bluuebunny/crossref_metadata_2025 huggingface dataset:
Dose-Intense BACOP for Treatment of High Risk Peripheral T-Cell NHL (PTCL)
Authors: Aurer Igor , Dujmovic Dino , Radman Ivo , Basic-Kinda Sandra , Nemet Damir , Ilic Ivana , Stern-Padovan Ranka , Santek Fedor , Sertic Dubravka
Published: October 2019

Abstract 4889

PTCLs, except ALK+ ALCL, have a bad prognosis with overall 5-year survival rates below 30%. Because of their relative rarity, no large randomized trials have been performed and the importance of individual cytotoxic agents, their dose density and intensity as well as that of autografting remains unproven. At our center we have been using dose-intense BACOP, a regimen modeled after the French ACVBP, for treatment of patients with high risk PTCL. The regimen consisted of cyclophosphamide 1500 mg/m2 day 1, doxorubicin 75 mg/m2 day 1, bleomycin 15 mg days 1 and 5, vincristine 2 mg days 1 and 5 and prednisone 50 mg/m2 days1-5 every 3 weeks for 4–6 cycles. Responding patients underwent stem-cell collection and autografting after BEAM conditioning. Areas of initially bulky disease and those not in CR prior to transplantation were irradiated posttransplant with 30–40 Gy. Patients with high risk for development of CNS disease received intrathecal methotrexate prophylaxis. Response evaluations were performed after 2–3 cycles and at the end of chemotherapy. Patients were considered high risk if they had any PTCL type with IPI 3 or more, bulky disease or failed to respond to 2–3 cycles of standard CHOP-21. We treated 28 patients, 20 men and 8 women, 17–67 years old (median 46). Thirteen had PTCL-NOS, 8 ALCL, 3 AILD, 2 nasal type, 1 panniculitis-like and 1 hepatosplenic PTCL. Two had stage IE bulky, 1 stage II bulky, 2 stage III and 23 stage IV disease. The toxicity of treatment was substantial, all patients experienced serious hematological toxicity and infections or neutropenic fever, 3 died of infection during chemotherapy and 1 during stem-cell transplantation, 2 had DVT/PE, 1 died of complications related to the necrosis of an extremely bulky tumor and one underwent emergency splenectomy after tumor necrosis caused splenic rupture. Three patients were unavailable for efficacy analysis due to early death, 4 progressed and 21 initially responded (75%); 14 achieved CR and 7 PR. None of the 3 patients with nasal type or hepatosplenic PTCL responded. Of the responders, 11 underwent autografting as intended, 3 relapsed prior to transplantation, treatment was modified in 3 due to toxicity, 2 were not transplanted due to patient refusal or physician decision, 1 underwent allogeneic transplantation and 1 failed to mobilize. After a median follow-up of survivors of 20 months, 9 patients are still alive, including 8 who were autografted as intended. Median response duration was 12 months, FFS 6 months and OS 9 months (Fig). In conclusion, a significant proportion of high-risk PTCL-NOS, ALCL and AILD patients respond to dose-intense BACOP followed by autologous stem-cell transplantation but the toxicity of treatment is substantial.

Disclosures:

No relevant conflicts of interest to declare.
Childhood Anaplastic Large Cell Lymphoma with Unusual Localization-A Case Report
Authors: Unuvar Aysegul , Uzunhan Tugce , Karakas Zeynep , Ozturk Gulyuz , Anak Sema , Agaoglu Leyla , Devecioglu Omer
Published: October 2019

Abstract 4903

Anaplastic large cell lymphoma (ALCL) accounts for approximately 10% of the childhood lymphomas. ALCL is characterized by peripheral, mediastinal or intra-abdominal lymph node involvement or extranodal involvement. We report a child with ALCL with unusual localization. An eleven-year-old boy had been admitted to the another university hospital with left knee pain and swelling for 15 days. The left knee joint punction had been done, and osteomyelitis had been diagnosed, and treated with antibiotics. However, there had been no recovery from his complaints, and left knee synoviectomy had been done. This operation had revealed anaplastic large cell lymphoma in the intraarticular soft tissue of the left knee. Then, he was referred to our hematology and oncology clinic for advanced therapy. There was nothing significant in the background and family history. Physical examination showed a 3 cm mass arising from the left knee joint with lobulated contours. PET-CT showed the left inguinal lymph node involvement, only. Chemotherapy according to ALCL 99 protocol was started because of the tumor was not resectable. After the first course of therapy, tumor mass was reduced apparently. By the end of his second chemotherapy course, tumor was not detectable. Now, he completed his therapy, and he is on follow-up without any complaint or recurrence for 18 months. In conclusion, to the best of our knowledge, this is the first reported case with ALCL originating from the intraarticular soft tissue which was treated successfully.

Disclosures:

No relevant conflicts of interest to declare.
LNA Anti-Microrna-155: a Novel Therapeutic Strategy In Waldenstrom Macroglobulinemia and Chronic Lymphocytic Leukemia
Authors: Zhang Yong , Roccaro Aldo M. , Broom Oliver , Kauppinen Sakari , Liu Yang , Morgan Brittany , Azab Abdel Kareem A. , Azab Feda , Maiso Patricia , Ngo Hai T , Sacco Antonio , Ghobrial Irene M
Published: October 2019

Abstract 4914

Background.

We and others have previously demonstrated that microRNA-155 (miR-155) is overexpressed in patients with Waldenstrom's Macroglobulinemia (WM) and Chronic lymphocytic leukemia (CLL) compared to healthy subjects; indicating that miR-155 may play a crucial role in regulating the pathogenesis of these diseases. We therefore evaluated the role of the novel locked nucleic acid (LNA) antimiR against miR-155 in WM and CLL-derived cell lines.

Methods.

