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1
Salem, M. Dr Rafid Abdel Kadhim. "Book of Mena Mannan in the defense of the Koran to Mr. Mohammed Sadr Reading in light of the reference method." ALUSTATH JOURNAL FOR HUMAN AND SOCIAL SCIENCES 227, no. 3 (December 5, 2018): 417–41. http://dx.doi.org/10.36473/ujhss.v227i3.790.
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His Eminence was not excluded from the academic study. He combined the study of Al-Hawzawi and Al-Jamaa'i. He even contributed to the introduction of the academic lessons of the estate and he obtained the rank of ijtihad. He was 34 years old and in the same year he studied arfan and used various sources and references. He was not keen on them, because most of theses contained in his book of the daughters of his ideas. In his book, he adopted an innovative approach in most of his writings, to develop the mind and the works of thought, especially the book in question. He contradicted his predecessor in his interpretive approach, beginning with the last line of the Holy Book. He emphasized the theory of inspiration, The number of signs recorded in the minyan (37) is a reference divided into several sections, including those related to the divinity, judgments, ethics, etc., as shown in the research table
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Rasheed, Dr Wesam Ahmed. "Map valuation of Map elements in the Geographic studies Comparative study of theses and distraction of department of Geography college of Education for girls and college of Education ( Ibn- Rushid ) University of Baghdad for the period ( 2000- 2015 )." ALUSTATH JOURNAL FOR HUMAN AND SOCIAL SCIENCES 219, no. 2 (November 9, 2018): 57–72. http://dx.doi.org/10.36473/ujhss.v219i2.515.
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This research aims to study elements of map through evaluation of using elements of map ofone hundred theses and dissertations of geography department in the college of Education for girls –university of Baghdad. The researcher makes a comparison between this department and the department of geography in the college of Education – IbnRushid – university of Baghdad that he would know which department is better in producing maps having accuracy in using elements of map especially if we know that the first department is in need of specialized professors, the other department has specialist in cartography. The research reaches to that some researchers fails in following some basic rules of maps. From other side, the research realizes that there is a clear development in producing a high quality map through using new technologies and programs by researchers. As to comparison which is made between two departments, the second department has a wide maintaining in elements of maps as there are specialized professors of cartography having the ability in producing accurate and scientific maps through supervision or giving advice .
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Guillemin, Marie-Claude, Emmanuel Raffoux, Dominique Vitoux, Scott Kogan, Hassane Soilihi, Valérie Lallemand-Breitenbach, Jun Zhu, et al. "In Vivo Activation of cAMP Signaling Induces Growth Arrest and Differentiation in Acute Promyelocytic Leukemia." Journal of Experimental Medicine 196, no. 10 (November 18, 2002): 1373–80. http://dx.doi.org/10.1084/jem.20021129.
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Differentiation therapy for acute myeloid leukemia uses transcriptional modulators to reprogram cancer cells. The most relevant clinical example is acute promyelocytic leukemia (APL), which responds dramatically to either retinoic acid (RA) or arsenic trioxide (As2O3). In many myeloid leukemia cell lines, cyclic adenosine monophosphate (cAMP) triggers growth arrest, cell death, or differentiation, often in synergy with RA. Nevertheless, the toxicity of cAMP derivatives and lack of suitable models has hampered trials designed to assess the in vivo relevance of theses observations. We show that, in an APL cell line, cAMP analogs blocked cell growth and unraveled As2O3-triggered differentiation. Similarly, in RA-sensitive or RA-resistant mouse models of APL, continuous infusions of 8-chloro-cyclic adenosine monophosphate (8-Cl-cAMP) triggered major growth arrest, greatly enhanced both spontaneous and RA- or As2O3-induced differentiation and accelerated the restoration of normal hematopoiesis. Theophylline, a well-tolerated phosphodiesterase inhibitor which stabilizes endogenous cAMP, also impaired APL growth and enhanced spontaneous or As2O3-triggered cell differentiation in vivo. Accordingly, in an APL patient resistant to combined RA–As2O3 therapy, theophylline induced blast clearance and restored normal hematopoiesis. Taken together, these results demonstrate that in vivo activation of cAMP signaling contributes to APL clearance, independently of its RA-sensitivity, thus raising hopes that other myeloid leukemias may benefit from this therapeutic approach.
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Smith, Daniel J., William F. King, Christine D. Wu, Bella I. Shen, and Martin A. Taubman. "Structural and Antigenic Characteristics of Streptococcus sobrinus Glucan Binding Proteins." Infection and Immunity 66, no. 11 (November 1, 1998): 5565–69. http://dx.doi.org/10.1128/iai.66.11.5565-5569.1998.
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ABSTRACT Three purified glucan binding proteins (GBP-2, GBP-3, and GBP-5) from Streptococcus sobrinus 6715 were compared structurally by mass spectroscopy of tryptic fragments and antigenically by Western blot analysis with rat antisera to each GBP or to peptides containing putative glucan binding epitopes of mutans streptococcal glucosyltransferases. Structural and antigenic analyses indicated that GBP-3 and GBP-5 are very similar but that both are essentially unrelated to GBP-2. None of theseS. sobrinus GBPs appeared to have a strong antigenic relationship with GBPs from Streptococcus mutans. Thus,S. sobrinus GBP-2 and GBP-3 appear to be distinct proteins with potentially different functions. S. sobrinusGBP-5 may be a proteolytic fragment of GBP-3, or, alternatively, the genes coding for these proteins may be closely related.
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Villanueva-Romero, Raúl, Irene Gutiérrez-Cañas, Mar Carrión, Selene Pérez-García, Iria V. Seoane, Carmen Martínez, Rosa P. Gomariz, and Yasmina Juarranz. "The Anti-Inflammatory Mediator, Vasoactive Intestinal Peptide, Modulates the Differentiation and Function of Th Subsets in Rheumatoid Arthritis." Journal of Immunology Research 2018 (August 1, 2018): 1–11. http://dx.doi.org/10.1155/2018/6043710.
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Genetic background, epigenetic modifications, and environmental factors trigger autoimmune response in rheumatoid arthritis (RA). Several pathogenic infections have been related to the onset of RA and may cause an inadequate immunological tolerance towards critical self-antigens leading to chronic joint inflammation and an imbalance between different T helper (Th) subsets. Vasoactive intestinal peptide (VIP) is a mediator that modulates all the stages comprised between the arrival of pathogens and Th cell differentiation in RA through its known anti-inflammatory and immunomodulatory actions. This “neuroimmunopeptide” modulates the pathogenic activity of diverse cell subpopulations involved in RA as lymphocytes, fibroblast-like synoviocytes (FLS), or macrophages. In addition, VIP decreases the expression of pattern recognition receptor (PRR) such as toll-like receptors (TLRs) in FLS from RA patients. These receptors act as sensors of pathogen-associated molecular pattern (PAMP) and damage-associated molecular pattern (DAMP) connecting the innate and adaptive immune system. Moreover, VIP modulates the imbalance between Th subsets in RA, decreasing pathogenic Th1 and Th17 subsets and favoring Th2 or Treg profile during the differentiation/polarization of naïve or memory Th cells. Finally, VIP regulates the plasticity between theses subsets. In this review, we provide an overview of VIP effects on the aforementioned features of RA pathology.
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Herrington, CS, and PA Hall. "Molecular and cellular themes in inflammation and immunology." Journal of Pathology 214, no. 2 (January 2008): 123–25. http://dx.doi.org/10.1002/path.2303.
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Y. Nasir, Dr Adel. "Identity and ideological reading paths in the conflict and developments." ALUSTATH JOURNAL FOR HUMAN AND SOCIAL SCIENCES 212, no. 2 (November 12, 2018): 243–68. http://dx.doi.org/10.36473/ujhss.v212i2.673.
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Ideology and identification is one of the main and big subjects nowadays, especially after the ideology specified the ideology and main nationality identification existence by process which leads to Replacement and paddlefish of ideology existence rather than identification , while ideology makes so much efforts ( intellectual and beliefs efforts ) to solve century problems and presents mental and intellectual developments which characterized by realism in such disturbed and Dispersion world in political, social, economic factions and dominated by cases of conflict and chaos . Identification seeks to connect the individual similarities participating in the land, history and real affiliation, which tend to personal fulfillment of humanity in its social and cultural frame and confirms his affiliation root. But while that facing the impact of ideological conflict which aims to achieve an intellectual image and adopts cultures and ideas from another civilization might disagree with identification characteristics existence. Ideology trying to dismantling in all Civilization and national directions, as well as seeks to Skepticism of identification with its ideological features while the weakness of the Factors affiliation , the local organizations , National parties and Religious institutions and their branches in its special directions . Which lead to the disability of resists the ideological Invasion weather is was local, regional or global. But despites of the some ideologies presents humanitarian and reformative theses .In different political shapes and frames still mainly defending to the Elements of existence which resulting the ideological conflict – identificational despite of that both of them have weakness in its Staff fundamental parts thus the ideology Remain to protect the benefits of the interests of the political elite and leaders while facing Cases of rejection and acceptance in social circles . Whereas the main nationality identification went through a conflict with sub-identification ,Sectarianism and loyalty instead of main identification or the national identification and from that description we can conclude that the conflicts and its pathways are continuous in such a world full of existence and hegemony and superiority .
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Roccaro, Aldo M., Xavier Leleu, Antonio Sacco, Xiaoying Jia, Molly Melhem, Anne-Sophie Moreau, Hai T. Ngo, et al. "Dual targeting of the proteasome regulates survival and homing in Waldenström macroglobulinemia." Blood 111, no. 9 (May 1, 2008): 4752–63. http://dx.doi.org/10.1182/blood-2007-11-120972.
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AbstractWaldenström macroglobulinemia (WM) is an incurable low-grade B-cell lymphoma characterized by high protein turnover. We dissected the biologic role of the proteasome in WM using 2 proteasome inhibitors, NPI-0052 and bortezomib. We found that NPI-0052 inhibited proliferation and induced apoptosis in WM cells, and that the combination of NPI-0052 and bortezomib induced synergistic cytotoxicity in WM cells, leading to inhibition of nuclear translocation of p65NF-κB and synergistic induction of caspases-3, -8, and -9 and PARP cleavage. These 2 agents inhibited the canonical and noncanonical NF-κB pathways and acted synergistically through their differential effect on Akt activity and on chymotrypsin-like, caspaselike, and trypsinlike activities of the proteasome. We demonstrated that NPI-0052–induced cytotoxicity was completely abrogated in an Akt knockdown cell line, indicating that its major activity is mediated through the Akt pathway. Moreover, we demonstrated that NPI-0052 and bortezomib inhibited migration and adhesion in vitro and homing of WM cells in vivo, and overcame resistance induced by mesenchymal cells or by the addition of interleukin-6 in a coculture in vitro system. Theses studies enhance our understanding of the biologic role of the proteasome pathway in WM, and provide the preclinical basis for clinical trials of combinations of proteasome inhibitors in WM.
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Posada, Ana C., Stacey L. Kolar, Renata G. Dusi, Patrice Francois, Alexandra A. Roberts, Chris J. Hamilton, George Y. Liu, and Ambrose Cheung. "Importance of Bacillithiol in the Oxidative Stress Response of Staphylococcus aureus." Infection and Immunity 82, no. 1 (October 28, 2013): 316–32. http://dx.doi.org/10.1128/iai.01074-13.
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ABSTRACTInStaphylococcus aureus, the low-molecular-weight thiol called bacillithiol (BSH), together with cognateS-transferases, is believed to be the counterpart to the glutathione system of other organisms. To explore the physiological role of BSH inS. aureus, we constructed mutants with the deletion ofbshA(sa1291), which encodes the glycosyltransferase that catalyzes the first step of BSH biosynthesis, andfosB(sa2124), which encodes a BSH-S-transferase that confers fosfomycin resistance, in severalS. aureusstrains, including clinical isolates. Mutation offosBorbshAcaused a 16- to 60-fold reduction in fosfomycin resistance in theseS. aureusstrains. High-pressure liquid chromatography analysis, which quantified thiol extracts, revealed some variability in the amounts of BSH present acrossS. aureusstrains. Deletion offosBled to a decrease in BSH levels. ThefosBandbshAmutants of strain COL and a USA300 isolate, upon further characterization, were found to be sensitive to H2O2and exhibited decreased NADPH levels compared with those in the isogenic parents. Microarray analyses of COL and the isogenicbshAmutant revealed increased expression of genes involved in staphyloxanthin synthesis in thebshAmutant relative to that in COL under thiol stress conditions. However, thebshAmutant of COL demonstrated decreased survival compared to that of the parent in human whole-blood survival assays; likewise, the naturally BSH-deficient strain SH1000 survived less well than its BSH-producing isogenic counterpart. Thus, the survival ofS. aureusunder oxidative stress is facilitated by BSH, possibly via a FosB-mediated mechanism, independently of its capability to produce staphyloxanthin.
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Kim, Hee Nam, Huong Thi Thanh Tran, Il-Kwon Lee, Yeo-Kyeoung Kim, Deok-Hwan Yang, Je-Jung Lee, Myung-Geun Shin, et al. "Polymorphisms of Thymidylate Synthase in the 5′- and 3′-Untranslated Regions Associated with Risk of Non-Hodgkin’s Lymphoma." Blood 108, no. 11 (November 1, 2006): 2394. http://dx.doi.org/10.1182/blood.v108.11.2394.2394.
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Abstract Thymidylate synthase (TS) catalyzes the conversion of dUMP by 5,10-methylenetetrahydrofolate to dTMP in DNA synthesis. Polymorphisms in the untranslated regions(UTRs) of TS, which may modulate TS transcription and expression, have been associated with susceptibility to several malignancies. In this study, to evaluate the association with TS the 28-bp tandem repeat(2R→3R) and the 6-bp deletion(6 bp-) and susceptibility to non-Hodgkin’s lymphoma(NHL), large-scale population-based case-control study was conducted in Chonnam National University Hospital between Mar 1997 and Feb 2006. 553 patients with histologically comfirmed lymphoma and 1,324 controls were evaluated. The cases consisted of 275 diffuse large B-cell lymphomas (DLBCL), 109 T-cell lymphomas and 169 unclassifiable lymphomas. TS 2R2R genotype was significantly associated with increased risk for NHL and T-cell lymphoma, but not for DLBCL. Using subjects with the TS 3R3R as a reference group, the OR of TS 2R2R was 2.00 (95% CI 1.21–3.31, p=0.007) for NHL and 3.30 (95% CI 1.52–7.17, p=0.003) for T-cell lymphoma. The association was 1.65 fold higher and more evident for T-cell lymphoma than NHL. However, there was no significant association of TS 6bp- with NHL. In conclusion, theses results suggest that TS (2R→3R) may play an important role in the pathogenesis of NHL, and that DNA synthesis may play a crucial roles in the pathogenesis of specific NHL subtypes.
