bluuebunny/crossref_metadata_2025 huggingface dataset:
Mobilization of Peripheral Blood Stem Cells with Rituximab Plus ESHAP Combination Regimen in B-Cell NHL - Preliminary Results.
Authors: Kim Min Kyoung , Kim Shin , Lee Seong Sook , Sym Sun Jin , Lee Dae Ho , Kim Sang We , Jang Seongsoo , Park Chan Jeong , Chi Hyun Sook , Huh Joo Ryung , Suh Cheolwon
Published: October 2019

Objectives: The mobilization chemotherapy should be directed toward the dual objectives of good antilymphoma activity and adequate PBSC mobilization. Previously, we reported the ESHAP plus G-CSF is an effective mobilization regimen in patients with relapsed and high risk NHL. The purpose of this study is to assess the efficacy and feasibility of autologous stem cell collection after mobilization with Rituximab plus ESHAP (etoposide, methylprednisolone, high-dose cytarabine, and cisplatin) combination therapy in B-cell Non-Hodgkin’s lymphoma.

Patients and Methods: CD20 + B-cell NHL patients with either high risk, relapsed or refractory patients were eligible for the study. The regimen consisted of: Rituximab 375mg/m2 given on day 1, standard ESHAP therapy (etoposide 40mg/m2 IV/2hr day 2–5, methylprednisolone 500mg/m2 days 2–6, cytarabine 2g/m2 IV/3hr on day 5 and cisplatin 25mg/m2 CIV days 2–5).

In all patients, the collection of PBSC was performed during recovery after giving G-CSF (10μ/kg/day) started on day 7. The first harvest was started only if the peripheral blood hematopoietic progenitor cell (HPC) count exceeded 5/μl.

Results: Fifty-five mobilization procedures performed on 20 patients with a median of three apheresis (range 2–5) per patient. Median age was 37 years (range 15–65). At the time of PBSC mobilization, Seventeen patients were considered to be responsive (CR, PR and sensitive relapse). Aphereses were started on day 16 (range 13–18). The number of total MNCs (×106/kg) collected was 5.4 (range, 1.4–14.5) and the number of CD34+cells (× 106/kg) was 10.6 (range, 4.9–52.6). The median days of G-CSF usage was 11(range 9–15). Most non-hematologic adverse events were mild and reversible. Nineteen patients (95%) were achieved optimal response, which was defined as ≥ 5 × 106 CD34 cells/kg. In all, sixteen of the patients underwent high-dose chemotherapy. The median time to ANC ≥ 0.5 × 109/L was 10 days (range, 8–17). The median days to platelets ≥ 20 × 109/L was 12 (range, 7–27).

Conclusion: Addition of Rituximab to ESHAP chemotherpy does not show any adverse effect in autologous stem cell mobilization and collection. R-ESHAP regimen is effective as a combined mobilization and second-line regimen for patients with pretreated B-cell lymphoma.

Figure 1. Days to achieve optimal response Figure 1. Days to achieve optimal response
Reduced Rate of Infectious Complications Following Bloodless Autologous Peripheral Blood Stem Cell Transplants (APBSCT).
Authors: Ford Patricia A. , Matthews Barbara A. , Brown Nicole M.
Published: October 2019

While blood transfusions can be life saving, the associated risk of transfusing allogeneic blood is significant. The most common patient fears are transfusion-transmitted diseases such as HIV, Hepatitis B and C and West Nile Virus; however, the known risk of transmitting these diseases is quite small. More common complications are due to immunosuppression which can cause an increased risk of cancer recurrence in the oncologic patient and a significantly increased risk of infection. In trauma patients, it has been shown that the risk of infection increases with each additional unit of blood transfused. Currently, about 2.2 units of platelets and 3.3 units of red blood cells are administered following high-dose chemotherapy and an APBSCT. At the Center for Bloodless Medicine and Surgery at Pennsylvania Hospital, many procedures are now being completed without the use of blood products. We have previously reported the ability to perform bloodless APBSCT (Ballen, et al. J Clin Oncol2004;22:4087-4094). Knowing that blood transfusions can increase the risk of infection, we wanted to evaluate this transfusion-free population to determine if there was a correlation between infection rates and blood transfusions in the high risk transplant population. We performed a retrospective chart review of 46 patients with multiple myeloma (22), lymphoma (22) and breast cancer (2) who underwent a bloodless APBSCT in our center. Prior to transplantation, all patients recieved standard transplant doses of cyclophosphamide, carmustine and etoposide (BCV) or Melphalan. A PubMed search was performed and the closest data set in terms of patient demographics was a study by Pereira, et al. who report the rate of infectious complications in 75 patients with myeloma (30), lymphoma (30) and breast cancer (15) who were transfused liberally following high-dose chemotherapy and APBSCT (Pereira, et al. Eur J Haematol2006;76:102-108). In our bloodless patients, 37% of the patients had at least one infection, compared to Pereira and colleagues’ rate of 68% (see table). Our results are also reported in the average number of infections per patient. This comparison demonstrates a substantial reduction in the rate of infection in the bloodless population. While immunosuppression and the resulting increased infection rates have been correlated to blood transfusions in other patients populations, to the best of our knowledge this is the first report that suggests decreased infection rates in transfusion-free transplant patients. This data provides further evidence to support the practice of blood management strategies in order to reduce or eliminate blood transfusions.

