National Repository of Grey Literature 21 records found  1 - 10nextend  jump to record: Search took 0.00 seconds. 
Pathophysiological development and differentiation of cells during hematopoiesis
Moudrá, Alena ; Štěpánek, Ondřej (advisor) ; Černý, Jan (referee) ; Kalina, Tomáš (referee)
In recent years, a great effort has been deployed towards a better understanding of the molecular changes in cells and in the bone marrow (BM) environment that contribute to the development and progression of myelodysplastic syndrome (MDS) to acute myeloid leukemia (AML). Among others, the aberrant hematopoietic stem cells in MDS often display increase in DNA double strand breaks, genomic instability with common loss or rearrangement of chromosomes and an ineffective response to DNA damage, a phenomenon that has been linked to the onset of cellular senescence. Additionally, the BM microenvironment can become more pro-inflammatory. In our effort to better understand the contribution of the BM microenvironment on MDS progression, we analyzed the expression profiles of cytokines in the BM microenvironment in all stages of MDS/AML and found several proinflammatory cytokines that increase with disease progression. Also, by repeated sampling of patients over the course of 5-azacytidine therapy, we were able to assess the changes in the proinflammatory cytokine milieu with the progression of the disease. Additionally, we aimed to identify the candidate markers for the improvement of MDS prognosis. We focused on naturally occurring germline polymorphism of NAD(P)H dehydrogenase (quinone 1) gene (NQO1*2)...
Cytometric assay of antigen-specific T cell response in monitoring of BCG vaccine therapy
Hadlová, Petra ; Drbal, Karel (advisor) ; Kalina, Tomáš (referee)
Bladder carcinoma (BCa) is among the most common carcinomas in the Western world. Despite the availability of effective therapies, there is currently an urgent need to develop a stratification method, which would enable the accurate identification of patients responsive to therapy. In the theoretical part of my diploma project I describe the heterogeneity of BCa and the currently applied immunotherapeutic approaches. I specifically focused on the Bacillus Calmette-Guérin (BCG) vaccine instillation. For decades another use of BCG has been a prophylactic vaccination against tuberculosis (TB) infection. BCG serves as a model treatment because it is highly efficient when prescribed to the responsive patient. However, an effective stratification is yet to be developed for BCa and latent tuberculosis infection (LTBI) diagnosis and/or monitoring. In the experimental part of my project, I developed and tested a 10-parameter panel for T cell- specific activation test (TAT) applicable for a stratification of BCa patients as well as for the detection of LTBI. I tested the panel on positive controls using flow cytometry (FCM) method because it allows for detection and measurement of dozens of markers at a single cell level. It is easily applicable to available urine and blood samples obtained from BCa...
Modern mathematical methods in the research of immune reconstitutions and immunodeficiencies
Stuchlý, Jan ; Kalina, Tomáš (advisor) ; Šinkora, Jiří (referee) ; Štěpánek, Ondřej (referee)
In this thesis we present new analytical and integrative approaches for broad spectrum of applications of flow cytometry. CVID Large (88 individuals) cohort of CVID patients was immunopheno- typed by flow cytometry. The "probability binning" algorithm was used to au- tomatically assess the immunophenotype which presented unprecedented stabi- lity. We were able to define sub-group of CVID patients with highly activated immunophenotype showing hallmarks of immunosenescence and common clinical characteristics (thrombocytopenia, lung fibrosis and bronchiectasis). The severity of clinical complications correlated quantitatively with the immunophenotype. Analysis of cellular proteome We have designed technics which allow for highly multiplexed (>1000 of antibodies) analysis of human proteome using af- finity proteomics. We analyzed the changes of proteome of human cell lines and characterized the proteome of acute leukemias. The yet undescribed stability of proteome with respect the sub-cellular localization is shown and theoretical and practical background for antibody standardization and validation for the use in a nity proteomics is presented. Topological analysis of cytometry data Multiparametric flow cytometry allows for description of complex topological relationships of the cells in di erent stages of...
