age, diagnosis, type of chemotherapy, presence of baseline fever or pneumonic consolidation in univariate analysis. The site of central access or body mass index as well as existence of CRBSI did not affect duration of fever. In multivariate analysis, early removal of central catheter was independently associated with short duration of fever (OR=0.619, 95% CI=0.46-0.83, p=0.001, Table 1). The effect of early removal of central catheter was highlighted when micro-organisms were documented (Figure 1B).

Summary: In patients with febrile neutropenia, early removal of central catheter can shorten the duration of fever particularly when BSI was documented.

Disclosures

No relevant conflicts of interest to declare.
Rare Presentation of Relapsed Extranasal Natural Killer/T Cell Leukemia Lymphoma Involving the Hypothalamus
Authors: Kim Byung Moon , Malik Mohsin , Roberts Todd F.
Published: October 2019

42 year old male was diagnosed with stage IV extranasal natural killer T cell leukemia/lymphoma in the liver in 2013. He was treated with 5 cycles of SMILE regimen and one cycle of gemcitabine based chemotherapy after which he achieved complete remission. He had 10/10 matched unrelated donor myeloablative allogeneic transplant on 2/26/2014. His Day 100 and 1 year work up showed complete remission and 100% donor chimerism. He had no chronic graft-versus-host-disease (cGVHD). In March 2015 he presented with change in mental status and electrolyte abnormalities. Patient was oriented to self only without focal neurologic deficits and initially presented with hypovolemic hypernatremia. His work up showed Central Diabetes Insipidous. CT head was unremarkable. MRI of brain showed bilateral hypothalamic, cerebellar and frontal hyperdensity concerning for tumor infiltration. Brain Biopsy of frontal lesion showed tumor cells diffusely infiltrating the neutrophil with perivascular predilection, tumor cells were positive for CD 3, CD 7, CD 56, bcl-2, and cytoplasmic positivity for granzyme B consistent with NK T cell leukemia/lymphoma. Cells were negative for EBV, CMV HSV, HHV-8, Polyoma virus and CD 20. This was similar to his pathologic finding at initial diagnosis. Further work up including bone marrow biopsy and aspirate, CSF analysis, and peripheral blood flow cytometry were unremarkable. He was treated with 6 cycles of high dose methotrexate. MRI brain at the end of treatment showed improvement in his brain lesion.

Mature T or NK/T-cell lymphoma makes up only 5-18% of all cases of non-Hodgkin lymphoma. In a case study among 1153 adult cases of peripheral T cell lymphoma, 11.8% were classified as extranodal NK/T cell lymphoma and only 26% of these were extranasal type. In published studies, NK/T-cell lymphoma extranasal type typically involve the skin, testis, cervix, intestine, liver, spleen, soft tissue, and orbit but CNS involvement is rare and has only been reported in only a select number of clinical cases. Primary CNS involvement of extranasal NK T cell lymphoma and direct intracranial extension and progression of nasal NK/T cell lymphoma have been reported as rare presentations however, to our knowledge, recurrence or relapse of extranasal NK/T cell lymphoma within the brain parenchyma and more importantly involving the hypothalamus after a year of disease remission has not yet been reported.

Disclosures

Roberts: Millenium: Speakers Bureau.
Prevalence and Clinico-Pathological Attributes of c-MYC Rearranged High-Grade B-Cell Non Hodgkin Lymphomas (NHL): Single Centre Experience
Authors: Sengar Manju , Epari Sridhar , Jain Hasmukh , Shet Tanuja , Gujral Sumeet , Menon Hari , Dangi Uma
Published: October 2019

Introduction: MYC rearrangement in high-grade B-cell NHL(non-burkitt), either as single hit or double hit, has significant prognostic and therapeutic implications. There is remarkable paucity of data on frequency and clinico-pathological features of MYC-rearranged large B-cell lymphomas from developing world.

Method: This study included de-novo high grade B-cell NHL cases (>15 years of age) registered at Tata Memorial Centre between January 2013- December 2014. Demography details, clinical features (stage at presentation, bone marrow involvement, presence and extent of extranodal involvement, B-symptoms, international prognostic index),original histopathological diagnosis, chemotherapy and radiotherapy details were recorded from electronic medical or paper case records. Response to therapy, relapses or death if any, status at last follow up, dates of relapse and/or death and last follow up were recorded. All cases were reviewed by expert hematopathologists to categorize the high grade B-cell NHL as per the WHO-2008 classification of hematopoietic malignancies. All cases with adequate formalin-fixed paraffin embedded (FFPE) blocks were evaluated by FISH for c-MYC , BCl-2 and BCl-6 using Vysis dual colour break apart probes. Cases which showed >10% cells with split signal were considered to harbor rearrangement.

Result: A total of 114 cases of de-novo high-grade B-cell NHL with adequate FFPE blocks were evaluated. Based on WHO 2008 classification, 112 cases were classified as diffuse large B-cell lymphoma (DLBL)) and 1 each as Burkitt's (BL) and B- cell lymphoma unclassifiable -intermediate between DLBL and BL(BCLU). A total of 9/112 (8%) cases of DLBL showed MYC rearrangement. One of these 9 cases had both MYC and BCl-2 rearrangement. BCLU did not demonstrate MYC or BCl-2 rearrangement. The Ki-67 index was variable (40-95%)in MYC rearranged cases. The median age of the c-MYC rearranged cases was 55 year (range-23-79 years). 88% were males. Advanced stage disease, bulky mass and extranodal disease was seen in 67%, 44% and 55% of cases respectively. All but one patient had high LDH, however none of the patients had elevation more than 2XULN. These patients received rituximab based chemoimmunotherapy (RCHOP-4,RCEOP-4, REPOCH-1) and 77% achieved complete response. At median follow up of 5 months, 1 year-overall survival and progression free survivals were 80% and 75% respectively.