Patients with at least one infection All infections per person Bacterial per person Viral per person Fungal per person Unknown per person Autologous Transplants (Pereira, et al.) 68% .64 .53 .01 .07 .03 Bloodless Autologous Transplants 37% .41 .39 0 0 .02
Multiple Displacement Amplification Enables Large Scale Clonal Analysis after Retroviral Gene Therapy.
Authors: Bleier Stephanie , Maier Patrick , Wenz Frederik , Zeller W. Jens , Laufs Stephanie , Fruehauf Stefan
Published: October 2019

Analysis of the fate of retrovirally transduced cells after transplantation is often hampered by the scarcity of available DNA. We evaluated a promising method for whole genome amplification named multiple displacement amplification (MDA) with respect to the even and accurate representation of retrovirally transduced genomic DNA. We were able to show that MDA is a suitable method to subsequently specify engraftment efficiencies by quantitative real-time PCR as the retroviral integrations are amplified the same way and by the same probability as all other parts of the genome. We validated the method by analyzing a dilution series containing retrovirally transduced DNA and untransduced background DNA and retroviral integrations found in primary material from a retroviral transplantation model by quantitative real-time PCR. The representation of the portion of retroviral DNA in the amplified samples was 0.9-fold (range 0.2 – 2.1-fold) of the portion determined in the original genomic DNA. Furthermore, the succession of the combination of MDA and integration site analysis by ligation-mediated PCR showed an increase in the sensitivity of the method as a specific integration site could be detected in a background of untransduced DNA, while the transduced DNA made up only 0.001%. These results show that MDA enables large scale sensitive detection and reliable quantification of retrovirally transduced human genomic DNA and therefore facilitates follow up analysis in gene therapy studies even from smallest amounts of starting material.
Quality of Life and Myelodysplasia.
Authors: Prommer Eric E.
Published: October 2019

The myelodysplastic syndromes (MDSs) are heterogeneous hematopoietic diseases associated with bone marrow failure, peripheral cytopenias, and a propensity for progression to acute myeloid leukemia (AML). In 2000, an International Working Group (IWG) of investigators proposed standardized response criteria which included quality of life as an important treatment goal. Quality of life becomes important when we are presented with a patient who is not a candidate for intensive curative therapy for myelodysplasia. The purpose of this literature review is to determine what quality of life instruments are optimal for this disease process and review is to examine the types of quality of life tools that have been used in published phase II and III studies of myelodysplasia. The focus is on adult hematology, and excludes studies with curative intent such as bone marrow transplantation, as well as phase I studies. Clinical trials from 1996—2006 were reviewed. 80 clinical trials were analyzed for the presence of attention to quality of life data. 7 trials used a quality of life instrument. All of the studies which used a quality of life instrument were published after 2003. Types of quality of life instruments used were the

(EORTC) QLQ-C30 FACT-An along with the Geriatric Depression Scale (GDS) Short Form-12 (SF-12).

The most common tool was the FACT-An. This was used in 5/7 studies that had quality of life tools. Of the 7 studies, 4 included the effect of therapy in all domains. The remainder focused on fatigue and/or anemia symptoms and their relation to quality of life.

Conclusion: While the IWG response criteria have prompted consideration of quality of life as response criteria, there must be better uniformity in quality of life assessment, as well as determination of the optimal tool to use. A disease specific quality of life instrument should be considered one solution to this problem.
Cyclosporin A, a P-gp Inhibitor, Augments the Anti-Central Nervous System Ph+ Leukemia Effects of INNO-406.
Authors: Yokota Asumi , Kimura Shinya , Masuda Satohiro , Ashihara Eishi , Urasaki Yoshimasa , Terui Yasuhito , Ruthardt Martin , Ueda Takanori , Hatake Kiyohiko , Inui Ken-ichi , Maekawa Taira
Published: October 2019

Central nervous system (CNS) relapse accompanying prolonged administration of imatinib mesylate, an Abl-specific tyrosine kinase inhibitor, has recently become apparent as an impediment to the therapy of Philadelphia-chromosome-positive (Ph+) leukemia. CNS relapse may be explained by limited penetration of imatinib into the cerebrospinal fluid due to presence of P-glycoprotein (P-gp) at blood-brain barrier. To overcome imatinib-resistance mechanisms such as bcr-abl gene amplification, point mutations within ABL kinase domain, and activation of Lyn, we recently developed a specific dual BCR-ABL/Lyn inhibitor, INNO-406 (formerly NS-187), which is 25–55 times more potent than imatinib in vitro and at least 10 times more potent in vivo (Blood106: 3948–3954, 2005). The aim of this study was to investigate the efficacy of INNO-406 in treating CNS Ph+ leukemia. The intracellular accumulation of [14C]INNO-406 in P-gp overexpressing LLC-GA5-COL150 cells was much less than that in parental LLC-PK1 cells. The addition of 10 mM cyclosporin A (CsA) increased the intracellular accumulation of [14C]INNO-406 in both LLC-PK1 cells and LLC-GA5-COL150 cells. The peak concentration of INNO-406 in the brain when 30 mg/kg INNO-406 was administered p.o. was 50 ng/ g (87 nM), representing only 10% of plasma drug level. These findings suggested that INNO-406 is also a substrate of P-gp, as is imatinib. However, the residual concentration of INNO-406 in the CNS was enough to inhibit the growth of Ph+ leukemic cells according to the in vitro data. To increase the concentration of INNO-406 in CNS, we next examined the combined effects of CsA. In the brain, the concentration of INNO-406 was doubled following prior administration of 50 mg/kg CsA. Since pharmacokinetic studies suggested the possible effects of INNO-406 against CNS Ph+ leukemia, we investigated in vivo anti-CNS Ph+ leukemia effects of INNO-406 alone and combination of INNO-406 and CsA using immunodeficient mice (nude or NOD/SCID) which received Ph+ leukemic cells into the cerebral ventricle. INNO-406 alone inhibited growth of leukemic cells harboring either wild type or mutated BCR-ABL such as E255K and M351T in CNS. Furthermore, CsA significantly enhanced anti-CNS Ph+ leukemia effects of INNO-406 in vivo not only against cells harboring wild type BCR-ABL but also against cells harboring BCR-ABL/M351T (Figure). In conclusion, INNO-406 was found to inhibit Ph+ leukemic cell growth in CNS in spite of efflux of the compound by P-gp, and CsA augmented the anti-CNS Ph+ leukemia effects of INNO-406. Phase I clinical study on INNO-406 was initiated in the U.S.A. in July 2006. The efficacy and safety of INNO-406 in the treatment of leukemias is expected to be verified by early-phase clinical trials.

Figure Figure
Rituximab Monotherapy for Cold Agglutinin Disease. Report on 16 Patients from a Single Institution.
Authors: Arriaga Francisco , Jarque Isidro , Paciello Mari-Liz , Cantero Susana , de la Rubia Javier , Sanz Guillermo , Sanz Miguel
Published: October 2019

Background: Cold agglutinin disease (CAD) is an acquired autoimmune hemolytic anemia mediated by cold-reactive autoantibodies that bind erythrocyte carbohydrate antigens, causing hemagglutination, complement-mediated hemolysis and C3d positive direct antiglobulin test. Conventional therapies for CAD are largely ineffective, but remissions after treatment with the anti-CD20 monoclonal antibody rituximab are increasingly being reported.

Patients and Methods: A total of 16 CAD patients (10 women, 6 men) with a median age of 48 years (range, 20–86 years), were treated in our center between May 2002 and January 2006. CAD was idiopathic in 7 cases and associated with other conditions in 9 (systemic lupus erythematous in 4, chronic lymphoproliferative disorder in 3, and unrelated donor cord blood transplant in 2). Hemoglobin concentration before treatment ranged from 4.9 to 10 g/dL. Median IgM anti-I titer was 512 (range, 128–100,000). Rituximab was given as single agent in doses of 375 mg/m2, at days 1, 8, 15 and 22.

Results: The overall response rate was 62.5%, with 9 patients achieving complete remission (56%). Median duration of response was 24 months (range, 5–48 months). Of the 6 non-responders, 5 died from disease progression and 1 remains alive with transfusion dependence. No serious infusion-related adverse events occurred with rituximab.

Conclusion: Rituximab is a safe and effective therapeutic option and should be considered as first-line treatment for patients with CAD.
Paroxysmal Nocturnal Hemoglobinuria (PNH): Long-Term Epidemiological Study.
Authors: de Latour Regis Peffault , Mary Jean Y. , Salanoubat Celia , Terriou Louis , Etienne Gabriel , Mohty Mohamad , Roth Sophie , de Guibert Sophie , Cahn Jean Y. , Socie Gerard
Published: October 2019

PNH is a rare acquired disorder. We aimed to analyze a large cohort of patients on the long term to better determine prognostic factors. We already reported the analysis on 220 patients in 1996 (Socié et al, Lancet). Data were updated and we collected additional data on 247 patients (total of 467 patients, 252 female), diagnosed from 1950 to 2005 from 58 centers. The date of diagnosis was based on flow cytometry analysis if there was no prior positive Ham’s test. The age at diagnosis was 34 years (±17.3). The median follow-up time was 6.8 years (±0.5). Kaplan-Meier 10-year survival estimate was 76.3% (± 2.6). Sixty seven (14.6%) patients developed pancytopenia during follow-up [10-year cumulative incidence (CIn) rate of 18.3% (14% – 22%)]. Deep-vein thrombosis occurred in 108 patients (23.5%) [10-year CIn 28% (23% – 33.1%)]. Nineteen patients developed myelodysplastic syndrome (MDS) [10-year CIn 4.6% (2.4% to 6.8%)]. Eight patients developed acute leukemia [10-year CIn 2.3% (0.4% to 3.9%)]. The following factors were related to poor survival: Older age at diagnosis; age between 40 and 55 years [relative risk RR=2.4 (95% CI 1.4–4.1); p value<10−3], age over 55 years [RR 3.6 (95% CI 2.1–6); p<10−3], no thrombocytopenia [RR 3.2 (95% CI 1.4–7); p<10−3 ], pancytopenia at diagnosis [RR 3.4 (95% CI 1.5–7.9); p<10−2], progression to aplastic anemia (AA) [RR 2.3 (95% CI 1.3–3.9); p<10−2], thrombosis as a complication [RR 8.9 (95% CI 5.7–14.4); p<10−3]; secondary MDS or acute leukemia [RR 13.7 (95% CI 6.1–30.5); p <10−3]. However, we also found a strong effect of time period. Diagnosis before 1986 among 114 patients with classical PNH (hemolytic anemia and/or thrombosis at diagnosis) was associated with worse outcome [RR 2.8 (95% CI 1.1–7.2); p: 0.025]. Similar results were found for 227 patients with marrow failure (AA-PNH syndrome) diagnosed before 1996 [RR 2.9 (95% CI 1–8.7); p: 0.04]. For the remaining 96 patients (here called Unclassified PNH), the cutoff time period was to be diagnosed before 1986 [RR 3.6 (95% CI 1–12.4); p: 0.046]. Fifteen years Kaplan-Meier survival estimates were 76% (± 6.6) for patients with unclassified PNH, 70% (± 4.6) for AA-PNH patients and 63.4% (± 1.4) for patients with classical PNH. Older age and evolution to thrombosis were found to be associated with bad prognosis in each of the 3 groups. Evolutions to AA or malignant disease (MD) were related to poor prognosis in 2 of the 3 groups (Classical PNH and AA-PNH for AA; AA-PNH and Unclassified PNH for MD). We also found a protective role of the treatment in the AA-PNH group [RR without treatment 3.1 (95% CI 1.5–6.5); p<10−2] while all 20 patients with unclassified PNH treated with immunosuppressive therapy were alive.
Acute Leukemia Over the Age of Fifty: A Study of Its Incidence and Natural History
Authors: GUNZ F. W. , HOUGH R. F.
Published: October 2019

(1) Analysis of the death certificates of 553 patients recorded as having died from leukemia showed that 57 per cent of deaths occurred over the age of 50, that more than 60 per cent of all leukemias were acute, and that 46 per cent of acute leukemias occurred over the age of 50.

(2) Investigation of a selected group of 97 patients over 50 dying from acute leukemia suggested that the clinical, hematologic and histologic features of the disease are distinctive and can be differentiated both from those of acute leukemia in younger patients, and from chronic leukemia.
The Direct Coombs Test in Lead Poisoning
Authors: SUTHERLAND DONALD A. , EISENTRAUT ANNA M. , MINSTER MARIE
Published: October 2019

1. The anemia produced by lead intoxication in humans as well as experimentally induced lead poisoning in dogs results in a positive direct Coombs test.

2. The direct antiglobulin test will become positive within twenty-four hours in cases of severe lead poisoning produced experimentally in dogs.

3. The blood from cases of severe lead poisoning in dogs will form a layer phenomenon when allowed to stand with the formation of a supernatant fraction of cells above the packed erythrocytes. These cells remain suspended in the plasma for many hours. This superior fraction on a column of blood has a high per cent of reticulocytes, and cells with coarse basophilic stippling. The direct Coombs test is positive in the superior fraction of such a column. The positivity decreases as the sampling approaches the bottom. In many instances the whole blood direct Coombs test may be negative and the cells from the superior portion of the column be strongly positive. The latter phenomenon was also found in asymptomatic workers exposed to lead in a smelting plant.

4. The possible significance of the correlation of the direct anti-globulin reaction and cell immaturity or chemical trauma to the membrane is briefly discussed.
The Anemia of Trauma
Authors: TOPLEY ELIZABETH , CLARKE RUSCOE
Published: October 2019

1. The anemia of civilian trauma (mainly limb injuries) has been studied in 57 patients.

2. In 22 patients the red cell volume, after primary blood loss has probably ceased, has been compared with the follow-up "normal" red cell volume. The red cell volume credit or deficit is taken as an index of the balance between primary blood loss and blood transfused, and was found to be the major factor contributing to the later polycythemia or anemia. The evidence suggests that an anemia of trauma due to unreplaced primary blood loss is still a common finding in civilian injuries today.

3. A further red cell volume estimate in 20 patients on the 4-14th day after injury showed on average a further fall of 11 per cent of the red cell volume. Evidence is presented for considering that there is a further disappearance of red cells after primary blood loss has ceased in all patients—even in those whose red cell volume did not fall.

4. The evidence suggests that the anemia of trauma is largely preventable by adequate blood transfusion in the majority of civilian injuries.
Hageman Factor (HF) Deficiency
Authors: RAMOT BRACHA , SINGER KARL , HELLER PAUL , ZIMMERMAN HYMAN J.
Published: October 2019

A patient with Hageman factor (HF) deficiency is described. This syndrome is characterized by complete absence of any hemorrhagic tendency in the presence of laboratory findings which, as a rule, are associated with severe disturbances in the hemostatic mechanism. The clotting time was markedly prolonged, the plasma prothrombin time was normal, but prothrombin consumption was decreased. The thromboplastin generation test revealed that HF is essential for blood thromboplastin formation at least in vitro.

Procedures for the differentiation of HF deficiency from AHF, PTC and PTA deficiency syndromes are outlined.

Transfusions of as little as 50 cc. of 20 day old blood normalized the abnormal clotting tests immediately for a period of about 36 hours.

The basis for the apparent lack of need for preoperative preparation with blood transfusions in HF deficiency is discussed.
Erythrokinetics in Pernicious Anemia
Authors: FINCH CLEMENT A. , COLEMAN DANIEL H. , MOTULSKY ARNO G. , DONOHUE DENNIS M. , REIFF ROBERT H.
Published: October 2019

Quantitative measurements of the erythropoietic activity of the marrow, of circulating red cell production and destruction have been made in patients with pernicious anemia in relapse and during response to vitamin B12 therapy.

Total erythropoietic marrow activity as reflected by turnover of heme components proceeds at a rate of approximately 3 times normal. The delivery of viable red cells to the circulating blood, however, does not increase above normal. This would indicate that the greater portion of marrow activity is ineffective in terms of blood production. This marrow dysfunction coupled with an increased rate of cell destruction of approximately 3 times normal is responsible for the anemia. Total erythropoiesis is somewhat less, and effective erythropoiesis considerably less, than that which may be expected of the normal marrow under the sustained stimulus of anemia.

The reticulocyte count is shown to be an unreliable index of blood production in untreated pernicious anemia due to loss of reticulum from cytoplasma of many red cells before their delivery into circulation.

During the response to vitamin B12 the ineffective erythropoiesis is converted to effective erythropoiesis, whereas total erythropoiesis remains unchanged. The rate of blood production during recovery is 3 to 4 times normal.
A Single-Centre Experience of Clinico-Pathological Features, Risk Factors and Treatment Outcome of Posttransplant Lymphoproliferative Disorders (PTLD) after Renal Transplantation.
Authors: Sondergaard Esben , Strandhave Charlotte , Pedersen Robert S. , Jorgensen Kaj A. , Bendix Knud , d’Amore Francesco
Published: October 2019

Background: PTLD is a lymphoid proliferation that develops as a consequence of immunosuppression and represents monoclonal B-cell, or rarely T-cell proliferations, occurring in a setting of decreased T-cell immune surveillance. PTLD is a major complication after solid organ transplantation. The disease is often unpredictable and devastating of nature resulting in a high rate of morbidity and patient mortality.

Methods: Between 1990 and 2005 872 renal transplantations in 793 patients were performed at Skejby University Hospital in Aarhus and 11 cases of PTLD were identified retrospectively. PTLD Patients were investigated according to transplantation procedure, patient characteristics, type of lymphoma, treatment and outcome.

Results: 11 of 793 renal transplant recipients (1,4%) developed PTLD. Patients (7 male, 4 female) ranged from 19 to 73 years of age at the time of diagnosis (mean age: 42 years). 8 had a cadaveric renal transplant while 3 received a transplant from related donor. Lymphomas were classified as diffuse large B-cell lymphoma in 10 cases and as Burkitt lymphoma in 1 case. No cases were of T-cell origin or Hodgkins lymphomas. Six cases were EBV positive (54%). Out of 11 cases, 5 were diagnosed within 1 year after transplantation, among which 4 were EBV positive, whereas 6 had a latency period of more than 1 year, among which only 2 were EBV positive. The mean latency time between grafting and diagnosis of PTLD among the early lesions was 7 months and among late lesions 6 years and 2 months (ranging from 1 month to 10 years and 8 months). Sites involved in PTLD were renal graft in 2 cases (both early lesions), lungs in 1 case, bowel system in 4 cases, CNS in 1 case, isolated lymph nodes in 1 case, widespread disease in 1 case and uncertain location in 1 case. All patients were treated by tapering of the immunosuppressive regimen. Two were treated with additional surgery and 3 with chemotherapy - all 5 of them are alive and in complete remission. Among those 5 patients 3 have preserved graft function (60%) and 2 returned to dialysis. One patient was treated with anti-CD20 monoclonal antibody, 3 had both conventional chemotherapy and anti-CD20 immunotherapy and 2 did not receive additional treatment. All 6 were dead at the end of the study period. With a median follow-up period of 5 years and 2 months (ranging from 6 months to 11½ years) 5 patients of 11 PTLD cases (45%) were alive with no sign of lymphoma relapse.

Conclusions: PTLD is a severe complication, usually running an aggressive course and the outcome remains poor. The incidence in our population-based regional study material is 1,4%. Overall survival after a median follow-up of 62 months was 45%, with 60% of survivors maintaining graft function.
Long Term Follow-Up and Retreatment of Patients with Low-Grade Lymphoproliferative Disorders (LPD) after 1 Year of Phamacokinetic (PK) Based Maintenance Therapy with Rituximab.
Authors: Duff Jennifer R. , Lynch James W.
Published: October 2019

Background: We previously published data from a PK based maintenance trial in patients with CD 20 positive LPD. The PK based schedule derived from that trial was 1 dose every 3 months and is currently being evaluated in a large randomized trial (ECOG 4402). There has been appropriate concern that offering maintenance therapy may select for tumor cells resistant to rituximab thereby compromising the chance of response if patients are retreated once relapsing after such therapy. We now present long term follow-up on patients with low grade LPD treated on that trial. We furthermore report the results of retreating 5 patients who initially responded but relapsed after one year of maintenance rituximab.

Methods: Patients with CD20 positive LPD (except SLL/CLL) were treated with four weekly infusions of rituximab 375mg/m2. All patients without PD were then monitored for 1 year and received a single infusion of 375mg/m2 when the level fell below <25mcg/ml. Among 22 patients with low grade LPD 10 (45%) achieved a complete response and 5 (23%) a partial response for an overall response rate with 68%. With a minimal follow-up of 69 months, all but 2 patients have sufficient data available to evaluate their subsequent treatment and response.

Results: Of the 22 patients, there were 18 with follicular lymphoma, 2 with lymphoplasmacytoid lymphoma and 1 each with MALT and NOS. The median progression free survival (PFS) for the entire group was 23 months, but for responders the PFS was 50 months. 6/22 (27%) remain in continuous complete remission with no further therapy with a median follow-up of 72.5 mo. (range 69–76). Nine patients who initially responded have subsequently relapsed and were treated at the discretion of the treating physician as follows: 2 received no treatment one of whom experienced a spontaneous CR lasting 59 months, one patient with an isolated CNS relapse received intrathecal and local radiotherapy and is currently in CR with no further therapy at 71mo, 1 patient died in PR of a presumed MI and 5 received retreatment with rituximab accompanied by 2 years of maintenance therapy given as 1 dose every 3 months. Of the 5 patients who were retreated with rituximab the outcomes are listed in table I.

Conclusions: Individualized PK dosing for rituximab for 1 year yielded 27% prolonged DFS in patients with LG LPD. Although the numbers of patients treated are very small, 80% of those who responded to rituximab and subsequently relapsed retained sensitivity to rituximab and have had durable benefits, comparable to their first course. Future trials which evaluate the efficacy of limited duration maintenance strategies should continue follow-up of patients after progression to determine whether subsequent treatment with rituximab offers clinical benefit.

Table I Patient # 1st PFS 2nd Response 2nd PFS 1 18mo CR 44mo+ 2 23mo NR NA 3 27mo CR 33mo 4 29mo CR 33mo+ 5 50mo PR 29mo+
The Lymphoid Cell Surface Receptor NTB-A: A Novel Monoclonal Antibody Target for Leukemia and Lymphoma Therapeutics.
Authors: Korver Wouter , Singh Shweta , Zhao Xiaoxian , Hsi Eric D. , Abo Arie
Published: October 2019

NTB-A is a CD2-related cell surface protein expressed primarily on lymphoid cells including B-lymphocytes from Chronic Lymphocytic Leukemia (CLL) and lymphoma patients. We have generated a series of murine and chimeric mAbs against NTB-A and assessed their therapeutic potential for CLL. Selective mAbs to NTB-A were further tested in functional Complement Dependent Cytotoxicity (CDC) and Antibody Dependent Cellular Cytotoxicty (ADCC) assays in cell lines and B lymphocytes freshly isolated from CLL patients. While lower levels of NTB-A were detected in T and NK cells, CDC and ADCC activity was demonstrated primarily in B cells isolated from CLL patients and B lymphoma cell lines. Knockdown of NTB-A by siRNA in target cells results in lower cytotoxicity, demonstrating the specificity of the mAbs. Furthermore, anti-NTB-A mAbs demonstrated anti-tumor activity against CA46 human lymphoma xenografts in nude mice and against systemically disseminated Raji human lymphoma cells in SCID mice. Taken together, these results demonstrate NTB-A as a potential new target for immunotherapy of leukemia and lymphomas.
Anti-Cancer Effects of Rapamycin on U937 Cell Line.
Authors: Jin Lina , Fu Chenchen , Cen Jiannong , Chen Peishuai , Wu De Pei
Published: October 2019

Purpose: Cytotoxicity of rapamycin alone or in combination with arsenic trioxide in lymphoma cell line U937 was assessed, for the study of Anti-cancer effects of rapamycin.

Methods: cell vability and proliferation was analyzed by MTT and cell counting, clone formulating ability was analyzed by semisolid medium, cell cycle was analyzed by Propidium Iodide/RN-ase stain, the phosphorylation level of mammalian target of rapamycin (mTOR) was detected after marked by Phospho-mTOR antibody (Ser2448) and FITC. Expression of P27 was assessed by western- blot.

Result: 1.MTT measurement showed with increasing concentrations of RAPA (10 nM 100 nM, 1000 nM) cell proliferation inhibition rate was follwed 16.2%, 25.5%, 47.8%. Clonogenic assay showed the U937 proliferation inhibition rate was 80.5% in a 7-day leukemia colony-forming assay with concentration of 10 nM RAPA.2 RAPA 10nM, As2O3 0.6uM, As2O3 0.9uM alone, RAPA 10nM in combination with As2O3 0.6uM and 0.9uM, cell proliferation inhibition rate was followed 13%, 26%, 35%, 43%, 54%. 3. Propidium Iodide/RN-ase measurement showed RAPA resulted in U937 cell arrested in G1-phase, and was blocked in S-phase. 4. FCM showed the phosphorylation level of mTOR down-regulated significantly after treated by rapamycin (10 nM),5.The expression of P27 enhanced after treatd by rapamycin (10 nM)

Conclutions: From the experiment we can see rapamycin alone can inhibit the proliferation and clone formulating ability of U937 cell line. mTOR kinase is involved in the regulation of cell growth and proliferation. Rapamycin can down-regulate the phosphorylation level of mTOR, which can induce increasing expression of P27, As a result U937 arrest in G1-phase and block in S-phase. Interestingly rapamycin exert additive effect in proliferation experiment when combind with As2O3, higher than rapamycin or As2O3 alone.
Interferon-alpha (IFN-a) Given Alone to CML Patients with Undetectable Disease after Imatinib (IM) Is Able To Maintain Their Complete Molecular Remission after IM Discontinuation.
Authors: Carella A.M.
Published: October 2019

Cytotoxic lymphocytes (CTLs) which recognize distinct antigenic peptides on CML cells, contributed greatly to the cure after allografting. Previously, BCR-ABL specific peptides and the PR1 non-apeptide, were found to elicit a CML-specific CTL response (Molldrem, 1996/1997; Bocchia 1996, Clark 2001). PR1 is a peptide derived from myeloblastin (MBN), also known as proteinase 3. MBN is abundantly expressed in azurophil granules of normal myeloid cells and is substantially overexpressed in certain immature myeloid leukemia cells where it may be important for the maintenance of a leukemic phenotype. PR1-specific CTLs (PR1-CTLs) lyses and inhibits the proliferation of CML cells but not of normal myeloid precursors. The association between the emergence of PR1-CTLs and the response to IFN-a in vivo strongly implied that IFN-a can induce remissions via induction of a CML-specific PR1-CTLs response (Molldrem et al, 2000). Recent data demonstrating a failure of IM to kill cell cycle-arrested CML precursors advice us to be attentive to arising escape mechanisms of even chronic phase CML; therefore the sequential combination therapy of IFN-a given alone to CML patient with undetectable disease achieved with IM, might be more effective than any current monotherapy. We have treated until now 4 CML patients who had undetectable residual disease for at least one year after IM therapy. The patients discontinued IM and started IFN-a at the dose of 3 M/U/m2 weekly. The median duration of IM therapy was 58 (range, 31–68) months before IM interruption. At a median follow-up of 5 months (range, 3–12 months) after IFN-a, all patients still have an undetectable level of BCR-ABL transcript. The details of patients characteristics will be discussed during the presentation. These preliminary results demonstrate that treatment with low dose IFN-a in CML patients with undetectable disease is feasible and did not lead to a molecular relapse. We are starting a large prospective randomized study to evaluate the role of IFN-a employed alone after 1 year of complete molecular remission achieved with IM (Arm A) vs contination of IM alone (Arm B). The main objective of this study is to verify the real effect of the immune modulation following IFN-a in CML patients with undetectable disease.
Optimal Treatment for Patients after Dasatinib or Nilotinib: Comparison of Preclinical Activities of Approved and Promising Investigational Kinase Inhibitors Against BCR-ABL Kinase Domain Mutations That Confer Clinical Resistance to Dasatinib or Nilotinib.
Authors: Kasap Corynn , Weier Christopher , Shah Neil P.
Published: October 2019