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Ji, Suk, Ji Eun Shin, Young Sook Kim, Ju-Eun Oh, Byung-Moo Min, and Youngnim Choi. "Toll-Like Receptor 2 and NALP2 Mediate Induction of Human Beta-Defensins by Fusobacterium nucleatum in Gingival Epithelial Cells." Infection and Immunity 77, no. 3 (December 22, 2008): 1044–52. http://dx.doi.org/10.1128/iai.00449-08.
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ABSTRACT We previously reported that infection by Fusobacterium nucleatum strongly induced the expression of both human beta-defensin 2 (HBD-2) and HBD-3 by gingival epithelial cells. The aim of this study was to characterize the pattern recognition receptors (PRRs) and regulatory mechanisms involved in the induction of HBD-2 and -3 expression by F. nucleatum in gingival epithelial cells. Immortalized human gingival epithelial HOK-16B cells were infected with live or heat-killed F. nucleatum, and the expression of HBDs and interleukin-8 (IL-8) was examined by real-time reverse transcription-PCR and enzyme-linked immunosorbent assay, respectively. Live, but not heat-killed, F. nucleatum invaded HOK-16B cells, as seen by confocal microscopy and flow cytometry. Live F. nucleatum induced both HBD-2 and -3 efficiently, whereas heat-killed bacteria induced only HBD-3 at a reduced level. Knockdown of NACHT-LRR- and pyrin domain-containing protein 2 (NALP2), the most abundant intracellular PRR in HOK-16B cells, by RNA interference (RNAi) significantly reduced the induction of HBD-3 but not HBD-2 and IL-8. In addition, knockdown of Toll-like receptor 2 (TLR2) by RNAi reduced the upregulation of HBD-2 and -3 but not IL-8. Heat-killed F. nucleatum was hindered in its ability to activate TLR2 and JNK signaling pathways. Theses data show that TLR2 and NALP2 mediate the induction of HBDs by F. nucleatum in gingival epithelial cells, but thresholds for the two HBD genes are different.
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Pavlova, Iglika V., Herbert W. Virgin, and Samuel H. Speck. "Disruption of Gammaherpesvirus 68 Gene 50 Demonstrates that Rta Is Essential for Virus Replication." Journal of Virology 77, no. 10 (May 15, 2003): 5731–39. http://dx.doi.org/10.1128/jvi.77.10.5731-5739.2003.
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ABSTRACT Gammaherpesvirus pathogenesis is dependent on the ability of these viruses to establish a lifelong latent infection and the ability to reactivate from latency. Immediate-early genes of theses viruses are thought to be critical regulators of lytic replication and reactivation from latency. The gene 50-encoded Rta is the only immediate-early gene product that appears to be conserved among all characterized gammaherpesviruses. Previous studies have demonstrated that, in Epstein-Barr virus (EBV), Kaposi's sarcoma-associated virus, and gammaherpesvirus 68 (γHV68, also referred to as murine gammaherpesvirus 68), ectopic expression of Rta in latently infected cell lines can lead to induction of the viral cycle. Recently, studies employing null mutants of EBV have provided a formal demonstration that both Rta and the BZLF1 gene product, Zta, the two EBV immediate-early gene products, are essential for EBV replication. Here we generate and characterize a gene 50-null mutant γHV68 and demonstrate that the gene 50 product Rta is essential for virus replication. Providing γHV68 Rta in trans was sufficient to restore replication of the gene 50-null virus. Notably, Rta expressed from the spliced form of the gene 50 transcript was sufficient to complement growth of the gene 50-null virus. In addition, we provide evidence that loss of Rta expression leads to a complete defect in viral DNA replication and a significant defect in late antigen expression. This work lays the foundation for characterizing the role of Rta in γHV68 chronic infection of mice.
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Vida, Zsófia Viktória, István Péter Járay, and Balázs Lengyel. "PhD students in life sciences can benefit from team cohesion." F1000Research 10 (July 29, 2021): 692. http://dx.doi.org/10.12688/f1000research.53743.1.
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Background: Scientific progress during doctoral studies is a combination of individual effort and teamwork. A recently growing body of interdisciplinary literature has investigated the determinants of early career success in academia, in which learning from supervisors and co-authors play a great role. Yet, it is less understood how collaboration patterns of the research team, in which the doctoral student participates, influences the future career of students. Here we take a social network analysis approach to investigate this and define the research team as the co-authorship network of the student. Methods: We use the Hungarian Scientific Bibliography Database, which includes all publications of PhD students who defended theses from the year 1993. The data also include thesis information, and the publications of co-authors of students. Using this data, we quantify cohesion in the ego-network of PhD students, the impact measured by citations received, and productivity measured by number of publications. We run multivariate linear regressions to measure the relation of network cohesion, and publication outputs during doctoral years with future impact. Results: We find that those students in life sciences, but not in other fields, who have a cohesive co-author network during studies and two years after defence receive significantly more citations in eight years. We find that the number of papers published during PhD years and closely after the defence correlates negatively while the impact of these papers correlates positively with future success of students in all fields. Conclusions: These results highlight that research teams are effective learning environments for PhD students where collaborations create a tightly knit knowledge network.
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Wiegering, Verena, Julia Taschik, Imme Haubitz, Matthias Eyrich, Beate Winkler, and Paul Gerhardt Schlegel. "TGFβ and IL10 Have an Impact on Risk Group and Prognosis in Childhood ALL." Blood 124, no. 21 (December 6, 2014): 5337. http://dx.doi.org/10.1182/blood.v124.21.5337.5337.
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Abstract Introduction: Considering that cytokines play an important role in immune response and that many infectious, autoimmune and malignant diseases are influenced by cytokine production, we hypothesized that genetically determined cytokine gene polymorphism might have an important influence on prognosis in pediatrics acute lymphoblastic leukemia (ALL). Methods: In this study, 95 pediatric ALL-patients were examined with regard to cytokine gene polymorphisms (TNFα, TGFβ, IL10 and IFNγ) and their potential association with prognosis. Moreover we analyzed the intracellular production of theses cytokines in patient T-cells. Results: TGFβ high-producer-haplotypes were associated with high-risk ALL-patients (Codon 10: T/T) and with the tendency of a reduced overall survival, whereas IL10 high-producer-genotypes were associated with a reduced relapse rate and a superior overall survival compared to IL-10-low-producer patients. Gene-polymorphisms of the pro-inflammatory cytokines IFNγ and TNFα did not show an impact on prognosis and risk-group of ALL in our cohort. On a functional basis TNFα and IFNγ expression of T-cells at initial diagnosis was significantly reduced in high-risk- and T-ALL-patients in comparison to healthy controls. Summary: Cytokine gene-polymorphisms of the regulatory/anti-inflammatory cytokines TGFβ and IL10, but not of the pro-inflammatory cytokines IFNγ and TNFα seem to have an impact on prognosis of pediatric ALL patients. Reduced capacity to produce pro-inflammatory cytokines at diagnosis may serve as a functional risk factor. These data may help in further risk stratification and adaptation of therapy-intensity. Disclosures No relevant conflicts of interest to declare.
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Branco, Patrícia Q., Cristina P. Abreu, Pedro Pessegueiro, Manuel Amoedo, Anabela Rodrigues, Antonio Cabrita, Margarida Bruges, Edgar de Almeida, and Mateus Martins Prata. "Anaemia Management in Patients on Peritoneal Dialysis in Portugal: A Nationwide Multicenter Survey." Blood 106, no. 11 (November 16, 2005): 3753. http://dx.doi.org/10.1182/blood.v106.11.3753.3753.
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Abstract Introduction: The European Best Practice Guidelines for the Anaemia Management (EBPG) recently published emerged as an international standard in the management of anemia in chronic kidney disease (CKD) patients. Objectives: To evaluate the impact of theses guidelines in the clinical practice in Portugal. Methods: This epidemiological, multicentric and cross-sectional study included patients on peritoneal dialysis that were under erythropoietin treatment in Portugal during 2004. Etiology of CKD, prevalence and anaemia treatment, comorbidity and side effects were evaluated. Results: 220 patients from 5 Units were evaluated. Mean haemoglobin was 12.34 g/dL and only 13,2% of patients had haemoglobin <11 g/dL. Ninety-nine percent of patients were treated with erythropoietin: 65% with beta erythropoietin (beta) and 35% with darbepoetin alfa (darbe). Subcutaneous route had been used in all cases: 5,4% with once-monthly administration (darbe), 19,55 % two times monthly (darbe and beta), 59% with once a week regimen (darbe and beta), 11,55% in two weekly doses (beta) and 4,5% in three weekly administrations (beta). Doses requirements were different according to administration frequency. More than 85% of patients had haemoglobin > 11 g/dl and no significant difference in haemoglobin levels was achieved in the two groups (beta and darbe). Erythropoietin doses were greater in the group treated with darbe (127 versus 113 UI/kg/week), but inflammation markers were significantly higher as well diabetic patients in this group. Conclusion: These results suggest that, according to the EBPG’s, mostly patients on PD in Portugal are already being well treated presenting heamoglobin levels in the therapeutic range.
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PIRC, Tadej. "CREATIVITY OF BIOTECHNOLOGICAL IMMUNOLOGY: INVENTION, NATURALNESS AND BEING." Creativity Studies 9, no. 2 (April 11, 2016): 126–40. http://dx.doi.org/10.3846/23450479.2016.1164254.
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The article discusses three intertwined issues posed by the modern biotechnological immunology and its creativity potential: invention, naturalness and Being. In the first part, the author reflects on evolutionary paradigm and Peter Sloterdijk’s theory of immunology, particularly in their relation to the biotechnological enhancement of human beings. The second part discusses Being’s own naturalness and the possibility of creative invention. In the third part, the author combines these viewpoints to highlight some metaphysical challenges of biotechnological immunology. Most importantly the forgotten question of what does it mean to be, especially after the postmetaphysical annulment of the possibility of death. The core thesis argues for the naturalness of the creative biotechnological practice for preventive and reparative purposes.
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Minami, Yosuke, Miho Minami, Yachiyo Kuwatsuka, Ryohei Tanizaki, Yuka Nomura, Akihiro Abe, Hitoshi Kiyoi, and Tomoki Naoe. "Treatment with mTOR Inhibitor, Everolimus (RAD001) Overcomes Resistance to Imatinib in Ph-Leukemia Quiescent or T315I-Mutated Cells." Blood 114, no. 22 (November 20, 2009): 3277. http://dx.doi.org/10.1182/blood.v114.22.3277.3277.
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Abstract Abstract 3277 Poster Board III-1 Recent studies suggest that leukemia stem cells (LSCs) are responsible for relapse of leukemia following conventional or targeted agents and that eradication of LSCs might be necessary to cure the disease. Aberrant activation of mTOR signaling has also been reported to be involved in LSCs. In order to examine mechanisms of drug resistance in Ph-positive (Ph+) LSCs and to seek strategies to overcome the resistance, we've previously established in vivo-murine and ex vivo-culture models using murine hematopoietic pluripotent progenitors transduced with BCR-ABL (Minami, et al., Proc Natl Acad Sci USA, 2008). Furthermore, Ph+ leukemia (including T315I-, F311I-mutated CML-BC, or Y253H-mutated Ph-ALL) patient cells were serially xenotransplanted into immunodeficient NOD/SCID/IL2rγnull (NOG) mice. Engrafted bone marrow and spleen cells were almost identical to the original leukemia cells as to phenotypes including karyotypes and distribution of primitive populations. Spleen cells derived from leukemic NOG mice were co-cultured with S17 stromal cells and treated with imatinib and the mTOR inhibitor, everolimus (RAD001, Novartis Pharmaceuticals). While quiescent (Hoechst-33342low/Pyronin-Ylow) CD34+ cells were insensitive to imatinib in spite of BCR-ABL- and CrkL-dephosphorylation, substantial cell death including CD34+ population was induced with nM level of everolimus. In imatinib-resistant Ph+ leukemia cell lines harboring T315I-mutation (Baf3p210/T315I and TCC-Y/T315I), everolimus induced cell death with low IC50 values in PI-exclusion assays. We are also investigating detailed biomarkers in the cell death (such as phosphorylation of 4E-BP1 or p70 S6K) and effects of theses drugs in the leukemic NOG mice systems. These results imply that treatment with everolimus can overcome the resistance to imatinib in Ph+ LSCs or T315I-mutated cells. Disclosures: Kiyoi: Kyowa Hakko Kirin: Consultancy. Naoe:Kyowa Hakko Kirin, Wyeth and Chugai: Research Funding.
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Silva, Alessandra Suelen Jardim, Juliana Mendonça Freire, Lenilton Silva DA Silva Júnior, Gustavo Henrique de Medeiros Oliveira, Antonia Eduarda Martins Oliveira Elói Silva, João Pedro Andrade Lima, Victor lima Soares, et al. "Study of Diagnosis, Classification and Prognostic Factors Determination of Acute Leukemia By Flow Cytometry Immunophenotyping." Blood 134, Supplement_1 (November 13, 2019): 5175. http://dx.doi.org/10.1182/blood-2019-127357.
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Introduction: Acute leukemias (AL) are clonal diseases classified into two large groups: acute lymphoid leukemia (ALL), more common in children, and acute myeloid leukemia (AML), more rare in childhood, besides the rare phenotype leukemias such as acute biphenotypic leukemia (ABL) and undifferentiated acute leukemia (UAL). Although the cytomorphology still be relevant in theses leukemias diagnoses, the immunophenotyping by flow cytometry (FC) have become essential in the diagnosis, classification and follow-up of these neoplasms, standing out as a modern and practical methodology, presenting characteristically as a method of multiparametric and quantitative analysis of the leukemic cells. Objective: The objective of this study was realize a retrospective study of immunophenotyping in 371 patients with AL. Methodology: Immunophenotyping was performed biological samples by FC after labeling with a panel of monoclonal antibodies specific for AL directed against lymphoid antigens (B, T and NK cells), myeloid, and markers related to other cell immaturity. At the same time, information was obtained regarding patients such as age, sex, clinical data related to the disease, and previous hematological analysis. Results: From 371 cases, 127 were ALL (71 B lineage and 55 T=ALL), 239 AML, 04 ABL and 2 UAL. In the ALL, it was observed a higher frequency in children, contrasting with the cases of AML, ABL and UAL more prevalent in adults. In ALL, clinical signs and laboratorial data related to disease were more present in the ALL-T and mature-B corroborating with information in the literature. In AML, ABL and UAL the most clinical parameters observed were splenomegaly, hepatomegaly and bleeding. The classification FAB subtypes of AML more predominant were M1, M2 and M4 and lower incidence of the M7 subtype. Conclusion: These data demonstrate the importance of FC technology in the diagnosis, classification and establishment of prognostic factors of these neoplasms. Disclosures No relevant conflicts of interest to declare.