There were no significant differences in clinical features (except higher proportion of males in the c-MYC rearranged subset), LDH levels, ki-67 index , response to therapy and survival between DLBL with or without c-MYC rearrangement

Conclusion: In our study 8% of all DLBL cases showed c-MYC rearrangement. The frequency of double hit lymphoma was less than 1% (0.8%).

Disclosures

No relevant conflicts of interest to declare.
Profound Hypereosinophilia Secondary to Sezary Syndrome
Authors: Davarifar Ardy , Julian Ricklie , Vipul Pareek , Yilmaz Emrullah , Verma Amit
Published: October 2019

Background: Sezary syndrome is a subtype of cutaneous non-Hodgkin's lymphoma that is characterized by erythroderma, the presence of atypical lymphocytes in the blood, and lymphadenopathy. The clonal cell in Sezary syndrome is thought to be derived from mature Th-2 cells that are able to express Il-4, Il-5, and Il-13 which can stimulate the replication of eosinophils. It is has been shown that higher levels of eosinophilia correlate with a worse prognosis. One explanation is that the degree of eosinophilia represents the level of imbalance between Th-1 and Th-2 cells. Here we report the first case of end organ damage caused by hypereosinophilia secondary to the Sezary syndrome. In rare cases Sezary syndrome can cause hypereosinophilia but has never been reported to cause hypereosionphilia that results in end-organ damage. To our knowledge this is the first case that reports end-organ damage resultant from secondary hypereosinophilia from the Sezary syndrome.

Case presentation: A 75-year-old man was admitted for severe rash and pruritis, massive lymphadenopathy, and extremely high eosinophil count. He had recurrent skin rash with pruritis for five years with previous skin biopsies without evidence of malignancy. He had never had hypereosinophilia until he presented to our institution. Peripheral smear showed marked eosinophilia as well as the presence of Sezary cells. A skin biopsy and lymph node biopsy revealed cutaneous T cell lymphoma. Bone marrow biopsy showed atypical large T-cells comprising 29% of the total population with loss of CD5 and CD7 and 22% eosinophils. The patient ultimately sustained multiorgan failure and expired from respiratory failure. Tissue eosinophilia was later demonstrated on autopsy.

Conclusion: Our report indicates that the Sezary syndrome can cause extreme hypereosinophilia that can result in end-organ damage.

Disclosures

No relevant conflicts of interest to declare.
Late Nephrotoxicity of Intensive Modified Program NHL-BFM-90 in Adult Poor Prognosis Patients with Diffuse Large B-Cell Lymphoma
Authors: Dorokhina Elena , Magomedova Aminat Umaraskhabovna , Kulikov Sergei M , Biryukova Ludmila , Vorobiev Andrey , Kravchenko Sergey , Savchenko Valeri
Published: October 2019

Aim. To evaluate late nephrotoxicity of intensive modified program NHL-BFM-90 (mNHL-BFM-90) in adult poor-prognosis patients with diffuse large B-cell lymphoma (DLBCL).

Materials and methods. The data of laboratory tests of blood and urine of 40 patients with DLBCL which received mNHL-BFM-90 protocol in the Hematology Research Center in the period from 2002 to 2009 years was analyzed. Group consisted of 20 men, 20 women, aged from 31 to 76 years, median age - 56.5 years at the time of the survey, the median period after therapy- 6 years. Individual cumulative dose of ifosfamide - 8-12 g / m2, cyclophosphamide - 2-3 g / m2, methotrexate - 6-9 g / m2. Comparison group included 19 patients (8 men and 11 women), aged from 39 to 78 years, median age - 70 years at the time of the survey, who received therapy to CHOP or R-CHOP. The median period after therapy - 5,5 years. Individual cumulative dose of cyclophosphamide - 3-6 g / m2. Laboratory tests of blood and urine were performed before chemotherapy and after 5 years after treatment.

Results. From all 40 patients with DLBCL received mNHL-BFM-90, signs of chronic kidney disease were found in 32 (80%) patients, only 2 (5%) of them showed a decrease in creatinine clearance daily. In comparison group signs of chronic kidney disease were found in 12 (63%) patients, 2 patients (10.5%) showed a reduction in the daily creatinine clearance. No significant differences in the manifestations of nephrotoxicity in the long term between comparison groups were found. No significant differences in the manifestations of nephrotoxicity in the long term period were found between two groups of patients (who underwent and not underwent acute renal failure during high-dose chemotherapy). Amount of nephrotoxicity signs was significantly dependent on the presence of hypertensive disease in history (p = 0.02).

Conclusion. Nephrotoxicity of high-dose mNHL-BFM 90 exceeds significantly nephrotoxicity program CHOP / R-CHOP. Conducting high-dose chemotherapy in 10 (25%) patients complicated by acute renal failure, 2 (20%) patients required hemodialysis in contrast to the group of standard chemotherapy without such complications (p=0,01). However, in the long term observations there weren't significant differences in the nephrotoxic manifestations between two groups. Impaired renal function in the long term period observed in 100% of patients undergoing acute renal failure during treatment and only in 71% of patients without acute renal failure (p=0,04). It was comparable repairing of renal function in the long term in patients undergoing high-dose therapy between patients with and without acute renal failure among those who showed signs of chronic kidney disease.

