bluuebunny/crossref_metadata_2025 huggingface dataset:
Relationship between Seroresponse to Epstein-Barr Virus and Survival in Patients with Lymphoid Malignancies.
Authors: Domingo-Domenech E. , Middeldorp J. , Gonzalez-Barca E. , Benavente Y. , Romagosa V. , Font R. , de Sevilla A. Fernandez , Meijer C. J. , de Sanjose S.
Published: October 2019

Introduction: Epstein-Barr virus (EBV) infection is a common life event that has been consistently associated to several lymphoproliferative malignancies. It is unclear to what extent EBV can be associated to clinical presentation and outcome of lymphomas.

Objectives: To evaluate the association between the seroresponse to EBV with the clinical presentation and survival in the different lymphoid malignancies.

Methods: As part of the EPILYMPH_Spain case-control study, 414 newly diagnosed lymphoma cases and 488 controls, age and sex matched to the cases, were systematically recruited in a single center during 20 months (starting in 1998). From all subjects, a blood sample was drawn at diagnosis and EBV serostatus was evaluated through ELISA assays to measure IgG reactivity against immunodominant epitopes of EBNA1(BKRF1) and VCA-p18 (BFRF3). Further, immunoblot analysis was performed to evaluate distinct antibody patterns to EBNA1, VCA18, VCA-p40 (BdRF1) and Zebra (BZLF1). These patterns were classified into two groups: reactive (with wide responsiveness to the different EBV proteins) and non-reactive (with an IgG reactivity restricted to a limited number of EBV proteins). Statistical differences between each group were analyzed using Chi-square test for categorical data and T-Student test for continuous data. Survival analysis was performed by Kaplan-Meier techniques and compared with the log rang test. Values of p<0.05 were considered to be statistically significant. All data were adjusted for age and sex.

Results: Twenty-four percent (101/414) of the cases and 17% (86/488) of the controls had an EBV reactive response pattern (p= 0.012). Among cases, 29% females versus 20% of males presented an EBV reactive pattern (p= 0.031), with no differences regarding median age. When we agrupated the different lymphoid malignancies into the 3 most important entities, differences in the prevalence of EBV seropositivity were found: B-cell neoplasms (25%), Hodgkin lymphomas (12%) and T-cell neoplasms (34%), p=0.05. When analyzing the different WHO entities, in myeloma patients, EBV reactive pattern was found to be associated to an early stage disease (29% Stage 1 vs 5% Stage 2–3; p= 0.038) and a longer median survival (not reached vs 38 months; p= 0.02).

Conclusions: EBV seroprevalence is higher in lymphoma patients, with a slight predominance in females. EBV reactive pattern seems to be associated to early stage myeloma, which implicates a better survival.
FDG-PET Demonstrates Different Metabolic Activities among Lymphoma Subtypes.
Authors: Kostakoglu Lale , Goldsmith Stanley J. , Chandramouly Anjaly , Vidan Erez , Verma Sumeet , Coleman Morton
Published: October 2019

Lymphomas may often present with more than one pathologic type in the same individual. This study assesses the variability of FDG uptake values as determined by SUV’s in various lymphoma subtypes to serve as a platform for differentiating and confirming the pathologic diagnosis in all disease sites.

Methods: We retrospectively evaluated the PET-CT studies of 184 lymphoma patients at initial staging or at relapse prior to therapy. Lymphoma subtypes were classified according to the WHO classification. SUV’s were obtained in a total of 1120 nodal or extranodal disease sites in 11 lymphoma subtypes. Mean SUV’s were calculated for each lymphoma subtype by dividing the total maximum SUV’s by the number of disease sites in all patients. Consequently, mean SUV +/− SD and SUV range were determined and correlated with histologic diagnosis as shown in the Table.

Results: Results are summarized in Table 1. The highest mean SUV’s were obtained in aggressive non-Hodgkin’s lymphomas (NHL) followed by Hodgkin’s disease (HD) and indolent NHL. Starting from the subtype with the highest mean SUV of the means and in a rank of decreasing order, the subtypes of lymphoma are as follows; Burkitts, DLCL, T cell rich B cell, natural killer T cell, HD, Anaplastic T-cell, mantle cell, marginal zone, follicular, T cell peripheral and chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL).

Conclusions: While SUV’s may be quite variable, a general pattern does exist for each lymphoma subtype. A major discrepancy between the SUV’s and histologic diagnosis should be assessed by repeat biopsy.

SUV Distribution Among Subtypes of Lymphoma* Type of Lymphoma Patient Number Mean+/−SD Range *Table does not cover all lymphoma subtypes studied in this study due to limited space, HD: Hodgkins Disease NHL: Non Hodgkins lymphoma, DLCL: Diffuse large cell lymphoma, CLL/SLL: Chronic lymphocytic leukemia/small lymphocytic lymphoma HD 45 7.5+/−3.7 2–22 Mantle cell 8 6.8+/−4.9 2.1–21 DLCL 58 11.3+/−7.3 2.2–58 T cell Rich B cell 5 9.1+/−5.5 4–25 Burkitts 9 11.8+/−10.5 2–44 Follicular 30 5.1+/−2.7 1.6–11 CLL/SLL 11 3.0+/−1.5 1.1–8.8 Marginal Zone 10 5.5+/−3.8 2.1–21
Angioimmunoblastic T- Cell Lymphoma by Flow Cytometric Analysis.
Authors: Agarwal Ashima , Fisher Stephen I.
Published: October 2019

Background: Angioimmunoblastic T cell lymphoma (AILT) is a peripheral T- cell lymphoma characterized by systemic disease, a polymorphous infiltrate involving lymph nodes with prominent proliferation of high endothelial venules and follicular dendritic cells. We present a novel technique for the presumptive diagnosis of AILT cases by flow cytometric analysis. The diagnosis of AILT was confirmed by histolopathologic features, immunohistochemical stains and molecular studies.

