bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL-18 Elicited Suppressor NK Cells with Immunoregulatory Functions
Authors: Ullrich Evelyn , Terme Magali , Aymeric Laetitia , Apetoh Lionel , Chaput Nathalie , Ryffel Bernhard , Mackensen Andreas , Schultze Joachim L , Zitvogel Laurence 
Published: April 2020

Abstract
Cancer-induced tolerance involves multiple immunosuppressive pathways, which subvert adaptive immune responses against tumor cells. Extensive phenotypic characterization of NK cells from tumor-bearing mice led us to the observation of a consistent expansion of c-kit-expressing NK cells (Kit+NK cells) compromising the NK cell arm of tumor immunosurveillance. In naïve animals, Kit+NK cells (CD3−NK1.1+CD117+) represent about 3.3±0.7% of blood, 4.8±1.4% of spleen and 13.2±2.4% of lymph node NK cells. In tumor-bearing animals, percentages and absolute numbers of Kit+NK cells increased to levels that mediate inhibitory effects on mature NK cells. Purified Kit+NK cells failed to produce IFNγ or GM-CSF in response to IL-2 and could not promote DC maturation in contrast to conventional Kit−NK cells. Moreover, adoptive transfer of Kit+ (but not Kit−) NK cells into mice injected with CT26 colon carcinomas or B16F10 melanomas promoted the establishment of lung metastases. Micro array comparison of CD3−NK1.1+Kit+ and CD3−NK1.1+Kit− cells revealed profound differences in their transcriptomes. Two major sets of genes involved in tolerance (B7-H1, CTLA4, Lag3, Hmox1) or tissue repair and bone marrow homeostasis (Cxcl2, CD81, CD63, Csf2) were markedly up-regulated in Kit+NK cells. Among these, B7-H1 appeared particularly intriguing as we found that Kit+NK cells killed Kit−NK cells in a B7-H1/PD-1-dependent manner. Moreover, IL-18 produced by CT26 or B16F10 tumors converts Kit− into Kit+NK cells endowed with immunoablative functions in lymph nodes. Upon tumor inoculation, Kit+ NK cells, which upregulate B7-H1, accumulate in lymphoid organs in an IL-18R/MyD88-dependent manner and directly kill conventional NK cells, thereby promoting the progression of NK-controlled cancers. Importantly, IL-18R−/− mice lost the development of Kit+NK cells during tumor progression. Neutralization of IL-18 by RNA interference in tumors or systemically by IL-18-binding protein (IL-18BP) reduced the expansion of Kit+NK cells, stimulated the NK cell-dependent immunosurveillance and significantly reduced tumor growth.
The inflammatory cytokine IL-18 is known to accumulate in cancer-bearing patients but its pathophysiological role still remains unclear. Here, we show that IL-18 is a major tumor-derived immunosuppressant affecting the innate arm of tumor immunosurveillance. In summary, these data suggest that IL-18 promotes the expansion of Kit+NK cells endowed with immunosuppressive functions that could act as negative regulator of general NK responses.
Molecular Characteristic of Variant and Classic Hairy Cell Leukemia.
Authors: Arons Evgeny , Suntum Tara , Sunshine Joel , Orthwein Anna , Margulies Inger , Stetler-Stevenson Maryalice , Kreitman Robert 
Published: April 2020

Abstract
10–20% of patients with hairy cell leukemia (HCL) have a variant (HCLv) and present with high tumor burden in spleen and peripheral blood, and are less responsive to purine analogs. HCLv cells lack CD25 and sometimes CD103. Differentiating HCLv from classic HCL (HCLc) is sometimes difficult and it is unclear whether HCLv and HCLc are different disorders. Detailed molecular distinctions between HCLv and HCLc have not been reported. Patients with HCL were studied by flow cytometry and PCR to sequence the monoclonal immunoglobulin heavy chain rearrangements. 50 and 21 VH-D-JH rearrangements were obtained from 49 HCLc and 19 HCLv patients, respectively. All rearrangements except 2 each for HCLc and HCLv were productive. The incidence of VH4 usage was higher in HCLv than in HCLc (57% vs 20%, p=0.0041). VH4-34 was the most common VH gene in HCLv and was more common in HCLv than in HCLc (33% vs 8%, p=0.012). The percentage of rearrangements which were unmutated (defined as &lt; 2% somatic mutations) was higher in HCLv than in HCLc (43% vs 18%, p=0.038), but % homology was similar in both groups by rank order (94.9 vs 94.3% p=0.22). However, in comparing 11 VH4-34 with 60 other rearrangements, homology was higher with VH4-34 (median 99.2 vs 95.2%, p &lt; 0.0001). In fact, the higher frequency of unmutated rearrangements in HCLv vs HCLc was due to VH4-34 cases, since the 7 HCLv VH4-34 rearrangements were all unmated and had higher homology than the other 14 HCLv rearrangements (median 99.6 vs 94.2%, p=0.006). Moreover, unmutated rearrangement incidence was similar between HCLv and HCLc for non-VH4-34 cases (14 vs 13%, p=1.0, median homology 94.2% vs 95.3%, p=0.5). Of the 4 VH4-34+ HCLc (CD25+) patients, 3 (75%) had unmutated rearrangements, and these 3 all had clinical features of HCLv, including presenting with lymphocytosis, large splenomegaly, absent cytopenias, and primary failure of purine analog treatment. Our data shown that homology of monoclonal immunoglobulin rearrangement to the germline sequence of HCL patients appears more related to VH4-34 status than to whether patients have HCLv or HCLc. In fact, no molecular distinctions between HCLv and HCLc were observed in VH4-34-negative patients. Our data suggest VH4-34-positive HCL is itself a variant of HCL affecting ~15% of our patients. Such patients have features of HCLv, but their HCL cells can be CD25+.
Treatment with Imatinib in Very Elderly (&gt; 75 Years) CML Patients.
Authors: Ata Roberto Latagli , Breccia Massimo , Sica Simona , Spadea Antonio , D’Addosio Ada , Carmosino Ida , De Matteis Silvia , Stefanizzi Caterina , Volpicelli Paola , Napoleone Laura , Vozella Federico , Petti Maria Concetta , Alimena Giuliana 
Published: April 2020

Abstract
Median age of patients with CML is about 65 years, according to SEER 2000–2005 data. However, the treatment with Imatinib in elderly has been seldom specifically addressed: in particular, there is a lack of data regarding very elderly CML patients. To highlight peculiar aspects of toxicity and efficacy of Imatinib in this subset which accounts for at least 10–15% of all CML cases, we revised retrospectively 28 CML patients treated with Imatinib when aged &gt; 75 years from 4 haematological Institution in Rome; there were 12 males and 16 females, median age at Imatinib was 78.7 years (IR 75.8 – 82.9), Sokal Risk at diagnosis was intermediate in 20 patients, high in 6 and not valuable in 2. One or more concomitant diseases requiring specific treatments were present in 26/28 patients (92.8%), with 15 patients (53.5%) assuming 3 or more concomitant drugs. Nine patients (32.1%) have been pretreated ≥ 6 months with HU before starting Imatinib; on the whole, median time from diagnosis to Imatinib was 1.8 months (IR 0.7 – 25.1). Starting dose of Imatinib was 400 mg/day in 23 patients (82.1%) and 300 mg/day in 5 patients (17.9%); overall, 15 patients (53.5%) (14/23 at 400 mg starting dose and 1/5 at 300 mg starting dose) needed a dose reduction and 6 (21.4 %) discontinued Imatinib for toxicity (early toxicity in 4 and late toxicity in 2). Excluding the 4 patients who discontinued Imatinib due to early toxicity, maximum tolerated daily dose during treatment was 400 mg in 8 patients, 300 mg in 13 patients and 200 mg in 3 patients. According to CTC-AE, grade 3 – 4 haematological and extra-haematological toxicities were observed in 7 (25%) and 12 (42.8%) patients, respectively; 2 patients (7.1%) presented a pleural effusion during Imatinib treatment. All patients were fully evaluable for response to Imatinib, with a median treatment period of 30.6 months (IR 15.7 – 49.2); six patients (21.4%) failed any response, (including 4 patients who discontinued Imatinib due to early toxicity) and 22 (78.6%) achieved a complete haematological response (CHR). Among these 22 patients in CHR, 4 refused any other karyotipic or molecular evaluation (3 are still alive in CHR, 1 died in CHR from unrelated cause after 44,5 months) and 17/18 (60.7% of all 28 patients) achieved a cytogenetic response (CyR), major in 1 patient and complete in 16 patients. In addition, 7 patients (25% of all 28 patients) achieved a complete molecular remission, with an undetectable BCR/ABL hybrid gene at qualitative nested PCR. In conclusion, Imatinib at reduced daily dose of 300 mg seems a relatively safe and quite effective treatment for very elderly CML patients; from our data, no upper age limit should be given for TKinhibitors treatment but also very elderly (and with concomitant severe diseases) patients should have this chance of cure.