WM and CLL-derived cell lines (BCWM.1, MEC1) have been treated with 20μM antisense LNA antimiR-155 or scramble probe (Santaris Pharma, Hørsholm, Denmark). Efficiency of delivered LNA oligos into cells was determined by using FAM-labeled LNA, followed by Flow cytometry (FACS). Survival and cell proliferation were assessed by MTT assay and thymidine uptake, respectively. Serum levels of miR-155 from peripheral blood of WM patients or healthy subjects and the level of miR-155-targeted genes were detected by quantitative RT-PCR (qPCR).

Results.

Efficiency of delivered LNA oligos into both WM and CLL-derived cell lines was higher than 90%. LNA antimiR-155 reduced proliferation of WM and CLL-derived cell lines, as compared to LNA scramble probe used as a control. In contrast, LNA antimiR-155 did not exert any effect on survival of them. We demonstrated increased expression of miR-155-targeted genes, such as CEBPβ, SOCS1, and TP53DINP1 in WM cells upon LNA antimiR-155 treatment. Moreover, we found miR-155 could be detected in peripheral blood serum of WM patients, but not the serum of healthy subjects.

These data provide preclinical evidence for using the novel antisense LNA antimiR-155 in WM and CLL. Moreover, serum levels of miR-155 could potentially be used as a biomarker in patients with WM.

Disclosures:

No relevant conflicts of interest to declare.
Acute Painful Radiculopathy After Intrathecal Rituximab
Authors: Bromberg Jacoline , Doorduyn Jeanette , Baars Johanna W. , Van Imhoff Gustaaf , Enting Roelien , van den Bent Martin
Published: October 2019

Abstract 4920

Within a prospective phase II study (HOVON 80) of patients with recurrent diffuse large B-cel lymphoma in the CNS, three of 13 patients treated with intrathecal rituximab developed an acute, transient, extremely painful lumbosacral radiculopathy. All were treated with systemic R-DHAP every 4 weeks with i.v. HD-MTX on day 15 for three cycles. In addition intrathecal Rituximab was administered twice weekly via lumbar puncture starting on day -1. According to protocol the first administration consisted of 10 mg of rituximab after premedication with acetaminophen, thereafter the dose was increased to 25 mg. No patient experienced side effects of the first intrathecal administration of rituximab. However, after the first administration of 25 mg rituximab three of 13 treated patients reported extremely painful tingling sensations in the buttocks, legs and feet immediately after administration and lasting 30–60 minutes. Concomitantly a temporarily increased bloodpressure was documented. Premedication with an antihistaminic in the third patient was ineffective. No neurologic deficits occurred and the pain resolved completely. The patients refused further treatment with intrathecal rituximab, and therapy was changed to intrathecal methotrexate, without any side effects. After these events the rituximab was diluted in saline to 5 mg/ml, the dose reduced to 10 mg per administration, and 4 mg dexamethasone was administered concomitantly in all subsequent patients. Twelve additional patients were thus treated and no further incidents of painful radiculopathy were observed. This serious, though completely transient, adverse effect of intrathecal rituximab precludes intrathecal administration of higher doses of rituximab via lumbar route. It has never been described after intraventricular administration.

Disclosures:

Bromberg: Roche: Research Funding. Off Label Use: rituximab administration intrathecally. Doorduyn: Roche: Research Funding. van den Bent: Roche: Consultancy, Research Funding.
Triptolide Induces Apoptosis In Acute Lymphoblastic Leukemia Cells by Inhibition of the Oncoprotein MDM2
Authors: Huang Mei , Gu Lubing , Zhou Muxiang
Published: October 2019

Abstract 4934

Triptolide, a nature product derived from the Chinese plant Tripterygium wilfordii, is reported to exhibit antitumor effects in a broad range of cancers. Recent studies indicate that the antitumor activity of triptolide is associated with its biological action to inhibit expression of many oncoproteins and anti-apoptotic or survival factors that were expressed in the cancer cells. Herein, we demonstrate that triptolide induces apoptosis in a subgroup of acute lymphoblastic leukemia (ALL) cells that overexpress MDM2 oncoprotein by inhibiting the MDM2 expression. In pediatric ALL, overexpression of MDM2 by leukemic cells is typically associated with a wild-type (wt) p53 phenotype and resistance to conventional chemotherapeutic drugs such as doxorubicin. In the present study, we evaluated the role of triptolide in regulating MDM2 and in inducing apoptosis, as compared to doxorubicin, using ALL lines and primary ALL samples. In contrast to doxorubicin, which induced p53 activation and a subsequent upregulation of MDM2, triptolide strongly induced persistent inhibition of MDM2 followed by a steady-state activation of p53, which resulted in potent apoptosis of the MDM2-overexpressing ALL cells tested, even if they were doxorubicin-resistant. We discovered that triptolide's inhibition of MDM2 in ALL cells occurred at the post-transcriptional level through inhibition of mRNA synthesis. Because p53 function is inhibited by MDM2 in chemoresistant/MDM2-overexpressing ALL cells, potent killing of these cells by triptolide suggests that this naturally-derived agent may be a novel therapeutic for refractory ALL.

Disclosures:

No relevant conflicts of interest to declare.
Vitamin E Does Not Reduce Cardiomyopathy In Doxorubicin treated Rats
Authors: Guerra Jhon A , Torres-Serrant Maribel , Santiago Pedro J
Published: October 2019

Abstract 4944

Introduction:

The clinical efficacy of doxorubicin is severely limited by its cardiotoxicity. Antioxidants represent the largest class of chemicals examined as potential protective agents and for which there is continuing interest. Dexrazoxane is the current agent used for the control of doxorubicin related cardiotoxicity. However, in large trials the incidence was reduced only by 50% (1). Vitamin E, a known antioxidant and free radical scavenger agent, has been evaluated in the past as a cardio protective drug in animal models receiving doxorubicin but contradictory conclusions regarding this effect have been reported (2, 3). We hypothesized whether Vitamin E has an effect in cardioprotection in rats receiving doxorubicin.