Method: Specimens: Fresh groin excision lymph nodes. Flow cytometric analysis utilizing side scatter versus CD45. Case 1 (figure 1): The gated population consists of 10–12% B cells and 56–61% T cells. CD3, CD4 and CD5 stain 56–61% of the gated cells, while CD7 stains 41% of the gated cells, consistent with aberrant partial loss of CD7 by T cells. A population of abnormal T cells expresses CD10 with CD2, CD3, CD4 and CD7 but not with CD19 (B cells) consistent with aberrant CD10 coexpression by a subset of the T cells (12–19% of the gated cells). Staining for CD34 highlights less than 2% immature cells staining while CD64 stains less than 2% monocytic cells. Surface kappa and lambda staining does not show restriction (kappa: lambda 1–2:1). Cytoplasmic kappa and lambda staining is not interpretable due to non-specific antibody binding. Case 2 (figure 2): The gated population consists of 69% T- cells and 28–33% B- cells. A population of abnormal T cells expresses CD10 with CD2, CD3, CD4 and CD7 but not CD19 consistent with aberrant CD10 expression by a subpopulation of T cells (8–11% of gated cells). Intracellular B cell kappa lambda light chain restriction is not present and there is no aberrant co-expression of CD5, CD10 or CD43 by B cells. The diagnosis of AILT was confirmed by standard morphologic, immunoperoxidase and molecular methods subsequently in both of these cases.

Discussion: Aberrant CD10 expression on neoplastic T cells has been shown with immunohistochemical staining of paraffin embedded tissue in AILT. Our cases show aberrant coexpression of CD10 by CD2, CD3, CD4 and CD7 positive T cells in more than 10% of gated cells by Flow cytometric analysis.

Conclusion: Flow Cytometric analysis is a useful and reproducible tool for immunophenotyping AILT cells and should be considered for presumptive diagnosis of AILT.

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Excellent Stem Cell Mobilization Using Escalated BEACOPP in High-Risk Patients with Hodgkin’s Disease.
Authors: Kandler Gabriele , Fillitz Michael , Moestl Michaela , Schloegl Ernst , Reisner Regina , Pittermann-Hoecker Elisabeth , Noesslinger Thomas , Pfeilstoecker Michael
Published: October 2019

Introduction: After intensive treatment regimens have been established, the survival rate for patients with advanced Hodgkin’s disease is approximately 91 % after five years and 13 % of the patients have a relapse or have primary progressive disease (2 %) within the first five years. For patients with relapse after conventional chemotherapy +/− radiotherapy, however, there is a real chance of achieving remission again. Since it is often difficult to harvest autologous stem cells following an intensive pretreatment, our center embarks on the strategy to harvest autologous blood stem cells in high-risk patients, defined according to the risk stratification of the German Hodgkin Study Group, already as part of the initial polychemotherapy.

Results: Between 9/2003 and 5/2005, we analyzed the results of the stem cell harvest of 12 consecutive patients with Hodgkin’s disease who were mobilized with the escalated BEACOPP regimen. There were 7 female and 5 male patients. Escalated BEACOPP was the primary therapy in ten patients and a relapse was treated in two patients; the previous treatment was 4 or 6 cycles of the ABVD regime + involved field radiation. The ten patients who did not receive previous treatment were classified as having Ann Arbor stage IIA/2 IIB/5, IIIB/2 and IVB/1 and all of them had a large mediastinal bulk as an additional risk factor. The two patients who did receive a previous treatment were classified as having an initial Ann Arbor stage IIA or IIB, without an additional risk factor. The stem cells were collected in 1 patient from cycle 2, in 8 patients from cycle 3 and in 3 patients from cycle 4 of the escalated BEACOPP regimen. A total of 11 patients received a standard dose of filgrastim, 5μg/kg body weight s.c., from day 8 up to the last apheresis and 1 patient received pegfilgrastim 6mg s.c. All aphereses were performed using an Amicus cell separator® (Baxter, MNC set, closed two-arm). 7 patients required only 1 apheresis and the remaining 5 patients required 2 aphereses. An apheresis result sufficient for a possible reinfusion could be achieved in all patients (4.26 – 14.4 x10e6 CD34 pos. cells/kg/body weight, mean: 7.7).

Summary: According to our experience, escalated BEACOPP regimen is very suitable for the harvesting of stem cells in high-risk patients with Hodgkin’s disease even though they are receiving procarbazine. A sufficient quantity of stem cells can also be collected from pretreated patients. The stem cell mobilization can be integrated into the escalated BEACOPP regimen safely and without a delay in treatment and thus creates, already at an early stage, the pre-condition for a high-dose therapy, which might be required in high-risk patients.
Study of Pegylated Liposomal Doxorubicin in First Line Therapy of Aggressive Lymphomas.
Authors: Mandala Eudokia , Lafaras Christos , Ilonidis George , Venizelos Ioannis , Grollios George , Tsioni Constantina , Bischiniotis Theodoros , Arvanitakis Constantinos
Published: October 2019

Purpose: Conventional doxorubicin is currently used in many treatment protocols for lymphoma. Pegylated liposomal doxorubicin has a different pharmacokinetic profile with prolonged half life, reduced toxicity, especially cardiotoxicity and possible enhanced antitumor activity. This study was designed to evaluate tolerability and efficacy of pegylated liposomal doxorubicin in patients with aggressive lymphomas.