Disclosures

No relevant conflicts of interest to declare.
Disruption of the Peripheral Lymphoid Niche Contributes to Lymphopenia in CML Patients Undergoing Imatinib Treatments
Authors: Thiant Stephanie , M.Moutuou Moutuaata , Laflamme Philippe , Sidi Boumedine Radia , Larochelle Fanny , Leboeuf Dominique , Roy Denis-Claude , Guimond Martin
Published: October 2019

PURPOSE: Chronic myelogenous leukemia (CML) is a disorder affecting early hematopoietic stem cells (HSC) and is characterized by excessive proliferation and accumulation of myeloid progenitors and progeny in the periphery. During the chronic phase of the disease, CML patients are normally at low risk of developing infections but such complications tend to rise during the progression of the disease. Gleevec (imatinib mesylate) is currently administered as first line therapy for patients with Philadelphia chromosome-positive CML. Despite the relative high specificity of tyrosine kinase inhibitor (TKI) treatment towards the BCR-ABL fusion protein, off-target multikinase inhibitory effects occur and can interfere with normal hematopoiesis. This study was conducted to evaluate how myeloid and lymphoid immune homeostasis are affected by Imatinib mesylate.

METHODS: Healthy volunteer donors (n=25) and CML patients were recruited during their first visit at our CML clinic. Seven CML patients were treated with Imatinib (400mg). The median time of Gleevec treatment was 2.9 years (range: 0.5-10.9). The median time of remission post TKI was 1.1 years (range: 0.3-3). Phenotypic analysis of dendritic cell (DCs) subsets: plasmacytoid (pDCs) and myeloid type 1, 2 and 3 (mDC1, mDC2, mDC3) were evaluated by flow cytometry. Percentage and absolute numbers of naive and memory CD4+ and CD8+ T cells, NK cells and B cells were also evaluated. DCs were differentiated from purified CD34+ cells culturedwith GM-CSF (800 U/ml) or Flt3-L (50ng/ml), IL-4 (10 U/ml) and TNFa (50 U/ml), in the presence of varying concentrations of Imatinib mesylate (0 to 5µM/mL). TCR and IL-7 signaling were evaluated based on ERK-phosphorylation (-p) and STAT5-p after incubation with 3µM of Imatinib.

RESULTS AND CONCLUSION: At diagnosis, several CML patients have a deficit in DCs resulting from a severe skewing affecting BM progenitor cells. After initiating Gleevec therapy, normalization of stem cell progenitors occurs but DC counts remain well below normal levels in all CML patients. We demonstrated a direct and dose dependent interference of Imatinib on GM-CSF and Flt3-L pathways for DC differentiation from CD34+ stem cells. For T lymphocytes, Imatinib interfered with TCR and IL-7 signaling through the inhibition of ERK and STAT5 phosphorylation respectively. The failure to maintain adequate numbers of DCs combined to diminished homeostatic response to cytokines and TCR stimuli explains T cell lymphopenia in these patients. Such immune dysfunction is at least in part responsible for infectious complications that are often increased in patients treated with Imatinib.

Disclosures

No relevant conflicts of interest to declare.
PDGF/PDGFR May be Involved in the Physiopathologic Mechanism of Essential Thrombocythemia
Authors: Zhou Li Xia , Ye Jieyu , Liang En Yu , Li Chunfu , Chong Beng H , Yang Mo
Published: October 2019

Our previous studies have demonstrated that PDGF (Platelet-derived growth factor) has a potential effect in the regulation of hematopoiesis and megakaryopoiesis (Yang et al, Thromb Haemastasis, 1997; Ye et al, Haematologica, 2010). Essential Thrombocythemia (ET) is characterized by persistently elevated platelet counts in the context of a normal red cell mass. However, the physiopathologic mechanism of ET is still under investigation. Here, we tested the bone marrow plasma levels of PDGF-BB in essential thrombocythemia patients (n=16) and normal control (n=8), and found an increased PDGF-BB levels in ET patients (2070.92±123.98 pg/ml), compared with normal control (1381.85±128.37pg/ml) (P=0.002). Furthermore, we have demonstrated the presence of functional PDGF receptors (PDGFR) in human megakaryocytes, and their ability to mediate a mitogenic response by bone marrow colony-forming unit-megakaryocyte (CFU-MK) formation assay (n=6). PDGF-BB stimulated in vitro megakaryopoiesis via PDGFR. It also showed a direct stimulatory effect of PDGF-BB on c-Fos expressions in megakaryocytic cells, CHRF. We speculate that these transcription factors might be involved in the signal transduction of PDGF on the regulation of megakaryopoiesis. PDGF also enhanced platelet recovery in mice model with radiation-induced thrombocytopenia. Studies showed that PDGF, like thrombopoietin (TPO), significantly promoted platelet recovery and the formation of bone marrow CFU-MK in this irradiated-mouse. An increased number of hematopoietic stem/progenitor cells and a reduction of apoptosis were found in the bone marrow histology sections. We also demonstrated that PDGF activated the p- Akt, p-Jak2 and p-Stat3 expression, while addition of imatinib mesylate reduced p-Akt, p-Jak2 and p-Stat3 expression in CHRF cells. Our findings suggested that the PDGF-initiated megakaryopoiesis is likely to be mediated via PDGF receptors with subsequent activation of the Akt and Jak2/ Stat3 pathways. These studies provide a possible explanation that PDGF/PDGFR may be involved in the physiopathologic mechanism of essential thrombocythemia.

