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The Role of Cytokines in Support of Malignant Properties of Glioblastomas
Fortelková, Nikol ; Hodný, Zdeněk (advisor) ; Nováková, Zora (referee)
The tumor microenvironment of Glioblastomas (GB) is very complex and transforms throughout radiotherapy and chemotherapy, posing a favourable environment for the malignant properties of GB including invasivity, which promotes aggressive recurrency of the tumor. These changes in the tumor microenvironment are partly induced by cellular senescence and cellular response to stress, accompanied by a specific secretome, so called senescence-associated secretory phenotype (SASP). The SASP is distinct in its secretion of growth factors, components of the extracellular matrix and remodelling enzymes, but above all proinflammatory cytokines, which may contribute to the malignant properties of GB. This thesis reviews the current state of knowledge of the potential role of selected cytokines of the SASP (IL-6, IL-1β, IL-8, IL-1α, TGF-β, CCL2, TNF-α) in support of the malignant properties of GB. Key words: glioblastoma (GB), senescence, SASP, malignant properties, cytokines
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Mechanisms of immune response against kidney allograft
Švachová, Veronika ; Stříž, Ilja (advisor) ; Mrázek, František (referee) ; Posová, Helena (referee)
Kidney transplantation improves the prognosis and quality of life of patients with end- stage renal disease. However, the occurrence of rejection still represents a major complication that can lead to failure of allograft function. In our work, we have studied the phenotypic changes of subpopulations of B lymphocytes and monocytes, which represent one of the key players involved in immune responses against kidney transplantation. In the part of the research devoted to B lymphocytes, we have observed a different redistribution of individual B cell compartments in the peripheral blood during the first year after transplantation without significant differences between used immunosuppression. We have demonstrated that sensitized and rejected patients had a lower level of IgMhigh CD38high CD24high transient B cells. These results support the hypothesis of a protective role of transient B lymphocytes. Activation of monocyte subpopulations in peripheral blood in the posttransplantation perion was also subject of research. We have compared a cohort of patients with normal graft function with a group of patients with acute rejection. The level of "transient" monocytes and HLA-DR expression in a subset of "non-classical" monocytes did not decrease in patients with rejection, compared to the group with a...
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Monitoring of sublingual allergen immunotherapy using serum cytokine levels
Brzáková, Adéla ; Řezáčová, Vladimíra (advisor) ; Andrýs, