bluuebunny/crossref_metadata_2025_split huggingface dataset:
Rituximab Mediated Depletion of B Cells Following Transplant of Human CD20 Transgenic Mouse Bone Marrow Results in Prolonged B Cell Depletion and Augmented Anti-Tumor Immune Response.
Authors: Zhang Yu D. , Nechushtan Hovav , Jones Monica , Chan Andrew , Shin Seung-uon , Pham Dien , Cho Hyung-Mee , Yi Kyung-Hee , Levy Robert , Rosenblatt Joseph D. 
Published: October 2019

Abstract
We previously demonstrated rejection of murine tumors due to enhanced anti-tumor cytolytic and Th1 cytokine responses in mice lacking B cells (BCDM) compared to immunocompetent mice. We wished to compare immune and anti-tumor responses in BCDM to mice functionally depleted of B cells using an anti-human CD20 antibody (Rituximab). We evaluated Rituximab effects on the outgrowth of human CD20 transgenic mouse (huCD20 Tg) B cells following the transplant of huCD20Tg CD45.2+ bone marrow (BM) into sublethally irradiated congenic CD45.1+ mice. CD45.1 mice were irradiated with 9.5Gy at day -1, followed by infusion of 3x106 huCD20Tg BM cells on day 0, and by treatment with or without Rituximab at 400ug/mouse given once weekly i.v. for 4 weeks. Without Rituximab treatment, B cell percentages in the peripheral blood (PB) were approximately 46.4%±1.3; spleen, 77.8%±1.5; and peripheral lymph nodes (pLNs), 43%±1.5 at day 47. With Rituximab treatment, CD19+ cells in the PB, spleen and pLNs were 1.8%±0.4, 4.3%±1.3, and 0.8%±0.2 respectively at 47days post transplant. Higher percentages of CD3+ T cells in PB, spleen and pLNs (66.1%±0.5, 51.7%±1.1 and 43.0%±3.8) were noted in the Rituximab treated group compared to controls (19%±0.9, 10.3%±1.0 and 32.3%±1.7 respectively). The percentage of remaining B cells in PB declined progressively from 5.6% at day 13 to 1.8% at day 47 post transplant indicating that Rituximab depletion of huCD20+ transgenic B cells was progressive, durable and very effective. In contrast, CD3+ cells in PB increased from 11.2% (day 13) to 66.1 % (day 47) following Rituximab treatment. Interestingly reconstitution with endogenous murine B cells in huCD20 transplanted mice was minimal. The huCD20 Tg donor-derived T cells in the Rituximab treated group maintained a CD44low naive phenotype.
The growth of MC38 tumors implanted 15 days following transplant was significantly slowed in the Rituximab treated group. Increased IFN-γ production and higher expression of the cytolytic marker CD107a by T cells was seen in the Rituximab treated group compared to controls. The combination of myeloablative irradiation, huCD20 transgenic bone marrow transfer and Rituximab depletion, was able to generate mice devoid of CD20+ B cells. Rituximab treated huCD20+ reconstituted B cell depleted mice have substantially higher percentages of T cells than control non-Rituximab treated mice, show sustained depression of B cell numbers and manifest an increased anti-tumor Th1 response. The huCD20 transplant model will be useful in studying effects of post-transplant B cell depletion on immune reconstitution and in the design of future clinical transplant and immunization strategies.
A Novel C-Type Lectin Receptor DCL-1 (CD302) Is a Potential Endocytic Receptor for Tumor Antigen Loading into Antigen Presenting Cells.
Authors: Kato Masato , Khan Seema , McDonald Kylie J. , Hart Derek N.J. 
Published: October 2019

Abstract
DCL-1 (CD302) is a novel C-type lectin receptor, discovered as a genetic fusion partner of DEC-205 (CD205), a C-type lectin that delivers antigen from the surface into DC antigen processing pathway (J. Biol. Chem.2003. 278:34035). We have now examined the molecular nature and expression of DCL-1. DCL-1 contains an external single C-type lectin-like domain followed by a transmembrane and cytoplasmic domain with putative signaling and endocytic motifs. The DCL-1 gene, consisting of 6 exons, was mapped to chromosome 2q24, approximately 5 kb downstream of the DEC-205 gene. Human DCL-1 has 76% protein identity to the mouse ortholog, suggesting its highly conserved function. Northern blot analysis showed that the 4.2 kb DCL-1 mRNA was present in myeloid cell lines, but not in T and B cell lines. Specialized phagocytic cells (i.e. monocytes, macrophages, granulocytes), monocyte-derived DC (MoDC) and blood DC (BDC) expressed DCL-1 mRNA, detected by RT-PCR. Immunoprecipitation/ Western blot (IP/WB) analysis using FLAG-tagged DCL-1 transfectants detected a broad band (modal size 35 kDa), which became a single 26 kDa band after N-glycosidase F digestion, indicating that DCL-1 is N-glycosylated. Flow cytometry studies with DCL-1 mAb (MMRI-20, developed in house) detected moderate levels of DCL-1 expression on granulocytes, monocytes, macrophages, MoDC and BDC, but not on lymphoid cells. Concomitant WB/IP analysis using the DCL-1 mAb detected 26 and 33 kDa doublets in monocytes, macrophages and MoDC. DCL-1 transfectants, macrophages, MoDC and BDC endocytosed cell surface DCL-1 after crosslinking with the DCL-1 mAb. DCL-1-Ig fusion protein bound to GlcNAc- and Man-conjugated beads, but not to GalNAc-, Fuc- and mannan-conjugated beads in a calcium-dependent manner. Thus, DCL-1 is expressed by phagocytes and DC, behaves as a C-type lectin, and is likely to be involved in endocytosis/phagocytosis of microbes. We are exploiting DCL-1 as a potential antigen-loading receptor for DC tumor immunotherapy.
Cytokine Gene Polymorphisms in Turkish Patients with Childhood Chronic Myeloid Leukemia.
Authors: Ozdilli Kursat , Oguz Fatma , Duvarci Yeliz , Bilgen Hulya , Anak Sema , Carin Mahmut , Gedikoglu Gunduz 
Published: October 2019

Abstract
Cytokines are necessary for normal hematopoiesis in the bone marrow and provide a means of fine-tuning bone marrow function in response to stimulation. Several of cytokines generated during both innate and adaptive immune responses stimulate the growth and differantiation of bone marrow progenitor cells. The vast majority of polymorphism found in cytokine genes and their reseptors are located in non coding regions. Cytokine gene polymorphism may be implicated in the pathogenesis of infections, autoimmun disease and malignancies via their effect on cytokine production and regulation. It is known that leukemic cells proliferate under the influence of cytokines.
Our aim is to analyze cytokine gene profiles in patients with chronic myeloid leukemia (CML)in order to clarify the pathogenesis of CML.
Genomic DNA from 26 CML patients were analysed. Genotyping was performed by polymerase chain reaction with sequence specific primers (PCR-SSP) using “ One lamda” kit in 26 childhood CML patients 60 unrelated healthy individuals. Genotype frequencies between CML patients and controls were compared using Chi-Square Yates, Fisher’s Exact Tests. In this study; 26 CML patients with a mean age of 15,6±5,3 and 60 healthy controls with a mean age of 18,2±5,4 were investigated.
In CML patients the frequencies of TGF-b(TC/GG) genotype (in chi-square p=0,01,odds ratio(OR)=3,46, 95% confidence interval (CI) 1,3–9,04) and IL-10 (GCC/ATA) genotype (in chi-square p=0,04,OR=3,3, 95% CI 0,9–11,1 were found higher in patients with CML compared to the control group. On the other hand IL-6 (CC) genotype (in chi-square p=0,012,OR=0,17, 95% CI 0,4–0,76 frequency was found higher in the control group compared to the patients with CML.
