bluuebunny/crossref_metadata_2025 huggingface dataset:
The Roles of Pten in Hematopoietic Stem Cell Regulation and Leukemogenesis.
Authors: Zhang Jiwang , He Xi , Jayasinghe Sach , Ross Jason , Haug Jeff , Li Linheng
Published: October 2019

Pten was the first phosphatase identified as a tumor suppressor and one of the most frequently mutated genes involved in human tumor/cancer. Pten, involved in regulation of both PI3K/Akt and MEK/Erk activity, is downstream of growth factor, cytokine, integrin and cadherin signaling pathways and therefore plays important roles in cell growth, survival, differentiation, metabolism and migration. Although Pten mutation is not common in leukemic cells, phosphorylated Pten (p-Pten), which represents the inactive form of Pten, has been observed in a majority of acute myeloid leukemias that are associated with poor clinical outcomes. To explore the role of Pten in hematopoietic stem cell (HSC) regulation and leukemogenesis, we generated an interferon-inducible Pten knockout mouse by crossing Mx1Cre mice with Ptenloxp mice. All of the mutant mice developed myeloproliferative disorder characterized by increased peripheral white blood cell counts, hyperproliferative macrophages and granulocytes in bone marrow and spleen, and multiple tissue infiltration by myeloid cells. The HSC number was decreased in the bone marrow but mobilized and expanded in the spleen. Extra-medullar hematopoiesis was shown by dramatically increased myeloid and erythroid progenitors in the spleen. B lymphocyte differentiation was blocked at the common lymphoid progenitor stage, while the T cell number was increased in all hematopoietic tissues. Compared to wild type, Pten mutant HSCs and progenitor cells were highly proliferative, forming larger colonies in an in vitro culture study. However, Pten mutant HSCs showed reduced competency in repopulation assay after in vivo bone marrow transplantation. Our study demonstrates that Pten plays important roles in restricting HSC activation, proliferation and mobilization. Pten also plays a role in HSC lineage decision by favoring myeloid differentiation at the expense of B lymphocyte lineage.
Evaluation of the Oncohaematologic Pathology of Two Oncologics Centers of Buenos Aires, Argentina in the Period between 1995–2005; a Total of 1884 Cases.
Authors: Dragosky Marta O. , Annetta Isabel , Alvarellos4 Viviana , Devotto5 Laura , Luchetta6 Patricia , Marquez7 Marisa , Alcaraz Susana
Published: October 2019

Introduction: In the absence of national and regional records of oncohaematologic pathology the information regarding the composition and characterstics of this patients population was evaluated in this two oncologic centers of Buenos Aires.

Results: Of the whole of 1884 patients, the distribution of the pathologies was: Non Hodgkin Lymphoma (NHL): 1040 cases (55,2 %), Hodgkin Lymphoma (HL) 358 (19 %), Leukemia 258 (13,7 %), (chronic lymphatic 105, chronic myeloid 93, acute 52, trycholeukemia 8) Myeloma 131 (6,95 %), thrombocythemia 24 (1,27 %), Myelodysplasia 22 (1,16 %), Polycythemia Vera: 17 (0,90 %), Plasmacytoma 14 (0,74 %), Myelofibrosis 8 (0,42 %), Histiocytosis. 5 (0,26 %), Macroglobulinemia of Waldenstrom 4 (0,2 %), Granulocytic Sarcoma 2 (0,1 %), Dendritics Cells Sarcoma 1 (0,05 %).

The sex distribution was: 982 (52.14 %) males, 902 (47,86 %) females. The range of ages was: 15 to 94 years, with an average of 52,3 years.

Of the whole of 1040 NHL 274 (26,34 %) presented extranodal locations, distributed as follows: skin 63, orbital 37, tonsil 35, oral cavity 17, stomach 16, central nervous system 16 (12 brain), breast 13, soft tissues 12, parothyd 11, cavum 11, bowel 9, testicle 7, paranasal sinuses and bones 6, rectum and lung 3, larynx, uterus and scalp 2, liver, ovary and thyroid gland 1. A whole of 29 patients presented pathology associated with the HIV: 22 NHL and 7 HL.

In the population with HL the average age was 36,7 years, with 38 % younger than 25 years, we do not observe the bimodal curve usually descript.

The predominant histologies: nodular sclerosis 40,7 % and mixed cellularity36,53 % (low frequency of subtypes lymphocitary predominance and lymphocitary depletion). Stage II 44,87 % and stage III 23,71 %. The most frequent locations: cervical 53,2 % and mediastinic 47,11 % (and low frequency of abdominal and inguinal locations).

Conclusions: The frequency of extranodal locations of NHL is similar as the international records. The distribution of the unusual locations is probably biased by the pathologies arrived from specific centers (for example: a center of ophthalmology). The population of HL is characterized by the young age and the supradiaphragmatic locations
Hepatitis C Viral Infection Is Not Associated with Waldenstrom’s Macroglobulinemia.
Authors: Leleu Xavier , O’Connor Kelly , Ho Allen , Santos Daniel D. , Manning Robert , Xu Lian , Hatjiharissi Evdoxia , Tournilhac Olivier , Chemaly Mariana , Branagan Andrew R. , Hunter Zachary R. , Patterson Christopher J. , Anderson Kenneth C. , Treon Steven
Published: October 2019

Infection with hepatitis C virus (HCV) has previously been associated with the pathogenesis of low grade B-cell lymphoproliferative diseases including Waldenstrom’s macroglobulinemia (WM). Most of these studies have utilized HCV antibody detection assays, which may not accurately assess HCV presence in patients with impaired humoral responses. In patients with WM, pronounced IgA and IgG hypogammaglobulinemia is present, which remains persistant despite therapy (Blood 104:306b). As such, we investigated the incidence of HCV in 88 randomly selected, and previously untreated patients with the consensus panel diagnosis of WM utilizing both HCV antibody detection and qualitative PCR assays (Quest Diagnostics, Cambridge, MA, USA). The median age for these patients was 61 (38 – 83 years) and the male: female sex ratio was 1.44. No patient had a known history of liver disease, hepatitis B, HCV or HIV infection. Liver function tests obtained at time of serum collection showed normal range SGOT and and SGPT levels for 86/88 patients. In contrast to previous reports, we failed to demonstrate HCV infection by both HCV antibody detection and the PCR assays. These studies therefore show no association of WM with HCV infection.
Follicular Lymphoma of the GI Tract - Distinct Entity or Extranodal Manifestation of Nodal Disease?.
Authors: Mehta Sharmila , Kulharya Anita , Kallab Andre , Mazzella Fermina M.
Published: October 2019

