bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibition of CKS1B Expression Affects Myeloma Cell Growth In Vivo.
Authors: Chen Bangzheng , Liu Ling , Mitchell Kristin L. , Wu Xiasong , Colla Simona , Zhan Fenghuang , Shaughnessy John D. 
Published: October 2019

Abstract
CKS1B is the regulatory component of the SCF-Skp2 ubiquitin ligase that ubiquinates the cyclin-dependent kinase inhibitor p27kip1 targeting it for proteasomal degradation. Overexpression of CKS1B is associated with poor prognosis in malignancies such as oral squamous cell carcinomas, colorectal carcinoma, breast cancer, prostate cancer, and others. Recently, high expression levels of CKS1B were associated with an aggressive course of Multiple Myeloma (Shaughnessy J. Hematology. 2005;10 Suppl 1:117). We investigated whether inhibition of CKS1B in myeloma cell lines with multiple copies of 1q21 and high expression of CKS1B affects in vivo tumor growth. Myeloma cell lines were transduced with lentivirus conditionally expressing (tet on) CKS1B shRNA and lentivirus constitutively expressing luciferase (for in vivo experiments), or with lentivirus constitutively expressing CKS1B shRNA (for in vitro work). Cells with inducible expression were injected into the human bones of SCID-hu mice, and, once tumors established, expression was induced with doxycycline in the drinking water. Changes in tumor sizes were followed by weekly luminescence imaging. Cells with constitutive shRNA expression were used to determine effects on cell cycle by propidium iodide flow cytometry. Effects of shRNA expression on cytoplasmic and nuclear CKS1B and p27kip1 levels were determined by western blot analysis. Induction of CKS1B shRNA expression in JJN3 and OCI-MY5 cells in SCID-hu mice inhibited tumor growth by 57% and 81% within one week, compared to non-targeting construct. As expected, viable cells recovered from these tumors had low levels of CKS1B and higher levels of p27kip1 proteins. Similar to observations in the MM cell lines JJN3 and OCI-MY5 (see Abstract by Zhan and colleagues), constitutive expression of shRNA in CAG and XG1 cells effectively reduced cytoplasmic and nuclear CKS1B protein levels. This was associated with increased levels of p27kip1 protein, marked (51%) increases of cells in the G1 phase of the cell cycle, and reduced S and G2 (33% and 52%, respectively), compatible with a G1 arrest, and a 76% increase in the number of sub G1 (apoptotic) cells. In contrast to these observations, cells constitutively over-expressing p27kip1 had no change in G1, 50% fewer cells in S, and a 150% increase in G2, indicating G1 and G2 arrests, with no change in the sub G1 fraction. Interestingly, cells over-expressing p27kip1 had lower levels of CKS1B proteins, compatible with the reported proteasomal degradation of the molecule during the later phases of the cell cycle (Hattori T et al, Genes Cells.2003;8:889). These observations suggest that the poor prognosis associated with 1q21 amplifications and high CKS1B expression in myeloma reflects an elevated proliferative state of the tumor cells that results in rapid regrowth of minimal residual disease.
CD4+ Peptide Epitopes from the WT1 Oncoprotein Stimulate CD4+ and CD8+ T Cells That Recognize and Kill Leukemia and Solid Tumor Cells.
Authors: May Rena J. , Pinilla Javier , Korontsvit Tatyana , Zakhaleva Victoriya , Zhang Ronghua H. , Dao Tao , Maslak Peter , Scheinberg David A. 