As a conclusion higher frequency in TGF-b(TC/GG) and IL-10(GCC/ATA) genotype polymorphism were significantly higher in the patients with CML so it may predispose to CML. On the other hand IL-6 (CC) genotype might be the preventive factor for CML.
The present study is rather significant that it is the study which assessed the relation of the cytokines in patients with CML compared to the control group taken from the genetic pool of The Turkish population, we believe studies like this will eventually help to understand the pathogenesis of CML and the role cytokines play in CML,but larger groups of studies must be done in future.
Short Nucleotide Insertions in the MCL-1 Promoter Affect Its Gene Expression.
Authors: Moshynska Oksana , Moshynskyy Igor , Bosch Mark , Qureshi Tania , Saxena Anurag 
Published: October 2019

Abstract
We have recently reported a short nucleotide (six and eighteen) insertion (6- and 18-nt) in the MCL-1 promoter (−190bp upstream of the major transcription start site, GenBank, AN: AF198614) in B-cell chronic lymphocytic leukemia patients. These insertions were found to be polymorphisms. CD38-negative CLL patients with these promoter insertions had higher mRNA levels, poor response to standard chemotherapy and shorter survival. The aim of the present study was to test the hypothesis that these insertions directly effect MCL-1 gene expression. K562 and HeLa cells were transfected with pGL3-Basic, pGL3-Control and four reporter plasmids were constructed with a +0/0, +6/6, +18/18, and −23/23 MCL-1 promoter linked to Luciferase. Firefly activity in each transfection was normalized to Renilla Luciferase activity. At 24-hours post-transfection cells were induced with phorbol 12-myristate 13-acetate (PMA), granulocyte-macrophage colony stimulating factor (GM-CSF), both PMA and GM-CSF, or with fresh media only. At 48-hours post-transfection cells were harvested and Dual-Luciferase Assay was performed. Compared to transfection with pGL3-(+0/0) plasmid, the Luciferase PMA-stimulated activity for K562 cells transfected with pGL3-(+6/6) and pGL3-(+18/18) was 1.7 (P=0.0277) and 1.5 times higher (P=0.0067) respectively, and for pGL3-(−23/23) was 3.2 times lower (P=0.0021). The Luciferase PMA-stimulated activity for HeLa cells transfected with pGL3-(+6/6) was 3.3 times higher (P=0.0038), pGL-3-(+18/18) was 2.3 times higher (P=0.0245), and did not differ significantly for pGL3-(−23/23), compared to pGL3-(+0/0). Similar results were observed for GM-CSF-stimulated activity for HeLa cells, where activity was 3.9 times (P=0.0068) for pGL3-(+6/6) and 2.5 times (P=0.0258) higher for pGL3-(+18/18). Unstimulated Luciferase activity was also 2.5 times higher (P=0.0115) for pGL3-(+18/18) compared to pGL3-(+0/0) plasmid. These results provide experimental evidence for the positive effect of these nucleotide insertions (6- and 18-nt) on MCL-1 gene expression in both hemopoietic (K562) and epithelial (HeLa) cells.
Gene Expression Studies Reveal a Potential Mechanism of Glivec Resistance in Patients with Ph+ ALL Treated.
Authors: Loi To Ha , Ma David D.F. 
Published: October 2019

Abstract
In Ph+ ALL, patients respond to Glivec but nearly always relapse due to acquired resistance. The biological processes associated with Glivec resistance in Ph+ ALL are yet to be fully elucidated.
The aim of this study is to use microarray and Q-PCR technology to dissect changes in signaling pathways of blast cells in Ph+ ALL patients treated with Glivec. Peripheral blood (PB) samples were taken before treatment and on consecutive days after administration of Glivec from two Ph+, c-ALLA+ and CD19+ ALL patients. Blast cells were isolated and their gene expression assayed using 19K cDNA microarrays.