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Nakashima, S., T. Kameda, H. Shimada, R. Wakiya, M. Mahmoud Fahmy Mansour, M. Kato, T. Miyagi, K. Sugihara, Y. Ushio, and H. Dobashi. "FRI0254 SERUM IL-17 AND IL-21 AFFECT THE HEMODYNAMICS IN CONNECTIVE TISSUE DISEASE ASSOCIATED-PULMONARY HYPERTENSION." Annals of the Rheumatic Diseases 79, Suppl 1 (June 2020): 711.2–711. http://dx.doi.org/10.1136/annrheumdis-2020-eular.5537.
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Background:Pulmonary hypertension associated with connective tissue disease (CTD-PH) has complicated pathology including immune abnormalities, cardiac and pulmonary lesions. Therefore, it is difficult for rheumatologist to manage CTD-PH. We hesitate to use immunosuppressants in addition to pulmonary vasodilators to treat CTD-PH. Although there is a report that the cytokine such as Interleukin (IL)-6, IL-17 and IL-21 are involved in the development of PH1), changes in the hemodynamics of theses cytokines with treatment are not clear.Objectives:We investigate serum cytokine profile and clinical appearance in CTD-PH, and clarify the relationship between change in cytokines and hemodynamics before and after treatment.Methods:14 cases of CTD-PH (8 cases in Scleroderma; SSc-PH group, 4 cases in Mixed connective tissue disease; MCTD-PH group, 2 cases in Systemic lupus erythematosus; SLE-PH group), 6 cases in Other-PH group, and 2 cases of SSc without PH as controls were included. The following clinical data were collected: age, gender, underlying disease, complication of interstitial lung disease, treatment (immunosuppressant and pulmonary vasodilator). Serum samples in pre- and post-capillary before and after treatment were collected during cardiac catheterization examination. Serum cytokines (MCP-1, IL-6, IL-17 and IL-21) of these samples were measured by ELISA (ABCAM, UK).Results:Serum MCP-1, IL-6, and IL-21 levels were higher in SSc-PH group than in the other groups. Conversely, serum IL-17 levels tended to be higher in non-SSc group compared to SSc-PH group. Additionally, serum MCP-1 levels in SSc-PH group decreased in post-capillary as compared to pre-capillary. Furthermore, patients with decreased serum IL-17 and IL-21 levels before and after treatment showed improved pulmonary hemodynamics.Conclusion:SSc-PH had a different cytokine profile compared with non-SSc-PH. We suggested that the serum IL-17 and IL-21 levels effect the hemodynamics in CTD-PH.References:[1]Hashimoto-Kataoka T. et al. Proc Natl Acad Sci U S A. 2015 May 19;112(20):E2677-86.Disclosure of Interests:None declared
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Andreozzi, Valeska, Björn Vandewalle, and Jorge Parreira. "Comparing Randomized Controlled Data with Grouped Data from the Literature to Inform Decision Making When Standard of Care Has Low Level of Evidence Available." Blood 132, Supplement 1 (November 29, 2018): 5920. http://dx.doi.org/10.1182/blood-2018-99-118601.
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Abstract Objectives: To apply indirect comparison methods to inform decision-making when standard of care has low level of evidence available. Methods: Simulated treatment comparison is one of the methods available to generate indirect comparison of data in different formats and levels of evidence. It is particularly helpful when patient level data exists for one set of data and there is the need to compare it with single-arm studies. In theses cases evidence is deemed ''unanchored'' due to a lack of a common comparator. We use the case of second line treatment of chronic immune thrombocytopenic purpura (ITP). Splenectomy has been historically recommended in guidelines mainly due to the lack of evidence for other second-line therapies for ITP. We use individual patient data from eltrombopag clinical trials and one of the only studies published in the literature reporting long term outcomes for splenectomy to generate an indirect comparison of eltrombopag and splenectomy in second line treatment of ITP. Results: A subset of patients (n=84) treated with eltrombopag but not subject to splenectomy in the randomised, phase 3 study RAISE (PMID:20739054) were indirectly compared with a large retrospective cohort of patients (n=233) who underwent splenectomy for ITP in the study by Vianelli et al (PMID:23144195). After controlling for gender, age, baseline platelet count and number of previous therapies, we estimate that the proportion of patients with ITP responding to second line therapy is higher with eltrombopag in comparison to splenectomy: complete response (platelet count >100x109/L) 68.6% vs 54.0%; partial response 18.6% vs 24.6%; no response (platelet count <30x109/L)) 12.8% vs 21.4%. Conclusions: Indirect comparison with simulated treatment comparison is useful for decision making when no head to head data exists between new treatment options and standard of care and also when standard of care has low level of evidence available. Disclosures No relevant conflicts of interest to declare.
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Sun, Boyang, Donglei Zhang, Huiyuan Li, Xueqing Dou, and Renchi Yang. "Detection and Analysis of Gene Mutations in Patients with Glanzmann's Thrombasthenia." Blood 134, Supplement_1 (November 13, 2019): 2373. http://dx.doi.org/10.1182/blood-2019-129446.
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Background: Glanzmann thrombasthenia (GT) is a rare inherited disorder of bleeding, and it is characterized by the impaired or absent platelet aggregation to multiple physiologic agonists such as collagen, adenosine diphosphate (ADP), arachidonic acid(AA), but normal reaction to ristocetin. There is qualitative or quantitative defect in platelet integrin αIIbβ3(GPIIb/IIIa). Pathogenic variants of either αIIb or β3 unit could cause GT. The database of gene mutations is continuously updated on the Internet (http://www.hgmd.org); it totally lists 236 variants of ITGA2B gene and 170 variants of ITGB3 gene. Aim: To characterize the clinical manifestation and molecular basis of GT patients in China, and update the pathogenic variants database. Method: Clinical features are evaluated in 104 patients with GT. New generation sequencing was performed with a custom designed panel for the bleeding and platelet disease involving 76 genes, while ITGA2B and ITGB3 were enrolled. Result: The initial bleeding occurred before 1 age in most patients. Incidence of consanguinity is 12.5%. Symptoms lessened with age in about 30% patients. Female patients suffered more severe bleeding than male patients. Fifty different mutations were detected, among which 15 were novel. Most patients were compound heterozygotes and most mutations detected were missense mutations. Among 15 novel mutations, there were 7 missense mutations, 2 nonsense mutations, 2 splicing mutations, 4 frameshift mutations. Pathogenicity of all novel mutations were evaluated according to the standards and guidelines of ACMG. All variants detected were pathogenic or likely pathogenic. Furthermore, c.1750C>T [p.R584X] and c.2333A>C [p.Q778P] in ITGA2B were detected in 10 and 16 unrelated families, strongly suggesting a founder effect. Conclusion: Our study reports the largest cohort of GT in China, describing the clinical, laboratory and genetic characteristics of 104 patients. We found 15 novel pathogenic mutations in ITGA2B and ITGB3 causing GT. Theses novel findings expand the GT mutation spectrum. Disclosures No relevant conflicts of interest to declare.
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Sriram, Swetha, Anargyros Xenocostas, and Alejandro Lazo-Langner. "A Systematic Review Of The Role Of Erythropoietin In The Pathophysiology Of Anemia In Elderly Patients." Blood 122, no. 21 (November 15, 2013): 3432. http://dx.doi.org/10.1182/blood.v122.21.3432.3432.
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Abstract Introduction Anemia is a significant issue in the geriatric population, having an impact on morbidity, mortality and quality of life. However, identifying the etiology of anemia is not possible in ∼30% of patients. Postulated mechanisms of anemia of unknown etiology (AUE) include a blunted response to erythropoietin (EPO) or inadequate EPO production in response to anemia. The latter mechanism is of particular interest since it might be amenable to pharmacological intervention. Therefore, in order to explore the relationship between EPO levels and hemoglobin in elderly individuals with AUE, we conducted a systematic review of observational studies. Methods We searched Medline, EMBASE, Web of Science, Biosis Previews and Dissertations and Theses using the terms erythropoietin, anemia, elderly and diagnosis as MeSH subject headings. Additional relevant articles were identified by hand searching the meeting abstracts of the European Hematology Association (2006 - 2012) and the American Society of Hematology (2004 - 2012). For inclusion into the final review, studies needed to report data on EPO levels in elderly individuals diagnosed with anemia of unknown etiology. No meta-analysis was conducted due to the heterogeneity of the retrieved studies. Results The search identified 4277 potentially relevant citations, of which 31 studies were reviewed in full and 7 cohort studies (2 retrospective, 5 prospective) were included in the final review. The included studies involved 2534 participants ( 1). In general, studies found: 1) lower EPO levels in AUE compared to iron deficiency anemia and other forms of anemia; 2) a lack of correlation between EPO levels and the severity of anemia; and 3) EPO levels in AUE that are in general higher than in non-anemic patients. Conclusion Our findings suggest that EPO levels are generally elevated in elderly individuals with AUE, but remain inappropriately low, particularly when compared to anemia of other etiologies suggesting either a relative EPO deficiency, an abnormal EPO response or an abnormal erythroid cell response to EPO. Further research is required to elucidate the mechanisms involved, and the value of pharmacological interventions. Disclosures: No relevant conflicts of interest to declare.
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Jang, Suhwa, Young Lee, Byoung Kook Kim, and Hee Yong Chung. "Characterization of Leukemia-Inducing Genes Using the Retroviral cDNA Library." Blood 112, no. 11 (November 16, 2008): 5336. http://dx.doi.org/10.1182/blood.v112.11.5336.5336.
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Abstract Acute myeloid leukemia is a disease caused by oncogenic change(s) of hematopoietic stem cells or progenitor cells. The purpose of this research is to characterize the gene involved in the leukemia induction processes by screening the retroviral cDNA libraries of cellular oncogenes, homeobox genes. The cDNA libraries were constructed by cloning the individual genes to a MSCV retroviral vector backbone. For convenience of detecting the transduced cells and their protein products, the MSCV retroviral vector was modified to include HA tag and GFP marker and ninety cellular oncogenes and 30 homeobox genes were individually cloned and the structural integrity was verified. To screen for the leukemia-inducing genes, 5-FU treated mouse bone marrow cells were transduced with retroviral mixtures of oncogenes, and the cells were transplanted into ten lethally irradiated mice. All ten mice developed acute leukemia between the eight and ten weeks post-transplantation. The oncogenes that were responsible for the leukemia induction were characterized by genomic DNA PCR of the leukemic cells of each mouse. Surprisingly, all ten mice had c-myc genes in their leukemic cells. However, except for three mice, all the mice have additional oncogenes within the leukemic cells. The list of the additional genes include; RAB3D, RAB7B, PDGF-beta, CRK, and PIM-2 and Ras. Theses results show that the c-myc is the major leukemia-inducing oncogenes in our system. In addition, since the initial transduction rate of 5-FU-treated bone marrow just prior to in vivo transplantation was 10.56%, it is highly unlikely that all these additional oncogenes were present in the transplantable leukemic cells just by chance. Therefore, the retroviral cDNA library-mediated leukemia induction system we developed may be an useful system in systematically screening the cooperating oncogenes in leukemia induction. We are currently verifying the cooperating potential of the genes co-transduced with c-myc in inducing leukemia in the same animal model.
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Yang, Yang, Ming Xia, Ming Tan, Pengwei Huang, Weiming Zhong, Xiao Li Pang, Bonita E. Lee, Jarek Meller, Tao Wang, and Xi Jiang. "Genetic and Phenotypic Characterization of GII-4 Noroviruses That Circulated during 1987 to 2008." Journal of Virology 84, no. 18 (June 30, 2010): 9595–607. http://dx.doi.org/10.1128/jvi.02614-09.
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ABSTRACT The predominance and continual emergence of new variants in GII-4 noroviruses (NVs) in recent years have raised questions about the role of host immunity and histo-blood group antigens (HBGAs) in NV evolution. To address these questions, we performed a genetic and phenotypic characterization of GII-4 variants circulating in the past decade (1998 to 2008). Ninety-three GII-4 sequences were analyzed, and of them, 16 strains representing 6 genetic clusters were selected for further characterization. The HBGA binding properties were determined by both saliva- and oligosaccharide-binding assays using P particles as a model of NV capsid. The antigenic properties were also examined by enzyme immunoassay (EIA), Western blot analysis, and receptor blocking assay, using P-particle-specific antibodies from immunized mice and GII-4 virus-infected patients. Our results showed that 15 of the 16 GII-4 viruses bound to saliva of all A, B, and O secretors. Oligosaccharide binding assays yielded largely consistent results, although the binding affinities to some oligosaccharides varied among some strains. The only nonbinder had a mutation in the binding site. While antigenic variations were detected among the 16 strains, significant cross-blocking on the HBGA binding was also noted. Sequence alignment revealed high conservation of HBGA binding interfaces with some variations in adjacent regions. Taken together, our data suggested that the ability of GII-4 to recognize different secretor HBGAs persisted over the past decade, which may explain the predominance of GII-4 over other genotypes. Our data also indicated that both the host immunity and HBGAs play a role in NV evolution. While host immunity may continue driving NV for antigenic change, the functional selection by the HBGAs tends to lock the architecture of the capsid/HBGA interfaces and allows only limited variations outside the HBGA binding sites. A potential outcome of such counterselection between theses two factors in NV evolution is discussed.
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Zapata, Nidia, Espinoza Ramiro, Eduardo Cervera, Judith Cruz, Diana Arcos, and Juan Labardini. "Molecular Screening of 28 Genes in Mexican Patients with Acute Myeloid Leukemia a Series of 70 Patients from the Instituto Nacional De Cancerologia, Mexico." Blood 132, Supplement 1 (November 29, 2018): 5197. http://dx.doi.org/10.1182/blood-2018-99-115300.
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Abstract Introduction: Acute Myeloid Leukemia is a clonal heterogeneous disease, where age is an important risk factor to develop theses disease, PCR studies and next generation sequence have proven the diversity of these disease. A lot of genes mutations have been identifying to play a role in the DNA metilation, epigenetics a transcription. We initiate a screening to all acute myeloid leukemias that where the novo or relapse with a 28 gene panel of HEMAVISION a 28q; DNA diagnostic, for the detection al ARN gene fusion and alternatives of the: PML-RAR ALFA (bcr2,V), CBF-MYH11, RUNX1-RUNX1T1, PML-RAR alfa(bcr1,L), KMT2A-MLT3, PML-RAR alfa (bcr3,S), KMT2A-ELL, FUS-ERG, ETV6-MN1, DEK-NUP214, KMT2A-EPS15, KMT2A-AFDN, TCF3-PBX1, ETV6-RUNX1, KMT2A-MLLT1, KMT2A-AFF1, TCF3-HLF, STIL-TAL1, BCR/ABL(p190), SET-NUP214, BCR/ABL(p210), BCR/ABL(p230), ZBTB16-RARalfa, ETV6-ABL1, ETV6-PDGFRB, KMT2A-MLLT10, KMT2A-MLLT11,KMT2A-FOXO4, KMT2A-MLLT6, RUNX1-MECON, NPM1-RAR alfa, NMP1-MLF1, RUNX1-MECON. FLT3 ITD mutation and D385 by PCR electrophoresis by Invivoscribe was also perform. And the regular cytogenetics and FISH mutations for BCR/ABL, PML/RAR alfa, Inv16, MLL, +8, ETO, BCR, ABL, monosomy 7, monosomy8 Objectives The main objective is the know the mutation in the Mexican population and the prognostic in these group of patients Results These study was perform at Instituto Nacional de Cancerologia, Mexico, randomized patient from 2016-2018 where screen. A total of 70 patients, 37 females and 33 males, ages from 18-85years old, 54 patients where newly diagnosis of acute myeloid leukemia, 4 where relapses and 12 where secondary leukemias, the most frequent FAB morphologic classification where M4:22 cases, M2:15 cases, M3:8 cases, M1:4cases, M0 and M5:3 cases each. Of the 70 patients: 56 patients where negative to all of the panel screen, FLT3 where only perform in 14 patients 12 where negative and 2 where insufficient to perform the test, the most common FISH translocation was PML/RAR alfa, follow by MLL, ETO and +8. For the cytogenetics we had 21 cases that didn´t have enough metaphases, 7 normal, 28 cases with more than 2 cytogenetics alterations and 9 with only 1. With a Cytogenetics risk: high risk 44, intermedium:10 and low12. Of the 70 patient, 14 have some genes mutations +: t(9;11)(p22;q23) KMT2A-MLLT3, t(6;11)(q27;q23) KMT2A-AFDN, t(5;12)(q33;p13) ETV6-PDGFRB, t(8;21)(q22;q22) RUNX1RUNX1T1, inv16(p13q;22q) CBFB-MYH11, t(6;11)(q27;q23) KMT2A-AFDN, t(3;21)(q26:q22) RUNX1-MECOM, inv16(p13q;22q) CBFB-MYH11, t(15;17)(q24;q21) PML-RARA (bcr2,V) t(15;17)(q24;q21) PML-RARA (bcr1,L) t(15;17)(q24;q21) PML-RARA (bcr3,S), t(8;21)(q22;q22) RUNX1RUNX1T1, t(8;21)(q22;q22) RUNX1RUNX1T1, t(15;17)(q24;q21) PML-RARA (bcr3,S) Out of 70 patients: 38 receive 7+3 (cytarabine + Daunorubicin) for first line of treatment, 41 received high doses of cytarabine at 3g /m2. Our first option for relapse treatment is MEC (mitoxantrone, cytarabine and etoposide) because of costs and the second line of rescued treatment is Flag- Ida (idarubicin, fludarabine and cytarabine) and not all patient can afford it. For the elderly patients the first line of treatment is low dose of cytarabine and only in those who can pay azacytidine it is use. The correlation between high risk cytogenetics with mortality is 12 cases out 70. And genes with morality only 4 patients with: t(9;11)(p22;q23) KMT2A-MLLT3, t(6;11)(q27;q23) KMT2A-AFDN, t(5;12)(q33;p13) ETV6-PDGFRB, t(6;11)(q27;q23) KMT2A-AFDN Conclusion We need to know our population characteristics, we don´t have the incidence and prevalence of the gene mutation in the Mexican population. In the market there are several screening panels with different genes. We need to have more genes and more patient to be analyzed to learn our molecular risk, to have a better approach to these patients and better techniques. There is no paper publish with the genetics and gene alteration in the Mexican Population, it is important to continuing working and to use panels with genes as ASXL1, FLT-TKD, CEBPA, KIT, KRAS, IDH1,2, TET2 and others. And other important issue that we found is the high number of patient that abandon treatment 4 cases, because of money issues. And the time of these population 24 patient where death. The incidence of FLT3 mutation ITD and D385 is low in theses population but it was performed only 14/70 patients, we need a large number of patient to know the real incidence. Table. Table. Disclosures No relevant conflicts of interest to declare.
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Piechota, Małgorzata, Barbara Kot, Aneta Frankowska-Maciejewska, Agata Grużewska, and Agnieszka Woźniak-Kosek. "Biofilm Formation by Methicillin-Resistant and Methicillin-SensitiveStaphylococcus aureusStrains from Hospitalized Patients in Poland." BioMed Research International 2018 (December 27, 2018): 1–7. http://dx.doi.org/10.1155/2018/4657396.
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Biofilm-mediated infections in the hospital environment have a significant negative impact on patient health. This study aimed to investigate biofilm production in vitro and the presence oficaABCDgenes in methicillin-resistantS. aureus(MRSA) and methicillin-sensitiveS. aureus(MSSA) strains isolated from hospitalized patients. MRSA (73) and MSSA (57) strains were evaluated for biofilm production by the microtiter plate method. The presence oficaoperon was investigated by PCR. Out of 130 strains, 99.2% were biofilm producers. Strong biofilms were formed by 39.7% of MRSA and 36.8% of MSSA strains. The highest percentage of strong biofilm producers was found among the strains isolated from sputum and tracheostomy tube (66.7%), nose and catheter (50%), throat (44.4%), and bronchoalveolar washings (43.8%). The strains isolated from bronchoalveolar washings produced significantly more biofilm than strains isolated from wound and anus. The ability of biofilm forming by fecal strains was significantly lower compared to strains from other materials. MRSA strains had significantly higher ability of biofilm formation than MSSA strains (P= 0.000247). The presence oficaoperon in MRSA was detected in all strains. Comparison of strong biofilm biomass of the strains withicaABCD,icaABD, andicaADrevealed that strains withicaABCDandicaABDproduced highly significantly more biofilm than strains withicaAD. Biofilm forming by both MRSA and MSSA strains indicates high ability of theses strains to persist in hospital environment which increases the risk of disease development in hospitalized patients.
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Erdfelder, Felix, Iris Gehrke, Rajesh Kumar Gandhirajan, Magdalena Hertweck, Regina Razavi, Julian Paesler, Alexandra Filipovich, et al. "Expression of the Transcription Factor Lymphoid Enhancer Binding Factor-1 (LEF-1) Is Correlated to Aberrantly Increased Fibromodulin Transcripts in Chronic Lymphocytic Leukemia (CLL)." Blood 114, no. 22 (November 20, 2009): 4589. http://dx.doi.org/10.1182/blood.v114.22.4589.4589.
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Abstract Abstract 4589 Chronic lymphocytic leukemia (CLL) is characterized by accumulation of monoclonal CD5+ B lymphocytes. Fibromodulin is a secreted extracellular matrix protein usually found in articular cartilage, bones, connective tissue and collagen rich tissues but has been shown to be excessively expressed in CLL. Moreover, we and others could show that WNT signaling is highly activated in CLL. The aim of our study was to investigate a possible connection between fibromodulin overexpression and the WNT pathway. Moreover, we wanted to explore possible relations between these parameters and the prognosis of CLL. Fibromodulin mRNA transcripts were correlated with the mRNA expression of the WNT pathway transcription factors lymphoid enhancer binding factor-1 (LEF-1) and T cell factor-4 (TCF-4) in primary CLL cells. Furthermore, we assessed correlations between the mRNA expression levels with ZAP-70 and CD38 protein expression. These parameters have been shown to be associated with a poor prognosis. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was used and calculated PCR-efficiency corrected relative expression values of Fibromodulin, LEF-1 and TCF-4 mRNA in primary CLL-cells determined. ZAP-70 and CD38 expression was determined by flow cytometry analysis. Based on 20 primary CLL samples we found a significant positive correlation between Fibromodulin and LEF-1 mRNA levels (Spearman's rho = 0.65, p = 0.009). Also, LEF-1 and TCF-4 were found to be strongly correlated (Spearman's rho = 0.61, p = 0.027). In contrast, fibromodulin and TCF-4 mRNA levels were only weakly correlated (Spearman's rho = 0.33, p = 0.271). CD38 and ZAP-70 did not correlate significantly to any of the other values. Both parameters did also not exhibit significant correlations with fibromodulin, LEF-1, and TCF-4 mRNA. The positive correlation of TCF-4 and LEF-1 was expected as LEF-1 has been shown to be a target gene of TCF transcription factors. The strong positive correlation of fibromodulin and LEF-1 indicates that fibromodulin might be a target gene of LEF-1 or part of the same (TCF-4 independent) transcription regulation pathway. Studies investigating theses functional relationships are underway. Disclosures: Hallek: BayerScheringAG: Honoraria, Research Funding.
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Zou, Dehui, Lugui Qiu, Yenan Li, Yaozhong Zhao, Mingzhe Han, and Sizhou Feng. "A High Long-Term Survival Were Produced by Early Sequential Intensive Consolidation Chemotherapy Followed by Autologous Stem Cell Transplantation in First Complete Remission in Patients with Adult Acute Lymphoblastic Leukemia." Blood 112, no. 11 (November 16, 2008): 3324. http://dx.doi.org/10.1182/blood.v112.11.3324.3324.
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Abstract Objective: To investigate the efficiency of our designed protocol with early sequential intensive consolidation chemotherapy followed by autologous stem cell transplantation (ASCT) in first complete remission (CR1)on adult acute lymphoblastic leukemia (ALL) patients who lack of matched sibling donors. Methods: The ALL patients were given a median 4 intensive/ consolidation therapy courses including high-dose of methotrexate, cytarabine and/or cyclophosphamide in CR1, followed by ASCT with total body irradiation (TBI)-based conditioning regimen. A 1 to 1.5 year maintenance chemotherapy with 6-mercaptopurine, methotrexate and vincristine-prednisone in combination with interleukin-2/IFN-α immunotherapy were administered after adequate hematological recovery. Results: A total of 83 patients were enrolled in this study. 78 patients reconstituted hematopoiesis except 5 patients died of early toxicity. By a median 47(3– 218) months follow-up after ASCT, the 5-year probabilities of leukemia free survival (LFS) and overall survival (OS) were 53.2%±5.9% and 54.4%±5.8%, respectively. The 5-year cumulative relapse rate (RR) was 35.7%±6.1%, and no patients relapsed more than 3 years after transplantation. According to the adverse features, age ≥35 years, WBC >30×109/L for B-cell ALL or >100×109/L for T-ALL, Pro-B and early T immunotyping, more than 4 weeks to attain remission, and high-risk karyotypes including t(9;22)/BCRABL, t(4;11)/ALL1-AF4 or t(1;19)/E2A-PBX1, the patients were identified in 3 prognostic risk groups of standard (24, 29.0%), intermediate(32, 38.5%), and high(27, 32.5%) with 0, 1, 2 or high-risk karyotypes adverse features, respectively. The 5-year probabilities of DFS were 80.36%±9.0%, 59.39%±9.2% and 24.78%±8.9% in these 3 group s, respectively. While, the 5-year cumulative incidences of relapse was 16.7%,37.5% and 70.4%, respectively.. The standard and intermediate -risk groups had significantly better OS and LFS than the high-risk group. Conclusion: Theses results indicated our designed protocol with early sequential intensive consolidation chemotherapy followed by autologous stem cell transplantation (ASCT) in first complete remission (CR1) could produced a relatively high log-term survival in standard and intermediate-risk group of adult ALL patients.
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Won, Sung Chul, Sun Young Rha, Seung Hwan Oh, Hwang Min Kim, and Chuhl Joo Lyu. "Clinical Implication of Bone Marrow Angiogenesis in Childhood Acute Lymphocytic Leukemia." Blood 106, no. 11 (November 16, 2005): 1455. http://dx.doi.org/10.1182/blood.v106.11.1455.1455.
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Abstract Purpose: Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are associated with increased angiogenesis, growth, and metastasis in solid tumors. But, until today, the importance of theses factors on leukemia, especially childhood acute lymphocytic leukemia (ALL) has received limited attention. Therefore, this study examined the bone marrow plasma VEGF and bFGF levels in ALL patients and normal controls. Methods: Bone marrow plasmas at diagnosis from 33 ALL patients (median age 5.97 years; range 1.8–13.9 years) were used for analysis. All ALL patients are divided into 3 groups; one group is standard risk ALL, other group is high risk ALL who did not experience relapse, and the other group is high risk ALL who have experienced relapse during treatment. The bone marrow levels of bFGF and VEGF were determined by enzyme-linked immunosorbent assay (R & D Systems) and compared with the bone marrow levels of 7 healthy control subjects (median age 11.98 years; 6 months–13.6 years). Results: The VEGF levels were higher in high risk ALL with relapse (N=7, 171.95 ± 202.54 pg/mL) compared with standard risk (N=9, 9.11 ± 21.47 pg/mL), high risk ALL without relapse (N=17, 37.09 ± 38.77 pg/mL) or normal control (N=7, 72.9 ± 68.22 pg/mL)(P=0.006). Figure Figure The bFGF levels were also significantly higher in high risk ALL patients who experienced relapse (high risk with relapse; 37.26 ± 35.73 pg/mL, standard risk ALL; 7.90 ± 12.99 pg/mL, high risk ALL without relapse; 7.27 ± 10.93 pg/mL, normal control; 6.39 ± 6.13 pg/mL) (P=0.003). Figure Figure The Pearson correlation coefficient between VEGF and bFGF was 0.775 (P < 0.01). Three patients who showed high levels of both angiogenic markers (VEGF; 366.80 ± 141.29 pg/mL, bFGF; 70.27 ± 5.57 pg/mL) are relapsed and died due to disease progression. The other relapsed patients except these 3 patients are alive without disease after salvage chemotherapy. Conclusion: Our data suggest that the increased levels of VEGF and bFGF in bone marrow may play an important role in estimating prognosis of childhood ALL.