The optimal management of patients with chronic myeloid leukemia (CML) is increasingly reliant upon molecular studies. Loss of response to imatinib in CML is most commonly associated with selection for a limited number of BCR-ABL kinase domain mutations that impair the ability of imatinib to effectively bind to BCR-ABL Molecular understanding of imatinib resistance mechanisms has led to the development of effective “second generation” BCR-ABL kinase inhibitors, such as dasatinib and nilotinib, which have clinical activity against most, but not all, drug-resistant mutations. Analysis of the BCR-ABL kinase domain in patients who develop resistance to second-generation inhibitors has implicated further selection of drug-resistant BCR-ABL kinase domain mutants in nearly all cases reported to date. Encouragingly, the number of resistant mutations capable of conferring clinical resistance to the most clinically-advanced second-generation agents, dasatinib (approved by the US FDA and EMEA) and nilotinib (approved in Mexico and Switzerland), appears to be restricted to a relatively small number of amino acid substitutions. As clinical experience with dasatinib and nilotinib grows, an understanding of the relative sensitivities of dasatinib- and nilotinib-resistant BCR-ABL mutants to other kinase inhibitors, both approved and investigational, is critical to optimize clinical outcomes in patients with resistance to dasatinib or nilotinib. At the present time, kinase inhibitor therapy options for patients with resistance to one of these agents include the investigational options bosutinib and MK-0457 (VX-680), as well as dasatinib and nilotinib (for patients not yet exposed to one of these agents) and re-exposure imatinib. It is likely that the success of therapeutic intervention in these cases can be predicted based upon the preclinical sensitivity of the mutation(s) involved with the agent chosen. We have therefore conducted a thorough biochemical and biological cross-analysis of the activities of each of these clinically-useful kinase inhibitors against mutations that confer clinical resistance to dasatinib or nilotinib. These studies provide clinicians with a useful reference for choosing an appropriate kinase inhibitor based upon the identity of the resistant BCR-ABL kinase domain mutation(s) detected at the time of relapse when faced with a patient who has lost response to dasatinib or nilotinib. It is hoped that the application of such “personalized medicine” strategies to the clinical management of CML cases will further improve outcomes in patients treated with kinase inhibitor therapy.
Expression Profiles in Chronic Lymphoid Leukemia by Hematochip Analysis.
Authors: Alvarez Patricia , Leon Virginia , Caballero Gonzalo , Rubio-Martinez Araceli , Pocovi Miguel , Giraldo Pilar
Published: October 2019

Aim: The gene expression analysis system is a good tool to identify biomarkers related to prognosis or response predictors of therapy in haematological malignancies. Low density microarrays are an useful technology to obtain genetic expression profiles in different neoplastic disorders. Recently we have designed an oligonucleotide biochip (Hematochip, Alvarez et al, Clin Chem2007;53:259) useful for study peripheral blood in hematological malignancies. Chronic Lymphoid Leukemia (CLL) is a frequent clonal lymphocyte disorder characterized by clinical diversity and that have known prognosis markers but a more accurate subclassification could be required. Our objective was to define CLL subgroups with differential genetic profile expression. Patients and

Methods: Analytical, prospective study in 49 consecutive CLL patients diagnosed since January 2003 to December 2004 in the Hematology Department of Miguel Servet University Hospital. Zaragoza. Spain. At diagnosis demographic, clinical stage, analytical, immunophenotype expression, genetic aberrations and mutational status of IgVH were collected at electronically data base. Simultaneously, peripheral blood from 20 healthy donors and all CLL patients were collected in PAXgene tubes (PreAnalytix). cDNA was generated from total RNA and double-strand cDNA was used as a template to generate biotinylated cRNA by in vitro transcription using the MEGA-script T7 High Yield Transcription kit (Ambion). Fragmented cRNA was denatured and hybridized using a Ventana Discovery Station (Ventana Medical Systems). Arrays were stained with Cy3-conjugated streptavidin (Amersham Biosciences) and scanned using a ScanArray 4000 scanner (Perkin-Elmer). The data were treated and analyzed by GeneSpring clusters method. The expression profiles were compared and stratified with clinical stability or progression, gender, ZAP 70 expression, associated neoplasia, time free of therapy and response or not to Fludarabine.

Results: The Hematochip has identified 9 probes obtained with filtered data and statistical significance (p<0.01), related to stable or progressive disease. The results had been validated by RT-PCR.

Comments: the study of genetic profiles by Hematochip could be a useful analysis to predict the stability or progression of CLL patients at diagnosis. Further studies analyzing more cases will be necessary in order to validate these results. This work has been partially sponsorized by grants: Mutua Madrilena del Automovil, Fundacion para el Estudio de la Hematologia y Hemoterapia de Aragon and I+CS.
Leukemic Meningitis as the Presenting Manifestation of Chronic Lymphocytic Leukemia (CLL) in a Young Man.
Authors: Sousa Aida B. , Veiga Joana , Ferreira Gilda , Rodrigues Antonio S. , Silva Madalena , Gouveia Joaquim
Published: October 2019

