bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparative Analysis of Outcomes of Allogeneic Peripheral Blood Stem Cell Transplantation From Related and Unrelated Donors for Adults with Hematologic Malignancies
Authors: Gao Chunji , Li Xiaohong , Li Honghua , Huang Wenrong , Wu Xiaoxiong , Ma Jian , Bo Jian , Wang Zhihong , Cao Yongbin , Zhao Yu , Yu Li , Da Wanming
Published: October 2019

Abstract 2302

Although allogeneic peripheral blood stem cell transplantation from a matched related donor (RD-PBSCT) presents the best curable opportunity for many patients with hematologic malignancies, only about one third of individuals have HLA matched sibling donors. Fortunately, from unrelated donor peripheral blood stem cell transplantation has been acceptable and expanded recently. In order to evaluate the effect of allogeneic peripheral blood stem cell transplantation from unrelated donor (URD-PBSCT), we compare results of URD-PBSCT and RD-PBSCT that were done for 172 consecutive adult patients with hematologic maligancies from Jan 2002 to Dec 2008 at a single-center. Among these patients, 62 cases underwent URD-PBSCT and 110 cases underwent RD-PBSCT. Myeloablative conditioning was adopted for all patients. In graft versus host disease (GVHD) prophylaxis, 49 URD-PBSCT recipients received CSA, MTX, MMF and ATG (URD-ATG group), 13 recipients were given simulect more in base of URD-ATG group (URD-ATG+CD25 group) while RD-PBSCT recipients (RD group) received CSA, MTX and MMF. Engraftment was achieved in 98.4% of URD-PBSCT patients and 98.2% of RD-PBSCT patients (100% for patients surviving beyond 28 days after transplant). The cumulative incidence of acute GVHD (grade II-IV) was 15.4%, 36.7% and 29.1% respectively in the URD-ATG+CD25, URD-ATG and RD groups (P = 0.275). The cumulative incidence of chronic GVHD was 0%, 45.6%, 39.6% respectively and significant lower in URD-ATG+CD25 group than the other two groups (P = 0.0134). Relapse incidence was 53.8%, 12.2%, 14.5% respectively and significant higher in URD-ATG+CD25 group than the other s (P = 0.0059), while there was no different between the URD-ATG and RD groups (P = 0.610). Estimated overall survival (OS) at 5 years was 30.8%, 69.4% and 67.3% respectively and no significant difference between URD-ATG group and RD group (p=0.898). Adverse prognosis factors for relapse and OS included transplant not in remission and GVHD prophylaxis with simulect. Our results indicate PBSCT from unrelated donor may be considered comparable to those from related donor.

Disclosures:

No relevant conflicts of interest to declare.
Restoring Cell Responsiveness to GM-CSF and IL-3 by PTEN Is Via Controlling the Regulation of Egr-1 In TF-1a Cell Line
Authors: Liu Y. Lucy , Malaviarachchi Priyangi , Emanuel Peter Dean
Published: October 2019

Abstract 2504

Cytokine independence is a common event in leukemia. The mechanism is often linked to deregulation of Ras/PI3K, Ras/MAP, and JAK/STAT pathways. PTEN, a major negative regulator of the Ras pathway, was initially isolated as a tumor suppressor in a variety of malignancies. We recently reported that PTEN is deficient in 67% of juvenile myelomonocytic leukemia (JMML) patients. Other groups have also found alterations of PTEN, PI3K, or AKT in 47.7% of T-cell acute lymphoblastic leukemia. Mice who are rendered pten-deficient develop myeloproliferative disorders and acute myeloid and lymphoid leukemias, and the leukemia-initiating stem cell in these mouse models could be distinguished from the normal hematopoietic stem cell by a differential response to rapamycin. The transcription factor, Egr-1, has been known to be a positive regulator of PTEN expression. It plays an important role in controlling proliferation as well as mobilization of hematopoietic stem cells. It is unknown whether PTEN had a feedback loop on regulating Egr-1 expression. Since poorly differentiated progenitors are often mobilized to the peripheral blood in leukemia patients, we hypothesized that PTEN deficiency alters the feedback loop of Egr-1 in response to cytokine stimulation. In this manner, PTEN deficiency might contribute to the cytokine independence observed in many types of leukemia. In order to evaluate this hypothesis we investigated the role of PTEN in cells responding to GM-CSF stimulation in the leukemia cell line, TF-1a. TF-1a is PTEN deficient as well as cytokine independent, and displays a constitutively high expression of Egr-1. We restored PTEN expression in TF-1a cells. This was accomplished by packaging the coding sequence of wild-type PTEN in a retrovirus vector, pBMN-GFP, that carried a gene encoding green fluorescent protein (GFP). TF-1a cells were transfected with pBMN-PTEN-GFP or pBMN-GFP by spinoculation. The pure population of transfected cells was sorted by FACS, followed by limiting dilution. The transfected single colonies were expanded for further evaluation of PTEN expression and biological function. Data from these experiments showed: 1) over-expression PTEN in TF-1a down-regulated the expression of Egr-1 and restored the responsiveness of Egr-1 to GM-CSF stimulation, 2) PTEN restored the responsiveness of TF-1a to GM-CSF and IL-3, and 3) these events are CREB-independent. In conclusion, our data demonstrates that PTEN contributes to the cell responsiveness to GM-CSF and IL-3, and has a negative feedback role to regulate Egr-1 expression in TF-1a cells. In addition to Egr-1 regulating PTEN in a positive manner, our data indicates the presence of a feedback loop from PTEN back to Egr-1. This appears to be an event independent of CREB.