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Shen, Shuhong, Yin Liu, JingYan Tang, and Long-Jun Gu. "FLT3, NPM1 and MLL Mutations Help Risk Stratification in Pediatric Acute Myeloid Leukemia." Blood 114, no. 22 (November 20, 2009): 1574. http://dx.doi.org/10.1182/blood.v114.22.1574.1574.
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Abstract Abstract 1574 Poster Board I-600 Introduction Acute myeloid leukemia (AML) is a heterogeneous disease which harbors various genetic alterations. Among theses genetic events, Mutations of FLT3, NPM1, MLL and other genes often predict prognosis, particularly in cases cytogenetic normal (CN-AML). Could these be criteria for risk stratification in Pediatric AML ? Patients and Methods 155 cases of de novo AML were diagnosed routinely according to morphology, immunology, cytogenetics, and molecular biology examination on bone marrow (BM) aspirates between Jan. 2002 and Dec. 2008. All patients received chemotherapy according to the AML-XH-99 protocol, which consist of Daunorubicin, Cytosine arabinoside, Etoposide, Homoharringtonine. For acute promyelocytic leukemia, all-trans retinoic acid and Arsenic trioxide were also included. Meanwhile, total RNA of leukemic cells form all diagnostic BM samples were extracted, and then reverse transcribed. MLL partial tandem duplication (MLL/PTD) fusion transcripts were screened by real-time quantitative polymerase chain reaction. FLT3 internal tandem duplication (FLT3/ITD), FLT3 tyrosine kinase domain mutation (FLT3/TKD) and NPM1 mutation were examined by High resolution melting analysis. Results Of the 155 children with de novo AML, 121(78.1%) had received chemotherapy for more than one week with data available for analysis. Among them, 55(45.5%) was cytogenetically normal (CN-AML). In this total cohort of patients 49(27.09%) had FLT3/ITD (32.70% in CN-AML), 14 (9.03%) had FLT3/TKD (7.30% in CN-AML), 62 (40%) had NPM1 mutation (49% in CN-AML), and additional 8 (5.16%) had MLL/PTD (5.50% in CN-AML). In this cohort of patients 98 (63.22%) had at least one mutation. The clinical outcomes were listed in table 1. Generally, patients with FLT3 mutation (ITD or TKD mutation) usually have worse results after chemotherapy, as reported previously by other researchers. Meanwhile, NPM1 mutations usually predict better prognosis in our cohort of AML patients. MLL/PTD always predicts the worst outcome in AML as other MLL rearrangements in leukemia. Among CN-AML patients, 5-year EFS and OS were similar to whole cohort of patients according to those mutations. Cox regression analysis in a univariate model revealed that the presence of FLT3/ITD and NPM1 was significant prognostic factor of EFS, (P<0.05). We therefore proposed a molecular-risk classification of pediatric AML patients based on the data we got in this study. For the newly classified groups of low, medium and high risk groups, EFS rate was 62.03%±8.42%, 45.42%±4.52%, and 14.85%±2.99%, respectively, P=0.00. CRD for the 3 groups was 27.69±21.34 months, 22.62±19.64 months, 13.26±11.95 months, respectively, p=.022. Our results indicate that combinations of these couple of molecular events may be the useful tool for further classify AML in children. Disclosures No relevant conflicts of interest to declare.
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Bousquet, J., T. Bieber, W. Fokkens, M. Humbert, M. Kowalski, B. Niggemann, and H.-U. Simon. "Themes in Allergy." Allergy 61, no. 1 (January 2006): 1–2. http://dx.doi.org/10.1111/j.1398-9995.2006.01020.x.
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Stoppa, Anne-Marie, N. Vey, C. Faucher, R. Bouabdallah, F. Viret, C. Chabannon, P. Ladaique, et al. "Stem Cell Transplantation in 281 Patients over 60 Years in Onco Hematologic Malignancies: A Single Center Experience." Blood 104, no. 11 (November 16, 2004): 5229. http://dx.doi.org/10.1182/blood.v104.11.5229.5229.
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Abstract From 06/1982 to 12/2003, 281 patients (163 male) received autologous (AuSCt) and/or allogeneic (AlloSCT) transplantations as part of first line treatment (61%) second line treatment(21%) or salvage therapy (17%). There were 140 multiple myelomas (including 3 amyloidosis), 62 lymphomas, 51 acute myeloid leukemias, 24 solid tumors and 4 chronic myeloid leukemias.Two hundred and sixty nine patients-median age: 64 y(60–77) - received Au SCT (28/261 received a second Au SCT in med of 2–7 months), 1patient (71 y) received a double syngeneic transplant,12 patients -median age: 62(60–68) received AlloSCT. Conditionning regimen (Cond reg) for the first Au SCT included Melphalan 140 mg/m² n=168; BEAM n=41; Melphalan 200mg/m² n=29; Mel Busulfan (8 to 16 mg/kg) or Mel TBI 8 grays n=18; Cytoxan 200mg/kg Melphalan1 40 n=9: TBI 8 Grays n=7; BCNU 600mg/m² n=3. Cond reg for the second Au SCT included Mel 200 n=20; Mel 140 n=4; BCU 600 n=4. Cond reg for the double syngeneic transplant were Mel 140 and Bu 8 Mel 140.Cond reg for the AlloSCT were Fludarabin/Busulfan/ATG (FBS) based reduced intensity regimen (RIC) in 11 cases. Ninety percent of the patients received peripheral blood stem cells-med 5,3.10*6CD34/kg (2–40)- 10% of the patients transplanted before 1994 received bone marrow cells. Tolerance was good with only 9/281 transplant related deaths: 5 sepsis/MOF in aplasia (D11–D21) 2 ARDS (D30–D80) 1 AGVH (D 60) 1 CGVH (3 years). Theses deaths occured in 4 ref MM( 2 renal instability,1 cardiac amyloidosis), 2 CR2 NHL, 1 OR1 MM, 1 stable MDS after FBS in 2 pts, BEAM in 2 pts, Mel 140 in 4 pts, Mel 200 (second transplant) in 1 pt. With a median follow up of 20 months from transplant,7/281 second cancers had occured: 1AML, 1ALL, 2 NSC pulmonary carcinoma, 1 colon cancer, 1uterin carcinoma, 1 melanoma; 2 of these 7 patients had died. One hundred and fifty eight patients had relapsed in a median of 11 months (1–92). Median survival for the 281 patients is 37 months without diference between pts older or younger than 65. Median survival for MM, NHL/HL, AML or ST are 40 months, 45 months, 18 months and 23 months respectively
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Vielreicher, Martin, Ralf Steinmeyer, Gregory Harms, Ulrich Walter, and Achim Obergfell. "Dynamic Regulation of Src by Csk in Integrin α IIb β 3 Mediated Signalling." Blood 106, no. 11 (November 16, 2005): 2661. http://dx.doi.org/10.1182/blood.v106.11.2661.2661.
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Abstract When platelets adhere to fibrinogen, they undergo profound actin rearragements and spreading. These responses require signals triggered by aIIbb3, a process known as outside-in signalling. Although aIIbb3 signaling is required for normal hemostasis, its molecular basis is incompletely understood. Recently we demonstrated by coimmunoprecipitation techniques that several Src family kinases are constitutively associated with aIIbb3 in platelets. Fibrinogen binding to aIIbb3 leads to rapid activation of Src and recruitement and activation of the Syk tyrosine kinase. Theses interactions are significant because murine platelets defiecient in multiple Src kinases or Syk fail to spread on fibrinogen. One of the earliest events upon binding of fibrinogen to the receptor is the assembly of a multimolecular protein complex inculding the tyrosine kinases Src and Syk. Phosphorylation of Src at Tyrosine 418 leads to Src activation, phosphorylation of Tyrosine 529 to inactivation. The inactivation of Src is mediated by Csk. Previous studies have shown that in resting platelets Csk forms a complex with integrin α IIb β3 and Src. After fibrinogen binding Csk leaves this complex. The aim of this study is to elucidate the dynamics of Src inactivation and the interaction with Csk in this process by using Fluorescence Resonance Energy Transfer (FRET). A5 CHO cells stably expressing integrin α IIb β3 were transfected with Csk-YFP and various Src mutatnts in a GFP vector. After cell adhesion on fibrinogen, a strong FRET signal could be observed at the edges of nascent lamellipodia of the spreading cell. The FRET signal was dynamic since it was transported in waves inside the cell within seconds. After completion of the adhesion process a weak FRET signal remained visible inside the cell. A different pattern could be observed when cells were transfected with Csk and the active Src mutant Y529F. The observed FRET signal was much weaker and less dynamic after cells were allowed to spread on fibrinogen suggesting that the muation of Tyrosine 529 to Phenylalanine inhibited Csk binding and inactivation of Src. This was not due to a spreading defect since the cells appeared to spread on fibrinogen normally. The use of the inactive Src mutant K295R revealed a FRET signal comparable to WT Src. These studies suggest that Src inactivation is dynamically regulated within seconds after cell adhesion on fibrinogen and that Tyrosine 529 is required for the interaction of Csk with Src in this process.
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Giraldo, Pilar, Esther Franco-Garcia, Ramiro Alvarez, Gloria Garcia-Carpintero, Mercedes Pascual, and Carmen Martos. "Primary Extranodal Lymphoma: Revised Incidence, Location, Histological Classification and Survival in Zaragoza (Spain), 1992–2002." Blood 112, no. 11 (November 16, 2008): 4674. http://dx.doi.org/10.1182/blood.v112.11.4674.4674.
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Abstract Background. Leukaemia and non-Hodgkin’s lymphomas (NHL) are the commonest haematological malignancies (HMs), accounting for about 10% of incident cases and 6% of all cancer deaths in the European Union (EU). There are few studies in order to estimate the differences in incidence and survival of primary extranodal Lymphoma (PEL). In Zaragoza (Spain) there is a population-based Cancer Registry (ZPCR), that includes all non-haematological and haematological (HMs) incident cases, conducted since 1960. The main aims of this study are: To review all cases with PEL diagnosis. To estimate the incidence of PEL in the ZPCR registered cases during the period 1992–2002. 2. To calculate the survival of PEL Methods. All PEL occurred in patients residing in Zaragoza during the period 1992–2002 were selected from ZPCR. All cases were reviewed in order to confirm the primary location and the morphology classification according to REAL. The population at risk was 9.266.609 person-years. The crude (CIR) and age standardized incidence rate (ASR) were calculated, using the European population as standard. Kaplan-Maier method was applied to calculate median survival time and their 95% confidence intervals, as well as 5-year survival. The end of follow-up was 31 December 2007 and Log-Rank was used to compare survival curves. Results. Among all 4,340 HMs, a total of 1,757 (40.0%) were NHL (CIR: 19×105 person-year), 252 (14.0%) of them were classified as PEL, yielding an ASR of 2.1×105 person-year (males: 140 cases (53.0%), mean age: 59.2 years, ASR: 2.7×105 person-year and females 112 cases (47.0%), mean age: 66.8 years ASR: 1.6×105 person-year. According to topography the most frequent sites were digestive tract (50.4%), skin (19.8%), gland tissue (10.0%), oral cavity-pharynx (7.9%), lung (2.9%), CNS (2.4%), orbit (2.1%) and others (4.5%). The median survival for PEL was 6.61 years (95%CI: 3.7–9.5) and for nodal Lymphomas 5.01 years (95%CI: 4.1–5.9). The 5-year relative survival was 53.5% and 50.0% respectively. There are not significant differences between both groups. Moreover, non significant differences in survival were observed between males and females. Theses results are similar to those found in the EU. Conclusions. The occurrence of PEL according to gender and mean age is similar to nodal NHL. No significant differences in survival were observed between nodal and extranodal NHL, probably the survival is more conditioned by the histology than topography.
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Bruns, Heiko, Dimitrios Mougiakakos, Mario Fabri, Shirin Pasemann, Regina Jitschin, Kerstin Amann, Maike Büttner, Andreas Mackensen, and Armin Gerbitz. "Vitamin D Triggers Killing of Burkitt Lymphoma Cells by Human Macrophages." Blood 120, no. 21 (November 16, 2012): 1035. http://dx.doi.org/10.1182/blood.v120.21.1035.1035.
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Abstract Abstract 1035 Introduction: Distinct macrophage subsets have been linked with either protective or pathogenic roles in cancer. M2 macrophages have been shown to suppress adaptive tumour-specific immune responses and promote tumour growth, while M1 macrophages eliminate neoplastic cells and activate tumour-killing mechanisms. Macrophages isolated from solid tumours (tumor-associated macrophages, TAM) reveal a suppressive and tumor promoting M2-like phenotype. However, it remains unclear which tumoricidal effector mechanisms are compromised in theses immune cells. While infilatration of macrophages is a characteristic morphological hallmark in Burkitts' lymphoma (BL), the phenotype and functional properties of TAM as part of the stroma are poorly understood. In this study we investigated the phenotype of lymphoma associated macrophages (LAM) and the mechanisms by which macrophages are able to modulate the growth of tumor cells. Methods: We included 19 patients diagnosed with BL and 20 patients with benign reactive lymphadenopathy as a control group. Phenotypic characterizations of LAM were evaluated by immunohistochemistry and by qPCR in paraffin embedded tissues. To investigate anti lymphoma cytotoxicity of distinct macrophage subsets we generated macrophages from PBMC either by GM-CSF to create the M1 phenotype, or by M-CSF to obtain the M2 phenotype. M1 and M2 macrophages were coincubated them with several BL cell lines and cytotoxicity was analyzed by flow cytometry, qPCR and immunofluorescence. Results: First, we could demonstrate that BL infiltrating macrophages displayed an anti-inflammatory M2-phenotype characterized by expression of surface markers such as CD68 and CD163. Second, we identified impaired vitamin D metabolism in LAM and M2 macrophages as demonstrated by low expression of vitamin D receptor and Cyp27B1, and increased expression of Cyp24A1. Third, macrophages revealed a reduced cytotoxic potential towards lymphoma cells which was dependent on the expression of the vitamin D dependent peptide cathelicidin. Finally, we could demonstrate that excess supplementation of calcitriol led to increased cytotoxicity of M1- and M2 macrophages towards BL cells. Conclusions: These results suggest a mechanism in which vitamin D is required for innate immunity to overcome the ability of lymphoma cells to evade macrophage-mediated antitumoral responses. The present findings underscore the importance of vitamin D for sustaining innate immunity and imply that the therapeutic activation of the vitamin D pathway may even result in triggering tumoricidal effector mechanisms of LAM. Disclosures: No relevant conflicts of interest to declare.