Design and Methods:

Three groups of 6 Rats each were used for the experiment. Group 1 received doxorubicin 1.5mg/kg intraperitoneally (IP) weekly for 9 weeks for a total cumulative dose of 13.5mg/kg. Group 2 received Vitamin E 100 IU/kg/weekly by IP injection, 48 hours prior to the doxorubicin. Group 3 received 0.5 cc of saline solution 0.9% IP weekly for 9 weeks as control group. Functional parameters including plasmatic nitric oxide (NO) levels and cardiac ejection fraction (EF) were determined on each group at the end of the experiment.

Results:

Doxorubicin treatment significantly increased plasmatic NO concentration when compared with controls (35.30 ± 5.63 mM vs. 14.72 ± 2.66 mM, n=6, P=0.016); however, in the group of rats receiving Vitamin E prior to doxorubicin, NO did not have a significant decrease (from 35.30 ± 5.63 mM to 31.77± 8.91 mM n=6, P=0.75). Regarding EF, doxorubicin treatment decreased EF significantly when compared with saline controls rats (59 ± 5.61% vs. 77 ± 3.89 %, n=6, P<0.05); however, in the group of rats receiving Vitamin E prior to doxorubicin, EF did not have a significant improvement (from 59 ± 5.61% to 69.17 ± 4.4, n=6, P=0.24).

Conclusion:

These results suggest that Vitamin E does not prevent or reduce cardiac injury in rats treated with doxorubicin. Different alternatives including other antioxidants could be explored in an attempt to decrease cardiotoxicity produced by doxorubicin and to improve the effect of the standard cardioprotective agent used Dexrazoxane. Further studies are necessary.

References:

1. Swain SM. Adult multicenter trials using dexrazoxane to protect against cardiac toxicity. Semin Onco 1998; 25 (4 Suppl 10):43-7

2. Breed JG, Zimmerman AN, Dormans JA, Pinedo HM. Failure of the antioxidant vitamin E to protect against adriamycin-induced cardiotoxicity in the rabbit. Cancer Res 1980; 40:2033-8

3. Myers CE, McGuire W, Young R. Adriamycin: amelioration of toxicity by alpha-tocopherol. Cancer Treat Rep 1976; 60:961-2

Disclosures:

No relevant conflicts of interest to declare.
Study on the Relevance of Inducing Apoptosis and Reversal Effect of Nilotinib In Combination with Tet on Multidrug Reststahoe of K562/A02 Cell Line
Authors: Chen Bao-An , Cui Ting-yun , Ding Jia-hua , Gao Chong , Shan Xue-yun , Cheng Jian , Bao Wen , Zhong Yue-jiao , Gao Feng , Xia Guo-hua , Schmitt Anita , Schmitt Michael
Published: October 2019

Abstract 4951

Objective

To investigate the relevance of Inducing apoptosis and reversal effect of nilotinib in combination with Tet on multidrug reststahoe of K562/A02 cell line and its mechanism.

Methods

Methyl-thiazol tetrazolium(MTT) assay is employed to examine the pharmacological effect of Nilotinib or Tet alone on K562/A02 cell line, to calculate the inhibitor concerntration 50(IC50) of daunorubicin(DNR) in K562/A02 cell line and the resistance multiple of DNR to K562/A02 cell line. Flow cytometry(FCM) was employed to comple the effect on the inducing apoptosis of K562/A02. The expression of bax/survivin mRNA was determined by RT-PCR, and the expression of bax/survivin protein was assessed by Western blot.

Results

After 48 h 5 nmol/L nilotinib or 1.0μmoL/L Tet treatment, IC50 of DNR to K562/A02 was 5.71mg/L or 6.52 mg/L respectively;While on combinative treatment, its IC50 decreased to 3.12 mg/L. Nilotinib or Tet alone was able to increase the DNR induced apoptosis rate of K562/A02 cell (P<0.05), while on combination treatment the apoptosis rate increased remarkably. After 48 h 5 nmoL/L nilotinib or 1.0μmoL/L Tet treatment alone, gray—scale vaule of bax mRNA Was 2.90±0.31 or 3.6±0.46, respectively Gwhile on combinative treatment the value increased to 5.9±0.98. The bax protein expression level in K562/A02 cells was 2.1±0.07 or 2.90±0.09 when treated with 5 nmoL/L nilotinib or 1.0μmoL/L Tet alone for 48 h, but on combination treatment, the level increased to 4.8±0.31. After 48 h 5 nmoL/L nilotinib or 1.0μmoL/L Tet treatment alone, gray—scale vaule of survivin mRNA was 0.70±0.01 or 0.55±0.02, respectively;while on combinative treatment the value decreased to 0.35±0.08. The survivin protein expression level in K562/A02 cells was 0.74±0.03or 0.61±0.04 when treated with 5 nmoL/L nilotinib or 1.0μmoL/L Tet alone for 48 h, but on combination treatment, the level decreased to 0.42±0.06.

Conclusion

Nilotinib or Tet alone can pattially reverse drug resistance of K562/A02 cells. There is resistance phenomena of DNR to K562/A02 cells, and Nilotinib or Tet alone alone is able to reserve the resistance of DNR and enhance the inducing apoptasis effect of DNR. The mechanism may be associated with the increase of bax mRNA and protein expression and decrease of survivin mRNA and protein expression and increase of the apoptosis rate. And there is a synergistic action with these two agants in combination.

