bluuebunny/crossref_metadata_2025_split huggingface dataset:
Augmentation of Therapeutic Tumor Vaccine Efficacy by Genetic Engineering of CD4+ T Cell Help for Tumor Vaccine-Reactive CD8+ T Cells.
Authors: Jalla Sanju , McCadden Erin , Wang Jie , Fuchs Ephraim J. , Whartenby Katharine A. 
Published: April 2020

Abstract
Since CD4+ T cell help has been proposed to be required for maintaining the activity of tumor-specific CD8+ T cells, tolerance in tumor-specific CD4+ T cells may seriously impair the efficacy of therapeutic tumor vaccines. To overcome this problem, we devised a strategy to “engineer” CD4+ T cell help by treating tumor-bearing animals with nonmyeloablative conditioning and transplantation of autologous hematopoietic stem cells (HSCs) that have been genetically modified, via lentiviral transduction, to express an antigen containing “foreign” CD4+ T cell epitopes. After hematopoietic reconstitution, animals received the combination of an autologous tumor cell vaccine and an infusion of primed CD4+ T cells specific for the expressed epitopes. Using influenza hemagglutinin (HA) as the model antigen, we first confirmed that transplantation of HA-transduced HSCs led to efficient expression of HA by antigen-presenting cells, as demonstrated by the clonal expansion of adoptively transferred, HA-specific CD4+ transgenic T cells in mice receiving HA-transduced HSCs but not in mice receiving nerve growth factor receptor (NGFR) gene-transduced HSCs. Next, BALB/c mice harboring 13 day old, metastatic 4T1 mammary cancer were treated with removal of the primary, nonmyeloablative conditioning and transplantation of HA-transduced syngeneic HSCs, and following hematopoietic reconstitution, with concomitant autologous tumor cell vaccination and adoptive transfer of in vitro activated, HA-specific transgenic CD4+ T cells. This therapy was successful in curing the majority of tumor bearing mice, and was superior to the same therapy given to mice transplanted with NGFR-transduced stem cells. Finally, we found that the anti-tumor effect of vaccination plus exogenous T cell help was abolished by the adoptive transfer of either CD4+ or CD8+ T cells from tumor-bearing mice, suggesting that tumor-bearing mice contain both potential effectors and suppressors of anti-tumor immunity, the latter of which are abolished by the non-myeloablative conditioning. These results highlight the importance of CD4+ T cell help in the induction of therapeutic anti-tumor immunity.
Possible Involvement of Histone Acetylation in Differentiation of Erythroid Precursors and Proliferation of Granulocytic Precursors.
Authors: Fujieda Atsushi , Katayama Naoyuki , Ohishi Kohshi , Masuya Masahiro , Shiku Hiroshi 
Published: April 2020

Abstract
The commitment and differentiation of hematopoietic progenitors are thought to be associated with the restriction of transcriptional accessibility for genes of unselected cell-lineage. Histone acetylation-mediated remodeling of chromatin structures is known to regulate transcriptional activity of genes, and the acetylation level of histones is controlled by the balance between opposing activities of histone acetyltransferases and histone deacetylases (HDACs). In this study, we investigated the role of HDAC activity in the proliferation and differentiation of erythroid and granulocytic precursors, using a HDAC inhibitor FK228 [depsipeptide]. CD36+ erythroid precursors were generated from cord blood CD34+ cells with SCF, flt-3 ligand, TPO, IL-3, IL-6 [5GF], and EPO. These precursors expressed low or undetectable level of glycophorin A (GPA) but mostly expressed CD45. When these precursors were cultured with EPO alone in the presence of FK228 (0.4~0.5 ng/ml), the resulting cells expressed GPA at a low level in comparison with the cells cultured without FK228, and a large population of the GPA+ cells remained to be CD45+. On the other hand, most of cells in cultures without FK228 expressed a high level of GPA and a majority of the GPA+ cells were negative for CD45. These data suggest that FK228 represses the EPO-dependent differentiation of erythroid precursors.