Methods: Twenty-nine patients entered the study, 12 male, aged 36–86 (median 66 years), 22 with newly diagnosed CD20 positive diffuse large B-cell lymphoma (DLBCL), elderly with median age 77 years, 4 MALT in transformation, 2 mantle and one anaplastic. Patients’ characteristics: IPI-1 10%, IPI-2 25%, IPI-3 10%, IPI-4 55%, stages III–IV 60%, bulky disease 40%, extranodal 55%. Combined immunochemotherapy with Rituximab 375 mg/m2, day1, added to standard CHOP was performed, 8 courses, with a follow up of nearly 3 years. Pegylated liposomal doxorubicin (Caelyx®) replaced doxorubicin at 30 mg/m2. Response was evaluated after 3 and 8 courses. Cardiac function was estimated at the beginning and after the end of therapy. LVEF, fractional shortening (FS) and myocardial performance index (MPI) according Tei et al, as most sensitive index for global LV function, were measured.

Results: All responded except for three, one with active SEL and one with CNS lymphoma (refractory 10%). Overall response 91%, CR 76%, PR 15%, overall survival 82%, relapse rate 4.5% (1/22). No synergistic toxicity was observed, as there were no remarkable hematological or extrahematological toxic events. No clinical deterioration of cardiac function was observed. The patient with anaplastic lymphoma had severe preexisting cardiovascular disease and died from cardiac complications, in lymphoma remission. Two patients experienced a single episode of atrial fibrillation with successful pharmaceutical conversion. None developed alopecia, nausea, vomiting, palmar-plantar erythrodysesthesia, local injury in extravasation area. Treatment related mortality was documented in none. LVEF and FS were not changed, but MPI was slightly increased, detecting subclinical myocardial dysfunction.

Conclusions: Pegylated liposomal doxorubicin is useful in the treatment of aggressive lymphomas, including elderly patients, well tolerated, producing durable complete remissions and offering advantages in combination with immunotherapy.
Membrane Derived Micorvesicles - Underappreciated Components of the Tumor Microenvironment That Modulate Tumor Growth, Vascularization and Metastasis.
Authors: Wysoczynski Marcin , Hayek Fadhi , Ratajczak Janina , Janowska-Wieczorek Anna , Ratajczak Mariusz Z.
Published: October 2019

Viable eukaryotic cells shed circular membrane fragments called microvesicles (MV) from the cell surface and secrete them from the endosomal compartments. These MV, which are different from apoptotic bodies, are enriched in lipids, proteins and mRNA. We postulate that MV play an important and underappreciated role in cell-cell communication by i) stimulating target cells with ligands that the MV express, ii) fusing with target cells and thus transferring various receptors to their surface, and iii) delivering mRNA, lipids and proteins. Since tumor cells secrete large quantities of MV we hypothesized that the latter are important constituents of the tumor microenvironment and their role in tumor progression merited investigation. First, we observed that human and murine lung cancer cell lines secrete more MV in response to non-apoptotic doses of hypoxia, irradiation and chemotherapy. The MV derived from human cancer cells chemoattracted bone marrow-, lymph node- and lung-derived fibroblasts and endothelial cells and activated in these stromal cells the phosphorylation of MAPKp42/44 and AKT. Furthermore, they also induced in bone marrow- and lung-derived fibroblasts expression of LIF, OSM, IL-11, VEGF and MMP-9. Moreover, conditioned media from marrow fibroblasts exposed to MV induced phosphorylation of STAT-3 proteins and chemoattracted lung cancer cells in a LIF- and OSM-dependent manner and, together with IL-11 and VEGF, activated osteoclasts and endothelial cells. Furthermore, MV from cancer cells embedded in Matrigel implants strongly stimulated angiogenesis. We also found that tumor-derived MV express tissue factor (TF) and activate platelets and as a result of this MV derived from activated platelets transfer several adhesion molecules from platelets to the tumor cell surface. This increases adhesiveness of lung cancer cells in endothelium and their metastatic spread in vivo after injection into syngeneic mice. Finally, we found that formation of MV depends on the formation of membrane lipid rafts. Thus we postulate that tumor- and platelet-derived MV are underappreciated constituents of the tumor microenvironment and play a pivotal role in tumor progression/metastasis and angiogenesis. As MV formation appears to be lipid raft-dependent, we suggest that inhibitors of membrane lipid raft formation (e.g, statins or polyene antibiotics) could decrease MV-dependent tumor spread/growth and we are currently testing this hypothesis in animal models in vivo.
A Phase III Clinical Study of Treatment of Patients with Chronic Lymphoproliferative Disorders (CLPD) with Fludarabine, Cyclophosphamide, and Rituximab (FCR) Combination.
Authors: Galanopoulos Athanasios G. , Tsakiridou Anastasia , Michalis Eurydiki , Marinakis Theodoros , Gortzolidis George , Zomas Athanasios , Xanthopoulos Vasilios , Savvidou Ioanna , Theodoropoulos George , Grigoraki Vasiliki , Anagnostopoulos Nikolaos I.
Published: October 2019

Background: The treatment of patients with chronic lymphocytic leukemia (CLL) with Rituximab in combination with fludarabine and cyclophosphamide was reported to be more efficacious, in terms of complete and molecular remission compared with historical data for fludarabine plus cyclophosphamide (S.O’Brien, Haematologica2002; 87:50–53).

Aims: Evaluation of the clinical efficacy and toxicity of the FCR combination in patients of our Haematologic Centre.

Methods: Seventeen patients, 8 males and 9 females with a median age of 69,5 years, with relapsed/refractory or de novo CLPD (9 CLL and 8 NHL patients) were enrolled in this study between February 2002 and August 2004. Fifty percent of CLL patients had Rai stage I/II and the rest 50% had Rai stage III/IV disease. Four NHL patients had an International Prognostic Index (IPI) 2, one patient IPI 3 and three patients IPI 4. All patients were treated with Rituximab 375 mg/m2 on day1 in combination with Fludarabine and Cyclophosphamide (25 mg/m2 and 250 mg/m2 respectively) for days 2 to 4, every 4 weeks, for 6 consecutive cycles. Nine patients had a history of a prior unsuccessful treatment.