Disclosures:

No relevant conflicts of interest to declare.
Multiple Myeloma: Clinical Implications of Cytogenetic Aberrations on Selected Plasma Cells on Conventional Cytogenetics and Interphase Flourescence In Situ Hibridization (FISH)
Authors: Verma Rakesh , Kumar Lalit , Halder Ashutosh , Sharma Atul , Gupta Ritu , Saxena Renu , Chauhan Shyam S , Sinha Subrata
Published: October 2019

Abstract 4987

Purpose:

We prospectively studied patients of multiple myeloma (MM) for presence of t(14q32), and del13q14 on FISH and numerical chromosomal abnormalities on conventional cytogenetics(CC).

Methods:

Between February 2007 and June 2009, 90 previously untreated patients of MM were enrolled, Patient's median age was 55 years (range, 34 to 75 years) and 67 were males (M: F: 2.9: 1). 24 patients had ISS stage I, stage II-46 & 20 patients had stage III. Purified bone marrow plasma cells, using magnetic activated cell sorter with CD138 micro beads were used. Conventional cytogenetics for numerical chromosomal abnormalities and FISH for deletion 13q14 (RB-1) and 14q32 (IgH) translocations was carried out on these purified plasma cells. 65/90 (72%) patients received therapy; using novel agents, n=54 (thalidomide-dexamethasone (n=50), Lenalidomide-dexamethasone (n=4)) and melphalan, prednisolone and thalidomide (MPT, n=11). Response was evaluated post 4 cycles using EBMT response criteria.

Results:

Metaphases could be obtained on CC in 53/90 patients (58%); with numerical abnormalities in 21: hyperdiploid -16 & hypodiploid in 5. Del 13q14 and t (14q32) by FISH was present in 51 (56%) and 75 (83%) patients, respectively. t (14q32) was present more in patients with ISS-III (p<.008). Presence of del13q14 correlated with BM plasma cells >40% (P<.007) and Hb ≤9.2G/dl (p=.06). Following therapy, 49/65 patients (75.2%) achieved significant response (CR+VGPR+PR). Patients aged ≤55 years (P = 0.07), those with ISS- I (P <0.05), IgG-kappa (P<0.04) and with absence of del13q14 (P = 0.04) responded better. The median overall survival (OS) has not reached yet, median event-free survival (EFS) is 33 months (95% CI 18–48). Estimated OS and EFS at 2 years is 77% and 50 %, respectively. Durie Salmon stage IIIB (HR 3.6) and response to therapy (HR 7.8) were significant predictors of OS. For EFS - del13q14 (HR 2.7, P <0.036), response to therapy (HR 2.7,P<0.01) and use of novel agents (HR 3.1, (P<0.008) were important predictors.

Conclusion:

Chromosomal abnormalities including IgH translocations and del13q14 with numerical chromosomal abnormalities are common in MM patients at diagnosis and can predict the treatment outcome and survival.

Disclosures:

No relevant conflicts of interest to declare.
African-American Multiple Myeloma Patients Exhibit Less Bone Disease Compared to Other Racial/Ethnic Groups
Authors: Dean Eric W. , Ziv Elad
Published: October 2019

Abstract 4994

Background:

Bone destruction remains one of the major complications in Multiple Myeloma (MM) leading to morbidity and mortality. African-Americans have a higher incidence of MM but exhibit longer survivals compared to Caucasians. We analyzed bone involvement in a cohort of patients with MM to determine if non African American (non AA) vs. African American (AA) race predicts the presence and severity of bone disease at presentation.

Methods:

Clinical data was gathered on 197 (176 non AA and 21 AA) MM patients at the University of California, San Francisco. Each patient had a skeletal survey at diagnosis and identified as having 0 lytic lesions, 1–2 lytic lesions or 3 or more lytic lesions. The presence of compression fractures was also documented for each patient as was age and sex.

Results:

The presence of compression fractures strongly correlated with the number of lytic lesions in both the non AA and AA groups, with no compression fractures observed in the patients with zero lytic lesions (p<0.001). Among the AA group, there were fewer (6 of 15) patients with compression fractures compared with patients from the non AA group (92 of 161) (p=0.02). There was also a trend towards fewer lytic lesions among the AA group (p=0.053). No significant difference was observed between the extent of bone disease and age or sex between the two groups.

Conclusions:

Within this cohort of patients, there is a significantly lower rate of compression fractures among African-Americans. These data supports the idea that African-American patients present with less bone disease which confers a survival advantage compared to other racial/ethnic groups.

Disclosures:

No relevant conflicts of interest to declare.
A Review of Multiple Myeloma Patients on Dialysis Treated with High Cutoff Hemodialysis
Authors: Podymow Tiina , Alam Ahsan , Vasilevsky Murray , Beauchemin Roch , Shustik Chaim , Sebag Michael
Published: October 2019

Abstract 5002

Background:

Kidney failure is a major cause of morbidity in patients with Multiple Myeloma (MM). While up to half of MM patients can be shown to have renal impairment, 10% will become dialysis dependent. The main mechanism of kidney failure is cast nephropathy, which is linked to high amounts of circulating free light chains. A proposed strategy to prevent this outcome is the use of extended high cutoff (HCO) dialysis. An ongoing randomized controlled trial is testing the Gambro HCO1100 filter. Here we report the use of another dialyzer (Bellco Phylther), which has a smaller pore size than the Gambro filter, but which may prove to be as effective and is less costly.

Methods:

We present three patients with symptomatic MM, elevated κ or λ serum free light chains (>500mg/L), biopsy confirmed cast nephropathy, and kidney failure (estimated GFR <15ml/min/1.73 m2) requiring dialysis. In all patients, dialysis consisted of 10 hemodiafiltration dialysis sessions in the first two weeks and three times a week thereafter using a high flux Bellco Phylther HF22SD dialyzer (Bellco, Modena, Italy). All patients received concurrent chemotherapy: bortezomib 1.3mg/m2 IV and dexamethasone 40mg po on days 1, 4, 8 and 11 of a 21 day cycle and liposomal doxorubicin at 30mg/m2 on day 4 of every cycle. Serum free light chains were determined pre- and immediately post-dialysis using a nephelometric immunoassay (FREELITE, The Binding Site, Birmingham UK).