Introduction: Primary GI follicular lymphoma is a rare, poorly studied entity. Current discussions in the literature debate whether or not it is a distinct extranodal entity, similar to cutaneous follicle center lymphoma. Like cutaneous follicle center lymphoma, it appears to have a good prognosis and low incidence of t(14;18).

Case History: This is a 74 year old white male who presented for routine screening colonoscopy in May 2003. On evaluation the patient was found to have nodularity of the colon. Biopsies demonstrated several submucosal and mucosal follicles with no distinct mantle zones, composed of a uniform population of centrocytes. No lymphoepithelial lesions or plasma cells were identified. These cells proved to be monoclonal B cells, with a phenotype of CD20+, CD10−/+, bcl-2+/−, bcl-6+, CD5−, CD43−, lambda+ and t(14;18) negative. CD21 stain highlighted the underlying follicular dendritic architecture. No adenopathy was noted at this time. A diagnosis of B cell lymphoma, consistent with extranodal follicular lymphoma, grade I was rendered. As the patient was otherwise asymptomatic, no treatment was given. Four months later, the patient developed retroperitoneal adenopathy and a large pelvic mass. A lymph node biopsy demonstrated numerous large follicles composed of a uniform population of centrocytes, with attenuated to absent mantle zones. The cells were CD20+, CD10+, bcl-2+, lambda+ and t(14;18) positive. There was a comcomitant transitional cell carcinoma of the urinary bladder, which required radical cystectomy. The patient was then initiated on chemotherapy with CVP, for three cycles. He was followed without treatment for 2 years when he developed a tonsillar mass. A biopsy demonstrated a CD10+ diffuse large B-cell lymphoma. Cytogenetic analysis revealed a complex karyotype with a t(14;18). The patient underwent a staging bone marrow, which was positive for follicular lymphoma. The cells were CD20+, CD10−/+, bcl-6+, lambda+, and t(14;18) negative. He was then started on R-CHOP for his B symptoms and extensive adenopathy, and is currently 2 months into therapy.

Discussion: This is an unusual case of almost concomitant presentation of t(14;18) negative extranodal follicular lymphoma of the GI tract and classical t(14;18) positive nodal follicular lymphoma. Possible explanations include that these are two manifestations of the same disease, two separate diseases coincidentally occurring around the same time, or a follicular lymphoma clone which has evolved into nodal and extranodal entities.
Collagen Type IV as Independent Prognostic Factor for Non-Hodgkin’s Lymphoma.
Authors: Hadzi-Pecova Liljana , Petrusevska Gordana , Panovska Irina , Stankovic Svetlana , Stojanovik Aleksandar
Published: October 2019

Many attempts have been made recently to improve the prognostic systems for non-Hodgkin’s lymphoma (NHL). There are no available data in the literature regarding the influence of intactness of the basement membrane of the blood vessels on the prognosis of NHL, when compared with the clinical characteristics of the disease. On the other hand, it is known that the integrity of the basement membrane is marker for the aggressiveness of the malignant disorders.

In order to analyze the association of the expression patterns of collagen type IV on the prognosis of the patients with NHL we have conducted a retrospective study.

We have evaluated 136 patients diagnosed and treated as NHL in the last ten years at the Clinic of Hematology, Medical Faculty -Skopje. All 136 patients with representative tumor tissues were diagnosed with NHL according to the REAL classification (71 patients were diagnosed as diffuse large cell lymphoma, 28 as follicular lymphoma, 24 as small lymphocytic lymphoma and 14 as marginal zone lymphoma). Median age of the patients was 49 years, with male: female ratio 1.5:1.0. The application of international prognostic index (IPI) identified four risk groups with predicted five-years survival rates of 87%, 82%, 18% and 0%, with statistical significance (p< 0.001).

Using immunoperoxidase staining we have analyzed the collagen type IV expressing patterns in our group of patients, and defined three tumor invasive grades: high tumor invasive grade as total lack of immunoreactivity for collagen type IV (68 patients), medium-presented as discontinuous expression (56 patients) and low-as a continuous linear expression (14 patients). The 5 year overall survival rate was significantly lower (p<0.01) in patients with high invasive grade than in patients with medium or low invasive grade (54% vs. 78% and 100%). Further, in multivariate analysis collagen type IV expression appeared as an independent survival-predicting factor together with the parameters of the IPI.

Our results indicated that the expression of a collagen type IV immunoreactivity could predict the outcome of patients with NHL. However, further studies are needed to define the role of collagen type IV in the future prognostic systems for NHL.
Expression of the BAFF-R (BR3) Is Positive in a Minority of Diffuse Large B Cell Lymphomas and Does Not Correlate with the Germinal Center or Non Germinal Center Origin of Lymphomatous Cells.
Authors: Proust Alexis , Adrar Idir , Rince Patricia , Mercier Virginie , Lazure Thierry , Raphael Martine , Garçon Loïc
Published: October 2019