Disclosures:

No relevant conflicts of interest to declare.
4-1BBL-Expressing aAPCs Attenuate IL-15-Induced NK Cell Expansion and Cytokine Production In Vitro but Induce NK Cell-Mediated Gvhd In Vivo
Authors: Capitini Christian M. , Meadors Joanna L. , Cho Monica M. , Orentas Rimas J. , Mackall Crystal L. , Fry Terry J.
Published: October 2019

Abstract 2536

Methods to expand natural killer (NK) cells ex vivo for adoptive cell therapy are being explored to improve outcomes after allogeneic blood and marrow transplant (alloBMT). Artificial antigen presenting cells (aAPCs) can present cytokines and/or co-stimulatory molecules that can potentially improve expansion and activity. 4-1BBL (CD137L) has demonstrated mixed results on murine and human NK cells, but the impact on murine NK cell biology after alloBMT has not been explored. NK cells were harvested from either C57BL/6 (B6) or CB6F1 spleens and cultured ex vivo with a recombinant interleukin (IL)-15/IL-15 receptor alpha (Ra) complex in the presence or absence of a CD137L+ aAPC. Because IL-15 is typically presented in trans by IL-15Ra, the complex was utilized to potently increase agonist bioactivity. NK cells cultured with IL-15/IL-15Ra alone showed a peak of 20-fold expansion, but this expansion was decreased with the addition of CD137L+ aAPCs if the ratio of aAPC to NK cells was greater than 1:1. In the presence of IL-15/IL-15Ra, the impact of CD137L+ aAPCs on expression of the inhibitory receptors, Ly49C+I and activating receptor Ly49H was variable and strain dependent, with increased expression in B6 NK cells, but decreased expression in CB6F1 NK cells. The expression of major histocompatibility complex (MHC) class I was not affected in NK cells from either strain by the presence of CD137L+ aAPCs. The production of gamma interferon and tumor necrosis factor-a was robust in NK cells expanded by IL-15/IL-15Ra alone, but attenuated with the addition of CD137L+ aAPCs. Animal experiments showed that administration of NK cells expanded ex vivo with IL-15/IL-15Ra alone was well tolerated after T cell depleted MHC-mismatched alloBMT (CB6F1–>B6), but surprisingly the addition of CD137L+ aAPCs to cultures caused NK cells to induce GVHD-associated weight loss. In summary, IL-15/IL-15Ra expanded murine NK cells demonstrate increased cytokine production and do not cause toxicity when infused after alloBMT. The presence of CD137L+ aAPCs attenuated cytokine production and increased Ly49 receptor expression in NK cells from B6 mice. Remarkably, NK cells expanded by IL-15/IL-15Ra in the presence of CD137L+ aAPCs demonstrate increased propensity to cause GVHD. Ongoing studies are exploring the anti-tumor efficacy of IL-15/IL-15Ra expanded murine NK cells cultured in the presence and absence of CD137L.

Disclosures:

No relevant conflicts of interest to declare.
The Efficacy and Safety of Recombinant Human Thrombopoietin in Patients with Hematological Malgnancies After Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Miao Miao , Depei Wu , Sun Aining , Wang Ying , Yan Lingzhi , Wu Qian
Published: October 2019

Abstract 4565

OBJECTIVE:

To evaluate the efficacy and sefety of recombinant human thrombopoietin(rhTPO) prior to engraftment in adults with hematological malignancie who received allogeneic haematopoietil stem cell transplantation(Allo-HSCT).

METHODS:

This sutdy was a randomized, controlled clinical trial,38 patients were hematological malignancie, inclulding acute and chrinic myeloid leukemia, acute lymphoblastic leukemia, lymphoma.They received Allo-HSCT and were randomly divided into groups(group A 19 cases, group B 19 cases).The group A was no-rhTPO as control, the group B were received rhTPO 15000U/d from +1 day, and continued until the untransfused platelet count was >70×109/L for two consecutived days. Patients received platelete transfusion when they developed severe thrombocytopenia<20×109/L. Efficacy and sefety of rhTPO on the outcome of Allo-HSCT were evaluated.

RESULTS:

In both group A and B, platelet decrease after Allo-HSCT had no sognificant difference. The platelet engraftment duration of group A and B was 15.68±1.36(range 11–31) days and 13.47±0.72(range 9–21) days, respectively. The amount of platelet transfusion of group A and B was 4±0.55(range 20–130) units and 2.89±0.36(range 0–50) units, respectively. The effects of rhTPO on neutrophil engraftment, hepatic function, renal function, alloergic reations and acute GVHD were not found.

CONCLUSION:

The platelet engraftment duration of group B was shorter than that of group A(t=27.2, p<0.001), the amount of need platelet transfusion was significently less than those in the group A.There was a statistically significant difference in platelet engraftment and platelet transfusion needed(t=2.523, p<0.05). Administration of rhTPO prior to platelet engraftment after Allo-HSCT seem to be efficacy and safe.

Disclosures:

No relevant conflicts of interest to declare.
Immune Tolerance Therapy in Children with FVIII Inhibitors
Authors: Vdovin Vladimir , Svirin Pavel , Shiller Ekaterina , Schwartz Bruce A.
Published: October 2019

Abstract 4656

In up to 40% of cases, Hemophilia A treatment is complicated by the development of FVIII inhibitors, rendering the patient resistant to replacement therapy and increasing the risk of unmanageable bleeding, particularly arthropathy and disability. Presence of an inhibitor dramatically increases overall treatment costs 3–5 fold. Bleeding episodes in inhibitor patients require hospitalisation and bypassing agents such as rFVIIa. Immune tolerance (ITI) therapy, involving repetitive and persistent treatment with FVIII replacement is the only proven strategy for inhibitor eradication.

