bluuebunny/crossref_metadata_2025_split huggingface dataset:
Chromosome 17 Abnormalities Are Associated with Worse Overall and Relapse Free Survival in Patients with Acute Myelogenous Leukemia and Poor-Risk Cytogenetics.
Authors: Borthakur Gautam , Tam Constantine , Kantarjian Hagop , Lin E. , Cortes Jorge , O’Brien Susan , Koller Charles , Pierce Sherry , Keating Michael 
Published: April 2020

Abstract
Purpose: Chromosome 17 abnormalities define a group of patients with acute myelogenous leukemia (AML) (Nahi, H. et al. Leukemia and Lymphoma2008;49:508) with poor outcomes. We analyzed the additional impact of chromosome 17 abnormalities (−17, −17p, −17q, der17) among patients with AML and cytogenetic abnormalities traditionally considered to be of adverse prognosis.
Patients and Methods: 1086 patients with AML [excluding inv 16, t (8;21), t (15;17), Diploid/-y abnormality] were included in this analysis. Based on cytogenetic abnormalities patients were grouped into: −5,−7,−5and −7, complex. The following parameters were included in uni and multi-variate analysis: age, performance status, WBC, hemoglobin, platelets, marrow blast percentage, bilirubin, creatinine, albumin, LDH, chromosome 17 abnormality (yes/no).
Results: Four hundred and fourteen (45%) patients achieved complete remission (CR) or CR with incomplete platelet recovery (CRp) and 267 (64.5%) patients relapsed. Two hundred seventy (24.9%) patients had abnormalities of chromosome 17. Abnormalities of chromosome 17 were associated with lower CR or CRp rate (p=0.02) and higher possibility of having cytogenetic abnormality of −5 or −7 (p&lt;0.0001). Multivariate analysis showed that patients with abnormalities of chromosome 17 had worse overall survival (OS) compared to patients without (p= 0.003)(Fig.1). Multi-variate analysis within cytogenetic subgroups showed that chromosome 17 abnormalities were associated with worse OS in patients with chromosome 5 abnormality(p=.02) (data not shown) and in those with complex cytogenetics (p=.04)(Fig.2) and not in patients with chromosome 7 (p=.17)or combined 5 and 7 abnormalities (p=.33). Similar analysis restricted to patients achieving CR/CRp after induction therapy showed that impact of chromosome 17 abnormalities on relapse free survival (RFS) mirrored their impact on OS.
Conclusion: chromosome 17 abnormalities are associated with worse OS and RFS in patients with AML and adverse cytogenetics and have additional negative impact on the outcomes in certain well-known adverse cytogenetic subgroups.
Figure 1: Kaplan-Meier estimates of overall survival by status of chromosome 17 Figure 1:. Kaplan-Meier estimates of overall survival by status of chromosome 17 Figure 2: Kaplan-Meier estimates of overall survival by status of chromosome 17 in subgroup of patient, complex Figure 2:. Kaplan-Meier estimates of overall survival by status of chromosome 17 in subgroup of patient, complex
Prevalence and Clinical Correlates of JAK2 Mutations in Pediatric Down Syndrome Acute Lymphoblastic Leukemia.
Authors: Gaikwad Amos S. , Rye Cassia L. , Devidas Meenakshi , Izraeli Shai , Plon Sharon E. , Basso Giuseppe , Pession Andrea , Rabin Karen R. 