BAFF, a member of the TNF family, has been shown to be implicated in B cell maturation and survival through an interaction with its three receptors, BCMA, TACI and BAFF-R. Recent data revealed that it also plays a role in B cell malignancies. For exemple, an autocrine activation of the BAFF receptors could participate to oncogenesis during multiple myeloma or chronic lymphocytic leukemia. BAFF-R, the sole BAFF specific receptor, is expressed in a subset of Diffuse Large B Cell Lymphomas (DLBCL). Since BAFF-R activation in B cells leads to cell proliferation and survival, expression of this receptor in DLBCL could correlate with more aggressive lymphomas. DLBCL can be divided into prognostically important subgroups with germinal center B cell-like (GCB), activated B cell-like (ABC) and type 3 gene expression profiles using a cDNA microarray. The germinal center origin of malignant B cells in DLBCL can also be characterized by immunohistochemistry (IHC) according to CD10, Bcl6 CD138 and MUM1 expression. In this work, we investigated whether expression of BAFF-R in DLBCL correlate or not with the GCB vs non GCB phenotype of lymphomatous cells. Lymph nodes biopsies from 23 DLBCL (among whom 3 post-transplantation lymphomas and 1 AIDS-related DLBCL) were analyzed. In a first step, we characterized by IHC the precise phenotype of each DLBCL: CD10+/Bcl6+ DLBCL were classified into the germinal center (GCB) group (n=9), whereas CD10-/BCL6-/MUM1+ lymphomas defined the non germinal center (NGCB) one (n=14). Each DLBCL was then analyzed for BAFF-R expression by IHC. In agreement with a recent report, most of the DLBCL were found to be negative for BAFF-R expression (16 out of 23). Interestingly, 4 of the 9 GCB DLBCL were found to be BAFF-R positive (44%), whereas only 3 out of 14 (21%) in the NGCB group. Although the difference did not reach statistical significance, we concluded from this observation that a consequent percentage of GCB DLBCL express the specific receptor BAFF-R. Thus, we actually analyze the clinical and biological characteristics of BAFF-R positive DLBCL and investigate whether expression of BAFF-R could represent a bad prognosis factor in term of clinical outcome and response to treatment.
Analysis of Nuclear Factor Kappa B in Mantle Cell Lymphoma.
Authors: Schrader Carsten , Klapper Wolfram , Janssen Dirk , Riis Paul , Meusers Peter , Brittinger Guenter , Siebmann Jens U. , Kneba Michael , Parwaresch Reza
Published: October 2019

Gene expression profiles revealed that proliferation associated genes are important prognostic factors in the clinical outcome in mantle cell lymphoma (MCL). Beside this well accepted markers also analysis of apoptotic proteins are now under investigation. We investigated immunohistochemically the expression of the apoptotic marker NF-kB in relation to the clinical course in 89 patients enrolled in two multicenter prospective trials. Biopsies were recut and stained with mandatory antibodies (CD20, CD5, CD3, CD23, cyclin D1) and NF-kB. The NF-kB expression was analyzed in three groups: negative, cytoplasmatic positive and nuclear positive (more than 1/HPF). The expression was compared with the overall survival data analyzed according to Kaplan and Meier. In 13 cases a negative NF-kB staining was detected. Fifty-seven cases were positive only in the cytoplasma and in 17 cases more than 1 cell per high power field showed nuclear activity. Patients with mantle cell lymphoma that had negative and cytoplasmatic positive expression had a median overall survival time of 35.7 months compared to 22.4 months for patients with a nuclear NF-kB expression. The Kaplan-Meier analysis showed a significant difference in the overall survival time (p=0.0121). The immunohistochemical detection of NF-kB in mantle cell lymphoma is possible and a tool to identify patients with a poor prognosis.
Value of Early Evaluation with FDG-PET for Follicular Lymphoma.
Authors: Gressin Remy R. , Vuilliez Jean Philippe J.P. , Molina Lysiane L. , Pegourie Brigitte B. , Courby Stephane S. , Garban Frederic F. , Bulabois Claude Eric C.E. , Orsini Frederique F. , Nicolas Emmanuelle E. , Makowski Caroline C. , Sotto Jean Jacques J.J. , Cahn Jean Yves J.Y.
Published: October 2019

Objective: evaluate the predictive value of the FDG-PET on the evolution of Follicular lymphomas.

Methods: FDG-PET was performed in 17 consecutive patients treated for follicular lymphoma during the course of their treatment (ttt): 8 were evaluate early (Ea) after the second course of chemotherapy (CT) and 9 later (La) after the third course (5 pts) or the fourth (4 pts). 9 were treated at diagnosis and 8 at relapse. 15 histologic grade 1–2 received 6 courses of Rituximab-VAD-Chlorambucil (n=13) or R-miniCHVP (n=2) and two grade 3 a R-CHOP like regimen. 12 were intensified with haematopoietic stem cell transplantation (10 auto, 2 allo). 6 were Flipi 0–1, 3 Flipi 2 and 8 Flipi 3–4.

Results:

Among the 8 Ea, only 3 negativate their FDG-PET, 4 had a minimal uptake and one remained positive (he relapsed 2 months after the completion of their ttt). 4/4 pts with minimal uptake became negative after the completion of the ttt. 7/7 negative pts remained relapse free (Median Fup 17 months).

Among the 9 La, 8 were negative (5/5 after the third course) and one had a residual uptake after the fifth course (he died disease free from non haematolgical cause). One/8 negative patient have relapsed 5 months after ttt, 7 are alive and disease free. The 2 histologic grades 3 were Ea. Negative. Flipi 0–1(n=6) or Flipi 2 (n=3) were all negative. The Ea. Positive and the La. Residual were Flipi 3.

Conclusion

The best time of evaluation for the follicular lymphoma might be later than for High grade and might be proposed after three courses of chemotherapy when associated with rituximab. The Flipi seems to be correlated to the FDG-Pet response. These preliminary results are encouraging to define the predictive value of FDG-PET for follicular lymphoma survival, especially for the flipi 3.
Hodgkin Lymphoma (HL), A Retrospective Analysis in Young Adults.
Authors: Koumarianou Anna A. , Xiros Nikolaos , Papageorgiou Efstathios , Pectasidis Dimitris , Economopoulos Theofanis
Published: October 2019

INTRODUCTION: Over the last four decades survival of patients with HL has improved. Various risk factors (stage III/IV, abnormal ESR, bulky mediastinal disease, >3 involved nodes, extranodal involvement) classify patients into prognostic groups for treatment accordingly.