Results: Overall, 14 out of 17 evaluable patients (82%) were responsive to the treatment [12 patients (70%) complete response (CR) and 2 patients (12%) partial response (PR)]. The remaining 3 patients had progressive disease (NR). Hematological toxicity was acceptable (grade 2–3 neutropenia in 6/17 patients, grade 2–3 anemia and thrombocytopenia in 2/17 patients). There were no septic episodes except one case with neutropenic fever. There were no adverse events like nausea or vomiting except one patient with a serious anaphylactic reaction due to Rituximab administration. Three CLL patients died because of progressive disease.

Summary/conclusions: this preliminary report suggests that the FCR regimen is an effective and safe treatment for CLPD patients, achieving higher CR rates than previous treatments. A longer follow up of a larger number of patients is required to confirm an improved survival in these patients.
Prospective Clinical Study of Surgical Resection Followed by CHOP in Localized Intestinal Lymphoma.
Authors: Lee Jeeyun , Kim Won Seog , Oh Sung Yong , Kwon Jung Mi , Kim Hyun Jung , Lee Hyo-Rak , Park Byeong Bae , Kim Kihyun , Jung Chul W. , Park Keunchil
Published: October 2019

Background: Due to the rarity of the disease, prospective clinical trials on optimal treatment of intestinal non-Hodgkin’s lymphoma have been scarcely performed.

Methods: Patients with; age > 18 years, pathologically proven diffuse large B cell lymphoma (DLBL), normal bone marrow, liver, renal and cardiac function were eligible. Patients with Burkitt’s lymphoma, lymphoblastic lymphoma, mantle-cell lymphoma, or infection with human immunodeficiency virus were excluded. All patients were staged after surgery according to the Ann Arbor classification modified by Musshoff et al. and Paris staging system (TNM system). Postoperative chemotherapy with CHOP regimen was administered for 6 cycles. CHOP consisted of cyclophosphamide 750 mg/m2 d1, doxorubicin 50 mg/m2 d1, vincristine 1.4 mg/m2 (max 2.0 mg/m2) d1, and prednisolone 100mg d1–5 repeated every 3 weeks.

Results: From 1999 to 2004, 41 patients were enrolled. The median age was 50 years (range, 26 – 84) and male:female ratio was 29:12. All patients had clinical stage I/IIE DLBL and underwent surgical resection. Of the 41 patients, 3 patients refused postoperative CHOP chemotherapy. The 5-year DFS and OS were 83.3% and 89.1%, respectively. Surgicopathologic staging of all patients according to the Ann Arbor classification revealed 34 patients with stage I, 11 patients stage II1, 1 patent stage II2 and 3 patients with stage IIE. The Ann Arbor stage significantly predicted the survival of intestinal DLBL (p = < 0.0001). The T staging according to Paris staging system did not correlate with survival (p = 0.1456) while the N staging showed significant predictability (p = <0.0001).

Conclusion: Surgical resection followed by 6 cycles of CHOP chemotherapy was a highly effective treatment scheme for localized intestinal DLBL patients. The Ann Arbor system seems to predict survival better than the TNM system.
Treatment of Primary Mediastinal Large B-Cell Lymphoma with Immunochemotherapy.
Authors: Tabacof Jacques , Yana Novis A.S.
Published: October 2019

Background: Primary mediastinal large B-cell (PMLBCL) non-Hodgkin’s lymphoma (NHL) is a relative rare disease and represents less than 2% of all newly diagnosed NHL. Analyses with DNA microarrays verified that primary mediastinal and diffuse large B-cell lymphomas are different diseases. The optimal treatment is still undefined. The aim of this study is to evaluate the response to immunochemotherapy in 10 cases of PMLBCL.

Patients and Methods: 10 previously untreated patients with confirmed diagnosis of PMLBCL were enrolled in 2 different institutions. All the patients were treated with rituximab (R) plus polychemotherapy. We retrospectively analysed their clinical data and response to immunochemotherapy.

Results: median age was 37 years-old (range 18 to 45 years), 5 were male and 5 were female, 6 had bulky disease, 4 had B symptoms and 4 presented HDL elevated. The immunochemotherapy regimens utilized for treatment were: 3 R+MACOP-B, 5 R+CHOP, 1 R+CBV and 1 R+MACOP-B+ICE+BEAM.

Complete remission was obtained in 9 patients (90%) and no relapse was observed in this group. One patient died 7 months after ABMT. Three out of 10 patients who were chemosensitive underwent Autologous Bone Marrow Transplantation (ABMT). Median disease free survival was 22 months. One patient got pregnant 4 months after the 8th cycle of R+CHOP, when she was in complete remission. The patient was delivered of a healthy child and is in complete remission for 34 months. It was observed toxicities related to chemotherapy. There was no report of toxicities during and after rituximab infusion.