Published: October 2019

Abstract
Wilms tumor protein 1 (WT1) is a transcription factor over-expressed in several types of leukemias and solid tumors, making it an ideal target for immunotherapy. A number of class I binding WT1 peptides have been identified and shown to stimulate CD8+ T cells. These peptides are being tested as potential cancer vaccine candidates in a variety of clinical trials. However, the induction and maintenance of a robust memory CD8+ cytotoxic T cell response requires CD4+ T cell help. Herein we report the identification of three HLA Class II peptide epitopes of WT1 using the SYFPEITHI and RANKPEP predictive algorithms. Peptides 328–349 and 423–441 are able to stimulate a peptide specific CD4+ response that can recognize WT1 positive tumor cells in multiple HLA-DRB*1 settings, as determined by IFN-gamma ELISPOT assays. Due to the highly polymorphic nature of the HLA class II alleles, such broad reactivity is critical in the development of a universal therapeutic. In addition, we identified a WT1 CD4+ peptide epitope (122–140) that lies within close proximity to a previously identified CD8+ peptide epitope (126–134). Residue 126 was mutated from an Arginine (R) to a Tyrosine (Y) thereby embedding a synthetic immunogenic analog CD8+ peptide that was previously designed to improve immunogenicity and induce a potent CD8+ response. Mutated peptide 122–140 is able to induce a CD4+ and cytotoxic CD8+ WT1 specific T cell response that can recognize the native WT1 epitopes on the surface of human CML and solid tumor cells. Cross-priming experiments demonstrated that APCs pulsed with either CML or mesothelioma tumor lysates can process and present each of the CD4+ peptides identified. These studies provide the rationale for using the three WT1 CD4+ peptides in conjunction with CD8+ peptide epitopes to vaccinate patients with WT1 expressing cancers.
A Novel Strategy for Generation of Human Tumor-Specific T Cell Clones for Adoptive Transfer.
Authors: Lee Seung-Tae , Liu Shujuan , Sukhumalchandra Pariya , Molldrem Jeffrey , Hwu Patrick , Liu Yong-Jun , Kwak Larry W. , Lizee Gregory , Neelapu Sattva S. 
Published: October 2019

Abstract
Adoptive T-cell therapy using donor lymphocyte infusions is a promising approach for treating hematological malignancies. But, efficacy is limited by the induction of graft-versus-host disease. Transfer of tumor-specific T-cell clones could enhance the graft-versus-tumor effect and eliminate graft-versus-host disease. However, isolating antigen-specific T-cell clones by the traditional limiting dilution approach is a time-consuming and laborious process. Here, we describe a novel strategy for rapidly cloning tumor-specific T cells. Lymphoma-specific T-cell lines were generated from two follicular lymphoma patients by repeated in vitro stimulation of lymphocytes isolated from tumor or blood with autologous soluble CD40 ligand-activated tumor cells. After four in vitro stimulations at 10-day intervals in the presence of IL-2 and IL-15, T-cell lines were found to be predominantly CD4+ T cells and produced significant amounts of TNF-a, GM-CSF, and IFN-γ in response to autologous tumor cells. The tumor reactivity was MHC class II restricted suggesting that it was mediated by CD4+ T cells. Staining with a TCR Vb antibody panel, a set of monoclonal antibodies against 24 human TCR Vb families, revealed that certain Vb families were overrepresented in each CD4+ T-cell line. In patient 1, 51% of CD4+ T cells were Vb1 positive, and in patient 2, 27% of CD4+ T cells were Vb8 positive. To clone lymphoma-specific T cells, CD4+ T-cell lines were labeled with CFSE and stimulated with autologous tumor cells. After 9 days of in vitro expansion in the presence of IL-2 and IL-15, CD4+ T-cell lines were stained with an anti-human CD4-APC monoclonal antibody and an anti-human TCR Vb-PE monoclonal antibody for each CD4+ T-cell line. Proliferating Vb1 cells from patient 1 and Vb8 cells from patient 2 were identified by their reduction in CFSE staining, and CD4+TCRV b +CFSEdim cells were sorted by flow cytometer. Monoclonality of the sorted cells was confirmed by PCR using a panel of optimized primers specific for 24 TCR Vb families, by TCR Vb spectratype analysis, and finally, by sequencing the TCR Vb gene used by each T-cell clone. Sorted tumor-specific T-cell clones could be expanded to large numbers using a 14-day rapid expansion protocol with allofeeder PBMCs, and confirmed to retain specificity against autologous tumor cells in a cytokine induction assay. This approach was also successfully used to isolate melanoma-specific CD8+ T-cell clones from two patients. We conclude that this approach is highly reproducible, rapid, and efficient for generating antigen-specific T-cell clones for adoptive T-cell therapy against human malignancies in the autologous or allogeneic setting.