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Bagge, Eva. "Summary of Thesis." Scandinavian Journal of Rheumatology 21, no. 1 (January 1992): 48. http://dx.doi.org/10.3109/03009749209095064.
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Kim, Hee Nam, Jeong-Hwa Choi, Ju-Hyun Yun, Yeo-Kyeoung Kim, Il-Kwon Lee, Deok-Hwan Yang, Myung-Geun Shin, et al. "Notch 4 Plymorphism Associated with Risk of Aplastic Anemia." Blood 108, no. 11 (November 16, 2006): 988. http://dx.doi.org/10.1182/blood.v108.11.988.988.
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Abstract Aplastic anemia (AA) is a heterogeneous disease characterized by bone marrow aplasia and peripheral blood pancytopenia, a profound deficit of hematopoietic stem and progenitor cells. Although the causes for aplastic anemia are multiple, the pathogenic mechanism leading to the disease might be limited to two main pathways: environmental factors and genetic susceptibility. The mammalian Notch family has four homologs, Notch1 to 4. Notch4 signaling plays a role in lineage commitment in hematopoiesis. The human Notch4 gene is located at the MHC on chromosome 6p21.3. Many single nucleotide polymorphisms (SNP) have been mapped to the Notch4 locus. Therefore, we hypothesized that some of these variants could potentially influence the expression level of Notch4 protein. To investigate the role of the two Notch4 rs2071282 (+2763 C>T, Pro204Leu) and 422951(+3323 A>G, Thr320Ala) polymorphisms in susceptibility to AA, a case-control study was conducted in Chonnam National University Hwasun Hospital between March 2002 and June 2006. We genotyped in 151 AA patients and 552 healthy control subjects using PCR-RFLP. Here, we report that rs2071282 CT genotype is significantly associated with increased risk for AA. Using subjects with the rs2071282 CC as a reference group, the odds ratio (OR) of CT is 1.77 (95% CI 1.05–2.98, p=0.032). The haplotype CG is significantly associated with the risk of AA. Using subjects with the haplotype CA, the OR of haplotype CG is 0.70 (95% CI 0.50–0.98, p= 0.037). In conclusion, we were able to identify two polymorphisms of Notch4, therefore, theses results suggest that Notch polymorphism may play an important role in the susceptibility to AA. Distribution of Notch polymophisms in aplastic anemia patients and controls Control (%) AA(%) OR (95% CI) p Rs 422951 AA 302(54.8) 94(62.3) 1 AG 216(39.2) 49(32.5) 0.72(0.478–1.098) 0.128 GG 33(6.0) 8(5.3) 0.54(0.23–1.279) 0.163 Total 551 151 Rs 2071282 CC 481(87.1) 120(81.1) 1 CT 68(12.3) 28(18.9) 1.769(1.049–2.982) 0.032 TT 3(0.5) 0 - - Total Haplotype frequency for AA and controls Haplotype Control (%) AA(%) P (Chi-square) OR (95% CI) p *Haplotype frequencies were estimated from genotype data using Gibbs- algorithm.*OR and 95%Cis were estimated using based logistc regression and adjusted for sex and age C-A 743(67.7) 206(70.0) 0.062 1 C-G 281(25.6) 60(20.4) 0.70(0.50–0.98) 0.037 T-A 72(6.5) 28(9.5) 1.49(0.90–2.47) 0.126
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Palmqvist, Lars, Nicolas Pineault, Bob Argiropoulos, Adrian Wan, and Keith R. Humphries. "FLT3 Expression Is Increased by MEIS1 and Collaborates with NUP98-HOX Fusion Genes in the Induction of Acute Myeloid Leukemia." Blood 104, no. 11 (November 16, 2004): 2552. http://dx.doi.org/10.1182/blood.v104.11.2552.2552.
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Abstract The TALE family member and HOX cofactor MEIS1 is important in leukemic transformation. MEIS1 has, although non-leukemogenic on its own, been shown to strongly collaborate with several HOX genes and NUP98-HOX fusions to induce acute myeloid leukemia (AML). We have recently described a novel in vitro culture system of cell lines established from murine primary bone marrow cells transduced with the AML-associated fusion gene NUP98-HOXD13 or an engineered NUP98-HOXA10 fusion. These pre-leukemic NUP98-HOX cell lines are transplantable and can efficiently be converted into AML-inducing cells upon MEIS1 transduction. Conveniently, the MEIS1 transduced cells can be purified and preserve their leukemogenic potential even after extensive in vitro expansion. Thus, the availability of the NUP98-HOX cell lines system provided the opportunity to investigate and characterize the mechanism of MEIS1-mediated AML-conversion. Potentially interesting target or candidate genes were screened for expression changes between the parental pre-leukemic lines and AML-inducing MEIS1 transduced cell lines with quantitative RT-PCR and Western blotting. Aberrant expression or mutations of the receptor tyrosine kinase FLT3 gene is a common finding in human AML. Interestingly, Flt3 was found induced 5 to 10 fold in MEIS1 transduced cell lines compared to the parental cell lines. The observed increase in Flt3 expression provided the MEIS1 transduced cells with Flt3 ligand driven growth. This was not seen in the parental cell lines, which could not proliferate with Flt3 ligand as single cytokine or with a MEIS1-homeodomain mutant expressing cell line. Importantly, the Flt3 inhibitor AG1295 could block the proliferative effect of the Flt3 ligand in the MEIS1 transduced cell lines. To test whether Flt3 could substitute for MEIS1-mediated induction of AML in NUP98-HOX pre-leukemic cells, a NUP98-HOXA10 cell line was transduced with an MSCV-Flt3-IRES-YFP construct. The resulting Flt3-transduced cells were shown to express Flt3 at levels similar to that of MEIS1 transduced cells without any significant increase in endogenous Meis1 expression. Transplantation of these cells into mice led to lethal and transplantable AML with a median disease onset of 116 days (n=8) compared to 59 days for MEIS1 (n=4), whereas control transplants remained healthy (n=2). In conclusion, this study demonstrates that MEIS1 can induce Flt3 expression and that Flt3 can collaborate with NUP98-HOX fusion genes in the induction of acute myeloid leukemia. Furthermore, theses results suggest a model in which the leukemogenic synergism of MEIS1 on HOX-mediated leukemia might in part be mediated through FLT3-dependent pathways.
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Chriti, N., M. Boudigou, E. Porchet, J. O. Pers, S. Hillion, and D. Cornec. "THU0034 AUTOREACTIVE B CELLS ESCAPE PERIPHERAL CHECKPOINT IN ANCA-ASSOCIATED VASCULITIS AND SJÖGREN’S SYNDROME." Annals of the Rheumatic Diseases 79, Suppl 1 (June 2020): 230.3–230. http://dx.doi.org/10.1136/annrheumdis-2020-eular.5748.
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Background:B cells play a central role in many autoimmune diseases (AIDs) including ANCA-associated vasculitis (AAV) and primary Sjögren’s syndrome (pSS). Most of the research that has been conducted on AID has focused on the production, secretion, and pathogenicity of auto-antibodies, but little is known on the characteristics of autoreactive B cells in humans.Objectives:This study aims at characterizing circulating autoantigen (PR3 ou SSA)-specific B-cells in patients with AAV and pSS compared to healthy subjects to better understand their role in thenatural and pathological autoimmunityand define the mechanisms leading to the breakdown of self-tolerance in patients with AID.Methods:First, we developed a new flow-cytometry method to detect circulating auto-reactive B cell based on the specificity of their B-cell receptor (BCR). To study surface phenotype of specific B cells by flow cytometry, blood samples were collected from patient with PR3-ANCA AAV, pSS and from healthy subjects. Functional analysis of antigen-specific B cells was also elicited by in vitro analysis of their capacity to secrete immunoglobulins against SSA or PR3 antigens by ELISPOTResults:Phenotype analysis showed that antigen-specific B cells in patients have a memory phenotype compared with healthy controls (5 to 9% are IgG-expressing memory B cells). It suggests that in AID, theses auto-reactive cells are able to differentiate into IgG isotype-switched cells and escape peripheral tolerance checkpoint but not in healthy subjects. Interestingly, Naturalauto-reactive Bcells are able to secrete only IgM isotype autoantibodies uponin vitrostimulation but not IgG class switched antibodies. In order to better understand what differentiates auto-reactive B cells and the mechanisms leading to pathological autoimmunity, agenomic analysisof the antibody repertoire as well as atranscriptional profilingof these cells by single-cell RNA seq is ongoing to understand further the differences of these autoreactive B cells between healthy subjects and patients with AIDs.Conclusion:We developed a technology to identify and isolate antigen-specific B cells from the peripheral blood of patients with AID. Our results suggest that autoreactive B cells escape peripheral tolerance checkpoint and are able to differentiate into IgG isotype-switched cells in patients with AIDs but not in healthy subjects.References:[1]D. Cornec, A. Berti, A. Hummel, T. Peikert, J.-O. Pers, et U. Specks, « Identification and phenotyping of circulating autoreactive proteinase 3-specific B cells in patients with PR3-ANCA associated vasculitis and healthy controls »,J. Autoimmun., vol. 84, p. 122–131, 2017[2]P. F. Kerkman et al., « Identification and characterisation of citrullinated antigen-specific B cells in peripheral blood of patients with rheumatoid arthritis », Ann. Rheum. Dis., vol. 75, no 6, p. 1170‑1176, juin 2016, doi: 10.1136/annrheumdis-2014-207182.Acknowledgments:with support of vasculitis foundation, CSL Berhing, SFRDisclosure of Interests:None declared
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Sivina, Mariela, Elena Hartmann, Diana Krupnik, Ruth LaPushin, Michael Keating, Thomas J. Kipps, Andreas Rosenwald, William G. Wierda, and Jan A. Burger. "CCL3 and CCL4 Plasma Levels Correlate with Established Prognostic Markers in Chronic Lymphocytic Leukemia: Towards a Simple, ELISA-Based Assay for Risk Assessment." Blood 114, no. 22 (November 20, 2009): 358. http://dx.doi.org/10.1182/blood.v114.22.358.358.
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Abstract Abstract 358 During the induction of normal immune responses, activated B cells secrete the chemokines CCL3 and CCL4 for recruitment of regulatory T cells (Nat Immunol. 2:1126-32, 2001). This may represent a mechanism enabling cognate interactions between rare antigen-specific T and B cells. We previously reported that CCL3 and CCL4 RNA and protein are induced in CLL cells by co-culture with nurselike cells and after B cell antigen receptor (BCR) cross-linking (Blood 113:3050-8, 2009). We found higher levels of CCL3 and CCL4 in ZAP-70 positive cases, and CCL3 and CCL4 secretion was abrogated by the Syk inhibitor R406. These findings suggest that CCL3/CCL4 secretion by CLL cells correlates with the signaling capacity of the BCR. Also, we previously noticed higher plasma levels of CCL3 and CCL4 in CLL patients, when compared to healthy controls. In this study, we measured CCL3 and CCL4 plasma levels by ELISA in 351 CLL patients, and correlated CCL3 and CCL4 levels with various prognostic markers, including RAI stage, immunoglobulin heavy chain variable region gene (IgVH) mutational status, ZAP-70, β2 microglobulin, CD38, white blood count (WBC), and cytogenetic subgroups. We found that the concentrations of CCL3 and CCL4 were significantly higher in plasma samples from CLL patients who had CLL cells that used unmutated IgVH genes or that expressed ZAP-70 or CD38, or who had an advanced stage of their disease (see Table 1). For example, the mean CCL3 plasma level was 56.4 ± 8.8 pg/ml in patients with CLL cells that used unmutated IgVH (mean ± SEM, n=123) versus 14.5 ± 2.4 in patients with CLL cells that used mutated IgVH (mean ± SEM, n=139, p<0.001). The mean CCL4 level was 171.4 ± 26.3 in patients that had CLL cells that used unmutated IgVH and 92.5 ± 17.5 in patients that had CLL that expressed mutated IgVH. On the other hand, the relative absolute white cell count or presence of chromosomal abnormalities did not correlate with high plasma levels of CCL3 or CCL4. Immunohistochemistry revealed that CCL4 was predominantly expressed in proliferation centers in approximately 50% of the cases. Currently, we are evaluating whether high plasma levels of CCL3 and/or CCL4 are associated with a relatively short time from diagnosis to requiring initial treatment by the IWCLL-working group criteria. Also, we are exploring whether high plasma levels of CCL3 or CCL4 are associated with high proportions of infiltrating T cells in proliferation centers. These studies suggest that patients can be stratified by their relative plasma levels of CCL3 and/or CCL4 and that high plasma-levels of theses chemokines define a characteristic that may be associated with aggressive disease. Disclosures: No relevant conflicts of interest to declare.
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Sakata-Yanagimoto, Mamiko, Fumio Nakahara, Etsuko Yamaguchi-Nakagami, Keiki Kumano, Toshiki Saito, Mineo Kurokawa, Seishi Ogawa, and Shigeru Chiba. "Balance of Transcription Factors Downstream of Notch Signaling Determines the Fate of Myeloid Progenitors toward Differentiation to Mast Cells or Immortalization without Differentiation." Blood 108, no. 11 (November 16, 2006): 676. http://dx.doi.org/10.1182/blood.v108.11.676.676.