Immunogenic cell death and it's relevance for biology and therapy of malignant diseases
Truxová, Iva ; Špíšek, Radek (advisor) ; Říhová, Blanka (referee) ; Kalina, Tomáš (referee)
Immunostimulatory potential of tumor cells depends on various factors, including primarily tumor antigen repertoire and the capacity to emit molecules associated with cellular stress or injury, so called DAMPs, during immunogenic forms of cell death. These molecules mainly act on dendritic cells (DCs), thus activating the antitumor immune response. Several immunogenic cell death (ICD) inducers have been described in the past years. The contribution of my PhD thesis into this topic was the characterization of the apoptotic pathways activated by high hydrostatic pressure (HHP). HHP induces rapid tumor cell death accompanied by DAMP release (mainly calreticulin (CRT), HSP70, HSP90, HMGB1 and ATP) that is characterized by the overproduction of reactive oxygen species (ROS) causing the establishment of integrated stress response. ROS-PERK-eIF2α-caspase-2-caspase-8 signaling pathway plays an essential role in CRT translocation to the tumor cell surface upon HHP treatment, thus influencing the immunogenic potential of these cells. Moreover, the importance of ICD concept was also confirmed in vivo. The results point out that the presence of CRT on the surface of malignant blasts from AML patients correlates with the activation of specific antitumor immune response and improved clinical outcome. Another...
Human lymphopoiesis and its examination via single-cell analysis
Novák, David ; Kalina, Tomáš (advisor) ; Drbal, Karel (referee)
Development of human B-lymphocytes is a convoluted process. A self-renewing stem cell progenitor in a primary lymphoid tissue commits to the lymphoid lineage. Subsequent B-lineage commitment entails somatic gene recombination processes which lead to the eventual expression of a surface antigen receptor. Functionality of the B-cell receptor, as well as successful testing for autoreactivity by the cell, are preconditions for the differenti- ation of a mature B-lymphocyte. Processes within this development are often investigated using single-cell analysis via flow cytometry, fluorescence-activated cell sorting and mass cytometry. Coupling these high-throughput methods with modern approaches to data analysis carries enormous potential in revealing rare cell populations and aberrant events in haematopoiesis. Keywords: B-lymphocyte, lymphopoiesis, flow cytometry, FACS, mass cytometry, clus- ter analysis, FlowSOM, PCA, t-SNE, Wanderlust.
Eight color flow cytometry test development for primary imunodeficiency patients
Šinkorová, Vendula ; Kalina, Tomáš (advisor) ; Brdička, Tomáš (referee)
Primary immunodeficiencies represent a heterogeneous group of hereditary immune system malfunctions with very variable causes and symptoms. Multiparametric flow cytometry has become an important tool in primary immunodeficiency diagnostics and research because it provides detailed information on the phenotype of individual immune cells and their proportions in circulation. We have developed a complex monoclonal antibody panel composed of five eight-color tubes which is designed for immunophenotyping of basic lymphocyte subsets and further analysis of B and T cell subpopulations. We have optimized and standardized the panels so they will identify any changes originating from primary immunodeficiencies and provide comparable data on the level of cooperation between more laboratories. This was achieved by cooperation of six European research facilities which are all parts of the Euroflow consortium. The panels have been validated both on peripheral blood samples from healthy donors and patients with either gentically defined primary immunodeficiency or common variable immunodeficiency. Keywords: T lymphocyte, B lymphocyte, primary immunodeficiency, flow cytometry, immunophenotyping, Euroflow, optimization, standardization
Genetic and molecular factors influencing the outcome of solid organ transplantation
Pavlova, Yelena ; Slavčev, Antonij (advisor) ; Kalina, Tomáš (referee) ; Mrázek, František (referee)
Since its beginning, graft rejection remains the key problem of solid organ transplantation. This reaction of the recipient's immune system against mismatched antigens of the transplanted organ causes graft damage and consequently loss of its function. Rejection involves cellular (lymphocyte mediated) and humoral (antibody mediated) mechanisms. Among the genetic factors which may have a prognostic value in rejection risk evaluation are the Human Leukocyte Antigens (HLA) genotype, the Killer Immunoglobuline-like Receptor (KIR) gene repertoir, cytokine and other gene polymorphisms. These factors could be screened for before transplantation to find the best possible combination of genetic characteristics of the donor and recipient and to reveal patients with "risky" genotypes, who may need more intensive immunosuppression and more careful post-transplant follow-up. Molecular factors, such as HLA and non-HLA antibodies, soluble CD30 molecule (sCD30), Hepatocyte Growth Factor (HGF) and other cytokines, measured before and/or after transplantation in the recipient's blood may be helpful for rejection risk estimation and may also be used as post-transplant rejection onset markers. In our study, we focused on some of the above mentioned factors. We found that ethnicity plays a significant role in the...

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