Results:

We observed significant (>50%) and rapid reductions in sFLCs. These reductions were sustained at 3 months in all cases, likely because of effective anti-myeloma treatment. Two of the three patients had recovery of kidney function and remain dialysis-independent at least 6 months after cessation of dialysis. Interestingly, the only patient that did not recover kidney function was also found to have tubulointerstitial disease on renal biopsy in addition to cast nephropathy.

Conclusions:

The HCO Bellco Phylther dialyzer appears effective in the rapid reduction of sFLCs in light chain MM patients with cast nephropathy requiring dialysis and concurrent anti-myeloma drug therapy. While the numbers are small, our review suggests that patients with uncomplicated MM cast nephropathy may benefit from aggressive hemodiafiltration with this HCO membrane. This dialysis membrane and protocol are worth considering for further study in patients with MM cast nephropathy.

Disclosures:

No relevant conflicts of interest to declare.
MARROW of Essential Thrombocythemia PATIENTS IS ENRICHED IN Effector Memory CD4+ and CD4+CD25++ Lymphocytes as Compared to BLOOD
Authors: Sedzimirska Mariola , Drabczak-Skrzypek Daria , Lange Andrzej
Published: October 2019

Abstract 5058

The role of T cells residing in the marrow is poorly understood. However, some literature data suggest that the presence of CD8+ lymphocytes in the marrow plays a role in keeping the tumor cells in a dormant state. In chronic myeloid leukemia a significant role of lymphocytes in controlling the malignant clones is well documented. The aim of this study was to define whether there is a specific enrichment of some CD4+ lymphocytes subpopulations in the marrows of newly diagnosed patents with essential thrombocythemia (ET).

Seven patients (F/M: 6/1, age 28–72 - median 6 yrs) with ET diagnosed according to the WHO criteria were studied. They had platelets 425–809 ×10∧3/ul (median 676 × 10∧3), NAP 74–138 score (median 112 score), 6 of them were positive for JAK 2 V617F mutation (heterozygotes). Marrow cellularity was from 18 – 64 ×10∧3/ul (median 44 × 10∧3/ul) Cytological evaluation revealed normal differentiation of erythroid and myeloid lineages, in contrast an increased number of megakaryocytes often enlarged, hyperlobulated and in clusters was seen in all patients. Trephine biopsy revealed activation of megakaryopoiesis with enlarged numerous megakaryocytes with normal representation of other lineages and a lack of an increase in the reticulin fibers. Lymphocytes in the marrows were in proportions from 4.5 – 15,7% (median 10,6%) what resulted with lymphocytosis from 2640/ul to 19405/ul (median 5645/ul). The whole populations of marrow and blood cells were labeled for a purpose of this study with the use CD4, CD25, CD45RO and CCR7 MoAb (Becton Dickinson, San Jose, CA, USA) followed by detection and analysis with the use of flow cytometry (FACS Calibur, Becton Dickinson) equipped with PCLysis programe.

We found:Proportions of lymphocytes in the marrows were rather in a normal value range but in numbers they prevailed over blood lymphocytes (2640-19405/ul, median 5645/ul vs 1300–3100/ul, median 1800/ul, p= 0,003 Wilcoxon Test for pairs),Among T cells subpopulations effector/memory CD4+ lymphocytes CD45RO+, CCR7-) were significantly higher in numbers in the marrows of ET patients as compared to blood (744-3241/ul, median 1126/ul vs 406–2437/ul, median 592/ul, p=0,04 Wilcoxon Test for pairs). There was no difference when numbers of naïve (CD45Ro-, CCR7+) and central memory (CD45RO+, CCR7+) CD4+ lymphocytes were analyzed.The marrows of ET patients had higher numbers of CD4+CD25++ lymphocytes as compared to blood (8,1-40,7/ul, median 11,4/ul vs 1,2-14,8/ul, median 7,3/ul; p=0,02 Wilcoxon Test for pairs).

It appeared that in ET patients there was a selective enrichment of marrow lymphocyte populations in CD4+EM cells and CD4+CD25++ lymphocytes what strongly suggests the presence of active immune response in the marrows affected by this myeloproliferative disorder.

Disclosures:

No relevant conflicts of interest to declare.
Waldeyer's Ring Marginal Zone B-Cell Lymphoma: Which Is Their Clinical and Prognostic Feature, Nodal or Extranodal?
Authors: Oh Sung Yong , Kim Won Seog , Kim Jin Seok , Kim Seok Jin , Lee Suee , Suh Cheolwon , Lee Dae Ho , Lee Soon il , Kang Hye Jin , Choi Chul Won , Jeong Seong Hyun , Park Jinny , Song Moo-Kon , Kim Hyo Jung , Kwon Jung Hye , Kwak Jae-Yong , Bae Sung Hwa , Park Byeong-Bae
Published: October 2019

Abstract 5084

Background:

Waldeyer's ring (WR) is circular band of lymphoid tissue located at the opening of the respiratory and digestive tract. There is controversy as to whether WR should be considered as a nodal or extranodal site. In this study, we conducted retrospective analyses of WR involving MZLs (WR-MZLs) to identify their clinical features, treatment, prognosis- favor nodal or extranodal. And we want to get additional information about specific organ relationship between WR-MZL and other MALT sites.

Patients and Methods:

From 1987 to 2010, 124 patients who were histologically confirmed as a MZL arising from head and neck (H&N) area was including WR (47 patients) were reviewed. Primary orbit and ocular adnexa MZL patient was excluded this data collection.