Since 2006, 30 children with inhibitors and poor-prognosis for ITI success have been treated with octanate® for ITI at seven Russian haemophilia treatment centers in the frame of the ongoing Observational Immune Tolerance Induction (ObsITI) study. The median age was 5 years.

The objective of this study was to evaluate the progress of prospective ITI with octanate® in children with inhibitors and a poor-prognosis for ITI success. In addition, joint status using a new musculoskeletal assessment tool before, during or after ITI was evaluated. A cost-of-care analysis to assess the consumption of octanate® in ITI in comparison with rFVIIa and to quantify the average direct medical costs for the treatment of young patients with FVIII inhibitors was also performed.

Despite the stringent success criteria, octanate®, along with the high-dose Bonn protocol, has led to a complete success rate of about 80% in children with inhibitors and poor prognostic factor for ITI success.

Successful tolerisation with octanate® allows resumption of fully effective replacement treatment with a consequent improvement in the patients’ musculoskeletal outcome and estimated savings over 5 years of € 600000. In addition, breakpoint analyses estimated that the number of years after which ITI with octanate® pays off compared with on-demand treatment with rFVIIa is 1.4 years.

In summary, octanate® is safe, well tolerated and effective in the induction of immune tolerance, even in poor-prognosis patients. The comparative cost-of-care analysis will help to optimize the therapy for the inhibitor patient and to ensure an effective allocation of financial resources.

Disclosures:

Schwartz: Octapharma: Employment. Off Label Use: This product Octanate is licensed in a number of countries world wide but not yet licensed in the US.
Frequency of Bleeding Disorders in Women Presenting Menorrhagia in the North of Iran
Authors: Janbabaei Ghasem , Borhani Samaneh , Rashidi Masoumeh , Farazmandfar Touraj , Shekarriz Ramin , Khademloo Mohammad , Ghavamzadeh Ardeshir
Published: October 2019

Abstract 4660

Objectives:

Menorrhagia is a common presentation of bleeding disorders, especially VWD in women. We decided to determine the frequency of these disorders in women with menstruation problems.

Materials and Methods:

208 patients in reproductive age with menorrhagia were investigated for bleeding disorders in two steps. Step one includes CBC, PT, PTT and BT, which performed for all patients, and for patients who had an abnormality in step one, step two was considered. Step two includes VWAg, RCO, factors level, RIPA and platelet aggregometry.

Results:

Among 208 patients who investigated for bleeding disorders 53 patients (25%) had abnormalities in coagulation tests or platelet count. In our survey frequencies of bleeding disorder was VWD=14(6.73%), thrombocytopenia=13(6.25%), deficiency of factor II= 2(0.96%), factor V=1(0.48%), factor VII=3(1.44%), factor VIII=2(0.96%), Factor XI =4(1.92%), factor XII=4(1.92%), Bernard Soulier=2(0.96%). Furthermore, we found 18 patients (8.65%) who had abnormal PT, PTT or BT with no definite diagnosis.

Conclusion:

In this study, the most common bleeding disorder was VWD and thrombocytopenia ranked second disorders. Although other bleeding disorders are rare, in our study a number of them were found. So, we recomment the above coagulation test for women with menorrhagia.

Disclosures:

No relevant conflicts of interest to declare.
Plasma VEGF Levels and Etiology of Thrombocytopenia, a Biomarker Study
Authors: Xiao Ying-Yi , Liu Yang , Zeng Thanthanaye , Khiani Komal , Burton Jack , Huang Yiwu , Xu Yiqing
Published: October 2019

Abstract 4684

Background:

Thrombocytopenia is a major medical problem associated with bleeding. Etiology of thrombocytopenia includes myelosuppression, splenic sequestration and increased peripheral consumption and/or destruction. Vascular Endothelial Growth Factor (VEGF) is produced in both endothelial cells and platelets and is stored in alpha granules in the platelets. It plays multiple roles in the regulation of vascular endothelial cell proliferation and vascular integrity.

Hypothesis:

VEGF release from the platelets in vivo due to platelet activation or destruction may result in changes in plasma and serum VEGF levels, and the level could be associated with various etiologies of thrombocytopenia.

Methods:

This was a prospective IRB approved on-going study. Eligibility includes platelet count less than 120,000/ul, no current diagnoses of solid tumor and thrombocytopenia is not due to chemotherapy. Blood was collected at the time of diagnosis, and VEGF was measured in serum and plasma specimens using a commercial ELISA kit from Ray Biotech (Norcross, GA). Plasma adjusted VEGF levels are defined as plasma VEGF value divided by the platelet count.

Results:

Forty seven patients were enrolled, the clinical diagnoses included AML or MDS (designated as A/M, n=6), hypersplenism (n=13), ITP (n=14), sepsis (n=11) and confirmed TTP (n=3). Blood was also collected from 4 healthy volunteers as controls. Plasma VEGF levels and plasma adjusted VEGF levels (defined as plasma VEGF value divided by the number of platelets) were selected as the main measurements of outcome as shown below

Conclusion:

Plasma VEGF adjusted with platelet count should be the measurement for in vivo release of VEGF from endothelial or platelet sources. Plasma adjusted VEGF level has a wide range of variation in patients with thrombocytopenia, about 50% of our test subjects had undetectable VEGF plasma levels. Although not statistically significant, patients with thrombocytopenia due to AML or MDS had lowest plasma VEGF values and least variation, while patients with sepsis had highest plasma VEGF readings and variations, probably due to increased endothelial cell production. The study is ongoing in large scales, and we are also interested in following the trend of serial plasma adjusted VEGF levels along with platelet recovery.