Lenalidomide Strongly Enhances Natural Killer (NK) Cell Mediated Antibody-Dependent Cellular Cytotoxicity (ADCC) of Rituximab Treated Non-Hodkin’s Lymphoma Cell Lines In Vitro.
Authors: Wu Lei , Schafer Peter , Muller George , Stirling David , Bartlett J. Blake 
Published: October 2019

Abstract
Lenalidomide (Revlimid® is approved for the treatment of transfusion-dependent patients with anemia due to low- or intermediate-1-risk MDS associated with a del 5q cytogenetic abnormality with or without additional cytogenetic abnormalities, and in combination with dexamethasone is for the treatment of multiple myeloma patients who have received at least one prior therapy. Encouraging early results suggest a potential for clinical efficacy in B cell non-Hodgkin’s lymphoma (NHL). Potential mechanisms of action include anti-angiogenic, anti-proliferative and immunomodulatory activities. Lenalidomide has been shown to enhance Th1-type cytokines and T cell and NK cell activation markers in patients with advanced cancers. Furthermore, lenalidomide has been shown to enhance rituximab-mediated protection in a SCID mouse lymphoma model in vivo. We have utilized an in vitro ADCC system to assess the ability of lenalidomide to directly enhance human NK cell function in response to therapeutic antibodies, such as rituximab (chimeric anti-CD20 mAb). Isolated NK cells produced little or no IFN-γ in response to IgG and/or IL-2 or IL-12. However, pre-treatment of NK cells with lenalidomide greatly enhanced IFN-γ production by NK cells in a dose-dependent manner. In a functional ADCC assay, NHL cell lines (Namalwa, Farage & Raji) were pre-coated with rituximab and exposed to NK cells pre-treated with lenalidomide in the presence of either exogenous IL-2 or IL-12. After 4 hours in culture the viability of the tumor cells was assessed. Lenalidomide consistently and synergistically increased the killing of tumor cells in a dose-dependent manner and up to >4-fold compared to rituximab alone. Rituximab alone had only a small effect in this model and there was no killing of cells in the absence of rituximab. The presence of either exogenous IL-2 or IL-12 was required to see enhanced killing by lenalidomide. In cancer patients lenalidomide has been shown to increase serum IL-12 levels and is also known to induce IL-2 production by T cells in vitro. Potential mechanisms for enhanced ADCC include increased signaling through NK FCγ receptors and/or IL-2 or IL-12 receptors. However, we found that these receptors are unaffected by lenalidomide, although downstream effects on NK signaling pathways are likely and are being actively investigated. In conclusion, we have shown that lenalidomide strongly enhances the ability of rituximab to induce ADCC mediated killing of NHL cells in vitro. This provides a strong rationale for combination of these drugs in patients with NHL and CLL.
The Effects of Iron Deficiency on Neutrophil/Monocyte Oxidative Burst Response in Children.
Authors: Berrak S.G. , Angaji Meryem , Turkkan Emine , Canpolat Cengiz , Timur Cetin , Demiralp-Eksioglu Emel Z. 
Published: October 2019

Abstract
Background: Microbial killing killing of by phagocytic cells by iron containing myeloperoxidase pathway is part of the immune system. We aimed to investigate the effects of iron deficiency anemia (IDA) on oxidative burst response of phagocytic cells and understand if the effect is reversible after iron supplementation.