The retardation of differentiation due to FK228 was, however, completely recovered when cells exposed to FK228 were incubated for additional 5 days without FK228. At these concentrations, FK228 did not affect the proliferation of erythroid precursors. Granulocytic precursors were also prepared from CD34+ cells with 5GF and G-CSF. In culture with G-CSF, FK228 did not affect the differentiation of granulocytic precursors but did inhibit their proliferation in a dose-dependent fashion. Histone acetylation induced by FK228 in precursors was confirmed using anti-acetylated histone antibodies. These data demonstrate a distinct and regulatory role for HDAC activity in the proliferation and differentiation of hematopoietic precursors.
High Frequency Gain-of-Function Mutation and/or Over-Expression of Proto-Oncogene C-KIT as Major Contributor to the Leukemogenesis in Human Acute Myeloid Leukemia with t(8; 21).
Authors: Zhou Guang-Biao , Wang Yue-Ying , Yin Tong , Bai Xue-Tao , Liang Wen-Xue , Xu Feng , Liu Yi-Wei , Chen Zhu , Chen Sai-Juan 
Published: April 2020

Abstract
To explore the genetic abnormalities that cooperate with AML1-ETO (AE) fusion gene in the pathogenesis of acute myeloid leukemia (AML) with t(8; 21), we undertook to screen for a number of candidate genes and identified a high frequency mutation of proto-oncogene C-KIT (25/53 cases, 47%). This event seems to be specific to t(8;21) leukemia since no mutation was detected in 57 patients with other types of leukemia. Totally eleven types of mutations were characterized and 6 ones including 3 point mutations in the tyrosine kinase (TK) domain and three types of internal tandem duplication (ITD) in the juxtamembrane (JM) or extracellular (EC) domains were discovered for the first time. Bioinformatics analysis revealed all these mutations might favor the activation of the C-KIT, as proven by TK activity assay. Patients with a mutant C-KIT (mC-KIT) allele had a higher white blood cell count and a poorer prognosis than those bearing wild-type C-KIT. The expression of C-KIT at both mRNA and protein levels was higher in t(8;21) leukemia patients than most other leukemia types. In addition, the TK inhibitor Gleevec suppressed the C-KIT activity, inhibited the proliferation and induced apoptosis of t(8;21) leukemic cells bearing C-KIT mutation or over-expression. Our study thus provides strong evidence to the model of leukemogenesis which requires a cooperation of cytosolic signaling and transcription factor for a full-blown leukemia phenotype to display.
Efficacy and Toxicity of FLAI-G-CSF and Mylotarg for Induction/Consolidation of AML Patients, Not Treatable with Conventional Chemotherapy.
Authors: Malagola Michele , Rondoni Michela , Russo Domenico , Piccaluga Pier Paolo , Paolini Stefania , Bosi Costanza , Gaitani Stavroula , Visani Giuseppe , Martinelli Giovanni , Baccarani Michele 
Published: April 2020

Abstract
Acute myeloid leukaemias (AMLs) usually share a high frequency and a high degree of Pgp-mediated multidrug resistance (MDR), one of the major causes of treatment failure. Patients older than 60 years, as well as patients with secondary AML and patients with relapsed/refractory disease do poorly with conventional chemotherapy. Gemtuzumab ozogamicin (Mylotarg) has shown some in vitro activity against CD33 positive AML blasts. As single agent it induces complete remission (CR) in almost 20 – 30% of cases with mild toxicity. Combinations with chemotherapy look promising. Moreover, several studies proved in vivo efficacy of fludarabine in combinations with cytarabine as anti MDR drug. Fifteen AML patients were treated with an induction regimen including fludarabine (25 mg/sqm/day for 3 days), cytarabine (1 g/sqm/day for 3 days), idarubicine (5 μg/sqm/day for 3 days), mylotarg (3 mg/sqm day +4) and G-CSF (5 mg/day starting on day +9, until hematological recovery). An identical consolidation cycle was administered if at least partial remission (PR) was achieved. Two patients were relapsed after standard induction chemotherapy. Median age at diagnosis was 66 (range 33 – 75). Median white blood cells count was 5.3 x 109/L (range 0.5 – 47.4). Seven out of 15 patients presented with at least 1 adverse karyotype abnormality. Eight patients presented with a secondary AML. Ten out of 15 patients responded to induction cycle (8 CR and 2 PR). The following data regard the induction cycle. The median time to recovery of neutrophils &gt; 1.0 x 109/L was 20 days (range17 - not reached) The median time to recovery of platelets &gt; 20 x 109/L and &gt; 100 x 109/L was 21 days (range 13 - not reached) and 34 (range 19 - not reached) respectively. Two patients developed a grade II WHO FUO and 5 patients developed a grade II-III Gram+ bacteremia. Two patients had radiologically documented fungal infection (lung) and 1 patient had microbiologically documented aspergillosis of maxillary sinus. Non-hematological toxicity was mild. Only one patient developed a grade III WHO hemorragic syndrome (melena). No veno occlusive disease were observed.