Conclusion: the addition of rituximab to polychemotherapy may be a good strategy to the management of primary mediastinal large-B cell lymphoma. In this small series there was a high rate of complete sustained response. However, larger randomized studies are needed to confirm these findings.
Retreatment with Gemcitabine Still Hold Efficacy in Relapsed Lymphomas.
Authors: Jajeh Ahmad , Tamkus Diemanti , Menini Perry , Osafo David
Published: October 2019

Lymphomas are highly sensitive to chemotherapy. Approximately half of the patients with aggressive Non-Hodgkin’s Lymphoma will survive for five years. Patients with diffuse large B-cell type NHL have an expected five years survival( OS) rate of fourty five percent (45%). Approximately more than twenty five percent (25%) of NHL and ten percent 10% of Hodgkin’s Lymphoma HD are refractory to conventional front line therapy. Fifteen to twenty five percent of hodgkin’s lymphoma will relapse after therapy or will not achieve complete response CR. Salvage chemotherapy, radiation and high dose chemotherapy with stem cells collection can cure and achieve durable response in some refractory or relapsed patients. High dose chemotherapy and autologous stem cells transplant is not a feasible options for many of our patients due to lack of medical coverage and co-morbide status. Gemcitabine has been used either as a single agent or in combination at our institute in the treatment of variety of hematological malignancies that failed many lines of therapy. Forty patients were treated; twenty two (55%)NHL, nine patients(19%) with HD, three patients with visceral mycosis fungoides, three patients with advanced chronic lymphocytic leukemia in richter transformation and three with peripheral T cell NHL. Mean age of 45 years. Thirty two males and eight females. Eighty percent were african american. All patients had advanced stage III-IV disease. Complete respose CR and near complete response nCR was seen in eleven patients. Partial response PR seen in nineteen patients, stable disease in six. Total response rate of eighty percent. Five patients relapsed after a year (two 25 years old females with HD, three males with high grade NHL age 52, 60 and 70 respectively). Two patients recieved four cycles and three recieved six cycles. Each cycle constitute of 1 gram per meter square given IV weekly for three weeks and one week rest. All five patients had response, three CR and two PR. In conclusion retreatment with gemcitabine is a valid and safe option. None of the patients had grade 3 or 4 toxicity.
Treatment of Systemic Primary Cutaneous Anaplastic Large Cell Lymphoma with High-Dose Methotrexate and Leucovorin Rescue.
Authors: Batra Reema , Boyer James , O’Connor Thomas , Tabbara Imad
Published: October 2019

A 59 year-old Caucasian male with a history of hepatitis C infection, presented in October 2003 with a painless skin lesion on his back. Physical examination revealed a 3 cm lesion on the back that resembled a sebaceous cyst, with similar lesions on the anterior aspects of both legs. A right inguinal lymph node was palpated as well, measuring approximately 2 cm. A wide surgical resection was performed on the back lesion due to its extensive nature at the time of surgery. Pathologic evaluation revealed a primary cutaneous anaplastic large cell lymphoma, measuring 12 cm x 5 cm in size. Immunohistochemical staining showed reactivity for CD30, CD3, UCHL-1, CD43 and CD45. Despite being asymptomatic, the patient had widespread disease on CT/PET scans, including diffuse lymphadenopathy, bony disease, and lung lesions. The patient was treated with weekly high-dose methotrexate at 2 grams/m2 followed by leucovorin rescue for cycle 1 and dose-reduced for cycles 2 through 5 to 1 gram/m2 secondary to myelosuppression. For cycles 6 through 10, the patient was able to tolerate the original dose of 2 grams/m2. After eight cycles, the patient had complete resolution of all disease by clinical examination and by CT/PET scans, and has remained in complete remission since February 2004. The patient tolerated the chemotherapy well with myelosuppression being the most significant toxicity. He also had a transient increase in his liver enzymes after cycle 1 that resolved spontaneously. He is currently being treated for reactivation of hepatitis C with pegylated interferon and ribavirin. To the best of our knowledge, this is the first case of primary cutaneous anaplastic large cell lymphoma with systemic involvement being treated successfully with weekly high-dose methotrexate and leucovorin rescue with prolonged complete remission. Weekly high-dose methotrexate with leucovorin rescue is an active and well-tolerated regimen with limited side effects.
Rescue with Rituximab ® Monotherapy in Resistant T - Cell Rich B - Cell Lymphoma (TCRBCL).
Authors: Burgos Ruben A. , Hanono Isabel A. , Foncuberta Maria C. , Domenichini Enzo , Huertas Eduardo , Sanchez Avalos Julio C. , Tartas Norma E.
Published: October 2019

TCRBCL is a large B - cell lymphoma with a prominent reactive T cell proliferation. More common in males in the fourth decade of life, usual clinical presentation includes generalized adenopathies and bone marrow involvement. More localized presentation is frequently misdiagnosed as Hodgkin’s (HL) or T - cell lymphoma. Cases resistant to anthracyclines (A) pose a medical dilemma. We present here a patient initially misdiagnosed as HL who showed nodal relapse after ABVD and was resistant to conventional chemotherapy. Case report: CP is a 45 years old gentleman who presented with left axillaries adenophaties. A lymph node biopsy was diagnosed as lymphocyte predominant HL (LPHL) stage IA. Treated with 3 cycles of ABVD plus involved field radiation therapy, he relapsed in the margin of the radiation field soon after treatment was completed. A new biopsy showed TCRBCL. A review of the first biopsy was also consistent with TCRBCL. Treated with Promace/Cytabom the patient experience a spleen relapse which was resistant to ESHAP. With a gross hypo dense image in the spleen, treatment with R was started at a dose of 375 mg sqm2 weekly for four weeks. Disappearance of the splenic mass on the CT scan prompted us to perform a splenectomy. The pathologic study revealed a residual 8 mm lesion. The patient is currently without evidence of disease 40 months after the splenectomy. In total he received 6 doses of R. Interestingly an additional patient with TCRBCL originally misdiagnosed as ABVD resistant HL, showed a durable partial remission after treatment with R. In conclusion: due to our experience we think that it might be worth including R in the front line treatment of TCRBCL.
Blastic Mantle Cell Leukemia Maitaining Remission for 3 Years after the Treatment with Rituximab.
Authors: Tsuji Masanori , Akiyama Hideki , Kobayashi Takeshi , Sakai Miwa , Yamashita Takuya , Ohashi Kazuteru , Sakamaki Hisashi
Published: October 2019

[Introduction] Blastic mantle cell leukemia is defined as CD5-positive, CD23-negative, cyclin D1-positive mature B-lymphoid tumor characterized by leukemic presentation, blastic blood and bone marrow features with aggressive clinical behavior. Its prognosis is usually dismal and overall survival was reported to be 3 months. We present the case of blastic mantle cell leukemia treated with rituximab, enjoying complete remission for 3 years.