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Abstract Notch signaling represents one of the fundamental communication channels in various types of cells. While Notch activation has been shown to inhibit myeloid differentiation in a subset of hematopoietic progenitors, the role of Notch signaling in mast cell differentiation is not clear. When common myeloid progenitors (CMPs) and granulocyte-macrophage progenitors (GMPs) purified from mouse bone marrow cells were stimulated with Delta1-Fc, a soluble form of Notch ligand, in the presence of stem cell factor, IL-3, IL-6, and thrombopoietin, granulocyte and macrophage differentiation, which is observed at day 7 of culture in the absence of Delta1-Fc, was markedly inhibited. Instead, Lin-c-Kit+FcεR+ mast cells dominated in the culture. Delta1-Fc did not increase mast cell generation from either CMPs or GMPs of the bone marrow of pI:pC-treated Mx-Cre x Notch2 flox/flox (N2-MxcKO) mice, in contrast to littermate Notch2 flox/flox mice treated with pI:pC, which suggests that Notch2 is responsible for the Delta1-Fc-augmented mast cell generation from CMPs and GMPs in culture. Retroviral transfer of constitutive active form of Notch2 (aN2) into CMPs and GMPs resulted in the complete loss of granulocyte-macrophage colony-forming cells and the emergence of basophilic granules-containing blast like cells, indicating the cell fate instruction. Real-time PCR analysis revealed that Delta1-Fc stimulation and aN2 introduction up-regulated the expression of Hes1, a transcriptional suppressor that is known to be a direct target of Notch activation in several cell types, within 12 h. Moreover, among GATA genes, Delta1-Fc stimulation and aN2 introduction resulted in increase of GATA3 mRNA, while expression levels of GATA1 and GATA2, which have been suggested to play a role in regulating mast cell differentiation, were unchanged. Next, we retrovirally expressed Hes1 and/or a GATA gene into CMPs and GMPs to see if the same effects were observed. Mast cells were increased only when both genes were expressed. On the other hand, when Hes1 alone was transduced, we observed rapid growth and immortalization of these cells without differentiation. C/EBPa, which is known to be suppressed in mast cell differentiation and upregulated in myeloid cell differentiation, was down-regulated within 48 h from the initiation of Hes1 retroviral transduction, suggesting that C/EBPa is a downstream target of Hes1 in this myeloid cell fate determination. Theses results indicate that, at the downstream of Notch activation, there are a C/EBPa down-regulation pathway that is Hes1-dependent and a GATA3 up-regulation pathway. Balanced regulation of these pathways should play a physiological role in myeloid and mast cell differentiation, while imbalance between these two pathways might provide a new model of myeloid transformation.
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Krishnan, Amrita, John A. Zaia, John Rossi, David DiGiusto, Michael Kalos, Larry Couture, Joseph Alvarnas, et al. "First in Human Engraftment of Anti-HIV Lentiviral Vector Gene Modified CD34+ Peripheral Blood Progenitor Cells in the Treatment of AIDS Related Lymphoma (ARL)." Blood 112, no. 11 (November 16, 2008): 2348. http://dx.doi.org/10.1182/blood.v112.11.2348.2348.
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Abstract Background: Autologous stem cell transplantation (ASCT) has become an accepted treatment option for high risk or relapsed ARL. Treatment related mortality is similar to the HIV negative setting. However, ultimately further improvement in ASCT will depend on both effective anti lymphoma therapy and better control of the HIV infection. Highly active antiretroviral therapy (HAART) can lower HIV viral loads to undetectable levels in the peripheral blood, but reservoirs of HIV are still present in the tissues and acquired resistance to HAART also remains a problem. A treatment strategy that would confer intrinsic resistance to HIV could circumvent theses issues. Herein we report on one such strategy using multiplexed RNA based anti-HIV gene transfer strategies to render autologous peripheral blood progenitor cells resistant to HIV. Patients with high risk ARL deemed candidates for ASCT were eligible. Seven subjects with NHL have been enrolled. (4PR, 2 REL, 1CR2), of whom 2 failed screening phase, 1 failed product release test, 2 are pending transplant, and 2 patients have undergone successful transplantation. Median age was 43 yrs at enrollment. Four pts to date were mobilized with chemotherapy plus GCSF and cells were collected for the clinical product (Fx1) and for CD34-selection (CliniMACSª, Miltenyi) and research treatment (Fx2). (see table ) UPN # Fx1 (CD34+/kg) Fx2 (CD34+/kg) Post Selection and transductionCD34+/kg 301 2.8X106 3.5X106 .26 X106 not infused 304 3.9X106 3.6X106 1.2 X106 305 3.4X106 3.8X106 1.4X106 306 5.6X106 8.8X106 pending Three days prior to the completion of CBV (cyclophosphamide 100mg/kg, BCNU 450mg/ m2, VP16 60mg/kg) conditioning, the Fx2 cells were thawed and transduced with a lentivirus vector (LV,rHIV7-ShI-TAR-CCR5Z) encoding 3 RNA elements including short hairpin RNA (shRNA) targeted to HIV tat/rev, a nucleolar localizing TAR decoy sequence, and a ribozyme targeted to CCR5. Cell viability post transduction ranged between 52–64% in three pts. On day 0 Fx2 is given and Fx1 is given 24hrs later (day+1). UPN301 did not receive the transduced Fx2 cells due to a low cell dose. For UPN304 and 305 who received the gene modified Fx2 cells, WBC engraftment occurred at day +11, platelet engraftment at day+16, and there have been no serious adverse events. Results to date at 30 and 60 days post ASCT reveal peripheral blood marking consistent with the ratio of gene modified to unmodified cells infused. Q-PCR analysis demonstrated distribution of genetically modified cells in myeloid and lymphoid lineages, and RT-PCR evidence of shRNA in progeny cells provided further evidence of successful transduction and engraftment of progenitor cells. Follow-up data for these and subsequent patients will be presented at the meeting. Conclusion: Lentiviral vector transduction of autologous peripheral blood progenitor cells with multiplexed RNA is feasible, well tolerated, and led to successful engraftment following high dose chemotherapy for ARL.
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Armellini, Adriana, Maria E. Sarasquete, Ramon Garcia-Sanz, Ricardo Lopez Perez, Ana Balanzategui, Maria C. Chillon, Jose M. Hernandez, et al. "ZHX2, CHC1L and RAN Gene Expression Levels Determine Different Prognosis Groups in Multiple Myeloma (MM) Patients." Blood 106, no. 11 (November 16, 2005): 4356. http://dx.doi.org/10.1182/blood.v106.11.4356.4356.
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Abstract Gene Expression Profiling through RNA arrays has provided new clues to Multiple Myeloma pathogenesis and prognostic pattern evaluation. Recently, ZHX2, CHC1L & RAN expression have been highlighted as key elements in MM. In the present paper, we have evaluated theses genes by real time quantitative RT-PCR (RQ-PCR) in purified plasma cells from 74 patients with plasma cell discrasias. MATERIAL AND METHODS: Purified Bone Marrow cells were obtained from the following patients: 6 MGUS, 7 smoldering MM, 59 newly diagnosed symptomatic MM patients and 2 Plasma cell leukemia (PCL). After RNA extraction, RQ-PCR of CHC1L(C), RAN(R) and ZHX2 (Z) genes was carried out using the standard protocol from TaqMan® gene expression Assays-on-Demand in an ABI-PRISM 7700 Sequence Detection System (Applied Biosystems, Foster City, CA, USA). Expression levels were normalized with ABL gene and expressed in n-fold times compared to the expression in a pool of RNA from mononuclear cells from healthy donors. The expression level of the different genes was evaluated for correlation with the diagnosis, clinical characteristics and prognosis of the patients. RESULT AND CONCLUSIONS: None of these genes displayed a clear relationship with the different stages of disease pathogenesis, although ZHX2 gene was slightly more expressed in the indolent forms of the proliferative disorders (MGUS and SMM). Within symptomatic MM patients, several interesting associations were observed. Thus, in hyperdiploid MM cases, CHC1L expressions observed were fewer than in those with a normal DNA index, confirming the participation of the gene product in chromosomal condensation during the mitosis. No other important associations were observed for this gene, although patients with the lowest expression displayed a very good prognosis, but without reaching statistically significant differences. As expected, RAN expression was related to S-Phase PC, since patients with high S phase values (>1.8 %) displayed higher levels of RAN transcripts. This, however, only resulted in a marginal impact on survival. ZHX2 provided the most interesting results, whereby decreased levels of ZHX2 were related to unfavorable prognostic indicators such as B2 microglobulin >4 mg/L and Hemoglobin levels <10.5 g/dl. This was significantly associated with a shorter survival. As such, patients with a level of ZHX2 expression lower than twice the expression of controls were associated with a very low survival (median of 6 months vs not reached, p=0.019). If we take into account the survival prediction value of these three genes, the following prognosis groups were defined: very good prognosis (Z+ C−), poor prognosis (Z− C+) and intermediate (the remaining patients). SUMMARY: From this study we can confirm that RAN, CHC1L and especially ZHX2 genes play a role in the pathogenesis and behavior of MM, this could be helpful in predicting survival of patients.
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Rajpal, Rajpal, Paul Dowling, Justine Meiller, Kenneth C. Anderson, Philip Murphy, Martin Clynes, and Peter O’Gorman. "Prediction of Thalidomide Response in the Newly Diagnosed Untreated Multiple Myeloma Patients Based on a Panel of Protein Biomarkers." Blood 112, no. 11 (November 16, 2008): 5018. http://dx.doi.org/10.1182/blood.v112.11.5018.5018.
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Abstract Background: Multiple Myeloma (MM) is an incurable plasma cell malignancy. Recently, there have been major therapeutic advances in the treatment of MM, including the use of immunomodulatory drugs. Thalidomide alone or in combination represents an effective treatment strategy for newly diagnosed, relapsed and refractory MM patients. The identification of novel biomarkers could lead to more effective, individualized therapeutic strategies with improved patient outcomes. Patients, Method & Material: Serum samples of sixteen newly diagnosed multiple myeloma patients, who had had initial treatment with thalidomide based regimens were analyzed. Based on D100 re-staging, 8 responders and 8 non responders to thalidomide were identified. Samples were analysed using 2D-DIGE, a technique based on pre-electrophoretic labelling of samples with one of three spectrally resolvable fluorescent CyDyes (Cy2, Cy3, and Cy5) allowing multiplexing of samples into the same gel. Initially serum samples were immunodepleted, which specifically removes 14 high-abundant proteins representing approximately 94% of total protein mass. This allowed for easier analysis of low abundance proteins, which are more likely to be a source of potential biomarkers. All 2D-DIGE images were scanned and collected on a Typhoon Fluorescent Imager. Pooled samples were used as an internal standard to quantify expression changes with statistical significance. Statistics and quantitation of protein expression were carried out initially using DeCyder Biological Variation Analysis (BVA) software before performing subsequent Extended Data Analysis (EDA). Results: 18 proteins have been identified to be differentially expressed in non-responders compared to responders: 13 were up-regulated and 5 were down-regulated (t-test ≤ 0.02). All 18 proteins were >1.25-fold differentially expressed, with changes up to 6.62-fold. For example, Fig.1 shows statistical analysis of protein 1 using DeCyder BVA software. This protein was increased 2.24-fold in the immuno-depleted serum from non-responders compared to responders, (t-test 0.0046). Once the 18 panel proteins were established, further statistical analysis was performed using DeCyder EDA software. Principal Components Analysis (PCA) was used to separate the responders from the non-responders based on the panel of 18 statistically significant differentially expressed proteins (Fig.2). Each dot on this plot represents a clinical sample; clinical samples from the same experimental groups are located in the same distinct areas, i.e. contained in one half of the plot, confirming consistency of results. Conclusion: Accurate prediction of an individual patient’s drug response is an important prerequisite of personalized medicine. Using a panel of proteomic biomarkers, we have demonstrated the feasibility of predicting sensitivity and response to thalidomide in previously untreated myeloma. We are in the process of identification of theses proteins and plan to confirm their predictive value in a larger group of patients. Figure Figure Figure Figure
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Schlund, G. H. "Thesen zur ärztlichen Aufklärung." Allergologie 26, no. 04 (April 1, 2003): 172–74. http://dx.doi.org/10.5414/alp26172.
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Tai, Yu-Tzu, Benjamin King, Cheng Li, XianFeng Li, Peter Burger, Weihua Song, Masood Shammas, et al. "Genomic Evolution of Multiple Myeloma In Vivo over Time." Blood 108, no. 11 (November 16, 2006): 3400. http://dx.doi.org/10.1182/blood.v108.11.3400.3400.
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Abstract Genetic instability is an important feature of malignant cells, specifically in multiple myeloma (MM). This instability, by activation of oncogene or deletion of tumor suppressor genes, plays an important role in oncogenesis. The ongoing genetic instability is responsible for tumor progression, acquisition of invasiveness, and development of drug resistance, as well as eventual mortality. We have previously demonstrated that MM cells have elevated homologous recombination activity that leads to acquisition of new genomic changes over time and is associated with development of drug resistance (Blood2004; 104: 3409). However, such genomic evolution in patient samples has not been documented. Here we have performed genome wide loss of heterozygosity (LOH) assay, using high-density oligonucleotide arrays capable of measuring up to 250K single nucleotide polymorphisms (SNP) loci and able to analyze small areas of gains or losses as an indicator of genomic instability to determine ongoing development of new changes that may reflect instability. We have evaluated nine MM patients with purified myeloma cells obtained at two time points, atleast six months apart. CD138-expressing myeloma cells from these patients were purified and their peripheral blood mononuclear cells were also obtained. Genomic DNA from these cells was digested with StyI, PCR amplified and hybridized to 250K SNP array. Results from CD138+ myeloma cells from two time points were compared with each other as well as with sample from normal peripheral blood mononuclear cells using the dChip software for LOH and copy number analysis and areas of deletions and amplifications. The changes at the first time point were considered as base line, and the subsequent sample was considered as test sample where acquisition of new changes was evaluated compared to base line. In two patients we had three samples available on a serial basis. We have observed that myeloma cells from the second time point demonstrated significant new areas of genomic change with acquisition of mean 4467 (range 79 to 18998) new LOH loci from base line sample. Of theses new areas of change, 41 SNP loci were found to be present in more than three patient samples indicating recurrent loci of interest in regards to progression and/or development of drug resistance. In 2 patients with 3 serial samples, we observed serial increase in acquisition of new SNPs, 4947 and 18998 at first follow up and 531 and 5275 at second follow up suggesting cumulative accumulation of new genetic change. The reproducible area of new acquisition interestingly involves the area with chromosome 1 and 8 that may have significant role in the pathogenesis and progression of the disease. We are currently obtaining the gene expression profile from these time points to identify expression changes correlating with the observed genomic changes on follow up samples. In summary our results confirm genomic evolution of myeloma in vivo over time and provide means to identify molecular targets associated with progression of MM that can be utilized to inhibit progression of the disease and possibly development of drug resistance.