Up to now 4 patients completed two cycles of chemotherapy. Three patients out of 15 died (1 progressive disease, 1 multi organ failure after allogeneic bone marrow transplantation, 1 myocardial stroke while in CR) and 7 patients out of 15 are in CR. In conclusion, these preliminary results suggest that FLAI-G-CSF-Mylotarg, as induction course, is an effective regimen with a limited hematological and non-haematologic toxicity and seems to be effective in this high risk subset of patients. A longer follow up and larger series of patients are indeed necessary to draw further conclusions.
Fluorescence In Situ Hybridization (FISH) for the Detection of the (8;14) Translocation in Archival Fine Needle Aspirates of Suspected Burkitt Lymphomas (BL).
Authors: Neto Lara F. , Mendonça Evelina , Cabeçadas José , Mendonça Andreia , Silva Maria G. 
Published: April 2020

Abstract
The detection of the chromosomal translocations involving the myc gene, characteristic of BL, is important in the diagnostic workup of agressive lymphomas given its impact on treatment strategies and prognosis. Recently this has been accomplished using FISH techniques either in paraffin-embedded tissue sections or fresh tumor aspirates. BL has an elevated cellular turn-over and a rapid diagnosis is highly desirable; this, and its high incidence in pediatric populations in whom organ biopsy needs general anesthesia, makes fine needle aspiration (FNA) samples attractive for diagnosis. In some cases, confirmation of the diagnosis may require the analysis of archival pre-stained samples. However, only recent samples have been reported as suitable for FISH analysis. We analysed 18 cytological samples (8 cytospins and 10 smears) obtained by FNA from 16 male and two female patients, median age 7.5 (3 to 48) years old, diagnosed with BL (15), atypical BL (1) or high grade lymphoma with a difficult differential diagnosis with BL (3) between February 2000 and July 2004 on the basis of morphology and phenotype of cytological and/or histological samples. In all cases the presentation and further clinical course suggested the possibility of BL and treatment was administered accordingly. Sixteen smears were May-Grunwald-Giemsa (MGG) stained whereas two were fresh-frozen; the samples had been stored for variable periods (1 day to 53 months). Cytology specimens (6 MGG-stained and 6 frozen) obtained from 12 reactive lymph nodes were used as controls, to determine the cut-off value for positive hybridization signals. After removal of the coverslips with xylene, the slides were destained in a series of water and ethanol washes. Pretreatment with pepsin was followed by the hybridization procedure. The dual fusion t(8;14) fluorescent probe was purchased from Vysis. Eleven out of 18 (61%) samples could be hybridized using this technique. The 7 cases without an interpretable signal were between 9 and 53 months old. The cut-off value for t(8;14) was 0%, meaning that in 12 negative controls the pattern of two fusion signals with or without a remaining signal from der(8) chromosome was never observed. In aggreament to previous publications 8 out of 11 specimens (72%) were positive for the (8;14) translocation, including 3 cases in which a definitive BL diagnosis could not be made by cytology/histology alone. The median percentage of positive cells was 80% (46 and 92%). Although the quality and age of the smears influenced the results, and better signals were observed with frozen specimens, FISH analysis of archival cytopathological material stored for prolonged periods was frequently possible and allowed the diagnosis of 3 difficult cases, whereas it confirmed the morphological results in another 5 patients.