Published: April 2020

Abstract
Approximately three-quarters of pediatric acute lymphoblastic leukemia (ALL) cases harbor recurrent, prognostically significant chromosomal abnormalities which affect assignment of risk group and therapeutic regimen, but these abnormalities occur far less often in children with Down syndrome (DS) and ALL. Recently, novel somatic activating mutations in Janus kinase 2 (JAK2) were reported to occur uniquely in DS ALL, affecting 18% of patients (Bercovich et al, in press). Here we report the identification and clinical correlates of JAK2 mutations in an independent cohort of cases. We sequenced JAK2 exon 16 and identified mutations in 10 of 53 DS ALL cases (18.9%). All were heterozygous point mutations affecting arginine 683 (Table 1). No JAK2 mutations were identified in the 25 available paired germline DS samples, including five germline samples associated with JAK2 mutated ALL cases, indicating that mutations were somatic in these cases. Eight of the JAK2 mutations (80%) occurred in males (p&lt;0.03), whereas the full cohort contained 24 males (45%). No significant correlation was observed between JAK2 mutation status and age, initial white blood count (WBC), or 5-year event-free survival or overall survival. Our results confirm the 18% prevalence of JAK2 mutations at arginine 683 in DS ALL, but differ from the prior report in finding a male predominance of JAK2 mutations, and no association with age or initial WBC, differences which may be attributable to sample size. These results confirm that activation of the JAK-STAT pathway via a specific JAK2 mutation occurs in approximately one-fifth of cases of DS ALL.
Sequence Amino acid Cases (n) AGA (wild-type) Arginine 43 GGA Glycine 6 AGT Serine 2 AGC Serine 1 ACA Threonine 1
Table 1. JAK2 mutation status in pediatric DS ALL
MRD Monitoring of Childhood ALL Using Hierarchical Clustering and Support Vector Machine Learning of Complex Multi-Parameter Flow Cytometry Data.
Authors: Fiser Karel , Sieger Tomas , Schumich Angela , Irving Julie , Dworzak Michael N , Vormoor Josef 
Published: April 2020

Abstract
Flow cytometry is an important tool both for research and diagnostics of hematologic malignancies - including monitoring of minimal residual disease (MRD). Recent progress and more widespread availability of 6 and higher color flow cytometry leads to complex, information-rich datasets which are very challenging to analyze. Here, we validate a novel approach to multi-parameter flow data analysis in MRD flow data sets from 39 ALL patients (including 23 patients from the I-BFM list mode data (LMD) ring trial). The approach combines hierarchical clustering (HCA) using a newly developed algorithm and support vector machine (SVM) learning. The algorithm employs a scale-invariant Mahalanobis distance measurement for merging clusters. This reflects the extended ellipsoid shape of the populations and is better suited for flow cytometric data compared with standard HCA metrics. The resulting hierarchical tree, combined with the heatmap of the CD marker expression allows visualization of hierarchically clustered data of all analyzed parameters displayed in a single plot. The clusters from HCA (representing the ALL blast population at diagnosis) were used to train SVM classifiers which were then applied to test for presence of a matching population in the test sample (follow-up sample). All work was carried out in MATLAB (MathWorks, Inc.). Using HCA, we have been able to detect the leukemic blast population in diagnostic and follow-up datasets (n=81) from three centers. The correlation (Pearson correlation coefficient = 0.98) between HCA and the standard gating approach was highly comparable to inter-laboratory comparisons within the I-BFM LMD ring trial (Dworzak MN et al; Cytometry B Clin Cytom. 2008 Jun 11.). To further improve sensitivity and exact quantification of low MRD levels and to automate MRD detection, we combined HCA with SVM learning. We have analyzed 21 samples from 5 patients with MRD levels between 0.004 to 57.54%. HCA plus SVM correlated better with standard gating results than HCA alone in particular in samples with low MRD levels (&lt;10−3). In summary, HCA in combination with SVM proved to be a strong analytical tool for flow cytometry with the potential for automated MRD detection. We validated this approach for use in ALL diagnostics and MRD monitoring by comparison with expert-based gating analyses of I-BFM LMD ring trial data.
The Leukemia-Associated Antigen PRAME Is Overexpressed in Myeloid Leukemias and Inhibits Cell Differentiation by Blocking the Receptor for Retinoic Acid (RAR)-Signaling in Vitro and Is Therefore a Interesting Candidate for Targeted Immunotherapies.