DNA-Based Monitoring of Minimal Residual Disease(MRD) in Chronic Myeloid Leukemia(CML).
Authors: Morley Alexander A. , Bartley Paul , Ross David M , Latham Sue , Martin-Harris Henry , Budgen Brad , Wilczek Vicki , Branford Susan , Hughes Timothy 
Published: April 2020

Abstract
Aim: To develop and evaluate a DNA-based method for monitoring of MRD in CML
Background: At present, monitoring of MRD in CML uses RNA and reverse transcription (RT-PCR). This leads to a number of disadvantages, including the potential for RNA degradation, requirement for reverse transcription, difficulty in standardisation and only an indirect relationship between assay result and cell number.
Methods: A highly multiplexed, short-range PCR using six BCR primers and a pool of 282 ABL primers was used to amplify across the BCR-ABL translocation breakpoint and was followed by 2–3 rounds of “bottleneck PCR”, a technique recently-developed in our laboratory, which adjusts primer concentration so as to minimise non-specific amplification and facilitate highly multiplexed PCRs. The breakpoint was isolated and sequenced, patient-specific primers were synthesised, and MRD was quantified using 10 μg of DNA and a 3 round nested quantitative PCR incorporating a Taqman probe in the third round. Samples from patients on treatment were divided and assayed both by this technique and by RT-PCR. 10 μg of DNA from a normal individual was used in each assay as a control for non-specificity.
Results: The BCR-ABL breakpoint was successfully isolated and sequenced in 28 of the 29 patients studied and MRD was assayed in 38 samples from 24 patients. Follow-up samples from 4 patients were unavailable MRD was detected and measured by both methods in 22 samples, detected and measured only by DNA-PCR in 10 samples and not detected by either method in 6 samples. The limit of detection of RT-PCR was in accord with previous results which indicated it to be a mean decrease of 4.5-logs below baseline. The mean limit of detection of DNA-PCR was an MRD of 7.2 x 10−7. Assay precision was determined by performing independent replicate assays on different days and by different individuals on the 32 samples with MRD detectable by DNA-PCR. The median SD of a single assay was 0.15 log units with the range being 0.00 – 0.64 log units.
Figure Figure
Conclusions
Monitoring of MRD by DNA-PCR is feasible in the great majority of patients with CML. In terms of possible clinical benefits, DNA-PCR is more sensitive than RT-PCR, and provides a direct measure of leukemic cell number in the individual patient. Use of DNA rather than RNA simplifies specimen collection and transport. In terms of laboratory benefits, DNA-PCR obviates reverse transcription and inter-laboratory standardisation. The principal disadvantage of the method is the initial cost, although this may be able to be amortised over several assays if monitoring is ongoing.
Monitoring of Adenovirus Load in Stool by Real-Time PCR Permits Early Detection of Impending Invasive Infection in Patients after Allogeneic Stem Cell Transplantation
Authors: Lion Thomas , Kosulin Karin , Landlinger Christine , Rauch Margit , Jugovic Dragana , Pötschger Ulrike , Lawitschka Anita , Peters Christina , Fritsch Gerhard , Matthes-Martin Susanne 
Published: April 2020

Abstract
Disseminated adenovirus (AdV) infection is associated with high mortality in allogeneic stem cell transplant recipients. We have demonstrated that molecular detection of AdV in peripheral blood (PB) precedes the onset of life-threatening virus disease (Lion et al., Blood 102:1114–20, 2003). In most instances, detection of AdV in stool preceded the onset of viremia, thus raising the possibility that intestinal infections represent a common source of virus dissemination. To address this question, we have investigated 170 consecutive allogeneic transplantations in 138 pediatric patients transplanted at our center. Patients were monitored for the presence and the load of AdV in stool and in PB by a real-time PCR approach covering the entire spectrum of known human AdV serotypes. Fifty one patients (37%) tested positive in serial stool samples, revealing adenoviruses of nearly all subgenera, with strong predominance of serotypes 2 and 1 of species C. The overall risk of developing viremia in patients displaying or lacking intestinal AdV infection was 31% and 0%, respectively (p&lt;0.0001). Among patients who had maximum AdV levels in stool ≤1×10E6 copies/g (n=29) neither experienced viremia during the post-transplant course, while the incidence of viremia in individuals with peak virus levels in stool above this threshold (n=22) was 73% (p&lt;0.0001). In contrast to earlier observations, the recovery of T-cells including the CD4 and CD8 subsets showed no statistically significant correlation with high AdV loads in stool, occurrence of viremia or disseminated disease (p-values for CD4 and CD8 cells: 0.5833 and 0.6313). The recovery of NK cells showed a borderline-significant correlation with the above parameters (p=0.0442). Our observations indicate that serial measurement of AdV levels in stool specimens by RQ-PCR provides a parameter permitting early diagnosis of impending invasive infection. The median time span between detection of AdV load in stool above 1×10E6 copies/g and first observation of viremia was 10 days (range 0–191). Quantitative monitoring of AdV loads in stool could therefore provide a rationale for early onset of antiviral treatment in attempts to prevent progression to life-threatening systemic infection.