Disclosures

Yang: National Natural Science Foundation of China(81270580): Research Funding.
Significance of Fecal Occult Blood Test Followed By Colonoscopy for Asymptomatic Dasatinib-Induced Hemorrhagic Colitis (CML1501 Study)
Authors: Nishiwaki Satoshi , Maeda Matsuyoshi , Yamada Masahiro , Okuno Shingo , Harada Yasuhiko , Suzuki Kotaro , Kurahashi Shingo , Urano Fumihiro , Okamura Shozo , Sugiura Isamu
Published: October 2019

Introduction: Positive fecal occult blood test (FOBT) was occasionally observed in some chronic myeloid leukemia (CML) patients treated with a tyrosine kinase inhibitor (TKI). Although there are several case reports of hemorrhagic colitis in patients treated with dasatinib, the incidence of TKI-induced hemorrhagic colitis and the screening efficacy of FOBT followed by colonoscopy are unknown.

Methods: We prospectively enrolled CML patients treated with a TKI. The first FOBT (FOBT1) was performed for all patients. The first CF (CF1) was performed for patients with positive FOBT1. When there were any lesions, tissue biopsy was performed for pathological analyses. After confirmation of TKI-induced colitis by pathological analyses, the TKI was interrupted for 2-4 weeks. And then, FOBTs were re-evaluated (FOBT2).

Results: Between February 2015 and September 2015, 30 patients were enrolled in this study. All patients were in chronic phase. FOBT1 was positive in 10 of 30 patients. All patients with positive FOBT1 were treated with dasatinib and revealed no symptoms. CF1 was performed for 10 patients with positive FOBT1. All patients showed abnormal endoscopic findings: colitis in 6 patients, polyps in 3, and hemorrhoid in 1. Dasatinib-induced hemorrhagic colitis was confirmed in 6 of 18 patients treated with dasatinib (33%). Median duration of dasatinib before FOBT1 was 16 months (range 3-35 months). Median dose of dasatinib was 85mg (range 50-100mg). It had been reduced in 3 patients due to pleural effusion. Its endoscopic feature was read flare and/or erosion. Immunohistological analyses showed CD3+, CD8+, CD56+ and Granzyme B+ cytotoxic T lymphocyte infiltration. Dasatinib was discontinued at a median of 27 days after CF1 in patents with dasatinib-induced hemorrhagic colitis. Discontinuation period was 14 days in 4 patients, 27 days in 1, and 28 days in 1. FOBT2 was negative in all but one patient who had concurrent colorectal polyps. No CML progression due to dasatinib discontinuation was observed.

Conclusion: Dasatinib-induced hemorrhagic colitis was observed in a third of asymptomatic patients treated with dasatinib. FOBT can be a useful non-invasive screening for the disease, and it can be confirmed by the following CF and pathological analyses. It was caused by cytotoxic T lymphocytes, and hemorrhage was resolved after dasatinib discontinuation.

Disclosures

No relevant conflicts of interest to declare.
Thrombospondin-1-Derived Peptide Rfyvvmwk Improves the Adhesive Phenotype of CD34+ Cells from Atherosclerotic Patients with Type II Diabetes
Authors: Cointe Sylvie , Rhéaume Eric , Martel Catherine , Blanc-Brude Olivier P. , Dubé Evemie , Sabatier Florence , Dignat-George Francoise , Tardif Jean-Claude , Bonnefoy Arnaud
Published: October 2019

Background: CD34+ progenitor cells are growingly used for vascular repair. However, in diabetic individuals with cardiovascular diseases, these cells have dysfunctional engraftment capabilities, which compromise their use for autologous cell therapy. The thrombospondin-1-derived peptide RFYVVMWK has previously been reported to stimulate cell adhesiveness through CD47 and integrin activation pathways.

Objectives: Our aim was to test whether RFYVVMWK preconditioning could modulate CD34+ cells phenotype and enhance its pro-adhesive properties in diabetic patients.

Patients/methods: Peripheral blood mononuclear CD34+ cells isolated from 40 atherosclerotic patients with (TIID; n=20) or without (NonTIID; n=20) type II diabetes were pre-conditioned with 30µM of RFYVVMWK (or truncated peptide RFYVVM. CD34+ cell adhesion was assessed on a vitronectin-collagen matrix and on a TNFa or IL-1b-stimulated HUVEC monolayers. Adhesion receptors, platelet/CD34+ cell conjugates, and cell viability were analyzed by flow cytometry and confocal microscopy.

Results: RFYVVMWK increased by 8 folds the adhesion of TIID CD34+ cells to the vitronectin-collagen matrix (p <0.001) corresponding to a 3-fold increase compared to unstimulated NonTIID CD34+ cells. The peptide induced the formation of platelet/CD34+ conjugates and increased the expression of TSP-1, CD29, CD51, and CD62P in both TIID and NonTIID cells. However, RFYVVMWK treatment did not affect the viability/apoptosis of CD34+ progenitor cells.

Conclusions: Priming CD34+ cells with RFYVVMWK may be instrumental to enhance their vascular engraftment during autologous pro-angiogenic cell therapy.

Disclosures

No relevant conflicts of interest to declare.
Inactivation of Zika Virus By Solvent/Detergent Treatment or Pasteurization
Authors: Schmidt Torben , Kuehnel Denis , Mueller Sebastian , Pichotta Alexander , Radomski Kai Uwe , Volk Andreas , Knaub Sigurd
Published: October 2019

BACKGROUND: Since 2015 Zika virus (ZIKV) has spread within the Americas and even before in other regions. In 2016, the World Health Organisation (WHO) declared the ZIKV as a "Public Health Emergency of International Concern" (PHEIC). Therefore, it is very important to confirm the pathogen safety of blood plasma products. These products generally demand an effective donor selection as well as robust pathogen elimination/reduction steps within the manufacturing processes. Here, we provide experimental evidence that both solvent/detergent (S/D) treatment and pasteurization effectively inactivate ZIKV.