Disclosures:

No relevant conflicts of interest to declare.
Neutropenia in Glycogen Storage Disease 1b (GSD1b)
Authors: Dale David C. , Bolyard Audrey Anna , Marrero Tracy M. , Phan Lan , Boxer Laurence A. , Kishnani Priya S. , Kurtzberg Joanne , Weinstein David A.
Published: October 2019

Abstract 4791

Neutropenia and recurrent infections are common manifestations of glycogen storage disease 1b (GSD1b), a disease caused by mutations in G6PT1, the glucose 6 phosphate transporter 1 gene. Patients with GSD1b have complex defects in neutrophil deployment and function. GSD1b causes recurrent infections – gingivitis, mouth ulcers, respiratory infections, abscesses and enterocolitis. Granulocyte colony-stimulating factor (G-CSF) has been used to treat these patients for more than 2 decades.

We reviewed the presentations, manifestations, complications and G-CSF treatment for 54 GSD1b patients (31 adults and 23 children) at Duke University, the University of Florida and through the Severe Chronic Neutropenia International Registry at the University of Washington. Observational data were available on most patients for at least 3 years (mean = 12 years, median = 13 years, range = 1 to 23 years). Available data showed that before treatment with G-CSF, all of these patients had mild to moderate neutropenia; the mean of the median absolute neutrophil count (ANC) was 0.839 × 109/L ± 0.165 SEM (median = 0.503 × 109/L, range = 0.038 to 7.665 × 109/L). All of the patients also had recurrent infections; the most common events were mouth ulcers, otitis and skin abscesses. There were 7 patients who had enterocolitis prior to G-CSF treatment.

G-CSF therapy was initiated at various ages ranging from birth to age 30 (mean = 6 years, median = 4 years). Initial daily doses of G-CSF ranged from 0.50 to 63.28 mcg/kg/day (mean = 5.86, median = 3.65) and were generally titrated upward to achieve a normal neutrophil count. The mean ANC on treatment was 2.598 × 109/L ± 0.136 SEM (median = 1.856 × 109/L, range = 0.0 to 12.645 × 109/L). G-CSF treatment was uniformly associated with an increase in blood neutrophils and reduced patterns of infection. In most patients the symptoms of enterocolitis improved within a few days to a few weeks after starting G-CSF therapy. An increase in spleen size also commonly occurred. In 630 patient-years of observation, there have been 3 deaths attributable to the following causes: 1 subject died of sepsis, 1 subject died of complications from hepatomegaly and neutropenia, and 1 subject died 6 years after a liver transplant. The subject developed AML 5 years after the liver transplant while receiving a combination of immunosuppressant drugs.

Patients with GSD1b associated neutropenia respond well to G-CSF with reduced infections and symptoms of enterocolitis. Long-term complications are infrequent and G-CSF is well tolerated in most patients.

Disclosures:

Dale: Amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding. Boxer:Amgen: Equity Ownership.
Autoimmune Hemolytic Anemia
Authors: CROSBY WILLIAM H. , RAPPAPORT HENRY
Published: October 2019

1. The records of 57 patients with autoimmune hemolytic anemia have been reviewed to learn if the hematologic data were of any prognostic importance. It appeared that higher mortality was associated with severe anemia, reticulocytopenia, thrombocytopenia and leukopenia. The presence of spherocytosis and erythroblastosis seemed to have no prognostic significance with regard to mortality.

2. Thrombocytopenia in several patients was associated with thromboembolic disease.

3. There was a definite seasonal incidence of hemolytic crises in AHA, more crises occurring during the winter months.
Syndrome of Hypogammaglobulinemia, Splenomegaly and Hypersplenism
Authors: PRASAD ANANDA S. , REINER E. , WATSON C. J.
Published: October 2019

Two cases of acquired idiopathic hypogammaglobulinemia associated with splenomegaly and hemolytic anemia due to hypersplenism have been reported. Splenomegaly and hypersplenism appears to be the result of reticulum cell hyperplasia caused by the lack of gamma globulin and resultant repeated infections. Both the cases had splenectomy with marked hematologic improvement.

A review of the literature indicates that this syndrome is relatively common, in comparison to the incidence of total number of acquired hypogammaglobulinemia that has been reported in the literature so far. It also seems very important to recognize it because of the obvious therapeutic implications. Serum gamma globulin level should be determined in cases with unexplained hepatosplenomegaly, hypersplenism and hemolytic anemia.
The Hemolytic Effect of Ionizing Radiations and Its Relationship to the Hemorrhagic Phase of Radiation Injury
Authors: STOHLMAN FREDERICK , BRECHER GEORGE , SCHNEIDERMAN MARVIN , CRONKITE EUGENE P.
Published: October 2019

Post-irradiation anemia is primarily the consequence of red cell aplasia and of hemorrhage secondary to thrombocytopenia. Data have been presented which indicate that in addition there is intravascular red cell damage. Of particular interest was the observation herein presented that the damage to the red cells produced by ionizing radiations is of an indirect nature. Using the chromium technic it was possible to demonstrate red cell damage as early as the 1st-3rd post-irradiation day. In these studies tagged cells were transfused 15-55 days prior to irradiation in order to establish the elution rate of Cr51. On the 1st-3rd day following irradiation there was an abrupt increase in the rate of Cr51 loss. When normal cells were transfused immediately after irradiation there was also a striking increase in the rate of Cr51 loss above that observed in normal animals, indicating that radiation damage to the red cell was indirect. When cells were transfused from irradiated donors into normal recipients, it was not possible to demonstrate red cell damage prior to the 10th post-irradiation day, indicating that the damage was progressive.