[Case presentation] A 68 year-old man was admitted because of leukocytosis. His CBC revealed WBC 31.0 x 10E9/L with 64% of blast. Hemoglobin level was 5.3 g/dl, and 78% of medium-sized blasts that were positive for CD5, CD19, CD20, and HLA-DR. Chromosomal study revealed normal karyotype. He was treated with adriamycin, vincristine, cyclophosphamide, and predonisone as for acute lymphoblastic leukemia, and then, with another two cycles of intensive chemotherapy including hyper-CVAD without any significant response.

He was transferred to our hospital for further treatment. The blastic cells showed bcl1/IgH re-arrangement on FISH analysis and bone marrow biopsy revealed CD23-negative and cyclin D-positive that confirmed the diagnosis of mantle cell lymphoma/leukemia. CT scan of the body showed no significant lymphadenopathy.

He was treated with fludarabine and idarubicin without any effect and then was treated with rituximab 375 mg/m2. After the initiation of rituximab treatment, blasts disappeared from peripheral blood within 24 hours completely and after 4 times of rituximab once per every week, CBC returned to normal and bone marrow examination revealed complete remission. He was treated with another 2 injection of rituximab and then followed without any further treatment. He has been in complete-remission status for 3 years since then.

[Discussion] Blastic mantle cell leukemia is usually chemotherapy-resistant as our case is and its prognosis is dismal. Our case represent the possibility that even in those patients, one may enjoy prolonged complete remission after the simple treatment with rituximab. This is, of course, only a single case experience and it is necessary to proceed for further investigation.
P210 Tyrosine Kinase Regulates the Expression of 14-3-3sigma in Hematopoietic Progenitors Expressing the bcr-abl Fusion Gene.
Authors: Santucci Maria Alessandra , Mancini Manuela , Zuffa Elisa , Brusa Gianluca , Barbieri Enza
Published: October 2019

The 14-3-3sigma is a member of a highly conserved family of acidic proteins that bind to serine/threonine-phosphorylated residues in a context specific manner analogous to the Src homology 2 (SH2)- and phospho-tyrosine- binding domains. Its central role in the subcellular distribution and transcriptional activity of partners molecules controlling cell cycle progression, survival, senescence, adhesion and metabolism made 14-3-3sigma isoform the most directly linked to cancer pathogensis and progression. Here we provide evidence for its involvement in deregulated proliferation and extended survival of Chronic Myeloid Leukemia (CML) hematopoietic progenitors. In LAMA cell line and CD34+ cells of CML patients at clinical diagnosis p210 bcr-abl dephosphorylation and enzymatic activity inhibition induced by in vitro exposure to 1 microM tyrosine kinase (TK) inhibitor Imatinib mesylate (IM) were associated with a significant reduction of 14-3-3sigma transcript and protein. 14-3-3sigma reduced expression arises from transcriptional mechanisms and involves epigenetic modifications of chromatin structure. In LAMA cell line PCR amplification of DNA from immunoprecipitated chromatin (ChIP) revealed histone H4 deacetylation at a discrete region of 14-3-3sigma promoter critical for gene transcription since 2nd up to 24th hour of IM treatment. Conversely, bisulfite-SSCP sequencing of PCR amplification of gene 5′ coding sequences encompassing the transcription start site did not show significant differences in CpG methylation status relative to p210 TK activity. Up-modulation and increased nuclear import of p27Kip1, downmodulation of cyclin D1 and Cdc25A levels, Bad dephosphorylation at Ser136 and Bax translocation to mitochondria proceeding from 14-3-3sigma reduction gave rise to cell cycle arrest in late G1-/early S-phase and committment to apoptotic death. Interestingly, higher levels of 14-3-3sigma protein were associated with the development of IM resistance in vitro and in vivo. To conclude, our study supports a role of 14-3-3sigma in the pathogenesis of CML hematopoiesis as well as in the disease progression towards a drug-resistant phenotype mostly due to its regulatory control on pro-apoptotic and growth-arrest proteins downstream of forkhead transcription factor Foxo3a and c-jun N-terminal kinase (JNK).
Inhibitory Effect of RNAi on Expression of bcr/abl Oncogene in K562 Cells.
Authors: Wang Chun , Ma Xiaoxia , Wei Ju , Qin Youwen , Yan Shike
Published: October 2019

The translocation t(9;22) can be found in 95% patients with chronic myeloid leukemia (CML). The resulting hybrid gene bcr/abl codes for a fusion protein with tyrosine kinase activity which activates signal transduction pathways, leading to uncontrolled cell growth. A promising anti-gene technology-RNA interference (RNAi) reported in recent years can disrupts the expression of the targeted cellular gene in a complicated manner in a variety of organisms and cell types.

To inhibit CML bcr/abl oncogene expression with RNAi, we used chemically synthesized anti-bcr/abl small interfering RNAs( siRNAs), mismatch control siRNAs having the three point mutations to transfect the K562 cells for different time through the electroporation. EGFP plasmid was used as a positive control and the amount of fluorescently stained cells was determined by FCM. Inhibitory effects of siRNAs were demonstrated by real-time quantitative RT-PCR and Western blots. Cell proliferation was measured by means of MTT assay and apoptosis was determined by Annexin V-FITC assay.