Induction of Hyperproliferative Fetal Megakaryopoiesis by An N-Terminally Truncated GATA1 Mutant.
Authors: Shimizu Ritsuko , Kobayashi Eri , Engel James , Yamamoto Masayuki 
Published: April 2020

Abstract
Two types of GATA1-related leukemias have been noticed. One is the erythroid leukemia develops in mice as a consequence of deficient expression of GATA1, while the other is acute megakaryoblastic leukemia (AMKL) that arises in Down syndrome (DS) children as a consequence of somatic N-terminally truncation (ΔNT) mutation of GATA1 (reviewed in Shimizu et al, Nat Rev Cancer 2008). We hypothesized that modeling of the human mutation in mice might provide novel insights into human TMD and DS-AMKL, since the GATA1 N-terminal (NT) function is apparently important in these disorders. A previous report showed that immature megakaryocytic progenitors in the yolk sac and early fetal liver of knock-in mice that expressed a mutant GATA1 protein lacking amino acids 3 to 63 (and not 1–83, as in the DS-associated AKML patients) developed a hyperproliferative phenotype, but also that these aberrant cells disappeared from fetal blood by E14.5. The same phenotype was detected in conditional knockout mice in which Gata1 exon 2 (which encodes the wild-type start codon) was deleted. Since this phenotype was substantially different from human TMD, the contribution of the GATA1 NT 83 amino acids to human TMD pathogenesis remained obscure. Therefore, we adopted a transgenic complementation/rescue approach to more directly analyze NT domain function. We generated mice that were rescued from GATA1-deficient lethality by transgenic expression of a GATA1 mutant lacking the N-terminal 83 amino acids, a protein that is identical to the mutant GATA1 factor encountered in human TMD and DS-AKML cases. We discovered that expression of this shortened GATA1 protein in mice phenocopies human TMD in DS children preceding AMKL; immature megakaryocytes accumulate massively in the perinatal livers of mutant mice. However, unlike the usual disease progression in humans, these mouse mutants never develop leukemia. These results unequivocally revealed that expression of the short form of GATA1 is sufficient to induce hyperproliferative fetal megakaryopoiesis, but additional mutation(s) to the GATA1/Gata1 N-terminal deletion is necessary to transform those hyperproliferative megakaryocytic cells into overt leukemic cells in both mouse and human. Thus, this model significantly clarifies our understanding of the distinct molecular mechanisms by which the GATA1 protein can contribute to the multi-step leukemogenesis.
NK-Like Homeodomain Proteins Regulate NOTCH3-Signaling in T-Cells.
Authors: Nagel Stefan , Venturini Letizia , Przybylski Grzegorz K. , Grabarczyk Piotr , Meyer Corinna , Kaufmann Maren , Battmer Karin , Schmidt Christian A. , Drexler Hans G. , Scherr Michaela , MacLeod Roderick A.F. 