PKC beta II Expression in Lymphoid Malignancies.
Authors: Decouvelaere Anne Valerie , Morschhauser Franck , Copin Marie-Christine , Dumontet Charles M. 
Published: April 2020

Abstract
Protein Kinase C (PKC) is an important enzyme family. Twelve different isozymes have been described, which are implicated in diverse cell responses. Recently, studies have begun to define isoform-specific functions of PKC, and PKC beta II seems to play an antiapoptotic role. PKC beta II expression was analyzed by immunohistochemistry in 119 tumor samples of patients with lymphoid malignancies. The median age of the group was 55 years (range 14–85 y). Immunohistochemical staining was performed on formalin-fixed paraffin-embedded tissue, using the labeled avidin-biotin immuno-peroxidase complex method on the Ventana Benchmark automated immunostainer (Ventana Medical Systems, S.A., Illkirch, France). The sections were incubated with primary antibody (polyclonal antibody cPKC ßII (C-1): sc-210, Santa Cruz Biotechnology, Inc.) for 32 min at 1/100 dilution. The intensity of cytoplasmic and membrane immunostaining was assessed semi-quantitatively by two pathologists independently. Cases were considered “negative” when &lt;10% of cells were stained, and “positive” when ≥10% of tumor cells showed cytoplasmic or membrane staining. Diffuse or heterogeneous staining was also specified.
Intense and diffuse staining was observed in all mantle cell lymphomas (13 cases, 100%), and chronic lymphocytic lymphomas (CCL) (6 cases, 100%). PKC beta II was positive too in most follicular lymphomas (10 cases out of 11, 91%), although the pattern and the intensity of the staining were variable in accordance with the degree of lymphomatous infiltration. Indeed, in the same lymph node, the follicular lymphomatous areas were constantly labelled, whereas residual germinal centers remained negative. A majority of angioimmunoblastic T-cell lymphomas, lymphoblastic T-cell lymphomas and Marginal Zone/MALT lymphomas were labelled with anti-PKC beta II antibody, but the pattern of expression was more heterogeneous in these subtypes. A minority of diffuse large B-cell lymphomas expressed PKC beta II (8 out of 22 cases, including 4 with weak staining). Nearly all plasma cell neoplasms were negative. Nevertheless, 2 cases out of 16 (13%) displayed a diffuse moderate staining. In peripheral T-cell lymphomas, staining for PKC beta II was less frequent, as well as in primary cutaneous B and T lymphomas and pseudo-lymphomas. None of the cases of Hodgkin’s disease (13 cases) and anaplastic large cell lymphoma (5 cases) expressed PKC beta II. However, most of the numerous reactive cells were strongly stained.
In conclusion, the intensity and the staining pattern of anti-PKC beta II antibody varied greatly according to the type lymphoid malignancy. These results highlight a significant correlation between PKC beta II expression and some particular type of lymphomas. In our series, the highest level of expression was found in mantle cell lymphomas and CLL.
Subset of T-Cell Prolymphocytic Leukemia Expresses CD117. Potential Target for Therapy.
Authors: Gorczyca Wojciech , Dong Henry Y. 
Published: April 2020

Abstract
Introduction
Excluding non-hematopoietic lesions, CD117 (c-kit) is expressed by erythroid, megakaryocytic and myeloid precursors, mature mast cells, subset of plasma cell neoplasm and occasional precursor lymphoblastic tumors. C-kit plays important role in T-cell development around birth, but the expression of CD117 in mature T-cell lymphoproliferations is rare. We present flow cytometry phenotypic data of 28 cases of T-cell prolymphocytic leukemia (T-PLL).