STUDY DESIGN AND METHODS: The ZIKV inactivation capacity of the pasteurization step and the S/D treatment for different products and process conditions were investigated. Therefore, in-process material from commercial batches was collected and spiked with ZIKV in a ratio of approximately 1:10. Viral loads were determined prior to the safety steps and at pre-defined intervals during the individual processes to investigate the inactivation kinetics. Each process was investigated in duplicate runs.

RESULTS: Sixty minutes after S/D treatment the viral titer was below detection limit during manufacturing of human plasma resulting in mean ZIKV reduction factor (LRF) of ≥ 6.78 log10. For immunoglobulin (IgG) after 240 min of S/D treatment the LRF was determined to be ≥ 7.00 log10. ZIKV was inactivated ≥ 6.18 log10 after 480 min by S/D treatment during the manufacture of factor VIII.

During pasteurization of human albumin ZIKV infectivity was already below the detection limit after the heat-up phase. This finding was confirmed by subsequent test samples up to 2 hours. A LRF of ≥ 7.48 log10 was demonstrated.

CONCLUSION: We showed that different S/D treatment procedures inactivate the ZIKV to below the detection limit. This effect was seen independently of various product matrices or the choice of S/D reagent at various concentrations and temperatures, while taking product-specific treatment times into consideration. We also showed that pasteurization is a very efficient inactivation step for the ZIKV. These results demonstrate the effectiveness of S/D treatment against even newly emerging lipid-enveloped viruses like ZIKV in the manufacturing of human plasma and derivatives thereof, such as IgG or factor VIII.

Disclosures

Schmidt: Octapharma: Employment. Kuehnel:Octapharma: Employment. Mueller:Octapharma: Employment. Pichotta:Octapharma: Employment. Radomski:Octapharma: Employment. Volk:Octapharma: Employment. Knaub:Octapharma: Employment.
Age-Related Changes in Methylome and Transcriptome of Hematopotietic Stem Cells Underlie Natural Genetic Variations
Authors: Liang Ying
Published: October 2019

The aging of hematopoietic stem cells (HSCs) contributes to the aging of blood system and perhaps the whole organism. The aging process is coordinately determined by both genetic and epigenetic factors, and demonstrates inter-individual variations. We used high-throughput sequencing methods to study the age-dependent changes of genome-wide DNA methylation and gene expression patterns in HSCs of C57BL/6 (B6) and DBA/2 mouse strains, which have shown natural variations in HSC aging process. We observed global age-associated decrease of DNA methylation in both strains, but D2 HSCs have a stronger loss of epigenetic control than B6 stem cells during aging. Majority age-related changes of DNA methylation occur from young to mid-aged stages. We identified stable strain-specific differentially methylated regions (DMRs) that overlap with cis-eQTLs. Moreover, transcription factor binding site motifs are more likely to be disrupted in the DMRs, suggesting the potential impact of genetic variations on epigenetic regulation of HSC aging. We further demonstrated that strain-specific DMRs have more profound effects on the aging of B6 HSCs than D2 stem cells. Transposons are differentially regulated by the DMRs in the two strains, in which D2 HSCs are prone to transposon insertion. This study comprehensively investigated the effects of natural genetic and epigenetic variations on HSC aging. Loss of DNA methylation is an epigenetic signature of stem cell aging, and DNA methylation variations correlates with genetic variations, both contributing to inter-individual differences in stem cell and perhaps organismal aging.

Disclosures

No relevant conflicts of interest to declare.
Autologous Hematopoietic Cell Transplantation in Patients with Multiple Myeloma: IMPACT of Age
Authors: Dhakal Binod , Nelson Ariel , Guru Murthy Guru Subramanian , Fraser Raphael , Eastwood Daniel , Hamadani Mehdi , D'Souza Anita , Hari Parameswaran
Published: October 2019

Background: In both the novel and pre-novel agents era, high dose therapy followed by autologous hematopoietic cell transplant (AHCT) has been shown to prolong survival in multiple myeloma (MM) in randomized trials, but prospective have only included younger patients (65 -70 years and younger). Given that the median age at diagnosis of MM is 66 years, it is important to know the outcomes of AHCT in older patients. Similarly, the definite outcomes of AHCT in very young patients (<50 years) is also lacking as they represent a very small proportion in clinical trials.

Methods: We analyzed a consecutive cohort of patients with MM receiving ASCT from 2000-2015 in two different age groups, old (>70 years; N=105) and young (≤50 years, N=86) and compared the outcomes. The primary objectives were to assess overall survival (OS), progression free survival, (PFS), and non-relapse mortality (NRM) in these two groups.

Results: Eighty-sex patients were young (≤50 years), while 105 patients were old (>70 years). Young patients had better performance status and lower co-morbidity index, while majority of older cohort received dose reduced high dose melphalan between 140-180 mg/m2. Median follow up of survivors in the younger cohort was 33 months (range, 2-164) compared to the 22.5 months (3-133) in the old group (p=0.02). The PFS at 1 year was 60% (95%CI 46-72) for young patients and 58 (95%CI, 45-69) for the old ones. The OS at 1 year was 92% (95% CI 84-96) for younger and 85% (95% CI 76-91) for the older cohorts. On multivariate analysis, age did not have any effect on survival (p =0.82), high-risk cytogenetics (HR 2.2, 95% CI 1.06-4.6, p=0.04) was associated with higher mortality. High-risk cytogenetics (HR 1.2, 95% CI 1.1-3.5, p=0.004) and non-responding or progressive disease at transplantation (HR, 5.0 95% CI 1.8-13.5, p=0.02) were significantly associated with inferior PFS.