Since there was a selective loss of Cr51 it is suggested that chromium produced an additive damage to the cell. The possible implication of this observation in the use and interpretation of Cr51 survival curves is discussed.

It was also determined that red cells which were recirculated following extravasation during the thrombocytopenic phase of radiation injury had a shortened survival, whereas normal cells, tagged and injected intramuscularly into normal recipients survived normally on return to the general circulation. Cells collected from dogs one day after irradiation, tagged with Cr51 and then injected intramuscularly into normal recipients, showed a striking increase in the rate of destruction. Cells collected on the 4th post-irradiation day were destroyed even more rapidly. These studies confirmed the early onset of radiation injury to the cells, its progressive nature, and demonstrated the damage incurred as a result of passage through the extravascular cycle. As might be expected there was also a shortened survival of red cells collected from the thoracic lymph of thrombopenic irradiated animals.

Conclusions: 1. Ionizing radiations produce intravascular red cell damage in addition to the known loss incident to hemorrhage. This damage is evident within 24-72 hrs. of irradiation.

2. The red cell damage is an indirect and progressive effect of irradiation.

3. Those cells which are returned to the general circulation, following passage through an extravascular cycle, are damaged. The extent of injury to normal cells is such that it is not detectable by current methods. However, the addition of a second injury, such as that following irradiation, enables the detection of this damage.

4. Chromium "elution" is altered by irradiation injury.
Preservation of Radiation Recovery Factor in Frozen Marrow
Authors: FERREBEE JOSEPH W. , BILLEN DANIEL , URSO IRENE M. , LU WAN CHING , THOMAS E. DONNALL , CONGDON CHARLES C.
Published: October 2019

Injection of mouse marrow preserved in glycerol at -80 C. for six weeks has been shown to be quantitatively similar to injections of fresh marrow in inducing recovery in mice after lethal radiation. The effect of the preservation procedure on subsequent rates of DNA synthesis has been determined in vitro for both preserved human marrow and preserved mouse marrow.

Since DNA synthesis is a satisfactory measure of rates of cell multiplication and since preservation is shown to affect DNA synthesis in human marrow to a lesser degree than in mouse marrow, it may be concluded that the procedure used (-80 C. plus 15% glycerol) is satisfactory for the maintenance of viability in human marrow.
Erythrocyte Glycolysis in Patients with Malignant Neoplasms and Other Chronic Diseases
Authors: ULTMANN JOHN E. , HYMAN GEORGE A. , HARVEY JANE L. , DENTE ANTHONY R.
Published: October 2019

The rate of erythrocyte glycolysis of blood from normal volunteers was 12.0 mg.% glucose consumed/hr with 5 x 106 Rbc/cu.mm. The mean glycolytic rate of blood from tumor patients (14.5 mg.% glucose consumed/hr. with 5 x 106 Rbc/cu.mm.) and from patients with other chronic diseases (17.4 mg.% glucose consumed/hr. with 5 x 106 Rbc/cu.mm.) was found to be greater to a statistically significant degree than that of normals in the absence of reticulocytosis. This suggests that the red cell population of the patients with neoplastic and chronic diseases may be younger than that of normal subjects and that only some of the young cells have demonstrable reticulum.
Alkali Resistant Type of Hemoglobin in Women with Molar Pregnancy
Authors: BROMBERG YEHUDA M. , SALZBERGER MACCABI , ABRAHAMOV ABRAHAM
Published: October 2019

In 4 patients with molar pregnancy the fetal Hb blood concentration was found to be significantly higher than that found in control groups of normal early pregnancy and in imminent and incomplete abortions. Following the molar abortion the fetal Hb decreased gradually and reached normal vlaues within three to five months.

It is assumed that the increased production of fetal Hb in these patients is due to the reactivation of fetal erythropoietic activity by either excessive amounts of chorionic gonadotropin, stress due to a rapidly growing tumor or to some other unknown factors.
Clot Firmness
Authors: WEINER MURRAY , WEISBERG LOUIS G.
Published: October 2019

1. The relative firmness of clots in different systems is not necessarily directly correlated with the speed of clot formation.

2. Thrombin will hasten the development of full clot firmness, but will not significantly alter the degree of clot firmness of citrated plasma specimens, although the firmness of "purified" fibrinogen preparations clotted by thrombin is increased with increasing thrombin concentration.

3. Citrate and/or calcium concentration influence clot firmness.

4. Fibrinogen preparations develop greater firmness on clotting with thrombin than do citrated plasma preparations.

5. A platelet factor influences clot firmness of recalcified plasma, but not of thrombin-activated or coagulase-activated plasma.

6. Clot firmness of recalcified citrated plasma is reduced by storage. This reduction in firmness is more rapid if the specimen is deep frozen as compared to unfrozen aliquots at 6 C.
The Diego System—Steps in the Investigation of a New Blood Group System. Further Studies
Authors: LAYRISSE MIGUEL , ARENDS TULIO
Published: October 2019

The distribution of the Diego factor has been studied in various Mongoloid, Caucasian and Negroid populations. In six South American Indian stocks, one Canadian Indian stock and two groups of Asiatic Mongoloids it was present in high incidence. In Caucasians tested, negative results were found. In two groups of Venezuelan Negroes with probable Indian admixture the factor was also present.

The Diego antigen was inherited in single dose (heterozygous) in 41 families studied and could be followed through several generations with no sex-linkage.