PURPOSE: This study was conducted to review the risk factors, treatments and outcomes in young adults with HL over the past 25 years in our Unit.

METHODS: Retrospective chart review of 55 young adults 16–23 years old (25 women, 35 men), treated from 1978–2003, was performed. Patients were divided according to risk factors into three prognostic groups: low (stage I/II without risk factors; 5 patients [9.1%]), intermediate (stageI/II with risk factors; 19 patients [34.5%]) and advanced HL (stage IIB with bulky mediastinal disease and extranodal involvement or stage III/IV; 31 patients [56.4%]). Patients were treated with established chemotherapy regimens available at each time period including MOPP (Group A, 1978–1987; 17 patients), MOPP/ABVD (Group B, 1988–1993; 21 patients), and ABVD or BEACOPP (Group C, 1994–2003; 17 patients). Patients with bulky disease at diagnosis (>5cm) or residual disease following chemotherapy received additional radiotherapy.

RESULTS: Histological diagnosis included 42 nodular sclerosis, 10 mixed cellularity and 3 lymphocyte predominant. Thirty five patients had mediastinal involvement (16 patients with bulky disease). Following chemotherapy completion 27 patients were referred for radiotherapy. A total of 21 patients experienced relapse (5 intermediate and 16 advanced stage), of whom 14 (66 %) within the first 2 years of diagnosis. Although the follow up periods were not equal between the three treatment groups, surveillance of evaluable patients showed significant improvement of the 5-year survival and 5-year relapse rate (Group A:64.7% and 47%, Group B:80.9% and 33.3%, Group C:94.1 % and 11.7% respectively) over the years. Eight out of 10 mixed cellularity patients died. One second primary malignacy (acute non lymphocytic leukaemia) was observed. In our study albumin less than 4 g/dl, and lymphocyte count less than 8% were identified as the most important risk factors in relation to relapse and survival regardless treatment.

CONCLUSION: Survival of young patients with HL is significantly improving over the last 25 years most probably due to improved treatment resources and understanding of risk factors. Tailoring treatment according to prognostic groups and inclusion of molecular markers is expected to refine further treatment strategies and improve survival.
Results of a Multi-Center, Phase II Study of Elsamitrucin (SPI-28090) in Patients with Relapsed or Refractory Non-Hodgkin’s Lymphoma (NHL).
Authors: Foon Kenneth , Allen Steven , Sen Kaushik , Mohrbacher Ann , Sanani Shamel , Lenaz Luigi , Rosales Rosamerlinda
Published: October 2019

Background: Elsamitrucin (SPI-28090) is a novel topoisomerase I and II inhibitor. An earlier small study with Elsamitrucin, conducted prior to the introduction of rituximab, showed anti-tumor activity in patients with relapsed or refractory NHL with an acceptable toxicity profile. The objectives of this current trial are to validate these earlier findings and to generate data in patients with relapsed or refractory NHL who had previously received rituximab-based chemotherapy.

Methods: Three groups of relapsed or refractory NHL were enrolled: G1: chronic or small B-lymphocytic lymphoma/leukemia; G2: indolent lymphoma (follicular/diffuse small cell or mixed); and G3: large cell or mantle cell lymphoma. Patients were treated with weekly Elsamitrucin 25 mg/m2 IV administered over 5–10 minutes. All patients were treated until disease progression or toxicity.

Results: 55 patients (34 males/21 females) have been enrolled; median age was 70 years (range 40–84). All patients had stage III or IV disease. Most patients were heavily pretreated with a median of 4 (1–8) prior chemotherapy regimens; 54 out of 55 patients had rituximab-based chemotherapy. Other study characteristics and results are shown on Table 1. One PR occurred in a mantle cell lymphoma patient who had received five prior chemotherapy regimens and lasting for 27 weeks. Minor responses have been documented in several patients still on therapy. Most side effects were mild to moderate in severity and consisted of nausea (51%), fatigue (42%), anorexia (22%), vomiting (16%) and cellulitis (11%). Myelosuppression was uncommon: grade 3 or 4 neutropenia was reported in 2 patients, grade 3 anemia and thrombocytopenia reported in 1 patient each. There was one patient who had moderate transient increase in transaminase level. No drug related serious adverse events have been reported.

Conclusions: Elsamitrucin appears to be safe and well tolerated over periods of up to 47 weeks in these patients lymphoma. There is evidence of anti-tumor activity in this very heavily pretreated patient population. With 2 PR, several minor responses and durable stable disease observed. In view of the minimal toxicities in particular myelosuppression, we plan to combine elsamitrucin with one or more active agents in less heavily pretreated patients.

Table 1 CLL/SLL (G1) Indolent (G2) Aggressive (G3) TTP means time to progression N 14 24 17 Median LDH (U/L) 244 253 242 Median # of doses (range) 8 (2–24) 15 (4–46) 4 (1–27) Best Response PR (%) − 1 (4%) 1 (6%) MR (%) 2 (14%) 6 (25%) 1 (6%) SD (%) 9 (64%) 12 (50%) 4 (24%) TTP (days) 98.5 138 34 Range 21–216 21–330 2–245
Rituximab - Inmunotherapy Combined with Chemotherapy, CHOP for the Treatment of Patients with Difusse Large B - Cell Lymphoma (DLBCL).
Authors: Milone Jorge H. , Bezares Fernando , Ardaiz Maria del Carmen , Riveros Dardo , Palmer Luis , Cerutti Ider , Basso Alfredo , Gelemur Marta , Cánepa Carlos , Kusminsky Gustavo , Zerga Marta
Published: October 2019

Several studies (GELA 98.5, MInT) have demonstrated the benefit of combination rituximab (R) with chemotherapy to improve event free and overall survival in patients with DLBCL.