Conclusion: Age by itself should not be a limiting factor in considering the modality of AHCT. However consideration could be given to augmenting therapy for young patients with additional novel treatments post transplant given their similar outcomes with the old patients based on our study.

Overall survival between the two age groups, 1: >70 years old and 2: ≤50 years old. Overall survival between the two age groups, 1: >70 years old and 2: ≤50 years old. Figure Figure.

Disclosures

Hamadani: Takeda: Research Funding. Hari:Merck: Research Funding; BMS: Honoraria.
Safety and Efficacy of Direct-Acting Antiviral Drugs in Patients with Haemoglobinophaties and Chronic Hepatitis C Infection
Authors: Piga Antonio , Rigano Paolo , Origa Raffaella , Cappellini M. Domenica , Pinto Valeria , Tartaglione Immacolata , Filosa Aldo , D'Ascola Domenico , Di Marco Vito , Forni Gian Luca
Published: October 2019

Background and Aim: Direct-acting antiviral drugs (DAAs) have a very high efficacy in patients with hepatitis C virus (HCV) infection, but they have not been extensively used in patients with haemoglobinophaties. To evaluate the safety and efficacy of DAA regimens in this subset we used the ITHACA-SITE dataset, which includes patients with haemoglobinophaties and chronic HCV liver disease treated in Italy.

Patients and methods: Between March 2015 and June 2016, 121 patients included in the ITHACA-SITE dataset started DAA regimens. Cirrhosis was defined by FibroScan®showing≥12 kPa performed within 6 months before the treatment. Regimen choice and use of ribavirin were based on viral genotype and stage of disease, according to guidelines. Negative HCV RNA at week 12 of post-treatment follow up was considered as Sustained Virological Response (SVR).

Results: The mean age of 121 patients was 42 years, 75 (62%) were males, 101 (83.5%) had β-thalassemia major, 9(7.5%) had β-thalassemia intermedia and 10 (8.3%) had sickle cell disease. Sixty-six patients (54%) had the diagnosis of chronic hepatitis and 55 (46%) hadthe diagnosis of cirrhosis. The prevalence of HCV genotypes (G) was: G1a 6 (5%), G1b 70 (57.8%), G2 31 (25.6%), G3 6 (5%), G4 8 (6.6%). Fifty-six patients (46.3%) were Peg-interferon (P) and Ribavirin (R) naïve and 65 (53.7%) were P/R experienced, 20 patients (16.5%) had diabetes and 29 (24%) had heart disease. By June 2016 63 patients (52%) had concluded the treatment and 33 (27.3%) had concluded the post-treatment follow-up. By ITT, the rate of response at the end of treatment (ETR) was 100% (63/63) and 94% (31/33) of patients achieved a SVR. No patients stopped treatment because of adverse events. No interference with chelation therapy was observed.

Conclusions: Use of DAAs regimens in practice is safe and effective in patients with haemoglobinophaties and cirrhosis or chronic hepatitis due to HCV. The therapy is indicated also in patients with co-morbidity. The lifetime utility of HCV eradication in terms of reduction of events and overall survival needs evaluation in long-term observational cohorts.

Disclosures

Piga: Novartis: Research Funding; Apopharma: Honoraria. Di Marco:Gilead: Research Funding. Forni:Novartis, Celgene: Research Funding.
Endothelin-1 Regulates Molecules of the Major Histocompatibility Complex: Role in Sickle Cell Disease
Authors: Rivera González Lorena , Inostroza-Nieves Yaritza , Lozano Alexandra , López Pablo J. , Rosado Alicea Jamie , Prado Gregory N. , Romero Jose R. , Rivera Alicia
Published: October 2019

Molecules of the Major Histocompatibility Complex (MHC), and in particular specific human leukocyte antigen (HLA) alleles, have been proposed in the pathophysiology of immune and vascular alterations leading to vasoocclusive crises (VOC) and stroke in Sickle Cell Disease (SCD). Endothelial cells express MHC molecules following exposure to cytokines. SCD is characterized, in part, by vascular endothelial cell activation, increased oxidative stress, sickle cell adhesion and excess levels of the potent mitogen, endothelin-1 (ET-1). ET-1 activates endothelial cells, induces oxidative stress and inflammation in the vascular wall and regulates erythrocyte homeostasis. However, the role of ET-1 on MHC regulation in SCD is not clear. We first characterized the effect of ET-1 on MHC expression in the human endothelial cell line, EA.hy926. We observed dose-dependent increases in the expression of MHC class I (HLA-A2 4.8 ± 2.1 folds p<0.01 n=4), MHC class II (HLA-DR 4.4 ± 1.7 folds p<0.01 n=4) and MHC transcription factor (CIITA 3.5 ± 1.8 folds p<0.05 n=4) in EA.hy926 cells. ET-1-stimulated expression of HLA-A2, HLA-DR and CIITA were significantly blocked by pre-incubation of cells with 10 µM BQ788, a selective blocker of ET-1 type B receptors (p<0.01, n=4). Chromatin immunoprecipitation (ChIP) studies in EA.hy926 cells showed that ET-1 increased Histone H3 acetylation of the promoter region within MHC molecules (HLA-A2 62% ± 5%, HLA-DRB 33% ± 18%, p<0.01, n=4); an event that was likewise blocked by BQ788. We then studied two sickle transgenic knockout mouse models of moderate to severe disease phenotype, βSAntilles and Berkeley (BERK) mice, respectively. We observed significant increases in MHC molecule, H2-Aa mRNA levels (n=7; p<0.01) in spleens from sickle transgenic mice when compared to transgenic knockout mice expressing human hemoglobin A (HbA). We then treated BERK, βSAntilles and HbA mice for 14 days with ET-1 receptor antagonists and observed significant reductions in H2-Aa mRNA levels in spleen tissue from sickle transgenic mice but not in HbA mice (n=7; p<0.05). These results implicate ET-1 as a novel regulator of MHC molecules and suggest that ET-1 receptor blockade represents a promising therapeutic approach to regulate both immune and vascular responses in SCD.