Authors: Greiner Jochen , Bullinger Lars , Giannopoulos Krzysztof , Schmitt Anita , Goetz Marlies , Kienle Lena , Döhner Hartmut , Schmitt Michael 
Published: April 2020

Abstract
The leukemia-associated antigen PRAME (preferentially expressed antigen of melanoma) is frequently expressed in several solid tumors. Earlier, we reported (Greiner et al., Blood 2006) that co-expression of leukemia-associated antigens including PRAME constituted a favorable prognostic parameter for AML patients. By microassay analysis and conventional RT-PCR, we detected over-expression of PRAME in more than 60% of AML and of CML patients. In contrast, PRAME is expressed neither in normal CD34-positive hematopoietic stem cells nor in normal tissues (other than testis and placenta). Here, we describe for the first time that retinoic acid-induced cell proliferation and differentiation of several AML cell lines is dependent on PRAME expression: PRAME negative cell lines treated with all-trans retinoic acid (ATRA) in cell culture showed lower cell proliferation than PRAME positive cells as assessed by cell counts and FACS analysis for BrdU, but an increase of cell differentiation detected by FACS analysis for CD66b in contrast with PRAME positive leukemia cell lines. The leukemia-associated antigen PRAME seems to be responsible for the resistance of PRAME-positive AML to ATRA treatment. We detected no differences in induction of apoptosis in PRAME-positive or PRAME-negative cell lines treated with ATRA in FACS analysis for annexin. Over-expression of the antigen and this critical role of PRAME in cell differentiation might be the reason for specific T cell induction against PRAME-derived peptides. To detect specific T cell responses against PRAME-derived peptides we examined samples from healthy volunteers and AML patients. Positive results in ELISPOT assays for IFN-g and Granzyme B secretion were observed in 70% of AML cases analyzed for the peptide PRAME-P3. In chromium release assays, we found a PRAME-specific cell lysis of PRAME-positive AML blasts. Taken together, PRAME is involved in a crucial mechanism for cell growth of leukemic cells, induces specific T cell responses in a high frequency of AML patients, and this constitutes an appropriate target structure for specific immunotherapies or other targeted therapies in AML.
The Critical Role of Iκbα Dependent Nuclear Export of NF-κb in B-Cell Development.
Authors: Yang David T , Wuerzberger-Davis Shelly , Chen Yuhong , Yu Mei , Zeng Hu , Bates Paul , Wen Renren , Wang Demin , Miyamoto Shigeki 
Published: April 2020

Abstract
Activity of the nuclear factor-κB (NF-κB) family of transcription factors is tightly regulated by its inhibitor, IκBα, through cytoplasmic localization of latent NF-κB: IκBα complexes. This arrangement is essential for efficient signal-inducible activation and regulation of biologic functions. Maintenance of cytoplasmic localization of latent NF-κB: IκBα complex requires continuous nuclear export that is dependent on the N-terminal nuclear export sequence (N-NES) of IκBα. While these mechanisms have been elucidated through in vitro studies, the biological significance of this “nucleocytoplasmic shuttling” has yet to be evaluated in vivo. To address this, we derived mice harboring germ-line M45A, L48A, and I52A amino acid substitutions in the N-NES of IκBα. In splenic B-cells, the disrupted N-NES caused constitutive nuclear accumulation of IκBα and inactive c-Rel containing complexes but surprisingly not IκBα: p65 complexes. Since p65 contains a NES sequence and c-Rel does not, nuclear export of N-NES mutant IκBα:NF-κB complexes appear to be NF-κB family member dependent. Functionally, NF-κB activity in splenic B-cells after stimulation with IgM or LPS was clearly reduced in the mutants compared to wild-type by electrophoretic mobility shift assay. B-cell development in the bone marrow of mice harboring the mutation was impaired, showing a preponderance of pro/pre B-cells and few mature B-cells compared to their wild type littermates (p &lt; 0.001). Concordantly, there were significantly fewer B-cells in the spleen (p &lt; 0.05) and lymph nodes (p &lt; 0.01) of the mutant mice. Additionally, populations of T2, follicular (FO), and marginal zone (MZ) B-cells, which represent mature B-cells in the spleen, were also reduced in the mutant mice (p &lt; 0.001). To demonstrate that this B-cell maturation defect in IκBα mutant mice was B-cell intrinsic, sublethally irradiated Jak3-deficient mice were transplanted with BM from either wild-type or mutant mice. B-cell development in mice transplanted with mutant donors was impaired relative to those with wild-type donors in a fashion identical to that of the primary mutants described above. Finally, severe phenotypes in inguinal lymph nodes and Peyer’s patch development were present, with mutant mice frequently lacking these secondary organs/tissues, the underlying mechanisms of which are currently being investigated. In conclusion, our findings uncover an in vivo mechanism controlling NF-κB localization and its essential role in the generation of mature B-cells and certain secondary lymphoid organs.