Material and methods
Multiparameter 4-color FC analysis was performed on peripheral blood and/or bone marrow aspirate samples using the following antibodies: CD2, CD3, CD4, CD5, CD7, CD8, CD10, CD11c, CD13, CD14, CD16, CD19, CD20, CD25, CD33, CD34, CD38, CD45, CD56, CD64, CD79a, CD117, TCRab/γδ, TdT, and HLA-DR. Fresh bone marrow aspirates and/or peripheral blood films and cytospin preparations from FC samples were stained with Wright-Giemsa.
Results
T-PLL was diagnosed 28 cases. The ages ranged from 43 to 92 years. Normal expression of pan-T cell antigens (CD2, CD3, CD5, CD7) was present in 57%. Abnormal expression of one, two and three antigens was noted in 21%, 11% and 11%, respectively. 4 cases showed aberrant expression of CD117. Those cases were always CD8-positive (50% of T-PLL were CD4+). There were no pan-myeloid (CD13, CD33) and blastic (TdT, CD34) markers present. Molecular studies confirmed clonal rearrangement of TCR.
Discussion
T-PLL is rare mature T-cell neoplasm characterized by marked lymphocytosis of small to medium-sized cells with conspicuous nucleoli. The response to conventional chemotherapy in T-PLL, similarly to other mature T-cell lymphoproliferations (except ALK+ anaplastic large cell lymphoma) generally is poor. While the use of alemtuzumab (Campath-1H) has improved remissions, the disease remains incurable. Novel treatment approaches, tailored to individual patients, and based on pathologic, phenotypic and molecular features will be crucial to improve survival for the patients with this disease.
Based on the flow cytometry analysis of 28 cases of T-PLL we identified 4 cases with co-expression of CD117 (14%). When present, it is restricted to CD8+ tumors. We speculate that the expression of CD117 (c-kit) in these patients apart from its diagnostic utility, may serve as a criterion for a combination therapy with Campath-1H and imatinib mesylate.
Serum Interleukin-18 Levels Are Associated with Response to Treatment and Survival in Patients with Aggressive Non-Hodgkin’s Lymphoma.
Authors: Goto Naoe , Tsurumi Hisashi , Takemura Masao , Hara Takeshi , Sawada Michio , Yamada Toshiki , Kasahara Senji , Kanemura Nobuhiro , Takahashi Takeshi , Seishima Mitsuru , Takami Tsuyoshi , Saito Koshiro , Moriwaki Hisataka 
Published: April 2020

Abstract
Interleukin-18 (IL-18), originally designated as an interferon (IFN)-gamma inducing factor, is a cytokine which induces cytotoxic NK cell activity and stimulates T cells to produce IFN-gamma, IL-2, and GM-CSF. Increased serum IL-18 levels have been found in patients with acute lymphoblastic leukemia, chronic myelocytic leukemia, multiple myeloma and non-Hodgkin’s lymphoma (NHL). The aim of the present study was to assess the prognostic significance of serum IL-18 in aggressive NHL. Consecutive 99 previously untreated patients with aggressive NHL (diffuse large B-cell lymphoma; 84 patients, peripheral T-cell lymphoma; 15 patients) prospectively participated in this study between 1997 and 2003. The patients were treated with 6–8 cycles of CHOP or THP (pirarubicin) -COP regimens. To evaluate serum levels of IL-18, venous blood samples were drawn from patients immediately before the initiation of treatment. In all patients with aggressive NHL, the mean ± SD of serum IL-18 level was 1200.8±151.5 pg/ml (range 96–6603.5) with a median of 556.75 pg/ml. Several poor prognostic features such as poor PS, multiple extranodal sites, advanced disease (clinical stage III/IV), existence of B-symptom, and high soluble interleukin-2 receptor (sIL-2R) were strongly associated with a high serum IL-18 levels. An increased LDH and gender were weakly associated with a high serum IL-18. Histology and age were not associated