The transfection efficiency was about 70%. The synthesized siRNAs inhibited CML bcr/abl oncogene expression at both mRNA and protein levels. The anti-bcr/abl siRNAs reduced the bcr/abl mRNA level by 67%–72% without an obvious effect on abl mRNA levels. The K562 cells electroporated with anti-bcr/abl siRNAs contained less BCR/ABL protein than cells electroporated with mismatch control siRNAs or without any siRNAs. P210 was reduced to a very low level in Western blots, whereas the wild-type ABL protein was not influenced by the anti-bcr/abl siRNAs. Depletion of bcr/abl leading to increased apoptosis and reduction of cell viability. The flow cytometric analysis showed that the percentage of dead and apoptosic cells induced by anti-bcr/abl siRNAs for 24hrs and 48hrs were 42.10% and 53.33%, respectively, including 15.05% and 19.47% of the preliminary apoptosis cells, respectively. A significant induction of apoptosis was observed after transfection compared with untreated control(1.00%)and the mismatch control siRNAs (2.98%) (p<0.05). Anti-bcr/abl siRNAs inhibited cell growth of the K562 cells and the inhibitory rate was 47% and 56%, 24h and 48h after transfection respectively, whereas the mismatch control siRNAs had no such effect on K562 cells.

At the cell level, inhibition of CML bcr/abl oncogene expression by chemically synthesized siRNAs provides the new method for anti-leukemia study. Therefore, siRNAs may be potent tools for gene-specific modality against CML.
Effective Treatment of Basophilic Leukemia of the Central Nervous System in a CML Patient with the Combination of Imatinib Mesylate and Valproic Acid.
Authors: Nahajevszky Sarolta , Magocsi Maria , Mikala Gabor , Andrikovics Hajnalka , Adam Emma , Masszi Tamas
Published: October 2019

Imatinib is effective in all stages of CML but the level of the drug in the cerebral spinal fluid is two logs lower than in the plasma. In November, 2000 a 35 years old man in CML chronic phase was treated with interferon (10MU/day) and achieved complete hematologic remission. Two years later he progressed to myeloid blastic crisis. The patient was treated with an acute leukemia protocol (60mg/m2 daunorubicin days 1–3, 200 mg/m2 ara-C days 1–7). After hematological recovery 600 mg/day imatinib was administered continuously. 9 months later headache and vomiting indicated CNS involvement. In the CSF presence of Ph+ basophilic leukemia cells (1,27G/l) was proven by FISH. Although the patient was given intrathecal methotrexate(15mg), cytarabine(40mg) and dexamethasone(4mg), the headache was a permanent problem with persistent basophilia in the liquor. In vitro culture of his bone marrow cells at the time of the blastic phase was tested for drug sensitivity to different doses of imatinib alone or in combination with other antitumor drugs. In the in vitro growth assay 5 uM imatinib alone, 5mM valproic acid alone and 0,5uM imatinib + 5mM valproic acid together caused 15%, 33% and 80% decrease in cell survival, respectively, suggesting that combinations with valproic acid, (an anticonvulsive agent) would be more effective. Based on the results of our previous growth assay the therapy was supplemented with valproic acid (4x150mg/day). The meningeal leukemia resolved in 3 weeks and the patient had no complaint for 3 months with the same therapy.

We find it noteble that basophilic CNS blast crisis was exquisitely sensitive to the administration of imatinib in combination with valproic acid, an anticonvulsive agent with antitumor activity.
A Rare Case of Primary Amyloidosis, Presenting with Severe Pulmonary Hypertension and Bilateral Pleural Effusion.
Authors: Agheli Aref , Arora Alka , Khalil Maged , Dumlao Theresa , Naik Seema , Steier William , Kalavar Madhumati
Published: October 2019

Pulmonary arterial hypertension secondary to vascular deposition is a rare presentation of Primary Amyloidosis. The natural history of pulmonary hypertension and concomitant right-sided cardiac failure is not very well understood and is associated with a very short median survival. This syndrome is a marker of advanced Amyloidosis Persistent pleural effusion is another much rarer presentation of pulmonary involvement in Primary Amyloidosis and is secondary to pleural infiltration, which sometimes is difficult to distinguish from the cardiomyopathic causes. The incidence of pleural effusion in primary Amyloidosis is 1–2%. This syndrome also has a very poor prognosis, and has limited response to treatment, however, pleurodesis has been temporarily useful in some cases.

Case report: A 74 year old African American female, non-smoker, with history of hypertension, who presented with shortness of breath and persistent productive cough and 18 pounds weight loss for the last six months. The initial chest x-rays showed bilateral pleural effusion and a transudate fluid on analysis. Cardiac workup for an atypical chest pain revealed moderate to severe pulmonary hypertension without myocardial infiltration on 2-D echocardiogram, and normal coronary arteries on cardiac catheterization. Extensive workup did not show any secondary cause of pulmonary hypertension, however, the patient gradually developed nephrotic range proteinuria and kidney biopsy showed infiltration of 10 nm fibrils, involving 3+ segmental mesangium, and 2+ segmental glomerular basement membranes. This was consistent with renal Primary Amyloidosis, AL Lambda type. A random urine and serum sample for immunofixation also revealed an M- spike of 23 mg/dl in gamma region. A concomitant bone marrow biopsy revealed presence of monoclonal IgG lambda plasma cell (CD38 bright) population, without any B-cell or T-cell markers.