Results:

The male/female ratio of the 47 patients was 23 to 24. The median age was 53 years (range, 17–77). The commonly involving sites were tonsil (53.2%) followed by nasopharyn (40.4%). Fourteen patients (29.7%) were accompanied with extra-WR area MALT site- gastrointestinal tract (14.9%), ocular and adnexa (12.8%). Ann Arbor stage I/II disease was present in 50% (23 out of 46). Thirty-nine patients were categorized into the low/low-intermediate risk group (84.8%) according to International Prognostic Index (IPI). Complete and partial remissions were achieved in 18 (78.3%) and 2 (8.7%) of the 23 stage I/II patients. In 23 patients with stage III/IV, CR and PR were achieved in 15 (65.2%) and 4 (17.4%), respectively. The median progression-free survival (PFS) was 3.6 years (95% CI, 2.4–4.8 years). The estimated 5-year overall survival (OS) was 88%. Compared with H&N MALT site MZLs, WR-MZLs were significantly poor in PFS and OS (P=0.005 and P=0.007).

Conclusion:

WR-MZLs were also an indolent disease like general MZL. But, WR-MZL patients presented with more advanced stage and poorer survivals than other site H&N MZL patients. Therefore, even though WR has been considered histologically extranodal MALT organ, clinically it is closer to nodal MZL.

Disclosures:

No relevant conflicts of interest to declare.
Associations of Aberrant T Cell Antigen Expression on B Cells of Diffuse Large B Cell Lymphoma Patients with Their Background and Prognosis
Authors: Suzuki Yuhko , Yoshida Tsutomu , Kimura Hirokazu , Ohsaka Manabu , Miyazaki Koji , Danbara Mikio , Horie Ryouichi , Satoh Yuhichi , Higashihara Masaaki
Published: October 2019

Abstract 5088

Aim:

With the exception of CD5, the clinical implications of aberrant T-cell antigen expression on B cells in diffuse large B cell lymphoma (DLBCL) have not been well studied. The present retrospective study aimed to determine associations between T cell antigen expression and patients′ characteristics and prognosis

Patients, Material and Methods: 249 cases with B-cell lymphoma (BCL) newly-diagnosed in our Institute between January 2002 and August 2009 were biopsied and also analyzed by flow cytometry. Biopsy specimens were fixed in formalin, stained with Hematoxylin-Eosin, and also immunostained. Histological subtypes were defined according to the World Health Organization Classification Version 3. Flow cytometric (F/C) analysis was performed following standard methods.

The aberrant expression of one or more T cell surface antigens on B cells (CD2, 3, 4, 7, 8) was assessed. Statistical analysis was conducted to seek associations between aberrant positive and negative cases and patients′ background laboratory data, and prognosis.

This study was approved by the Ethics Committee of Kitasato University School of Medicine.

Results:

150 DLBCL, 68 Follicular lymphoma (FL), 17 Marginal zone lymphoma (MZL), 6 MCL and 4 CLL patients were tested. Of the DLBCL cases, 12 (8%) showed aberrant T cell antigen expression with 4, 1, 5, 1 and 1 patients′ B cells positive for CD2, CD4, CD7, CD8 and CD7 + 8 respectively. Aberrant expression among the FL, MCL, MZL and CLL cases was seen in 0, 1, 1 and 1 patient, respectively.

Among the DLBCL patients, T antigen-negative cases tended to have higher WBC counts and greater CD20 expression by F/C analysis. No statistically significant associations with gender, age, IPI, clinical stage, laboratory data, expression of CD5 and other markers, ABC/GCB, or karyotype were observed between the positive and negative groups.

96 of the 150 DLBCL patients received rituximab-chemotherapy. Again, there were no statistically significant differences between the two groups in overall survival and response to treatment.

Discussion:

1) CD2 and CD7 are relatively common among aberrant T cell antigen- positive DLBCL. 2) No statistically significant differences were observed between the two groups in terms of background and prognosis. 3) Both CD8-positive DLBCL patients experienced a very aggressive clinical course and rapidly succumbed to their disease. CD8-positive DLBCL cases may in general have poorer survival. 4) Further analysis will be necessary to confirm these results.

Disclosures:

No relevant conflicts of interest to declare.
Rare Splenic Intravascular Diffuse Large B-Cell Lymphoma with Massive Hypersplenism and Concomitant Minor Liver Infiltration and Supra-Mesenteric Nodes.
Authors: De Salvo Dalila , Salas-Auvert Rodolfo J , De Salvo Cardullo Luigi
Published: October 2019

Abstract 5101

We report a 23 yr female patient which relates beginning of current illness three months prior diagnosis; characterized with increased abdominal volume, concomitant active hemorrhagic manifestations consisting of ecchymosis and petechiae. Upon clinical, bone marrow biopsy, immunohistochemistry and subsequent surgery, case corresponded to Diffuse Large B-Cells Lymphoma(DLBCL) of splenic intravascular origin. Organ weight and size after splenectomy 2.53 Kg size 30×22×10 cm). Microscopic diagnosis description corresponds with presence of monomorphic lymphoid cells with predominance of small neoplasic lymphoid cells with small round nucleus, granular chromatin and small nucleolus accompanied with eventual cytoplasmic membrane irregularities. Presence of reactive plasmocytes and sever splenic congestion. The unusual case reported corresponds to an invasive lymphoma of splenic vascular origin. By our knowledge this is the first case register in Venezuela.