Published: April 2020

Abstract
Three NK-like (NKL) homeobox genes, TLX1/HOX11, TLX3/HOX11L2 and NKX2- 5/CSX, have been implicated in T-cell acute lymphoblastic leukemia (T-ALL). Here we screened further NKL genes in 24 T-ALL cell lines by RT-PCR and identified common expression of MSX2, highlighting this homeobox gene as a potential physiological family member in T-cells. Subsequent quantification of MSX2 confirmed expression in primary hematopoietic cells demonstrating higher levels in CD34+ stem cells when compared to peripheral blood cells or mature CD3+ T-cells. Analysis of core thymic factors in T-ALL cell lines, including IL7, BMP4, TGFbeta, NOTCH and T-cell receptor signaling, suggests their involvement in MSX2 regulation during T-cell differentiation. Chromosomal and genomic analysis of the MSX2 locus (at 5q35) uncovered deletion in t(5;14)(q35;q32) positive T-ALL cell lines associated with low expression levels of MSX2 and ectopic activation of TLX3 or NKX2-5, respectively. For functional analysis we lentivirally transduced T-ALL cells for overexpression of either MSX2 or oncogenic TLX1 and NKX2-5. These cells displayed transcriptional activation of NOTCH3-signaling, as indicated by expression array profiling and real-time PCR analysis of NOTCH3, HES1 and HEY1. The sensitivities to gamma-secretase inhibitor analyzed by MTT-assay of cells overexpressing MSX2, TLX1 or NKX2-5, respectively, were consistently decreased. Furthermore, in addition to MSX2, both TLX1 and NKX2-5 proteins interacted with repressor proteins of the NOTCH-pathway, SPEN/MINT and TLE1/GRG1, as shown by co-immunoprecipitation, probably representing one mechanism of (de)regulation. Elevated expression of NOTCH3 and HEY1 mRNA was detected in TLX1/3 positive T-ALL patients, confirming data obtained from cell lines. In conclusion, we have defined expression patterns, regulation and targets of MSX2 in hematopoietic cells, to reveal a novel modulatory activity in T-cell differentiation operating via NOTCH-signaling, and in leukemogenesis when replaced or supplemented by oncogenic NKL homeodomain proteins.
Results of the First International Round Robin for the Quantification of Urinary and Plasma Hepcidin: Need for Standardization
Authors: Kroot Joyce JC , Kemna Erwin HJM , Bansal Sukhvinder , Busbridge Mark , Campostrini Natascia , Girelli Domenico , Hider Robert C , Koliaraki Vasiliki , Mamalaki Avgi , Olbina Gordana , Tomosugi Naohisa , Tselepis Chris , Ward Douglas G , Ganz Tomas , Hendriks Jan CM , Swinkels Dorine W. 
Published: April 2020

Abstract
The hepatic peptide hormone hepcidin plays a central role in body iron metabolism. Quantification of hepcidin concentrations in urine and plasma holds promise as a biomarker for diagnosis and monitoring of disorders of iron metabolism. To date, various mass spectrometry (MS, time of flight and liquid chromatography-MS/MS) as well as immunochemical (IC, competitive radio-immunoassay and ELISA) and functional methods exploiting an internal or external standard, have been developed to measure hepcidin levels in both plasma and urine. Only a few of them have been published. The hepcidin levels quantified by these methods are likely to differ substantially in analytical characteristics. Therefore, comparative studies are imperative for progress in the field by allowing meaningful exchange of hepcidin results derived from various laboratories. In this study we compared the analytical characteristics of several available urinary and plasma hepcidin assays as a first step towards standardization of hepcidin quantification methods. Six and 8 laboratories participated in an international inter-laboratory evaluation of hepcidin levels from a common human urinary (n=8) and plasma (n=7) sample set, respectively, exploiting MS (n=5) and IC (n=3) based methods. Participating laboratories were requested to perform 3 hepcidin measurements for each sample on 4 different days, making a total of 96 urine and 84 plasma measurements per laboratory. The methods differed widely in mean urine and plasma hepcidin level, i.e. range 2.9–427.1 nmol/mmol creatinine and 11.4–124.6 nM, respectively. Coefficient of variation (CV)-ranges for the between-sample and withinsample variation were 126.1–191.5 % and 10.8–33.8%, respectively, for urine and 40.4–96.6 % and 5.5–31.0%, respectively, for plasma. The ratio between-sample CV/within-sample CV varies between 15.3 and 4.1 for urine and 15.1 and 2.4 for plasma. Hepcidin results of most methods mutually correlated (plasma: R range 0.82–1.00; urine: R range 0.73–0.99). We conclude that for the various methods
absolute hepcidin values differ widely, but generally correlate and the ratio between-sample and within sample CV vary greatly, suggesting relevant differences in ability to detect small differences in hepcidin concentrations.
To allow future comparison, exchangeability and transferability of hepcidin data, we recommend
publication of all hepcidin results according to STARD (www.stard-statement.org) to ensure complete reporting of analytical characteristics and to develop initiatives for standardization of assays.