The first samples of anti-Diego serum reacted only in the indirect anti-globulin test to a titer between 1:256 and 1:512. The enzymes papain and trypsin gave no advantage. It is more active at room temperature (25 C. and 37 C.) than at cold temperature (4 C. and 18 C.). The most suitable pH range is from 5.0 to 8.0.

The Diego factor can be considered as part of a new blood group system (the Diego system) which could be classified as the tenth firmly established blood system.
Viability and Distribution of Leukocytes Following Cross-Circulation Experiments Between Leukemic and Normal or Irradiated Rats
Authors: HOLLINGSWORTH J. W. , FINCH STUART C. , CHALTAS MARY
Published: October 2019

1. Recipient rats were observed following periods of circulatory mixing with normal or leukemic donor animals.

2. The normal rat tail appears to store leukocytes in its vascular channels, as evidenced by a progressively falling leukocyte count from a tail incision as the tail is "milked."

3. Comparison of irradiated rat tail and arterial blood leukocyte counts during and after cross-circulation revealed the tail values to be consistently higher than the simultaneous arterial levels. This observation excludes a major removal and destructive mechanism by some organ such as the lung as a major cause of the rapid decrease in arterial blood leukocyte levels noted in irradiated rats after the cross-circulation procedure.

4. Tail blood leukocyte counts of irradiated, cross-transfused rats remained higher than those of control irradiated rats for at least three days.

5. After cross-circulation with leukemic donor rats, leukemic cells were identified in the tail blood of normal rats for at least three days.

6. Prolonged observations revealed that 2 irradiated Sprague-Dawley strain recipients of leukemic cells from Sherman strain leukemic donors survived the severe radiation damage but developed leukemia. This observation suggests that transfused leukemic white blood cells protect against radiation damage.

7. Five normal Sherman strain rats developed leukemia after cross-circulation with leukemic Sherman donors. This incidence (50 per cent) of transmission to adult animals is much higher than has been reported by conventional methods of transfer.

8. The observations reported support the concept of viability of leukocytes transfused in rats by this method, and suggest that at least one site of noncirculating transfused leukocytes is within sluggish vascular channels.
Recent Advances in the Knowledge of Total Red Cell Volume, Production and Destruction
Authors: BERLIN NATHANIEL I. , LAWRENCE JOHN H. , ELMLINGER PAUL J.
Published: October 2019

Since anemia or polycythemia can result only from a change in rate of formation of red cells or a change in the life span of the red cells, a determination of the total red cell volume, Fe59 studies to estimate the rate and site of production of red cells, and C14-labeled glycine or Fe59 studies to determine the life span of red cells permit a description of the pathogenesis of any alterations in the erythropoietic state. This has been of great value in clinical hematology from the stand-point of understanding the basic nature of the various diseases and, in particular, the selection of patients with unusual anemias possibly related to hypersplenism who might benefit from splenectomy. Splenic erythropoiesis can be easily differentiated from splenic red cell destruction.

Since two of these methods—the determination of the blood volume and the Fe59 studies—are now possible in general commumnity hospitals and since modifications of the Fe59 method permit the determination of the life span of the red cell, adequate evaluation of the erythropoietic state is now possible in general hospitals.
Reed-Sternberg Cells in the Peripheral Blood
Authors: LUDMAN HAROLD , SPEAR PAUL W.
Published: October 2019

A case of Hodgkin’s disease is presented, in which Reed-Sternberg cells were observed in the peripheral blood.
Chronic Myelosis Following the Use of Thorotrast
Authors: NETOUSŠEK M. , BORESŠ J. , DVORŘAÁK K.
Published: October 2019

Following hepatolienography which was carried out in a woman aged 52 in 1931, for a tumor of the right kidney which was removed at operation, chronic myelosis developed in 1954. This was remarkable for the absence of splenomegaly, a relatively mild course and a paradoxic reaction to x-ray irradiation of the spleen, when instead of the expected decrease there was a doubling in the number of leukocytes. A causal relationship of myeloid leukemia to the thorotrast administered 23 years previously cannot be excluded.
Morphology and Cytochemistry of the Granule-Vacuole Body of Leukemic Cells
Authors: ACKERMAN G. ADOLPH
Published: October 2019

A rare and hitherto undescribed vacuole-like structure, the granule-vacuole body, found in the cytoplasm of certain cells of granulocytic and monocytic leukemia has been described morphologically and cytochemically. This inclusion has been compared with the cytoplasmic granules and Auer bodies occurring in leukemic cells.
Some Observations on the New Human Blood Factor Dia
Authors: LEVINE PHILIP , ROBINSON ELIZABETH A.
Published: October 2019

Evidence is presented that the Diego antigen is independent of 15 other low-incidence blood factors and is not genetically related to 4 high-incidence factors. In all cases of low incidence antigens, similar comprehensive studies with a panel of essential test cells are required to exclude possible serologic or genetic relationship to other blood factors.
Microdrepanocytic Disease in Greece
Authors: CHOREMIS C. , ZANNOS L.
Published: October 2019

Among the 57 patients with the hematologic picture of sickle cell anemia, examination of the parents revealed that 44 were suffering from the microdrepanocytic disease and 13 from sickle cell anemia. The much more frequent occurrence of microdrepanocytic disease was expected since the microcytemia trait appears to be much more frequent in Greece than the sickle cell trait.