Over 400 differentially expressed genes were identified with at least a 1.5-fold up- or down-regulation in treated cells compared to cells collected pre-treatment. Based on these gene expression results, three main gene ontology groups were further evaluated: Apoptosis, Proliferation and B cell differentiation. The up-regulation of Bim and Bcl-6, and down regulation of Cyclin D2, confirms the induction of apoptosis via the FOXO3a pathway in cells treated with Glivec in vivo. Interestingly, both the proliferation genes, Tcl1-A and PKCe, and B cell differentiation associated genes, including CD79a, ETS1 and a cohort of IGH and Igl and k genes, were up-regulated during therapy.
These gene expression changes observed in vivo were confirmed by Q-PCR in the Ph+ cell lines K562 (derived from CML blast crisis) and SUP-B15 (derived from ALL) treated with Glivec. The unexpected finding of increased expression of pro-proliferative genes and B cell differentiation genes by microarray revealed potential links with early B cell development and B cell receptor (BCR) signaling.
Evidence for apoptosis and proliferation of Ph+ cell lines treated with Glivec were then examined by FACs. After 5 days of treatment with Glivec, 90% of K562 and 50% SUP-B15 cells underwent apoptosis. Furthermore, cell cycle analysis revealed the existence of a population of cells in G2 phase even after 6 days of Glivec treatment in SUP-B15 but not K562 cells, thus providing evidence of a population of cells undergoing proliferation during Glivec treatment in vitro.
In summary, our in vivo observations supported by in vitro experiments suggest that Glivec induces the majority of Ph+ ALL blasts to undergo apoptosis. However, as treatment is prolonged, a population of ALL cells escapes death and undergoes proliferation and differentiation. We hypothesise that Glivec induced differentiation and proliferation of Ph+ cells may result in the clonal enrichment of cells resistant to Glivec.
Antitumor Genotoxic Agents, Such as Fludarabine*, Doxorubincin or Cis-Platine, Induce Activation of STAT1 in a p53 Protein Dependent Manner.
Authors: Youlyouz-Marfak Ibtissam , Le Clorennec Christophe , Najjar Imen , Baran-Marszak Fanny , Gachard Nathalie , Remenieras Liliane , Jaccard Arnaud , Bornkamm Georg W. , Fagard Remi , Feuillard Jean 
Published: October 2019

Abstract
Introduction:
Chemotherapeutic drug such as Fludarabine*; doxorubicin or cis-platine induce cell cycle arrest and apoptosis via activation of p53. Convergent studies suggest that p53 and STAT1 cooperate in the induction of apoptosis, and that STAT1 favors p53 activation. However, to our knowledge, the role of p53 in the activation of STAT1 is not documented. We present our results suggesting that (i) genotoxic agents are STAT1 inducers, (ii) STAT1 activation depends on the presence of p53 protein, and (iii) this phenomenon is modulated by the tyrosine kinase inhibitor STI571.
Materials and Methods:
To analyse the role of p53 in STAT1 activation, we have used different cellular models with different p53 status: PRI (p53wt), BL2 (p53wt), BL41 (p53 mutated on Arg248, resulting in the loss of p53 DNA binding activity (p53mut)), Jurkat, HL60 and MEF (the 3 latter being p53 null). The following cDNAs were used for functional studies: p53wt, p53mut, MDM2 and MTBP (MDM2 transforming protein). These cDNAs were cloned either in a pcDNA3 vector or a pRT-1 inducible vector (in the latter, the gene of interest is expressed from a bidirectional doxycycline regulatable promoter allowing simultaneous expression of truncated NGF receptor, used as a surrogate marker of inducibility).