Material and methods: There were 57 IDA patients and 31 healthy children as the control group that were aged between 6 months-12 years with similar demographic status. IDA patients were classified according to the severity of their anemia as follows; group 1 (Hb<8 gr/dl), group 2 (Hb≥8 gr/dl and Hb≤10 gr/dl), and group 3 (Hb>10 gr/dl). Neutrophil and monocyte oxidative burst response of each three groups of IDA and control group were compared by flow cytometry. IDA group were given oral iron supplementation (3 mg/kg-Ferrosanol) therapy. On day 15 of iron supplementation therapy neutrophil and monocyte oxidative burst response of each three groups of IDA were rechecked and compared to the control group. In order to minimize the discrepancies observed in functional studies, the oxidative burst index was calculated as the proportion of oxidative burst response in IDA group to the simultaneous oxidative burst response in the control group.
Results: The number of infection episodes observed in the last three months was significantly higher in the IDA group (0–13 episodes) than the control group (1–2 episodes) (p=0.05). Neutrophil (p=0.1) and monocyte (p=0.02) oxidative burst responses in IDA group were found to be less than the control group. On day 15 of iron supplementation therapy, these values were observed to increase to the control group’s values. Evaluation of neutophil and monocyte burst indexes at the diagnosis of IDA revealed that the neutrophil burst indexes in the Hb ≤10 gr/dl group (p=0.006), and monocyte burst indexes in the Hb <8 gr/dl group (p=0.01) were significantly low. The initially observed significant differences in oxidative burst responses disappeared at day 15 of iron supplementation therapy in all hemoglobin groups.
Conclusion: Correction of the differences in oxidative burst response after iron supplementation therapy implies the fact that IDA might be the reason for changes in neutophil and monocyte burst indexes.
Analysis on the Correlation of RPS19 Expression Level to Diamond-Blackfan Anemia; Insights from the Expression Level of RPS19 Point Mutants in Erythroid Cell Lines.
Authors: Kuramitsu Madoka , Hamaguchi Isao , Takuo Mizukami , Masumi Atsuko , Momose Haruka , Takizawa Kazuya , Motizuki Masayo , Naito Seishirou , Yamaguchi Kazunari 
Published: October 2019

Abstract
Mutations in ribosomal protein S19 (RPS19) gene is closely related to Diamond-Blackfan anemia (DBA). We have found point mutation, deletion and insertion in RPS19 gene in DBA patients (Blood, 100: 2724–31, 2002). In our analysis of DBA patients, gene expression of RPS19 from these mutated genomes were down regulated. Retroviral transduction of RPS19 gene to patient bone marrow cells restored the ability to differentiate into erythroid in vitro (Mol Ther., 7: 613–22, 2003). However, the mechanisms of how these mutations in RPS19 associate with anemia remains unknown.
To analyze the mutated genome, we have made constructs of mutated RPS19 gene-expression vectors. From reported 56 different RPS19 mutations, we focused on the missense mutants and tried to characterize them. Flag-tagged twelve missense mutants (V15F, L18R, P47L, W52R, R56Q, S59F, A61Q, R62W, R101H, G120R, G127Q, and G131R) were exogenously expressed in several cell lines by retroviral vectors and were analysed by western blotting or FACS. When these 12 mutants were expressed in erythro-leukemic cell lines K562 and HEL, almost all the mutant proteins (except for G120R) were expressed at significantly low level compared to that of wild type. However, some of these mutants were expressed in human kidney epithelial cell line, 293T cells. These findings suggest that the expression level of RPS19 protein derived from mutated RPS19 genome is dependent on cell types, whether erythroid-lineage cells or others.
In order to analyze the function of RPS19, doxycycline (Dox)-inducible expression of siRNA against RPS19 was induced in K562 cells and observed growth arrest at day 4 after induction of siRNA by Dox. Expression level of RPS19 decreased to one tenth of the normal level. Cell cycle analysis and quantitative-PCR analysis revealed that the growth arrest of K562 cells expressing siRNA against RPS19 was due to the induction of cell cycle arrest at G1 phase (G0/G1 54 % vs 36% ) coincide with the up-regulation of p21 and p57 mRNA level. From these results, we could possibly suggested that, in DBA patients, RPS19 protein expression is not stable and the decreased level of RPS19 protein prolongs G1 phase of cell cycle especially in the course of differentiation on erythroid cells, which may contribute to the retarded production of red blood cells.