Disclosures:

No relevant conflicts of interest to declare.
Folic Acid Food Fortification and Incidence of Anemia In General Population
Authors: Bansal Goldy , Fernandez Gloria
Published: October 2019

Abstract 5140

Introduction:

Folic acid (FA) deficiency is considered one of the important causes of anemia and of neural tube defects in infants. In 1996 FDA issued a regulation that by 1998 all enriched grain products contain 140 mg of folic acid per 100 grams. This process began in 1996 and was essentially completed by mid-1997. This program intended to increase folic acid intake among women of childbearing age has decreased the incidence of pregnancies affected by neural tube defects. This has also led to a decrease in incidence of FA as a cause of anemia. In 2000 we reported the impact of folate food fortification in our community hospital. In 1994 to 1996, of 4403 patients sample tested 60 were found to be folate deficient. From 1997 to 1999 only 3 of 6326 patients tested were folate deficient. The first patient was 73 year old and had severe CHF and COPD, the second patient was 62 year old alcoholic admitted with delirium tremens, and third patient was 90 year old with dysphagia who required PEG replacement. All three appeared to have stopped eating in the days prior to admission. On the basis of this experience in our community hospital we concluded at that time that folate food fortification has largely eliminated FA deficiency and that the routine determination of FA as part of anemia work up in this era is not indicated. We now report our continuing experience from 2003 to 2006.

Methods:

We studied retrospectively the patients who presented in our community hospital with anemia and folate level ordered as a part of anemia work up. Serum folate levels were measured using Bayer Center Chemiluminescent Assay. Serum FA level <1.1 mg/ml is considered indicative of deficiency. We evaluated 1861 patients from year January'2003 to May'2006. We also assessed the cost effectiveness of ordering this test routinely as part of anemia workup.

Results:

Among the 1861 patients, we did not find any patient who had folate level of less than 1.1, which is the lower limit for folate level in our laboratory. Medicare cost for serum folate level test for our institute is $20.54 (code 82746) and phlebotomy cost for one test is $3.00 (code 36418). Total cost for this test for one patient is $23.54. Total health cost for these tests in 3 years and five months was $43807.94.

Conclusion:

Our results which are similar to our previous report suggest that food fortification and possibly also vitamin supplementation has substantially improved blood folate levels and has largely eliminated folic acid deficiency as a cause of anemia in the population that we tested. Routine folate level determination in the workup of anemia is not cost effective and may be best limited in patients who are malnourished as in patients with poor nutrition status, malabsorption, alcoholics and psychiatric disorders.

Disclosures:

No relevant conflicts of interest to declare.
KLF1 Dependent Pathways In Developmental Globin Gene Switching
Authors: Felice Alexander E.
Published: October 2019

Abstract 5162

We provide additional data on members of the family from Malta with Hereditary Persistence of Fetal Hemoglobin (HPFH) due to KLF1 haplo-insufficiency. The data indicated a possible role of additional loci in the pathway of globin gene control. We showed that KLF1 functions as a master regulator of erythropoiesis and developmental globin gene switching (Borg et al., Nature Genetics doi: 10. 1038/ng.630, 2010), at least partly through BCL11A. Given the phenoytpes of the HPFH heterozygotes, the truncating KLF1 p.K288X was best described as a dominant mutation with variable penetrance; most likely due to interplay with other regulatory factors that we have been seeking.

Genome-wide association analysis, in the context of genome-wide expression profiles from cultured Human Erythroid Progenitors (HEPs) of critically informative family members, revealed additional loci of potential interest.

The effect of the Hb F inducer Hydroxyurea on the gamma globin profiles of the KLF1- (p.K288X) HPFH HEPs was enhanced compared to the wild type, and 74 loci were differentially expressed.

It is anticipated that extensive re-sequencing of these new targets may reveal the extent of the molecular pathways under control of KLF1 in erythropoiesis and globin gene switching, and in particular those that may be targeted for therapeutics in patients.

Disclosures:

No relevant conflicts of interest to declare.
Severe Fetal and Neonatal Anemia Due to Heterozygosity for a 198 Kb Deletion Removing the Entire β-Globin Gene Cluster
Authors: Verhovsek Madeleine , Shah Nirmish , Thornburg Courtney , Koenig Sara , Wilcox Ibifiri , Barros Tiago , Steinberg Martin H. , Luo Hong , Chui David H.K.
Published: October 2019

Abstract 5171

Infants who are heterozygous for a number of (γδβ)0-thalassemia deletions are known to present with neonatal hemolytic anemia (Koenig et al, Am J Hematol 84:603, 2009). The breakpoints for most of these large deletions have not been identified. Molecular diagnoses of these deletions therefore can be challenging. We now report an infant girl of Irish/Scottish descent with self-limited fetal and neonatal hemolytic anemia, in whom we have defined the extent of the large deletion removing the entire β-globin gene cluster. At 32-week of gestation, fetal tachycardia prompted percutaneous umbilical cord blood sampling, which revealed a hematocrit of 14%. Umbilical cord transfusion was undertaken followed by Caesarean section delivery. Brisk hemolysis continued in the first few days of life (reticulocyte count of 22 – 24%), with no other causes of hemolysis identified. In the following weeks, the infant was transfused on three occasions. After 2-month of age, she became transfusion-independent with a stable microcytic anemia. The infant's mother also had a history of hemolytic anemia requiring transfusion in the neonatal period, and subsequently became transfusion independent with a microcytic anemia. The mother and several of her family members were extensively investigated, and shown to have a novel (γδβ)0-thalassemia deletion of over 100 kb (Pirastu et al, J Clin Invest 72:602, 1983). Multiplex ligation-dependent probe amplification (MLPA) was carried out in the genomic DNA from the present neonate. The infant was heterozygous for a large deletion spanning at least from 5′ to the HS 5 of the LCR to 3′ of the β-globin gene. Sequential gap-PCR reactions and nucleotide sequencing were done. The deletion was characterized with its 5` breakpoint at nt 5,376,341 and 3` breakpoint at nt 5,178,572 (GenBank NT_009237). The deletion measures 197,770 bp, removing the β-globin LCR, all of the β-like globin genes, and several olfactory receptor genes. A diagnostic gap-PCR test was established for detection of this deletion. This case illustrates the syndrome of neonatal hemolytic anemia caused by large deletions removing the entire β-globin gene cluster. MLPA is a useful tool to screen for these deletions. The pathophysiology of these self-limited and sometimes severe fetal and neonatal hemolytic anemias is presently not understood. We speculate that expression of α-hemoglobin stabilizing protein (AHSP) and/or the proteolytic capacity to degrade excess α-globin chains within erythroid cells might be diminished during fetal and neonatal development, accounting for increased red cell membrane damage and hemolysis in affected fetuses and neonates.