Conclusion: Pulmonary hypertension and pleural effusion are two rare and lethal presentations of pulmonary involvements in Primary Amyloidosis, and it should be considered in the differential diagnosis of any patient presenting with pulmonary symptoms. There is no definite treatment of these syndromes, although combinations of steroids and melphalan, along with colchicines, vincristine, adriamycin, and thalidomide have been used with minimum satisfactory outcomes. Autologous stem cell transplantation (SCT) provides the opportunity to give higher doses of chemotherapy and may be of benefit in some cases.
T Cell Acute Lymphoblastic Leukemia Originates from a Differentiation Block in T Cell Development Due to Aberrant Expression of LMO2 and SCL(TAL1).
Authors: Layh-Schmitt Gerlinde , Crable Scott , Crable Kathleen , Kraft Elizabeth , Bailey Jeff , Klanke Charles , von Kalle Christof
Published: October 2019

T cell acute lymphoblastic leukemia (T-ALL) is frequently associated with overexpression of the oncogenes LMO2 and SCL(TAL1) which are normally down regulated following the double negative stage of T cell development. Our goal is to decipher the molecular and cellular mechanisms leading to the onset of LMO2 associated T-ALL. We were able to isolate a complex containing the transcription factors LMO2, SCL(TAL1) and E47 from primary human T-ALL cells with proven aberrant expression of LMO2 and SCL(TAL1) by applying immunoprecipitation and Western blotting techniques. This protein complex regulates the transcription of a truncated form of RALDH2 (retinaldehyde dehydrogenase) in T-ALL cells as shown by gene transcription profiling in conjunction with RT-PCR and siRNA approaches. To monitor the effect of LMO2 expression on T cell development and leukemogenesis, lethally irradiated mice (C57BL/6) were transplanted with bone marrow cells that had been transduced with a retrovirus carrying LMO2 as the transgene. One year later, 88% of the cells in the thymus expressed LMO2 and a shift towards CD3−/CD44+/CD25+ cells was observed (an 88% increase compared to normal thymocytes), suggesting a differentiation block caused by LMO2 leading to an accumulation of immature T cells. To test and identify cooperating genes in T-ALL development, bone marrow cells of LMO2 double transgenic mice in which tet-inducible LMO2 is controlled by a thymic specific promoter, were retrovirally transduced with SCL(TAL1). So far, none of the control animals, transplanted with bone marrow cells transduced with a vector only containing EGFP, developed T-ALL. However, six out of the seven test animals developed T-ALL exhibiting enlargement of the spleen, liver and thymus between seven and nine months after transplantation. Organs and blood of the diseased animals were infiltrated with T-ALL cells of the immature phenotype CD8+/CD4+ in five cases and of the CD3−/CD44+/CD25+ phenotype in one case. This indicates that the differentiation block caused by a lack of down-regulation of LMO2 and SCL(TAL1) in maturing T cells leads to a block in T cell differentiation and precedes T-ALL. These models will be used to examine the involvement of other cooperating genes in T-ALL development as well as downstream target genes of LMO2/SCL(TAL1), such as RALDH2, in the onset of T-ALL. We conclude that aberrant expression of LMO2 in T cells leads to a block in T cell maturation and, in conjunction with up-regulation of secondary genes like SCL(TAL1), triggers deregulation of genes in immature T cells leading to impaired T cell development and the onset of T-ALL. The described model will help to identify cooperating genes in LMO2 associated T-ALL as well as the chain of events leading to malignancy.
Inositol 1,4,5-Trisphosphate Receptors Mediating Spontaneous Ca2+ Oscillation Favors Proliferation in Human Mesenchymal Stem Cells from Bone Marrow.
Authors: Tao Rong , Lau Chu-Pak , Li Gui-Rong
Published: October 2019

Although human mesenchymal stem cells (hMSCs) constitute a very small population of cells in bone marrow, they play an important role in the regulation of hematopoietic microenvironment. The self-renew and/or proliferation of hMSCs is believed to be particularly important in maintaining bone marrow niche; however, the regulation of this process is not fully understood. The present study was designed to investigate whether spontaneous Ca2+ oscillation mediated by inositol 1,4,5-trisphosphate (IP3) receptors participates in the proliferation regulation in cultured hMSCs from bone marrow using RT-PCR, cell proliferation assay, Western-blotting analysis. It was found that no gene expression for ryanodine-sensitive receptors was detected in undifferentiated hMSCs, while three subtypes of IP3 receptor genes (i.e. IP3R1, IP3R2, and IP3R3), and genes for sarco/endoplasmic reticulum Ca2+ ATPases (SERCA) (ATP2A1, ATP2A2, and ATP2A3) were expressed in these cells. The proliferation of hMSCs was reduced by inhibiting Ca2+ oscillation with the IP3 receptor antagonist 2-aminoethyl diphenylborinate (2-APB) or the SERCA inhibitor cyclopiazonic acid (CPA). In addition, inhibition of Mek/Erk and PI-3K/Akt signaling also decreased hMSCs proliferation. The relation of Ca2+ oscillation to the activity of these kinases was revealed by Western blotting analysis. Erk1/2 (Thr185/Tyr187) phosphorylation level was found to be reduced by directly inhibiting Ca2+ oscillation with 2-APB or CPA either in the presence or absence of serum. However, Akt (Thr308) phosphorylation was decreased only in the presence of serum, and serum-free starvation (2 h) eliminated Akt (Thr308) phosphorylation. Finally the calmodulin inhibitors W-7 and SKF-7171A were employed to further investigate whether this ubiquitous Ca2+ sensor is involved in the Ca2+ signal-mediated effect. Similarly, the phosphorylation level of Erk1/2 (Thr185/Tyr187) and Akt (Thr308) reduced upon the inhibition of calmodulin. In conclusion, our results demonstrate that IP3 receptors-mediated spontaneous Ca2+ oscillation and/or Ca2+/calmodulin signaling play(s) a crucial role in the regulation of cell proliferation mediated by multiple pro-proliferation signaling pathways (e.g. Mek/Erk and PI-3K/Akt). Importantly, Erk1/2 phosphorylation is sensitive to spontaneous Ca2+ oscillation, and therefore is responsible for the proliferation induced by Ca2+ oscillation in hMSCs.