Disclosures

No relevant conflicts of interest to declare.
Increased T Follicular Helper Cells in Patients with Aplastic Anemia
Authors: Verigou Evgenia , Galanopoulos Athanasios , Kattamis Antonios , Symeonidis Argiris , Solomou Elena E.
Published: October 2019

Introduction:T-follicular helper cells represent a sub-population of CD4+helper T cells (TFH), and they are characterized by the surface expression of CXCR5, ICOS, and PD1, the transcription factor Bcl-6, and produce mainly IL-21, but also IL-17, IL-4, and IFN-γ. They represent the major population that helps B cells to turn into plasma cells and they are implicated in the pathogenesis of different autoimmune diseases. Peripheral CXCR5+CD4+ T helper cells (p-TFH) are the circulating component of TFH. p-TFH cells have been extensively studied in the context of inflammation and autoimmunity. Patients with systemic lupus erythematosus and rheumatoid arthritis have increased p-TFH. T follicular regulatory T cells (Tfr) express CXCR5, ICOS and PD1, CD25 but also FOXP3, and they can inhibit B cell responses.

Immune dysregulation characterizes patients with aplastic anemia. We and others have previously shown that aplastic anemia patients have decreased levels of regulatory T cells (Tregs) at presentation and increase if they respond to treatment. Additionally, these patients display increased Th17 cells at presentation.

Specific aim: In this study we wanted to examine the expression of p-TFH and Tfr cells in patients with aplastic anemia. We isolated peripheral blood mononuclear cells (PBMCs) from patients with aplastic anemia (n=8) and ten healthy, age- matched controls. Written informed consent was obtained from all study subjects. PBMCs were left untreated or activated with PMA and Ionomycin, in the presence of Brefeldin A, for 5 hours. Subsequently, cells were stained with the surface markers CXCR5, CD4, CD57, CD25, CD127, CD278, CD279, IFN-g,IL-21, and IL-17, and subsequently analyzed by flow cytometry.

Results: Patients with aplastic anemia showed increased expression of CD4+CXCR5+ cells (Tfh) compared to healthy controls (3,03±0,55% vs 1,3±0,52% respectively, p=0.08). Additionally, these patients also showed decreased expression of Treg cells compared to healthy controls (0,74±0,27% vs 2,25±0,70% respectively, p=0.03). When analyzed for Tfr cells, patients with aplastic anemia showed minimal levels of these cells compared to helathy controls (0,05±0,02% vs 1,14±0,16% respectively, p<0.001) (Fig. 1).

Conclusion: Although the number of patients analyzed is limited, our preliminary results suggest that aplastic anemia patients show increased CXCR5+CD4+ cells compared to age-matched control subjects, and decreased Tfr cells. Further analysis of a larger pool of subjects is underway along with the examination of the specific transcription factor Bcl-6, to reveal the role of p-TFH and Tfr cells in the immune pathogenesis of aplastic anemia.

Figure Figure.

Disclosures

No relevant conflicts of interest to declare.
Assessing Patient Preferences for the Benefits and Risks of Treating for Acute Myeloid Leukemia (AML): A Pilot Study
Authors: Seo Jaein , Smith B. Douglas , Estey Elihu H. , Voyard Ernest S. , O'Donoghue Bernadette , Bridges John F. P.
Published: October 2019

Introduction:Acute Myeloid Leukemia (AML) is a blood cancer which progresses rapidly in the absence of treatment. There have been few advances in the treatment of AML over the last three decades. In 2015 the Leukemia and Lymphoma Society (LLS) initiated a research program to assess patient preferences for AML treatments. The aim of this research was to promote patient-focused drug development and inform future regulatory decisions. We sought to develop and pilot a patient-centered survey instrument to assess patient preferences for the benefits and risks of AML treatments.

Methods: Development was informed by a targeted literature review and engagement with an expert stakeholder committee (n=12) to guide the clinical accuracy and relevance of the survey instrument. A community stakeholder committee, consisting of patients with AML and caregivers (n=15), provided information about their experiences with AML and various treatments. They also engaged in pretest interviews to test comprehension and ensure it captured the patient experience. A discrete-choice experiment (DCE) was developed spanning 5 benefits and risks, including event-free survival (EFS), complete remission (CR), time in hospital, short-term side effects, and long-term side effects. This DCE consisted of 16 pairs of hypothetical treatments, with participants being asked to identify which treatment they would prefer in each pair. Results of a pilot study with AML patients and caregivers were assessed by Z-score that were derived from a conditional logistic model regressing each attribute upon their choices.