We analyzed retrospectively the safety of combination R-CHOP for 6 cycles (rituximab 375 mg/m2 day 1; cyclophosphamide 750 mg/m2 day 1; doxorrubicin 50 mg./m2 day 1; vincristine 1.4 mg/m2 day 1 and prednisone 100 mg/m2 day 1 to 5) the tolerance and adverse effects. We evaluated the response (R), event free survival (EFS) and the overall survival (OS).

Between March to December 2004, 28 patients with DLBCL were evaluated, 17 men and 11 women, with a median age 57 years old (range 28 – 84).

They were IPI low 21,4 %, low - intermediate 25%, high - intermediate 35,7 % and high risk 17,9 %.

Elevated LDH was present en 14 patients, bulky disease > 7 cm in 50% of cases.

During the treatment they presented hematologic toxicity grade III 21,4 % and grade IV 25% of cases; 1 patient had anaphilactic reaction; 2 patients pneumonia; 1 patient sepsis; 4 patients neutropenia and fever; and gastric bleeding 1 patient.

Response was achieve in 71,4 %: complete response (CR) in 57,1%, parcial (PR) in 14.3 %, and there was no response in 8 patients (28.6%).

With follow up of 10 months (range 2 to 18.5) 15 patients were in CR 53,6%, 8 patients died, 7 of then primary no responders. Analyzed by IPI, 60% of CR in intermediate high and high was obtained.

The R-CHOP combination is a feasible and safe treatment in our hospitals, and 71,4 % of response was obtained in all patients and 60% of CR in high risk group.
Treatments for Marginal Zone Lymphoma.
Authors: Boxer Michael , Nasr Rashad Aboul
Published: October 2019

Marginal Zone Lymphoma (MZL) is a low grade malignant B cell lymphoproliferative disorder that comprises 2% of all lymphomas. Often an enlarged spleen and villous lymphocytes are present. Many patients however may appear to have chronic lymphocytic leukemia or other lymphoproliferative disorders. Immunophenotyping can differentiate these low grade lymphoproliferative disorders. MZL frequently is CD 20 + but CD5, CD 10, and CD23−. Treatment regimens include observation, chemotherapy, splenectomy, and immunotherapy with or without chemotherapy. 20 patients over the past six have been identified with MZL based on pathologic and immunophenotypic characteristics. 6 patients have had no treatment and have had stable disease from 9 months to over 4 years. 2 patients underwent splenectomy from 3 months to 2 years from diagnosis and have stable disease. 1 patient was diagnosed at splenectomy and relapsed 5 1/2 years later and remitted with rituxin. 1 patient received weekly rituxin for 4 weeks and has been in remission for 40 months.1 patient received weekly rituxin for 4 weeks, relapsed 9 months later, and remitted with rituxin and then maintenance rituxin. 1 patient responded to weekly rituxin for 4 weeks and is receiving maintenance rituxin. 1 patient responded to weekly rituxin for 4 weeks and then had residual disease irradiated. 1 patient remains in remission after 50 months after a cervical node was irradiated. 1 patient with blastic MZL remitted with CHOP-rituxin. 2 patients progressed after rituxin and required chemotherapy. A 92 yo patient has had a partial response to rituxin. An 82 yo patient died from complications from MZL. 15/20 patients have so far avoided chemotherapy. Rituxin may be an appropriate treatment for MZL, but clinical trials will be necessary to confirm this approach.
Patient-Specific Three Dimensional (3D) Dosimetry for 90Y Ibritumomab Tiutexan (Zevalin®) in Patients with Follicular Lymphoma.
Authors: Assié Karine , Dieudonné Arnaud , Gardin Isabelle , Buvat Irène , Tilly Hervé , Vera Pierre
Published: October 2019

Objectives: Dosimetry calculations for radioimmunotherapy are usually performed using a two dimensional (2D) protocol based on planar whole body (WB) scintigraphy and fixed organ masses derived from the MIRD anthropomorphic model (Wiseman GA et al., J Nucl Med2003:44;465–74). We compared the absorbed dose obtained with this method with an approach involving patient-specific 3D dosimetry based on quantitative SPECT and patient organs mass estimation.

Methods: Five patients with follicular lymphoma in partial or complete remission underwent a CT scan, and five successive WB and SPECT scans at day 0 (D0) 1 hour, D0 4 hours, D1, D4 and D6 after injection of 111In ibritumomab tiutexan (185 MBq). WB scans were used to estimate the activity in the liver, spleen and kidneys. SPECT images were reconstructed using a quantitative processing including corrections for scatter, attenuation, and partial volume effect. The organs mass and activity values were estimated in 3D volumes of interest manually drawn on the CT scan and registered to the SPECT scan. Absorbed doses after administration of 15 MBq/kg of 90Y ibritumomab tiutexan were derived from both 111In WB and SPECT data accounting for the fractions of injected activity measured in each organ at the five time points.

Results: The mean differences between the organs mass estimated from the patient CT and the MIRD anthropomorphic model were 8%, 105% and 12% for liver, spleen and kidneys respectively. Measurements on an abdominal phantom filled with 111In showed that errors in organ activity estimates ((estimated activity - true activity)/true activity) for WB and SPECT protocols, were 37% and −6% respectively in the liver, 20% and −10% in the spleen, −26% and −23% in the left kidney, and 47% and −9% in the right kidney. Patient liver, spleen and kidney activity values determined from the SPECT scans were on average 35%, 60% and 39% less than those found from the WB scans. The absorbed doses calculated with the 3D patient-specific protocol were less than those calculated with the 2D protocol, by 46±14%, 77±14% and 70±22% in the liver, spleen and kidneys respectively, except in one patient kidney where SPECT dose exceeded WB dose by 2%.