Suppression of Allogeneic Immune Responses by Human TCRαβ+ CD4− CD8− Double-Negative T Cells.
Authors: Voelkl Simon , Gary Regina AM , Mackensen Andreas 
Published: April 2020

Abstract
Regulatory T lymphocytes play an important role in the maintenance of immune tolerance to self antigens and are involved in downregulating immune responses in autoimmunity, transplant rejection and tumor immunity. Numerous studies have demonstrated the existence of distinct T cell subsets with immunoregulatory properties. Recently, a novel subset of TCRαβ+ CD4− CD8− (double-negative, DN) T cells has been characterized to specifically suppress immune responses in both mice and humans. Here we demonstrate for the first time that human DN T cells are highly potent suppressor cells of allogeneic CD4+ or CD8+ T cell responses after priming with allogeneic antigen presenting cells (APC). A prerequisite for the immunosuppressive activity is the repetitive priming with allogeneic dendritic cells whereas stimulation with artificial APCs has no effect. Using a transwell system we could show that the suppressive activity against allogeneic immune responses, mediated by DN T cells, requires cell contact. In contrast to murine DN T cells, which eliminate effector T cells via a fas/fasL or perforin/granzyme pathway, human DN T cells suppress the proliferation of alloreactive T cells in an active manner. Taken together, our data indicate that human DN T cells possess strong immunosuppressive effects on alloreactive CD4+ or CD8+ T cells. DN T cells may serve to limit clonal expansion of alloantigen-specific T cells after allogeneic peripheral stem cell transplantation.
T Cells Demonstrate Enhanced Specificity for CD19+ Malignancies When Stimulated with IL-21.
Authors: Singh Harjeet , Huls Mary Helen , Dawson Margaret J. , Mi Tiejuan , Dotti Gianpietro , Lee Dean A. , Kebriaei Partow , Shpall Elizabeth J. , Hackett Perry B. , June Carl H. , Champlin Richard E. , Cooper Laurence J.N. 
Published: April 2020

Abstract
T cells genetically modified to express CD19-specific chimeric antigen receptors (CARs) are being evaluated in early-phase clinical trials in patients with B-lineage malignancies. Therapeutic success is predicted by ability of infused CAR+ T cells to both persist and kill in an antigen-dependent fashion. The first of these dual goals can be achieved by altering the CAR molecule to provide T-cell survival signals through a chimeric CD28 endodomain (designated CD19RCD28). We now report that altering the culturing microenvironment with IL-21 improves antigen-dependent cytolysis of T cells when propagated on CD19+ artificial antigen presenting cells (aAPC) derived from K562. To test whether IL-21 acts in conjunction with CD28 signaling to support acquisition of redirected effector functions we electro-transferred quiescent T cells from peripheral blood with Sleeping Beauty system DNA plasmids to introduce CD19RCD28 CAR transposon. Selective outgrowth of CAR+ T cells was achieved on CD19+ aAPC that provide co-stimulation with the addition of exogenous IL-2 and/or IL-21. When IL-21 was present there was preferential numeric expansion of CD19-specific CD8+ T cells which lysed and produced IFN-g in response to CD19 (Figure). Furthermore, the CD8+CAR+ T cells displayed a central memory (CM) cell surface phenotype characterized as CD62L+ and CD28+. In contrast, genetically modified T cells propagated with exogenous IL-2 resulted in predominately CD19-specific CM CD4+ T cells. Thus, cytokines can be used to tailor the CD8/CD4 ratio of CAR+ T cells derived from peripheral blood. These data demonstrate that the dual goals of persistence and lysis can be achieved by altering CAR and the cytokine milieu and have implications for infusing CAR+ T cells in next-generation immunotherapy trials.