Most patients with sickle cell anemia originated from areas where the trait has been found in a very high frequency. In contrast, the geographical distribution of patients with microdrepanocytic disease is much wider.
A Comparative Evaluation of the Sensitivity of the L.E. Cell Test Performed Simultaneously by Different Methods
Authors: DUBOIS EDMUND L. , FREEMAN VIVIAN
Published: October 2019

1) A group of at least three types of L.E. cell tests should be performed in order to adequately screen strongly suspected cases of SLE. This battery consists of the sieved two hour clot, the Snapper-Nathan ring method and the Zinkham-Conley rotary glass bead technic. Utilizing these three methods in studying a group of 47 proven cases of SLE both treated and untreated, 63 batteries were performed. In 42 of these batteries there was at least one positive test. In 5 batteries only the rotary method was positive, in 4 the clotted method and in 2 the ring method. The largest numbers of L.E. cells and hematoxylin bodies were found by the rotary method. If only one test is to be performed this latter technic is the method of choice.

2) The use of excessive amounts of heparin over 0.75 mg./10 cc. of blood resulted in a depression of the L.E. phenomenon. The technic of adding "small amounts" of heparin to blood or "heparinizing a syringe" to draw specimens for L.E. studies is to be avoided.

3) The explanation for the augmentation of the L.E. phenomenon by clotting or rotation with glass beads seemed to be the production of leukocyte trauma by either method. This explained the poor results obtained when the clotted blood specimen is broken up with wooden applicator sticks rather than macerated through a fine sieve. The clotting mechanism itself probably has little to do with the increased numbers of L.E. cells.

4) With adequate treatment the L.E. cells disappeared in many patients. Forty patients with positive L.E. cell tests treated two months or more had a simultaneous battery performed on them as described above. All the tests were negative in eighteen patients, seven patients had fewer than 5 L.E. cells per 500 leukocytes and fifteen patients had more than that number.
An Explanation of the Discrepancy between Direct and Indirect Platelet Counts
Authors: FITCH HARRY J.
Published: October 2019

The indirect platelet count is higher than the direct count because the red cells, which are used as a point of reference in the indirect method, are not randomly distributed beneath the coverslip. The red cells are concentrated at the edge of the coverslip so that the true ratio of red cells to platelets cannot be accurately established. Indirect platelet counts based on the ratio in the central areas of the coverslip are too high.
In Vitro Incorporation of C14—DL-Leucine into Normal and Leukemic White Cells
Authors: BAKER W. H. , ZAMECNIK P. C. , STEPHENSON M. L.
Published: October 2019

Normal white blood cells and cells from patients with leukemia are able to incorporate C14-DL-leucine into proteins in vitro. Cells from individuals with chronic myelogenous leukemia were able to incorporate significantly greater amounts of the amino acid, and they were able to maintain this incorporation for a longer period of time than the control cells.

Both types of cells had the same in vitro survival time of four days, following storage at 3 C.

The incorporation was partially decreased in the presence of dinitrophenol, as well as in an atmosphere of nitrogen.
AML Bone Marrow Microenvironment Impairs Dendritic Cell Maturation and Antigen Presentation
Authors: Wagner Bettina , Rebmann Vera , May Tobias , Duehrsen Ulrich , Göthert Joachim Rudolf , Horn Peter A , Opalka Bertram
Published: October 2019

Abstract 1491

In hematopoietic malignancies the bone marrow (BM) stroma has been reported to be in dynamic contact with neoplastic cells in a way that malignant cells are supported by various mechanisms including blocking of apoptosis, induction of resistance to therapeutic drugs, and suppression of potential anti-tumorigenic immunity. Here, we studied the immunomodulatory properties of cellular factors derived from BM stroma cells and their influence on maturation of dendritic cells (DC). To circumvent the inter-individual heterogeneity often observed with primary stroma, novel BM stroma cell lines derived from BM stroma of two AML patients (before and after chemotherapy) and one healthy donor were established. The surface marker profile of the cell lines is in accordance with an MSC pattern. Furthermore, differential expression patterns were observed for molecules such as HLA-G, MICA, MICB, ULBP2, ILT-4 and intracellular IL-10 known to be implicated in the regulatory network of the immune system. To monitor DC maturation in presence and absence of BM stroma cell fragments, obtained by ultra-sonication, surface markers including CD40, CD80, CD83, CD86, and CD274 were investigated before maturation on monocytes, three days after BM stroma fragment addition and on terminal differentiated DCs. In addition, the expression of classical (HLA class I, HLA-DR) and non-classical immunoregulatory molecules (HLA-G, HLA-E) were determined. For terminal differentiation of DCs on day seven, the medium was exchanged and 1000 U/ml IL-4, 1000 U/ml GM-CSF, 2.0 U/ml IL-1β and 0.2 U/ml TNFα were added to complete the maturation process by giving a “danger” signal. A markedly decreased expression of differentiation (up to −36%) and antigen presentation molecules (up to −68%) of DCs cultured in presence of AML BM stroma cell fragments was observed compared to DC maturation without AML BM stroma cell fragments. In particular, the decreased HLA-DR expression could not be restored by the addition of the pro-inflammatory cytokines at day seven. In comparison, the addition of BM stroma cell fragments derived from AML stroma had a higher impact on the decrease of HLA-DR expression than BM stroma cell fragments of the healthy donor cells. This indicates that molecules derived from the AML stroma cells are implicated in mechanisms impairing DC maturation and function, which may promote immune escape during leukemia development.