Conclusions: Accounting for patient-specific organ mass and using SPECT activity quantification have a great impact on estimated absorbed doses of 90Y ibritumomab tiutexan if compared with the 2D protocol. This method could improve the correlation between dosimetry and clinical consequences of this treatment, especially in view of dose escalation with stem cell rescue.
High pH Lysis Allows Immunoprecipitation of p210BCR-ABL in Primary Chronic Myeloid Leukaemia.
Authors: Patel Hetal , Marley Stephen B. , Gordon Myrtle Y.
Published: October 2019

Chronic myeloid leukaemia (CML) is a myeloproliferative disorder, which is characterised by the presence of the Philadelphia translocation (Ph) (t (9; 22)). The translocation occurs when parts of the c-ABL gene, located on chromosome 9 fuse with the BCR gene, located on chromosome 22. BCR-ABL fusion results in the production of p210BCR-ABL, a chimeric protein with the molecular weight of 210kda. p210BCR-ABL is located in the cytoplasm and has constitutive protein tyrosine kinase activity. Numerous publications have reported that p210BCR-ABL forms complexes with multiple cytoplasmic proteins which affect several signalling pathways. However this published data exists for cell lines expressing p210BCR-ABL and not for primary CML cells. The lack of information based on primary CML cells is due to the presence of a degradative activity which is released when cells are lysed under standard conditions, destroying p210BCR-ABL rapidly and permanently. This prevents quantification or investigation of proteins co-precipitating with p210BCR-ABL in primary CML cells. The degradative activity can be assayed by co-lysing K562 cells, a Ph+ cell line that does not contain the activity, with primary CML cells. Using this assay we show that the degradative activity found in the primary CML cells destroys the p210BCR-ABL present in the K562 cells. The same assay was used to show that the activity is present in both primary CML and normal mononuclear cells (MNC). Conventional protease inhibitor cocktails have not proven effective in inhibiting this degradative activity. We show here that the degradative activity in primary cells is associated with the cell lysosomes and may be an acid dependent hydrolase. By lysing primary CML cells at pH12 we have shown that the degradative activity can be significantly inhibited thereby allowing us to detect p210BCR-ABL in primary CML cells by western blotting. Furthermore, p210BCR-ABL can be immunoprecipitated from primary CML cells when cells are lysed at high pH; however immunoprecipitation is more effective when the pH of lysate is reduced to pH8. We demonstrate that the alterations in pH during the immunoprecipitation procedure do not disrupt the co-immunoprecipitation of p210BCR-ABL-Cbl and Crkl - Cbl associations. Thus, this method is capable of confirming or refuting in primary cells the molecular associations that have been demonstrated in CML cell lines.
Assessing the Benefit/Risk Balance of Long-Term Anagrelide Hydrochloride (Xagrid®/Agrylin®) Treatment for Essential Thrombocythemia.
Authors: Petitt Robert M.
Published: October 2019

Essential thrombocythemia (ET) is a myeloproliferative disorder (MPD) associated with thrombohemorrhagic complications. The disease uncommonly progresses to myelofibrosis or acute leukemia; the risk of transformation may be increased by cytoreductive treatments. Anagrelide (Xagrid®/Agrylin®) is a non-cytotoxic agent that selectively reduces platelet production. A retrospective analysis of an open-label, multi-center trial including MPD patients treated by long-term anagrelide was performed.1 The total safety population comprised 3660 MPD patients (2251 with ET) with a maximum follow-up of 11.4 years. Prior myelosuppressive therapy had been administered in 81% of patients; 33% of these patients were changed to anagrelide because of previous toxicity and 31% because of poor platelet control. Efficacy and safety data hereafter are described for the ET population. There were 934 ET efficacy evaluable patients: with a response seen in 78.7% of them (complete and partial response: 67.2% and 11.5%, respectively). This was similar to that of patients who failed previous therapy (78.8%) or were intolerant (75.6%). After the first year, platelet count decreases were well maintained. Results were similar for both sexes, different age groupings and ethnic origins. AEs occurred in 40.2% of the patients and were generally mild. Anagrelide was discontinued in 38.6% of patients (adverse events accounting for 29.2% of patients stopping treatment). At baseline, 163/934 (17.5%) patients reported ET-related symptoms including GI and other bleedings, arterial or venous thrombosis, angina, pulmonary embolism, transient ischemic attacks, peripheral ischemia and paresthesia. This had reduced to 7.9% (63/796, P<0.001) after 12 weeks and was maintained during follow up (4.3%: 15/470 at 1 year, 3.2%: 6/239 at 2 years, P<0.001.2 Transformation to AML/MDS occurred in 47/2251 (2.1%), but only in subjects who had previously been exposed to cytoreductive treatment. Mortality rate in the safety population was as to be expected in ET patients: 8.8%. Most common reason for death (> or =1%) were CML, reason unknown or unspecified and sepsis. Anagrelide (Xagrid®/Agrylin®) effectively reduces platelet counts and thrombohemorrhagic complications in patients with ET. This is independent of gender, age, ethnic origin and prior therapy. The drug demonstrates a recognized safety profile. Benefits are maintained during long-term follow-up without an increase in disease transformation.
The Role of Histone Acetylation and Death Receptor 5 (DR5) Expression in the Treatment of Chronic Lymphocytic Leukemia (CLL).
Authors: Graham Bonnie A. , Kuschak Brenda , Johnston James B. , Gibson Spencer B.
Published: October 2019