Our findings allow to understand the entire feature of DBA-associated RPS19 mutations especially on the erythorid differentiation and may help defining the therapeutic target to improve DBA.
Clinical Characteristics of Patients with Fanconi Anemia in Complementation Group J.
Authors: Dasi-Carpio Angeles , Casado Jose A. , Sanz Guillermo , Rio Paula , Santos Maria-Teresa , Bueren Juan A. , Surralles Jordi , Monteagudo Emilio , Carreras Maria C. 
Published: October 2019

Abstract
Fanconi anemia (FA) is an inherited disorder characterized by chromosomal instability, developmental abnormalities, progressive bone-marrow failure, and cancer susceptibility. To date, twelve complementation groups have been described for FA (A, B, C, D1, D2, E, F, G, I, J, L and M) and eleven associated genes have already been identified (FANCA, FANCB, FANCC, FANCD1/BRCA2, FANCD2, FANCE, FANCF, FANCG/XRCC9, BRIP1/FANCJ, FANCL and FANCM/Hef). The FA complementation group J (FA-J) was first reported in 2004. FA-A is the most common group, accounting for approximately 65% of all affected individuals while FA-J is a rare group, comprising only 1.6% of all FA patients in the International Fanconi Anemia Registry (IFAR). To the best of our knowledge 11 FA-J patients have been reported in the IFAR. Patients with Fanconi anemia show extreme clinical heterogeneity.
Thus, it is of interest to associate the phenotype with the complemantation group, since this could provide predictive information on clinical outcome.
So far, in the Hospital La Fe, 4 patients (two siblings and two unrelated patients) belong to scarce FA-J complementation group. This represents the 4.5 % of the FA patients included in the Spanish Fanconi Anemia Registry. Subtyping was done by viral complementation. Cells from these patients were not complemented with a battery of vectors with FA genes (FANCA, FANCC, FANCG, FANCE, FANCF, and FANCD2). However, the characteristic cellular hypersensitivity to mitomycin C was reverted when a lentiviral vector encoding for BRIP1/FANCJ was used. Moreover, mutational studies confirmed the subtyping of FA-J in these Fanconi patients. The clinical course of the 4 FA-J patienst are shown in the Table.
In conclusion, its noteworthy the high frequency of the FA-J genotype in Spain. The 75% of our patients have major congenital malformations and severe endocrinopathy. All our patients had short stature and 50% were mycrosomic. None of our patients had skin abnormalities, although this is present in 80% of the patients in other registries. Although two of these patients developed aplasia very early (5 years old), FA-J siblings developed severe aplasia at the age of 20 years but none of them developed leukemia or solid tumors.
Clinical course of patients Patient 1* 2* 3 4 * Sibling sex male female male female Aplasia onset (years) 24 20.5 4.3 3.5 Survival (years) 28 31.5 6 alive Short stature Yes Yes Yes Yes Microsomy No Yes Yes No Skin pigmentation No No No No Eyes abnormalities No No microphthalmia Epiphora Skeletal malformations Syndactyly No Abnormal thumbs Abnormal thumbs Kidneys Agenesis, left No No Horseshoe Endocrinopathy failure puberty not available hypotiroidism hypopituitarysm Gastrointestinal malformations No No Esophageal atresia Onphalocele Malignacies No No No No Family cancers Not available Not available Liver Lung, brains and breast
The Changing Map of the Sickle Cell Gene in Kenya.
Authors: Aluoch Joash R. , Aluoch Louise H.M. 