with serum IL-18 levels. The median serum IL-18 levels of the different IPI risk groups were; 321 pg/ml for the L risk; 442 pg/ml for the LI risk; 557 pg/ml for the HI risk; 1532 pg/ml for the H risk, respectively (P&lt;0.005). The CR rate of patients with an IL-18 level of less than 700 pg/ml was better than that of 700 pg/ml or higher (77% and 50%, P&lt;0.01). Patients with high IL-18 (700 pg/ml and over) at onset had a significantly lower survival rate (5-year: 25%) than those with low IL-18 (below 700 pg/ml) (68 %) ( p&lt;0.0001). Multivariate analysis employing IL-18 and some conventional prognostic factors demonstrated that IL-18, performance status and the number of extranodal sites were significantly poor prognostic factors for both overall survival (OS) and event free survival (EFS). The results suggest that a high serum IL-18 level predicts a poor prognosis in aggressive NHL and may be a useful biomarker for selecting appropriate treatment.
Cellular Immunodeficiency in Untreated Lymphoma Patients.
Authors: Jentsch-Ullrich Kathleen , Gerber Annegret , Reppin Regina , Koenigsmann Michael , Mohren Martin , Franke Astrid 
Published: April 2020

Abstract
The association of lymphoma with a plethora of secondary immune defects is well established. However, most investigations were performed many years ago and little is known about disease specific alterations of the immune system. The availability of new diagnostic methods and a new classification of hematologic diseases (WHO 2000) makes a newly designed study of the immuno-dysfunction in this patient group worthwhile. From January 2003 to August 2004 50 patients with newly diagnosed malignant hematologic diseases - diffuse large cell lymphoma (DLCL) (n = 15), acute myeloid leukemia (AML) (n = 11), mantle cell lymphoma (n = 6), follicular lymphoma (FL) (n = 5), Hodgkin’s disease (HD) (n = 8) and chronic lymphocytic leukemia (CLL) (n = 5) - were analyzed. Parameters of cellular and humoral immunity, including CBC, FACS analysis of lymphocyte subpopulations, cytokines sIL2-R, IL-6, IL-10, TNF-a, serum complement components C3, C4, CH50 and total concentrations of serum immunoglobulin IgG, IgM, IgA were measured. At diagnosis patients with AML had severe cellular immunodeficiency (neutro-, monocyte- and lymphopenia). Infectious complications occurred in 8 of 11 AML patients (73%). Patients with DLCL and MCL had predominantly increased CD3+/CD8+ lymphocyte subsets (median 0,26/m l) with an infection rate from 30%. However, the rate was significantly higher if CD4+ depletion (43%) or CD19+ depletion (40%) were additionally present. High IL-2, IL-6, IL-10 and TNF-a levels were mostly observed in FL and MCL. Elevated TNF-a levels were also associated with infections (43%) and advanced stage or B-symptoms. IL-10 was found to correlate with mortality and relapse risk in all patients (overall: 18% and 20% respectively, increased IL 10 level: 39% and 33% respectively). Serum complement levels were normal to slightly elevated in all patients. Very high levels (CH50 &gt; 160) were found in patients with Hodgkin’s disease (5/8 patients, 63%) and decreased immunoglobulin levels occurred in CLL patients only, none of whom had infections during follow up. These results indicate that untreated patients with DLCL and MCL have a significant lower CD8+T-lymphocyte cell count. In contrast to previous studies we did not found a predominant depletion of CD4+T-lymphocyte. A combination of CD4+/CD8+ T-cell deficiency or T-/B-cell deficiency was strongly associated with infections, thus antibiotic prophylaxis seems warranted in this patient group. Additionally IL-10 may serve as prognostic marker particularly in MCL and FL.