Results:
Treatment of the different cell lines with the 3 genotoxic drugs Fludarabine*, doxorubicin or cis-platine induced STAT1 activation in p53wt BL2 or PRI cells and in p53mut BL41 cells, but not in Jurkat cells neither in HL60 or MEF cells. Induction of STAT1 was also obtained in presence of the RNA synthesis inhibitor Actinomycin D or in presence of secretion inhibitor Brefeldine A. Over-expression of p53wt or p53mut markedly increased STAT1 activation in PRI cells. This effect was reversed by over-expression of MTBP. Complementation of both HL60 and MEF cells with both p53wt and p53mut cDNA induced constitutive STAT1 activation, an effect that was increased by treatment with doxorubine in transfected HL60 cells. This effect was reversed by over-expression of MDM2 in HL60 cells. Finally, we found that treatment of cells with the inhibitor STI 571 of c-Abl tyrosine kinase, a kinase known to be associated with ATM during p53 activation, decreased STAT1 activation by genotoxic drugs.
Conclusion:
Our results show that genotoxic agents are inducers of STAT1, that p53 protein but not p53 transcriptional activity is responsible for this STAT1 activation, and suggest a possible involvement the cABL tyrosine kinase.
The TCL1 Gene Is Involved, Other Than in Lymphoid Differentiation and Early Embryogenesis, Also in the Development of the Hair Follicle.
Authors: Ragone Gianluca , Bresin Antonella , Picchio Maria Cristina , Lazzeri Cristina , Cristofoletti Cristina , Kang Sang-Moo , Narducci Maria Grazia , Russo Giandomenico 
Published: October 2019

Abstract
The TCL1(T-cell leukaemia lymphoma1) oncogene plays a causative role in the onset of T and B cell leukaemia of mature phenotype, such as T-prolymphocytic leukema T-(PLL) in human and mice and B-cell-chroni lymphatic leukaemia (B-CLL) in mice due to its overexpression. Moreover, Tcl1 −/− mice show that this gene is also important during the early embryogenesis and lymphocyte differentiation. An undiscovered role was found for TCL1 gene in the skin. In this work, we show evidences that give Tcl1 a new, important role in mouse hair follicle (HF) and skin homeostasis. In fact, Tcl1 −/− adult mice develop premature hair loss leading to alopecia followed by extensive skin lesions, starting from the dorsal region. The expression pattern of Tcl1 in the skin throughout the different stages of hair and skin differentiation, by in vivo immunofluorescence and fluorescent in situ hybridization experiments, shows a high level of the mRNA and protein in HF mainly in the secondary hair germ cells and hair bulge but not in dermal papilla (DP), both in early anagen and late catagen phases, and to less extent in the epidermis in all the stages examined.
Loss of Tcl1 expression in the skin, although initially does not show any apparent morphological defects in the HF and epidermis, nonetheless leads to an increased rate of apoptosis and to a parallel reduction of proliferation through an unknown pathway, independent from the phosphorylation of akt proteins. Indeed, we show evidences that loss of Tcl1 affects CD34 high expression level in the bulge stem cells that is necessary, to these population, to maintain slow-cycling and stem characteristics along the time. Finally, the higher expression of the Tcl1 ortholog Tcl1B2 in epidermis and hair bulb can further explain the late onset, in adult mice, of alopecia and skin lesions. Our findings suggest that Tcl1 gene(s) have important roles in cycling, self-renewing stem cell populations in skin. This observation, coupled with other previous work showing that TCL1 gene is expressed in B-cell entering the germinal follicle in the lymph-node and in the preimplantation embryo until the 8-cell stage, points out to an interesting role for this gene for cells that at some point in their life must re-enter cell cycle.
Duffy Antigen/Receptor for Chemokines (DARC): Is There a Role in Prostate Cancer?.
Authors: Anyona Samuel B. , Zheng Shiyan , Wheeler Thomas , Ittmann Michael , Hayes Teresa G. , Mims Martha P. 