Meningeal infiltration is extremely rare in CLL. Moreover, part of the cases found in the literature occurred in fact in patients with prolymphocytic leukemia. In the few CLL cases, this complication was reported at late stages of the disease, in elderly patients. A previously healthy 30-year-old man was referred to the emergency department for evaluation of blurred vision and headache that developed 2 weeks earlier. Bilateral papilledema was observed, with no other neurological abnormality. CT and MRI of the head and orbits disclosed thickening of both optic nerves. Hemoglobin and platelet count were within normal limits. The WBC count was 41800/μl with 83% small lymphocytes, which were SmIg+ λ(weak) CD19+, CD5+, CD23+, CD20+, CD79b+, CD11c+, bcl2+, CD38+, ZAP70+ and CD10 negative. Moderately enlarged cervical lymph nodes were present. A diagnosis of Binet stage A, Rai stage I B-CLL was made. Bone marrow aspiration and biopsy showed 95% lymphocytes, with a diffuse pattern of infiltration. No metaphases were obtained for karyotyping. FISH disclosed a rearranged IgH, and was negative for trisomy 12 and for deletions of 11q, 13q and 17q. Abdominal CT showed retroperitoneal lymphadenopathy. Serology for HIV and EBV was negative. The cerebrospinal fluid (CSF) revealed >1000 lymphocytes/μl which were immunophenotypically identical to the peripheral blood cells. Systemic corticosteroids and intrathecal chemotherapy (methotrexate and dexamethasone) followed by cranial irradiation led to resolution of headache, but blurred vision persisted. Clearance of leukemic cells from the CSF was slow, a normal CSF being achieved only after 6 alternate day intrathecal injections. Systemic chemotherapy and stem cell transplantation are planned. To our knowledge this is the first reported case of CLL presenting as meningeal infiltration. Two possible reasons for the apparent rarity of CLL meningitis can be put forward: meningeal invasion would frequently be clinically silent, a hypothesis that is strengthened by autopsy findings; and it would usually occur in very advanced stages of CLL (with misinterpretation of neurological symptoms in erderly patients) making underdiagnosis easy.
Distinctive IgVH Gene Segments Usage and Mutation Status in Chinese Patients with Chronic Lymphocytic Leukemia.
Authors: Chen Lijuan , Zhang Yaping , Zheng Wenjuuan , Li Jianyong , Ruan Changgeng
Published: October 2019

Chronic lymphocytic leukemia (CLL) is characterized by the relentless accumulation of monoclonal B cells with the appearance of small mature lymphocytes and a characteristic CD5 and CD19 co-expression immunophenotype. The incidence of CLL in Asian countries is lower than that in the Western ones, where CLL is the most common leukemia. To evaluate the frequency and mutation status of immunoglobulin (Ig) variable heavy chain gene (IgVH) expression in Chinese patients with CLL. We investigated IgVH gene segments usage and mutation status by multiplex RT-PCR in 52 CLL patients, and analyzed the relationship between IgVH somatic mutation status and the expression of CD38, ZAP-70 and CLLU1. 38 patients had mutated IgVH, and 14 had unmutated IgVH. The most frequently expressed VH gene family was found to be VH3 (46.2%) followed by VH4 (40.4%), VH1 (5.8%), VH2 (5.8%) and VH7 (1.9%), with no expression of VH5 and VH6 gene families. VH1-69 and VH3-21 which commonly overused in Western CLL weren’t detected in our cohort. The frequency of IgVH gene families indicates significant difference in Chinese CLL patients compared with Western patients, suggesting involvement of ethnic and/or environmental factors in CLL disease initiation. IgVH gene mutation status was significantly associated with the expression of CD38 and CLLU1. The expression of them may be simple and reliable surrogates for the identification of IgVH mutations.

VH gene family usage and mutation status VH family n Mutated VH gene Unmutated VH gene VH1 3 3 0 VH2 3 2 1 VH3 24 19 5 VH4 21 16 5 VH5 0 0 0 VH6 0 0 0 VH7 1 0 1
Symptomatic Waldenstrom’s Macroglobulinemia (WM) in Young Patients: Disease Characteristics and Outcome.
Authors: Kastritis Efstathios , Eleftherakis-Papaiakovou Evangelos , Migkou Magdalini , Gika Dimitra , Terpos Evangelos , Zervas Konstantinos , Kyrtsonis Marie Christine , Anagnostopoulos Nicolaos , Delimpassi Sossana , Symeonidis Argiris , Dimopoulos Meletios Athanasios
Published: October 2019

Introduction: WM is a disease of the elderly with a median age of 70 years in most series. Advanced age is recognized as an adverse prognostic feature and age > 65 years has been included among the 5 adverse covariates in the recently reported International prognostic Scoring System for WM (Morel et al, ASH 2006). There is little information regarding the incidence of disease features and outcomes after treatment of young patients (≤ 50 years of age) with symptomatic WM.

Patients and methods: Our database includes 220 patients with previously untreated, symptomatic WM. These patients were separated in two groups according to age ≤ 50 years and >50 years at the time of initiation of treatment. Clinical and laboratory characteristics, response to treatment, overall survival (OS) and disease-specific survival (DSS) were compared among the two groups.

Results: Twenty-two patients (10%) were ≤ 50 years of age at the time of initiation of treatment. The incidence of clinical and laboratory features was similar among young and older patients: gender (p=0.3), B-symptoms (p=0.3), splenomegaly (p=0.9), lymphadenopathy (p=0.1), anemia (p=0.3), median serum IgM level (p=0.6), hyperviscosity syndrome (p=0.7), elevated β2-microglobulin (p=0.3) and IgM related disorders (p=0.6). primary treatment consisted of chlorambucil, nucleoside analogs or rituximab-based regimens with equal distribution among young and older patients. At least partial response was observed in 73% of young patients and in 57% of older patients (p=0.16). The median OS was not reached in young patients and it was 106 months in older patients (p=0.07). The median DSS was not reached in young patients and it was 116 months in older patients (p=0.2).

Conclusion: 10% of patients with WM are ≤50 years at the time of treatment initiation. Their clinical and laboratory features and response to treatment are similar to those of older patients. Overall survival tend to be longer in young patients, but when deaths unrelated to WM or complications of treatment are excluded, survival times among young and older patients are similar.