Chronic lymphocytic leukemia (CLL) is a disease in which two-thirds of patients will require therapy, usually with the alkylating agent, chlorambucil (CLB) or the nucleoside analog, fludarabine (FLU). TRAIL is a death receptor ligand that has shown selective cytotoxicity towards a variety of malignancies. CLL cells, however, are relatively resistant to this agent. We have previously demonstrated that CLB- and Flu-induced apoptosis is partially mediated through activation of the TRAIL apoptotic pathway and this is related to the up-regulation of the TRAIL death receptors, DR4 and DR5 (Oncogene, 22:8356–8369, 2003). Combining CLB or Flu with TRAIL produced a synergistic apoptotic response in CLL cells. In contrast, the upregulation of DR5 and the synergistic apoptotic response was not observed in normal lymphocytes. We have subsequently demonstrated that the up-regulation of DR5 is mediated by transcription factors, nuclear factor κB (NFκB) and p53, and histone acetylation. Using chromatin immunoprecipitation assays, we have found that the p65 subunit of NFκB and p53 are bound to the DR5 gene in CLL cells following either CLB or Flu treatment. In addition, histone 3 is acetylated following CLB and Flu treatment corresponding to histone acetylase p300 binding to the DR5 gene. Histone deacetylase 1 (HDAC1) also binds to the DR5 gene following CLB or Flu treatment, but generally at later time points. Overall histone acetylation was also found to be increased in CLL cells, as compared to normal lymphocytes. Treatment with HDAC inhibitors, which are being evaluated in clinical trials, resulted in increased binding of p53 and NFκB, but not HDAC1, to the DR5 gene and increased DR5 mRNA levels in cells. Furthermore, enhanced apoptosis was also observed in CLL cells treated with combinations of SAHA (another HDAC inhibitor) and TRAIL. These findings suggest that histone acetylation is important in regulating DR5 expression and that HDAC inhibitors increase DR5 expression mediated by p53 and NFκB. This could provide a mechanism to sensitize CLL cells to TRAIL-induced apoptosis.
Transactivation of CCL20 Gene by CD30 in Hodgkin’s Lymphoma.
Authors: Mori Naoki , Okudaira Taeko , Yamamoto Kazuo , Horie Ryouichi , Watanabe Toshiki , Matsuyama Toshifumi
Published: October 2019

Hodgkin’s lymphoma (HL) is characterized by a minority of neoplastic cells, the so-called Hodgkin/Reed-Sternberg (H/RS) cells, and an admixture of reactive cells including lymphocytes, plasma cells, eosinophils, and histiocytes. Chemokines produced by H/RS cells play a major role in leukocyte trafficking. These molecules with specific receptor affinities contribute to the maintenance of a favorable environment for survival of H/RS cells. The C-C chemokine CCL20 is expected to play a crucial role in the initiation of immune responses and tumor growth. However, expression of CCL20 in HL has not been studied. We found that in lymph nodes from patients with HL, CCL20 was expressed in H/RS cells by immunohistochemistry. CCL20 was expressed in HL cell lines KM-H2 and L428 established from B-cell type HL patients, but not in cell lines L540 and HDLM-2 established from T-cell type HL patients. CD30 was identified in H/RS cells of HL and has attracted much interest as a molecular marker of HL. All four H/RS cell lines expressed CD30, but not CD30 ligand. CD30 induced CCL20 mRNA expression. Analysis of the CCL20 promoter showed the importance of NF-κB binding site for CD30-induced CCL20 expression. Coexpression of IκB, IKK, NIK, and TRAF dominant-negative constructs with CD30 inhibited CD30-induced activation of CCL20 promoter, suggesting that CD30 induces CCL20 via NF-κB signaling. Electrophoretic mobility shift assay showed the NF-κB complexes that consisted of p50, p65, and c-Rel in KM-H2 and L428 cells. In contrast, the NF-κB complexes contained p50 homodimer that was transcriptionally repressive in L540 and HDLM-2 cells. The CCL20 promoter was activated by the C-terminal region of CD30, which activated NF-κB signaling. A decoy CD30 lacking the cytoplasmic region inhibited CCL20 promoter activity in KM-H2 and L428 cells. Thus, in B-cell derived H-RS cells, ligand-independent activation of CD30 signaling activates NF-κB and this leads to constitutive CCL20 expression.
Pleiotrophin Secreted from Human Breast Cancer MCF-7-Ptn Cells Activates Stromal Fibroblasts, Induces Epithelial Island Formation, and Remodels the Microenvironment.
Authors: Zuka Masahiko , Chang Yunchao , Wang Zhaoyi , Berenson James R. , Deuel Thomas F.
Published: October 2019

Pleiotrophin (PTN, Ptn) is an 18 kD cytokine that is expressed in many human breast cancers and its gene is inappropriately expressed in cell lines derived from these breast cancers. To study the siginificance of inappropriate expression of Ptn in human breast cancer cells on surrounding stromal cells, we first compared nude mouse xenografts of MCF-7 and MCF-7-Ptn cells. MCF-7-Ptn cells lack the Receptor Protein Tyrosine Phosphatase (RPTP)b/z, the PTN receptor, and thus are not responsive to PTN through autocrine or paracrine stimulation. The MCF-7-Ptn cell xenografts grew rapidly whereas MCF-7 cells xenografts were barely detectable 6 weeks after injection. MCF-7-Ptn cells that were co-injected with equal numbers of NIH3T3 cells grew even more rapidly in the flanks of the nude mice. Surprisingly, the MCF-7-Ptn cell explants developed a morphological phenotype remarkably similar to that of the human invasive ductal carcinoma. We then co-cultured MCF-7 cells that express Ptn (MCF-7-Ptn cells) with NIH 3T3 cells. Secretion of PTN from MCF-7-Ptn cells induced formation of sharply defined clusters of MCF-7-Ptn cells, termed “epithelial islands”, that were surrounded by dense fibrous bands interspersed with NIH 3T3 cells that morphologically closely resemble carcinoma associated fibroblasts (CAFs). A striking increase in tropoelastin and expression of type IV procollagen mRNA was identified in NIH3T3 cells co-cultured with MCF-7-Ptn cells. Furthermore, different markers often resulting from stromal cell-carcinoma cell interactions in breast cancer, including protein kinase C (PKC)-d, and both human and murine matrix metalloproteinase (MMP) 9 were identified either in cells or in the culture media taken from MCF-7-Ptn/NIH3T3 cell co-cultures. The induction of these biochemical and morphological features in the co-cultures of MCF-7-Ptn and NIH3T3 cells was demonstrated to be Ptn expression dependent, PTN-secretion dependent, and NIH3T3 cell dependent. The data suggest that PTN secretion alone from human breast cancer cells with inappropriate expression of Ptn is sufficient to markedly remodel the microenvironment of the breast cancer cell and induce a morphological transition of the MCF-7-Ptn cells and NIH3T3 cells to patterns resembling breast carcinomas through activation of the PTN/RPTPb/z signaling pathway in NIH3T3 cells and reciprocal signaling between the carcinoma stromal cells and the PTN secreting breast cancer cells.
Serum Adiponectin Compared with ZAP-70, CD38 and Immunoglobulin Heavy-Chain Mutation Status as a Predictor of Time to First Treatment in Early B-Cell Chronic Lymphocytic Leukemia.
Authors: Molica Stefano , Vitelli Gaetano , Cutrona Giovanna , Digiesi Giovanna , Mirabelli Rosanna , Giannarelli Diana , Sperduti Isabella , Ferrarini Manlio
Published: October 2019