Figure Figure
BCX-4208 (RO5092888), a Purine Nucleoside Phosphorylase (PNP) Inhibitor, Is a Novel, Potent Orally Active Anti-T-Cell and B-Cell Agent.
Authors: Bantia Shanta , Parker Cynthia , Upshaw Ramanda , Kilpatrick John Michael , Cunningham Amanda , Zhang Jianwen , Kotian Pravin , Morris Philip , Chand Pooran , Babu Yarlagadda S. 
Published: April 2020

Abstract
The profound suppression of T-cell immunity seen in patients with an inherited PNP deficiency supports the potential application of inhibitors of this purine salvage enzyme in the therapy of T-cell malignancies and T-cell mediated autoimmune diseases. About thirty percent of PNP deficient patients also show evidence of B-cell dysfunction. BCX-4208 is a novel potent transition state analog inhibitor of PNP (IC50 ~ 0.0005 μM) and in the presence of 10 μM deoxyguanosine (dGuo), inhibits human lymphocyte proliferation induced by MLR, IL-2 or Con-A with IC50s of 0.159, 0.26 and 0.73 μM, respectively. The IC50 for dGuo in the same assays in the presence of 1 μM BCX-4208 ranges from 1–3 μM. Neither BCX-4208 alone nor dGuo alone inhibits proliferation of lymphocytes. In the presence of PNP inhibitor, dGuo is converted to dGMP and then to dGTP. Accumulation of dGTP results in the alteration of deoxynucleotide (dNTP) pools, causing death of cells via a mechanism characteristic of apoptosis. In vitro data demonstrates that following exposure to BCX-4208 and dGuo, dGTP in human lymphocytes is elevated and a 5–8 fold increase in dGTP results in 50% inhibition of lymphocyte proliferation. Flow cytometric analyses of human lymphocytes using annexin-V staining reveal that BCX-4208 in the presence of dGuo induces cellular apoptosis not only in T cells (CD3+), but also in B cells (CD20+; CD19+) (Table 1). BCX-4208 is orally bioavailable in mice, can achieve maximal inhibition of PNP, and elevates plasma dGuo levels to 3–5 μM (predose levels &lt; 0.004 μM), which is similar to levels seen in PNP deficient patients and to levels needed to cause apoptosis in T and B-cells. These data support the evaluation of BCX-4208 in the treatment of not only T-cell mediated diseases but also B-cell mediated diseases. BCX- 4208 is currently undergoing early clinical investigation in patients with psoriasis.
Table 1.
Cell subsets % Apoptotic cells mean ± SEM (n = 4–7) Vehicle Treatment group *p ≤ 0.01 compared to vehicle CD3+ 7.2 ± 0.9 18.3 ± 3.7* CD20+ 24.0 ± 3.6 41.8 ± 6.2* CD19+ 25.0 ± 3.1 51.6 ± 7.4*
Role of Human NK Cells in Dentric-Cell Based Vaccine.