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Baron, Frederic, Michael B. Maris, Brenda M. Sandmaier, Barry E. Storer, Mohamed Sorror, Razvan Diaconescu, Ann E. Woolfrey, et al. "Graft-versus-Tumor Effects after Allogeneic Hematopoietic Cell Transplantation with Nonmyeloablative Conditioning." Blood 104, no. 11 (November 16, 2004): 184. http://dx.doi.org/10.1182/blood.v104.11.184.184.
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Abstract We have used a nonmyeloablative conditioning regimen consisting of 2 Gy total body irradiation +/− fludarabine, 30 mg/m²/day x 3 days, to condition elderly or ill patients (pts) with hematological malignancies for allogeneic hematopoietic cell transplantation (HCT). This approach relies almost exclusively on graft-versus-tumor (GVT) effects for control of malignancy. Here, we analyzed GVT effects in 322 pts with hematological malignancies given grafts from HLA-matched related (n=192) or unrelated (n=130) donors. Grades I, II, III and IV acute GVHD were seen in 26 (8.1%), 141 (43.8%), 34 (10.6%) and 11 (3.4%) pts, respectively. Extensive chronic GVHD was seen in 181 (56.2%) pts and of these, 64 (19.9%) cases had de novo chronic GVHD. Putative GVT effects were evaluated using time-dependent Cox regression models. Of the 221 pts with measurable disease at HCT, 126 (57%) achieved complete (n=98) or partial (n=28) remissions. Multivariate analysis identified chemosensitivity for B-cell malignancies (p=.02), and tandem autologous/allogeneic HCT (p=.04) as pre-transplant factors associated with higher probabilities of achieving complete remissions (CR) after HCT. After adjusting for these factors, acute GVHD of any grade was not found to be associated with an increased probability of achieving CR. There was a trend for a higher probability of achieving CR in pts with chronic GVHD (p=.07). Progression/relapse was observed in 108 pts. Multivariate analysis identified that lower disease-risk (p=.0004), tandem autologous/allogeneic HCT (p=.02) and adapted Charlson comorbidity index (CCI) score at transplant < 3 (p=.002) resulted in significantly decreased risk of progression/relapse. After correcting for these factors, extensive chronic GVHD was associated with a decreased risk of progression/relapse (p=.006). Pts with grade 1 acute GVHD tended to have less progression/relapse (p=.07). Conversely, grade II–IV acute GVHD did not significantly affect the risk of progression/relapse. Nonrelapse mortality was observed in 70 pts. Multivariate analysis showed that lower disease-risk (p=. 001), tandem autologous/allogeneic HCT (p=.002) and CCI score at transplant < 3 (p<.0001) significantly decreased nonrelapse mortality. After adjusting for these variables, grade II (p=.04) and grade III–IV (p<.0001) acute GVHD increased nonrelapse mortality while extensive chronic GVHD did not. The 3-year probability of progression-free survival (PFS) was 38.5%. In multivariate analysis, lower disease-risk (p<.0001), tandem autologous/allogeneic HCT (p=.0008) and CCI score at transplant < 3 (p<.0001) resulted in significantly better PFS. After adjusting for theses variables, grade 1 acute GVHD (p=.02) and chronic extensive GVHD (p=.003) were both associated with significantly better PFS, while grade III–IV acute GVHD (p<.0001) was associated with decreased PFS. In summary, chronic GVHD in pts given nonmyeloablative conditioning was associated with substantial GVT effects which led to improved PFS. Conversely, any potential GVT benefits from grade II–IV acute GVHD were offset by higher nonrelapse mortality resulting in worse PFS. Efforts should be directed at reducing the risk of grade II–IV acute GVHD while allowing de novo chronic GVHD for best PFS after allogeneic HCT with nonmyeloablative conditioning.
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Loberg, Robert D., Shen-Wu Wang, Scott D. Patterson, and Ian McCaffery. "C-Mpl Is Not Expressed or Functional At Detectable Levels on Primary Chronic Lymphocytic Leukemia (CLL) Tumor Samples." Blood 118, no. 21 (November 18, 2011): 2849. http://dx.doi.org/10.1182/blood.v118.21.2849.2849.
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Abstract Abstract 2849 The development of thrombocytopenia is associated with progression of CLL to a more advanced stage and is commonly used to classify CLL into the higher disease risk categories. It has been estimated that 5% of CLL patients will present with associated immune thrombocytopenia purpura (ITP) and immune mediated destruction can exacerbate thrombocytopenia related to bone marrow infiltration. Thromopoietin (TPO) mimetics stimulate platelet production though activation of the thrombopoietin receptor (c-Mpl) on megakaryocytes and are currently indicated for chronic ITP. In principle, this approach could be considered for treatment of ITP in CLL patients. In order to rule out the possibility that c-Mpl is functionally expressed in CLL tumor cells; we analyzed expression and TPO-dependent function in primary patient samples. We have developed assays for the analysis of c-Mpl expression on the surface of B-cells in CLL patient samples and to assess functional response to ex-vivo stimulation of theses primary samples with TPO. Peripheral blood was collected from CLL patients and mononuclear cells isolated by Ficoll separation. This cohort included treatment naïve CLL samples (n=57) and CLL samples collected from subjects undergoing active treatment (n=30). CLL cells were analyzed by flow cytometry and identified based upon viability and CD5+/CD19+ expression. c-Mpl expression was measured using a novel c-Mpl-specific monoclonal antibody that was shown to be specific for c-Mpl by flow cytometry. Residual platelets associated with individual PBMC aliquot served as an internal control for c-Mpl expression. To investigate c-Mpl function, CLL cells were stimulated with rhTPO (10 ng/mL for 10 min) and induction of pSTAT5 was measured to assess a functional response to ex-vivo stimulation of TPO. In addition c-Mpl mRNA expression was surveyed in CLL by using mRNA microarray analyses to correlate c-Mpl mRNA expression with protein and functional expression of c-Mpl. Robust c-Mpl protein expression was observed in platelets from normal and CLL patients (fold over control -normal: 31.90 ± 6.39, CLL: 26.76 ± 6.57; mean ± 95%CI), no significant expression of c-Mpl was observed on CD5+/CD19+ CLL cells (fold over control −1.06 ± 0.021; mean ± 95%CI). No induction of STAT5 phosphorylation was detected in B-cell CLL cells in any of the samples stimulated with TPO (fold over control - normal: 0.90 ± 0.02, CLL: 1.04 ± 0.034; mean ± 95%CI). Microarray analysis of the CLL samples demonstrated c-Mpl mRNA expression levels equivalent to background across all CLL samples analyzed (intensity score −10.3 ± 3.0; mean ± 95%CI). In conclusion, we demonstrated a lack of cell surface c-Mpl protein expression in CLL cells. The additional data suggesting the lack of evidence for significant expression of c-Mpl mRNA expression supports the hypothesis that CLL cells do not express c-Mpl and unlikely to be stimulated in patients treated with TPO mimetics. This hypothesis will need to be tested in an appropriate clinical trial to assess the potential benefits of TPO mimetics for the treatment of thrombocytopenia in CLL patients. Disclosures: Loberg: Amgen: Employment, Equity Ownership. Wang:Amgen: Employment, Equity Ownership. Patterson:Amgen: Employment, Equity Ownership. McCaffery:Amgen: Employment, Equity Ownership.
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Le, Quan, Bipin N. Savani, Aarthi Shenoy, Eleftheria Kozanas, and A. John Barrett. "Hepatitis B Reverse Seroconversion in Long Term Survivors of Allogeneic Hematopoietic Stem Cell Transplantation." Blood 110, no. 11 (November 16, 2007): 1978. http://dx.doi.org/10.1182/blood.v110.11.1978.1978.
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Abstract Reverse seroconversion (RS) known as reactivation of resolved hepatitis B virus (HBV) infection is a complication after allogeneic hematopoietic stem cell transplantation (SCT) in patients exposed to hepatitis B virus. Four hundred thirty six (436) patients with hematological disorder received SCT from an HLA identical sibling in our institute between September 1993 and June 2004. Of these, 103 patients who are 3 or more years post-transplantation have been enrolled in a long-term evaluation protocol. Sixteen had pre-transplant resolved HBV infection (negative HBsAg, positive anti-HBs and anti-HBc antibodies). We investigated the serologic markers of HBV infection in this cohort (median follow-up 59.5 months, range 40–113). Diagnoses included chronic or acute myelogenous leukemia and myelodysplastic syndrome (14), acute lymphoblastic leukemia (1), and severe aplastic anemia (1). Median age at SCT was 32 years (range 10–56). Thirteen patients were positive for anti-HBc and anti-HBs. One had anti-HBc, one had anti-HBs alone, and 1 had no antibodies to hepatitis B. None had detectible HBsAg (HBV surface antigen) at transplant. Twelve patients (median age 39.5 years) received a 12–13.6 Gy total body irradiation (TBI)-based myeloablative SCT followed by a T-cell depleted peripheral blood stem cell transplantation (PBSCT). Four patients (median age 31 years) received a reduced-intensity regimen of fludarabine 125 mg/m2 and cyclophosphamide 60 mg/kg, followed by a PBSCT. All received cyclosporine as graft-verse-host disease (GVHD) prophylaxis. Nine developed acute GVHD, and 13 developed chronic GVHD. Nine (56%) patients received immunosuppressive therapy (IST) for chronic GVHD beyond 3 years from SCT. Fifteen (94%) patients were alive with a median follow-up of 59.5 months. One patient with RS of HBV died at 59 months after SCT. Six (38%) patients developed RS of hepatitis B with reappearance of HBsAg, 21–101 (median 29.5) months after SCT. Four (67%) of the 6 developed clinical hepatitis and received Lamivudine treatment with a decrease of HBV viral load. There was no significant difference in median age of 6 patients with, and 10 patients without RS (38 vs 32 years). RS of hepatitis B was associated with prolonged immunosuppressive therapy for cGVHD. Theses results show that RS of HBV post SCT is a significant long-term complication of patients with pre-transplant resolved HBV infection. This emphasizes the importance of long-term hepatitis B serologic monitoring, post-transplant prophylaxis with Lamivudine, HBV vaccine, and prompt initiation of Lamivudine treatment if RS of HBV occurs. Variable With RS of HBV Without RS of HBV Cases 6 (38%) 10 (62%) Age in years (median, range) 38 (10–46) 32(13–56) Myeloablative 4 8 Reduced intensity 2 2 aGVHD 2/6 (33%) 7/10 (70%) cGVHD 5/6 (83%) 8/10 (80%) IST for cGVHD >3 years 4/6 (67%) 5/10 (50%) Lamivudine Treatment 4/6 (67%) Lamivudine Prophylaxis 5/10 (50%) HBV Vaccine after HSCT 5/10 (50%) RS of HBV in months (median, range) 29.5 (21–101) Follow-up in months (median, range) 71(55–107) 57.5 (40–113)
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Mueller, Martin C., Philipp Erben, Giuseppe Saglio, Enrico Gottardi, Thomas Schenk, Thomas Ernst, Stephanie Lauber, Michael Emig, Ruediger Hehlmann, and Andreas Hochhaus. "Harmonization of BCR-ABL mRNA Quantification Using an Uniform Control Plasmid in 36 International Laboratories." Blood 106, no. 11 (November 16, 2005): 1991. http://dx.doi.org/10.1182/blood.v106.11.1991.1991.
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Abstract Quantitative determination of residual BCR-ABL transcript levels has been accepted as integral part of the management of CML patients. However, heterogeneity of molecular approaches results in a lack of comparability between different studies. Thus, there is an unmet need for harmonization of both procedures and expression of results. In a series of consensus meetings within the European LeukemiaNet a list of prerequisites to achieve an optimal sensitivity and standardization has been elaborated: use of at least 10ml peripheral blood processed within 36 hrs; bedside RNA stabilization for multicenter trials; standardized PCR protocols optimized for each platform; use of a single plasmid containing target and housekeeping genes to avoid dilution errors; use of total ABL and/or beta glucuronidase (GUS) as internal controls. To substantiate these theses an international multicenter trial within 36 labs in 14 countries was initiated. The aim of the study was to assess the variability of results obtained from different labs using the PAXgene Blood RNA System® (PreAnalytiX, Hombrechtikon, Switzerland) for RNA extraction, individual protocols for cDNA synthesis, 3 different PCR platforms (TaqMan®, TM, n=24, LightCycler®, LC, n=14, Rotorgene® n=1), and optimized quantitative RT-PCR conditions. In order to standardize results, b3a2 BCR-ABL and GUS sequences were cloned into a pCR 2.1-TOPO vector® (Invitrogen, Carlsbad, CA), which was distributed to all participants in serial dilutions as external control for quantification of BCR-ABL, total ABL, and GUS mRNA transcripts. Ten samples containing dilutions (10, 2, 1, 0.1%) of b3a2 or b2a2 BCR-ABL positive cells in normal leukocytes and negative controls were prepared, blinded, and shipped to the participants. Transcript numbers were determined in triplicates, ratios BCR-ABL/ABL and BCR-ABL/GUS were calculated and expressed in %. Median ratios BCR-ABL/ABL for b3a2 samples were 8.9, 1.7, 0.85, and 0.11%; for b2a2 samples 9.1, 1.6, 0.82, and 0.10%. Median ratios BCR-ABL/GUS for b3a2 samples were 3.4, 0.77, 0.37, and 0.042%; for b2a2 samples 2.8, 0.48, 0.29, and 0.031%. Four of 36 participants (11%) detected low BCR-ABL copy numbers in negative control samples. The coefficients of variation (CV) for all participants, TM, and LC users were 0.62, 0.57, and 0.59 for ratios BCR-ABL/ABL; 1.03, 0.85, and 1.22 for ratios BCR-ABL/GUS, respectively. Standard errors to the regression line were significantly lower evaluating ratios BCR-ABL/GUS (median 0.075, range 0.0046–0.90) compared to ratios BCR-ABL/ABL (median 0.18, range 0.022–2.2, p<0.001). Overall, mean TM ratios were 1.7times higher than LC ratios indicating a difference of the amplification efficiencies between target and control genes. We conclude, that harmonization of BCR-ABL mRNA quantification is feasible employing a common plasmid for BCR-ABL, total ABL, and GUS. However, the remaining variability of results indicates minor differences of the PCR efficiencies using individual protocols. We therefore suggest (i) the use of a common standard plasmid, (ii) the introduction of a calibrator, and (iii) regular control rounds to achieve and maintain comparability of results between individual laboratories.