We analyzed the correlation between well-established biological parameters of prognostic relevance in B-cell chronic lymphocytic leukemia [CLL] (i.e, mutational status of the immunoglobulin heavy chain variable region [IgVH], ZAP-70- and CD38-expression) and serum levels of adiponectin by evaluating the impact of these variables on the time to first treatment [TFT] in a series of 69 previously untreated Binet stage A B-cell CLL patients. Higher levels of adiponectin inversely correlated with peripheral blood lymphocytosis (r=−0.254; P=0.03), male gender (P=0.0002), percentage of ZAP−70-positive (r=− 0.285; P=0.04) and CD38-positive (r=−0.294; P=0.04) B-CLL cells.

After a median follow-up time of 32 months (range, 2–120 months) 28 (40.5%) out of 69 patients experienced a need for chemotherapy. Kaplan-Meier estimates of patients’ TFT, plotted after searching the best cut-off for adiponectin (i.e., 5.88 μg/mL ), demonstrated that low adiponectin concentration was associated with a shorter TFT (median TFT 32 months; P=0.01). The relationship among the various bio-pathological parameters, analyzed by the multiple correspondence analysis (MCA), showed two different clinico-biological profiles. The first, characterized by higher adiponectin serum levels (i.e., > 5.88 μg/mL), higher platelet count (> 174 x 109/L), lower β2-microglobulin [β2-m] (< 2.35 mg/L), presence of mutation in the IgVH and low percentage of either CD38-positive (< 20%) or ZAP-70-positive (< 20%) B-CLL cells, was associated with a stable pattern of disease generally not requiring therapy. The second, defined by lower adiponectin levels, lower platelet count, high β2-m concentration, absence of mutation in the IgVH, high percentage of CD38- and ZAP-70 positive B-cells, was associated with a more progressive pattern of disease and a shorter TFT. The univariate Cox proportional hazard model demonstrated that in addition with lower serum levels of adiponectin (Hazard Ratio [HR], 2.936; P=0.01), the absence of IgVH mutational status (HR, 6.378; P=0.002) and ZAP-70-positivity (HR, 3.314; P=0.02) were associated with a shorter TFT. However, in multivariate analysis only ZAP-70 positivity emerged as predictor of the TFT (HR, 5.187; P=0.008).

Our results indicate that in early B-cell CLL clinico-biological profile including among other parameters adiponectin may provide a useful insight into the complex interrelationship of prognostic variables and semplify their interpretation. However, adiponectin may not replace the need for the determination of ZAP-70 and IgVH mutational status.
Vaccine Properties of a Novel Marker Gene-Free Recombinant Modified Vaccinia Ankara (MVA) Expressing Immunodominant CMV Antigens.
Authors: Diamond Don J. , Wang Zhongde , La Rosa Corinna , Srivastava Tumul , Zhou Wendy , Li Zhongqi , Ly Heang , Akbarnejad Tamineh , Krishnan Aparna , Britt William J.
Published: October 2019

Human CMV continues to be a major risk factor in patients undergoing allogeneic hematopoietic stem cell (HSCT) and solid organ transplant (SOT), despite advancement of antiviral therapy. CMV tegument protein pp65 and CMV immediate early gene product IE1 are both considered immunodominant targets of cell-mediated immunity (CMI) and potentially capable of controlling CMV infection. Similar to healthy adults, CMV tegument protein pp65 and IE1 are frequently recognized by human CD4 and CD8 T cells in HCT and SOT recipients. To better assess their role in host defense, we have constructed a novel attenuated poxvirus (MVA) transfer vector named pZWIIA and generated a recombinant MVA (rMVA) expressing both full-length pp65 and exon4 of IE1 (pp65-IE1-MVA) at high levels, followed by the genetic removal of the bacterial marker gene used to distinguish recombinant forms during the screening process. Immunogenicity evaluation indicates that pp65-IE1-MVA not only can induce robust primary CMI to both antigens in HLA A2.1, B7 and A11 transgenic mice, but also can stimulate vigorous expansion of memory T lymphocyte responses to pp65 and IE1 in PBMC of CMV-positive donors. The recent discovery that additional early antigens of CMV are also well-recognized by healthy adults prompted us to include the IE2 antigen in the vaccine. Evaluation of in vivo immunogenicity of the vaccine expressing IE2 in both transgenic mice and in vitro antigenicity in human peripheral blood lymphocytes of HSCT and SOT recipients will be presented. A novel overlapping peptide library spanning full-length IE2 has been constructed in our laboratory, and used in conjunction with the vaccine expressing IE2, along with IE1 and pp65. Since MVA has a foreign gene capacity of over 30 kilobases (Kb), we have developed a strategy to maximize coverage of haplotypes through insertion of 6 additional antigens into novel insertion sites of the virus, for a total of just 12.0 Kb of the CMV genome. The selected antigens cover 83% of the population, while representing over 22% of the combined CD4 and CD8 T cell repertoire in seropositives. Our studies of the recognition of these antigens expressed in MVA and monitored with our own synthesized over-lapping peptide libraries in both HSCT and SOT recipients will be presented. These properties make the MVA-based vaccine ideal for the dual role of priming and boosting CMV-specific T cell immunity as a means to control CMV disease in recipients of HCT or SOT. pZWIIA alone or in combination with other MVA transfer vectors can be used to generate MVA based multiple-antigen vaccine which have application in vaccine development for a wide spectrum of infectious diseases and cancer.