Published: October 2019

Abstract
Introduction: The Duffy blood group antigens function as chemokine receptors and as receptors for several species of malarial parasites. Approximately 70% of African Americans are Duffy negative as a result of a single nucleotide polymorphism in the promoter region of the Duffy antigen/receptor for chemokines (DARC) gene in a consensus-binding site for GATA 1. This mutation abolishes the promoter activity in erythrocytes, impairing the expression of DARC on red blood cells. Other than resistance to certain species of malaria, the functional consequence of Duffy negativity is unclear; however, it has recently been proposed that the Duffy antigen acts as a biological ‘sink’ to clear pro-inflammatory chemokines from tissue microcirculation. Further it has been suggested that since the incidence of prostate cancer in African Americans is 60% higher than in Caucasians, absence of the Duffy antigen might predispose African Americans to prostate cancer by impairing downregulation of proinflammatory cytokines. We tested the hypothesis that lack of expression of DARC on erythrocytes predisposes African American men to develop prostate cancer.
Methods: We conducted a case control study of 89 African American men with confirmed prostate cancer and 51 age matched healthy African American men. The samples were genotyped for the promoter polymorphism using an allele specific real time PCR assay (Taqman assay) developed for this study.
Results: The frequency of the Duffy negative allele was 75.8% among the patients and 81.4% among the controls (odds ratio = 0.72, p value = 0.28). The distribution of heterozygous and homozygous Duffy negative patients did not differ between the cases and controls.
Conclusion: It has been suggested that individuals lacking erythrocyte expression of DARC have higher prostate levels of angiogenic chemokines that might promote more rapid development of prostate cancer. The data presented here found no difference in the frequency of the Duffy negative allele between African American men with prostate cancer and healthy age matched African American controls. Since several studies suggest that polymorphisms associated with differential production of IL-8, IL-10 and VEGF are risk factors for prostate cancer, it remains possible that a polymorphism associated with differential cytokine production in combination with Duffy negativity increases the risk for prostate cancer.
The Tyrosine Kinase src Is Highly Activated in AML and Plays a Crucial Role in Leukemic Cell Proliferation.
Authors: Schuch Gunter , Schafer Ellen , Eggert Katharina , Loges Sonja , Jucker Manfred , Hossfeld Dieter K. , Bokemeyer Carsten , Fiedler Walter 
Published: October 2019

Abstract
The tyrosine kinase pp60src is involved in several signal transduction pathways such as signalling of hematopoetic growth factors and cytokines. The viral form v-src was the first oncogene described and mutations of regulatory tyrosine residues in cellular src (c-src) have been linked to malignant transformation. However, no mutations in the gene of c-src have been described in leukemia so far, although some data of src mutations in solid tumors have been reported. The current study was undertaken to examine the role of src in acute myeloid leukemia (AML).
Blood and bone marrow specimen of patients with newly diagnosed or recurrent AML treated at our institution were sampled. AML cell lines or CD34 positive cells of healthy donors served as positive and negative controls, respectively. RNA was isolated, and RT-PCR was performed using 4 different primer pairs spanning the coding region of c-src. Protein expression and phosphorylation was studied after protein extraction and Western blot analyses using src and phospho-src specific antibodies. The effect of src inhibitors PP1 and PP2 on leukemic cell proliferation was studied in human and murine cell lines. Mutational analyses of the coding region were performed using SSCP/heteroduplex and bi-directionally sequencing.
In all 60 patients analyzed expression of c-src mRNA was detected by RT-PCR. Western blot analyses confirmed strong expression of src on the protein level and revealed a robust activation of the protein as determined by tyrosine phosphorylation. Inhibition of src phosphorylation by src-specific inhibitors PP1 and PP2 was detected by Western blot using an antibody specific for phospho-src. Incubation of leukemic cells with PP1 and PP2 caused significant inhibition of proliferation in a dose dependent manner. Mutational analyses as performed by SSCP/heteroduplex and bi-directionally sequencing revealed wildtype sequence in all cell lines and 60 clinical samples.
In summary, pp60src is highly expressed and activated in cell lines and clinical samples of human AML. Moreover, phosphorylation of src is essential for leukemic cell proliferation. Mutations in the coding sequence of c-src causing constitutive activation could be excluded by mutational analyses of primary AML samples. These data suggest that pp60src plays a crucial role in AML and src inhibition by targeted therapy might offer a useful new approach in the treatment of AML.