Published: October 2019

Abstract
In a ten year prospective records survey we received reports of sickle cell anemia (SCA) from ethnic groups which initially had no sickle cell trait (SCT) in Kenya. A follow up study was done in 2000 using alkaline Hb-electrophoresis to reassess the current status of SCT in comparison with a study reported in the 1950’s. We found that of the initially unaffected groups, SCT was 8% in the sabaot, 6% in the Taita, 3% in the inland (Mt. Kenya) Kikuyu, 1% in the Turkana, but still absent in the Somali. There was also a shift in %-ages from the 1950 survey in tribes with SCT: 33% in the Suba in stead of 28%, and 22% in the Kambe instead of 35%. We also noted a remarkable difference between cosmopolitan and rural populations in two SCT endemic areas: 15% in Kisumu city against 28% rural neighborhood and 15% in Bungoma town against 21% in its rural population. It appears that sickle cell (HbS) gene distribution and spread is changing in Kenya and that this trend most likely applies to the rest of Africa as well. The findings might be useful in genetic counseling as well as for diagnosis of SCA on clinical grounds in ethnic groups among whom SCA was hitherto considered to be absent.
Cell Free Fetal and Total DNA Levels in Pregnancies at Risk of Sickle Cell Disease and Significant Ethnic Variation.
Authors: Gerovassili Ageliki , Nicolaides Kypros H. , Thein Swee Lay , Rees David 
Published: October 2019

Abstract
Cell free (cf) DNA in maternal circulation is increasingly investigated in pregnancy. We aimed to determine if sickle cell trait women had quantitative differences of cfDNA with controls and if there was an ethnic difference between the cfDNA levels of Northern European and African/African-Caribbean populations. Non-invasive prenatal diagnosis through quantification of fetal and total cfDNA was tested in 33 pregnant women at risk of carrying a fetus affected with sickle cell disease and 124 control pregnancies. Fetal cfDNA assays were based on two Y chromosome specific markers (SRY and DYS14) and total cfDNA was based on the β-globin gene on chromosome 11. Maternal age (MA), gestation age (GA), fetal sex, storage time prior to extraction, maternal weight and body mass index (BMI) before pregnancy were compared to the fetal and total cfDNA levels. No significant difference in the fetal or total cfDNA levels was found between any of the control pregnancies and the sickle cell trait mothers carrying HbAA, HbAS and HbSS fetuses. However, higher levels of total cfDNA, but lower fetal cfDNA levels were observed in the African/African-Caribbean population compared with the Nothern Europeans. A significant variation in cfDNA was found between ethnic groups, which should be taken under consideration in future studies measuring cfDNA.
Bone Density Measurements in Thalassemia Patients with Iron Overload.
Authors: Grosse Regine , Frieling Isolde , Kuehl Bettina , Janka Gritta E. , Fung Ellen B. 
Published: October 2019

Abstract
Since the introduction of an improved blood transfusion and iron chelation therapy in patients with beta-thalassemia major (TM) and intermedia (TI), changes in bone structure and bone deformities are significantly diminished. However, older patients with thalassemia have a higher prevalence for fractures caused by osteoporosis. Additionally, most of these patients suffer from chronic bone pain.
In 13 patients (10 TM und 3 TI, age 8 – 32 years), we have measured the bone mineral density (BMD) by dual-energy x-ray absorption (DXA). All patients, except one, regularly received blood transfusions and iron chelation treatment (deferoxamine, deferiprone, or deferasirox). In the young patient group (n = 5, age: 8 -20 y), a mean Z-score of −1.53 was found (Z-score was calculated from age related BMD). In patients > 20 years (n = 8, age: 21 – 32 years), a mean T-score of −1.66 was observed (T-score was calculated from BMD related to 30 years). Except two patients, all patients had a decreased BMD in comparison to the normal population. One female patient with normal BMD, already received osteoporosis treatment since 5 years due to initially low BMD. Moreover, all patients had a reduced vertebra height in comparison to the normal population.
First results from monitoring the bone density by DXA and the trabecular bone structure by micro-CT, will be reported in all patients after 12 to 18 months under oral supplementation treatment with calcium and vitamin D.