Authors: Bos Gerard M.J. , Van Elssen Janine CHMJ , Vanderlocht Joris , Senden-Gijsbers Brigitte LMG , Germeraad Wilfred LMG 
Published: April 2020

Abstract
Figure Figure
Besides their prominent role in the destruction of altered self-cells, natural killer (NK) cells have been shown to potentiate T cell responses by interacting with dendritic cells (DC). In mouse models as well as in a recent human study it has been demonstrated that DC might activate NK cells. In the context of dendritic cell-based vaccines – i.e. optimising the optimal maturation cocktail - it remains to be determined if and how NK-DC interactions depend on differential DC maturation and what factors influence the NK activation.. By comparing differential DC differentiation (IL-4/GM-CSF and IL-13/GMCSF) and maturation cocktails (IFN-γ/FMKp and PGE2/TNF-α), we show that the ability of human DCs to attract NK cells is imprinted during DC maturation. Only FMKP/IFN-γ (stimulation Toll like receptor 2 and 4) maturated DCs have the capacity to actively recruit NK cells in vitro and our data indicate that CCR5 is the dominant chemokine receptor in this recruitment (Figure 1). Furthermore, in contrast to PGE2/TNF-α matured DC, FMKP/IFN-γ maturated DCs activate NK cells to produce IFN-γ in a IL-12/IL18 dependent manner, of which we show it contributes to strong TH1 polarization. In addition upon contact with these DCs NK cells upregulate their lymph node homing receptors, possibly inducing secondary migration to the lymph nodes.
In conclusion, besides the identification of a superior DC maturation cocktail which contributes to NK-DC interactions, we identified a novel recruitment mechanism for peripheral human NK cells which may contribute to secondary, central DC-NK interactions and strong TH1 polarization.
IL-10 Present in the B Lymphoma Microenvironment Drives Macrophages to Efficient Tumour Suppressing Cells with High Phagocytic Activity in Presence of Rituximab.
Authors: Golay Josee , Leidi Marzia , Gotti Elisa , Palumbo Giuseppe , Introna Martino 
Published: April 2020

Abstract
Since macrophages have been implicated as major players in the mechanism of action of Rituximab (Mabthera®) in vivo, we have investigated the factors that modulate their tumour cell killing potential in vitro. Human macrophages expressing CD16, CD32 and CD64, were differentiated from CD14+ peripheral blood monocytes by culture for 5–7 days in presence of M-CSF. Binding of rituximab opsonised target cells was measured after 5 minutes incubation of macrophages with CFSE labelled B-CLL cells and FACS analysis. Phagocytosis was quantified after 2 hours at 37°C by microscopic count of stained slide preparations. ADCC was measured by standard release assays. Rituximab induced specific binding of CD20+ target cells to macrophages and this was followed by phagocytosis, but not ADCC. Phagocytosis was maximal at 0.1 μg/ml rituximab and was not significantly affected by CD20 expression levels on target cells. The CD16A polymorphism at amino acid 158 (Val/Phe) that affects IgG binding did not significantly modify the efficacy of phagocytosis at different rituximab doses, possibly due to the role of additional Fcγ receptors. Indeed phagocytosis was blocked by excess human immunoglobulins. Since macrophages can be differentiated to M1 type or M2 type cells with either GM-CSF or M-CSF, respectively, and can be classically activated by pro-inflammatory stimuli (IFNγ + LPS) or undergo alternative activation with cytokines such as IL-4 or IL-10, we have analysed the effect of these different polarisation programs on the phagocytosis mediated by rituximab. Macrophages differentiated in presence of M-CSF showed a 2–3 fold greater phagocytic capacity compared to GM-CSF induced cells. Furthermore addition of IL-10 significantly increased, whereas IL-4 decreased phagocytosis by both M-CSF and GM-CSF differentiated macrophages. LPS/IFNγ had little effect. Expression of CD16, CD32A/C and CD64 correlated with the phagocytic capacity of the different polarised populations, suggesting that M-CSF and IL-10 induced maximal phagocytosis through upregulation of several activating FcγRs. Several B lymphoma cell lines were observed to secrete 400–1300 pg/ml IL-10 in vitro and co-culture of human macrophages with lymphoma supernatant increased significantly their phagocytic capacity. These data suggest that IL-10 produced in the lymphoma microenviroment may activate macrophages to become M2 type cells with high phagocytic capacity. Thus usually tumour promoting M2 type macrophages within the lymphoma would become tumour suppressing in presence of rituximab.