Disclosures:

No relevant conflicts of interest to declare.
Complement Enhances Foxp3+ T Regulatory Cell Induction by B Cell Lymphoma
Authors: Veeramani Suresh , Dahle Chris , Weiner George
Published: October 2019

Abstract 5176

The effect of complement on the immune system is complex and includes inhibition of aspects of the immune response. For example, we previously reported that C3b can inhibit the ability of mAb to activate NK cell-mediated antibody-dependent cellular cytotoxicity and that polymorphisms in C1q associated with lower C1q levels correlate with better therapeutic outcome. Polymorphisms in C5 and C9 complement components have also been shown to be associated with lymphoma risk. The current studies were designed to assess whether complement components may exert their inhibitory effects through the immune regulatory network, such as that mediated by T regulatory (Treg) cells.

Peripheral Blood Mononuclear Cells (PBMCs) from healthy donors were co-cultured with Raji B cell lymphoma cells for 48 hours in the presence of unmodified autologous human serum or complement-depleted serum [heat-inactivated (HI) or cobra-venom factor (CVF)-treated serum]. Cells were stained with anti-CD3-APC-Cy7, CD4-PE-Cy7, CD25-PerCP-Cy5.5 and Foxp3-APC and the percent of CD4+ T-cells that were CD25highFoxp3+ (henceforth referred to as Tregs) determined. The ability of cells cultured in this manner to inhibit granzyme B expression in PHA-activated CD8+ autologous T cells was also determined.

Tregs were not induced when PBMCs were cultured without Raji cells regardless of whether the cells were cultured in serum (data not shown). However, PBMCs co-cultured with Raji B lymphoma cells showed significantly higher percentages of Tregs in the presence of unmodified human serum (*p=0.0387) when compared to HI or CVF-treated human serum (**p=0.0370 and 0.0227, respectively) (Fig. 1). Tregs enriched from PBMCs cultured with unmodified human serum and Raji cells also showed an increase in immunosuppressive function, as determined by their inhibitory effect on PHA-induced GrzB expression by autologous CD8+ T cells (*p=0.0029) (Fig. 2).unmodified or complement-inactivated humanunmodified or complement-inactivated human

We conclude that complement enhances generation of Treg cells induced by B lymphoma cells as indicated by both phenotypic and functional studies. Ongoing studies are exploring the role of individual complement components in Treg induction, and the possibility of using complement depletion as a strategy to boost immune responses against lymphoma.

Disclosures:

No relevant conflicts of interest to declare.
Protected by Nature: Effects of Exercise In Non-Severe Haemophilia Patients
Authors: Uijl Ingrid E.M. den , Groen Wim G , van der Net Jan Jaap , Grobbee Diederick E , de Groot Philip G , Fischer Kathelijn
Published: October 2019

Abstract 545

Background:

An increase of clotting factor VIII (FVIII) within 10 minutes after physical exercise has been reported in healthy subjects and patients with von Willebrand disease. The mechanism of FVIII increase through exercise is thought to be similar to that of desmopressin, which increases FVIII. Therefore, moderate and mild haemophilia A patients should also be able to increase their endogenous FVIII levels after exercise. A proof of principle study was performed to test this hypothesis.

Methods:

Changes in coagulation associated with exercise of 15 haemophilia A patients, aged 20–39 years, were measured. Eight had moderate haemophilia (FVIII 1–5 IU/dl), 7 had mild haemophilia with endogenous FVIII activity levels at the lower end of the range (FVIII 6–16 IU/dl) Patients were encouraged to cycle until volitional exhaustion, on an electronically braked cycle ergometer. Blood samples were drawn before and 10 minutes after the incremental exercise test. Blood was frozen and analyzed for FVIII, von Willebrand factor (vWF) and von Willebrand propeptide (vWpr), in one batch to avoid inter-assay variation. In mild haemophilia patients, changes in coagulation profile were compared to DDAVP tolerance tests.

Results:

All 15 patients had an increased FVIII level after exercise. Median relative increase was 2.5 times (interquartile range (IQR)= 1.2–4.0). The relative increase did not vary across severities (median relative increase of FVIII was 2.4 times (IQR= 1.3–4.0) for moderate haemophilia patients compared to median 2.5 times (IQR=1.2-4.0) in patients with mild haemophilia Mann-Whitney: p =0.91). However, the absolute FVIII levels were very different with a median FVIII level of only 9 IU/dl; IQR= 8–11 IU/dl) in patients with moderate haemophilia, compared with a much higher median FVIII level of 21 IU/dl; IQR=19-44 IU/dl) attained in patients with mild haemophilia. vWF and vWFpr also increased independent of severity (p=0.3-0.6) in all patients, median 50%; IQR 30–79% for vWF and median 165%; IQR 130–244% for vWFpr, reflecting acute release of coagulation factors. DDAVP response tests were available only for patients with mild haemophilia. Relative increase after DDAVP administration was median 3.5 times (IQR= 2.5–4.5), similar to increase in FVIII after exercise (Wilcoxon: p=0.14).

Conclusion:

Although normal levels were not achieved, the increase of FVIII activity during exercise may reduce bleeding risk and therefore help promote participation in sports and other strenuous activities for moderate and mild haemophilia A patients with modest endogenous FVIII activities who might be considered at high bleeding risk.

Disclosures:

No relevant conflicts of interest to declare.