Disclosures:

May: InSCREENeX GmbH: Employment, Equity Ownership, Patents & Royalties.
T(8;9)(p22;p24)/PCM1-JAK2 Activates SOCS2 and SOCS3 Via STAT5
Authors: Ehrentraut Stefan , Nagel Stefan , Scherr Michaela , Schneider Bjoern , Quentmeier Hilmar , Geffers Robert , Prochorec-Sobieszek Monika , Ketterling Rhett P , Knudson Ryan A , Feldman Andrew L. , Kadin Marshall , Drexler Hans G. , MacLeod Roderick AF
Published: October 2019

Abstract 1567

Fusions of the tyrosine kinase domain of JAK2 with multiple partners occur in leukemia/lymphoma where they are believed to promote JAK2-oligomerization and autonomous signalling although the underlying mechanisms remain unclear. Affected entities are candidates for therapy with JAK2 signalling inhibitors. Among 200 peripheral T-cell lymphomas surveyed, we identified only two with JAK2 amplification and none with JAK2 translocations, confirming their rarity in T-cell neoplasias. Here we describe the genomic, transcriptional and signalling characteristics of PCM1-JAK2 formed by t(8;9)(p22;p24) in cell lines established at indolent and aggressive phases of a cutaneous T-cell lymphoma. To investigate signalling, PCM1-JAK2 was subjected to lentiviral knockdown which inhibited 7 genes most upregulated in t(8;9) cells, notably SOCS2/3. SOCS3, but not SOCS2, was also upregulated in a chronic eosinophilic leukemia bearing PCM1-JAK2. Conversely, expression of GATA3, a key T-cell developmental gene silenced in aggressive phase cells, was partially restored by PCM1-JAK2 knockdown. Activation of the tumor suppressor SOCS3 by PCM1-JAK2 may follow structural alteration of JAK2 affecting the ternary complex it forms with SOCS3 and receptor proteins. Treatment with JAK2 inhibitor (TG101348) to which MAC-1/2A/2B cells were conspicuously sensitive mimicked knockdown results, highlighting JAK2 as the active moiety. PCM1-JAK2 signalling required pSTAT5 as reported for JAK2V617F or ETV6/TEL-JAK2, thus extending the paradigm of STAT5 activation by JAK2 alterations in hematopoietic malignancies. MAC-1/2A/2B are the first JAK2– translocation cell line models. Our data support further investigation of SOCS2/3 as signalling effectors, prognostic indicators and potential therapeutic targets in cancers with JAK2 rearrangements.

Disclosures:

No relevant conflicts of interest to declare.
Alisertib Added to Rituximab and Vincristine Is Synthetic Lethal and Curative of Double Hit Diffuse Large B-Cell Lymphoma in Mice
Authors: Mahadevan Daruka
Published: October 2019

Abstract 159

About one-third of diffuse large B-cell lymphoma (DLBCL) patients develop relapsed/refractory disease, a major cause of morbidity and mortality. Of DLBCL, 5% are double-hit lymphomas (DHLs) with translocations of MYC and BCL2 characterized by poor outcomes with R-CHOP. Auroras (A and B) are a family of mitotic oncogenic serine/threonine kinases required for high fidelity regulation of the mitotic phase of the cell cycle. Aberrant over-expression of Auroras leads to genetic instability, polyploidy and resistance to microtubule targeted agents. Previously we showed Aurora inhibitor (MLN8237, alisertib) [M] plus vincristine [V] plus rituximab [R] is synthetic lethal and a potential curative therapy in a mantle cell lymphoma model (Granta-519) with several cell cycle genetic defects [t (11;14)], ATM (haploinsufficient), p16/p14 (Del)] but p53 (WT). We evaluated ABC-DLBCL cell lines TMD-8 and U-2932 where the BCR/NF-kB pathway is constitutively active. In TMD-8, CD79B is mutated and in U-2932, Bcl2 [3q27] and Bcl6 [18q21] are amplified, p53 (mutated) and c-Myc over-expressed [DHL]. Triple therapy with MVR abolished proliferation, disrupted cell cycle processes and induced apoptosis compared to single agent and doublet treatments. A Mouse xenograft model of U-2932 showed modest activity for M (30 mg kg, PO, QD, 3 weeks), V (0.375 mg/kg, IV, Q1Wx3) and MV with tumor growth inhibition (TGI) of ∼10% (p=0.01) respectively. However, R (10 mg/kg, IV, Q1Wx3) alone induced ∼ 50% TGI (p=0.01) due to effective targeting of Bcl2. Doublets MR and VR led to tumor regression (TR), but relapse soon after stopping therapy. In contrast, MVR demonstrated TR with no relapses >40 days after stopping therapy. All treatments were well tolerated (no weight loss) and Kaplan-Meier analysis showed that mice treated with MR, VR and MVR had a statistically significant improvement in overall survival compared to control, M, V and MV (p<0.0001). Moreover, with MVR, OS was 100% (Figure 1). Gene expression profiling, RT-PCR, IHC and Western blotting of harvested tumors at the end of treatment (3 weeks) has been interrogated to ascertain the mechanistic role of Aurora inhibition when added to R [R Vs MR] and VR [VR Vs MVR]. Thus, addition of R to MV represents a novel therapeutic strategy that is currently under clinical trial evaluation in aggressive B-NHL including double hit lymphomas sponsored by Millennium Pharmaceuticals (C14011).

Figure 1: Kaplan-Meier Survival by Treatment: MLN8237 (M), Rituximab (R), Vincristine (V), MR, MV, VR, MVR in a U-2932 ABC-DLBCL SCID Mouse Xenograft Model Figure 1:. Kaplan-Meier Survival by Treatment: MLN8237 (M), Rituximab (R), Vincristine (V), MR, MV, VR, MVR in a U-2932 ABC-DLBCL SCID Mouse Xenograft Model

Disclosures:

No relevant conflicts of interest to declare.