Results: The pilot included 18 patients and 8 caregivers with a mean age of 50 years (range=24-81). Most participants were college educated (n=22), Caucasian (n=19), privately insured (n=21), and employed (n=13). Participants valued CR the most (Z-score=7.95, p<0.001), followed by EFS (5.32, p<0.001). They were most averse to time in hospital (-3.41, p=0.001), followed by long-term side effects (-3.03, p=0.002) and short-term side effects (-1.99, p=0.047), which was marginally significant.

Conclusions: This study demonstrates the value of rigorous community engagement in developing survey instruments to measure patient preferences. The results of this pilot study demonstrate the ability of our DCE to measure treatment preferences of AML patients and caregivers. Given this success, we are currently engaged in a nationally study where we will recruit a larger and more diverse sample. The results of this national study will inform drug developers and regulatory decision makers.

Disclosures

Seo: The Leukemia & Lymphoma Society: Research Funding. Voyard:The Leukemia & Lymphoma Society: Employment. O'Donoghue:The Leukemia & Lymphoma Society: Employment. Bridges:The Leukemia & Lymphoma Society: Research Funding.
Does Normal Reticulocyte Hemoglobin Rule out Iron Deficiency As a Cause of Normocytic Normochromic Anemia?
Authors: Yu Zhou , Blankenship Leann Michelle , Ibrahim Mohammed , Gbadamosi Bolanle , Stender Michael , Anderson Joseph , Jaiyesimi Ishmael , Huang James
Published: October 2019

Background: Normochromic, normocytic anemia is the most frequently encountered type of anemia with a wide differential diagnosis. Iron studies are frequently ordered as part of the anemia work up as iron deficiency anemia could be potentially normocytic and normochromic. Recently reticulocyte hemoglobin has emerged as a marker to help identify early stages of iron deficiency at a time that other traditional biochemical studies for iron deficiency are non-informative. We hypothesize that normal reticulocyte hemoglobin level is a sensitive negative predictor of iron deficiency.

Aim:To evaluate whether a normal reticulocyte hemoglobin level can be used to exclude iron deficiency anemia in patients with normocytic normochromic anemia.

Methods:We reviewed the results of all hematological tests obtained as a part of hematopathology consults which were requested between January to April of 2016. Electronic medical records were screened for inclusion criteria. We included patient with hemoglobin less than 12g/dl, normal MCV (80-100) and normal reticulocyte hemoglobin level (>31 pg). We calculated the percentage of patients who had normal iron store suggested by ferritin, total iron binding capacity and iron saturation studies.

Results:We included 41 patients who has normocytic normochromic anemia with normal reticulocyte hemoglobin level. 39 out of 41 patients have normal iron studies indicating iron deficiency anemia was not the cause of anemia. The overall negative predictive value is 95.1%. Among the two patients whose ferritin levels are mildly decreased, suggesting possible iron deficiency, one patient had a blood transfusion prior to checking reticulocyte hemoglobin level. The other patient donates blood frequently and was started on oral iron supplementation trial without any hemoglobin improvement after two weeks. It appeared that these two patients might not have true iron deficiency.

Conclusion: Measurement reticulocyte hemoglobin level can serve as a useful marker to rule out iron deficiency as a cause of normocytic normochromic anemia. Serum Iron and ferritin studies are not indicated in patients with normocytic normochromic anemia with normal reticulocyte hemoglobin.

Disclosures

No relevant conflicts of interest to declare.
Clinical Characterization of Staphylococcus Septicemia-Associated Hemophagocytic Lymphohistiocytosis
Authors: Qiu Hongxia , Song Meng , Ni Fang , Wang Jujuan
Published: October 2019

Abstract Objective: To analyze the epidemic characters, pathogenesis, clinical manifestations, laboratory data, therapeutic response, and prognosis in patients with staphylococcus septicemia-associated hemophagocytic lymphohistiocytosis(HLH). Methods A r

Objective To analyze the epidemic characters, pathogenesis, clinical manifestations, laboratory data, therapeutic response, and prognosis in patients with staphylococcus septicemia-associated hemophagocytic lymphohistiocytosis(HLH).

Methods: retrospective study was performed on 7 patients with staphylococcus septicemia-associated HLH who were admitted to The First Affiliated Hospital of Nanjing Medical University from January 2010 to December 2014.

Results Seven out of 633patients(1.11%) with staphylococcus septicemia were confirmed to have HLH. The incidence of HLH in staphylococcus subspecies were:S.caprae, 12.50%;S.warneri, 12.50%;Methicillin-resistant Staphylococcusaureus(MRSA), 3.23%;S.hominis, 1.40%;S.haemolyticus, 1.25%;S.epidermidis, 0.59%, respectively. The main clinical manifestations included persistentfever(100%) and splenomegaly(85.71%). The most prominent laboratory abnormalities werehemocytopenia, abnormal hepatic function, and elevated ferritin. Bone marrow aspirations were performed on 6 patients and all of them hadhemophagocytosis shown in bone marrow smears. Among 7 patients, 4 patientssurvived(57.14%) and 3 patients died(42.86%). All the 4 cured patients were treated with antibiotic plus corticosteroid. And 2 of them used gamma globulin, while one used the COP chemotherapy regimen. Treatment with antibiotic plus corticosteroid was effective in our study.

Conclusion Treatment with antibiotic plus corticosteroid was recommended for staphylococcus septicemia-associated HLH, and gamma globulin may also be useful in reducing the risk of death.

Disclosures

No relevant conflicts of interest to declare.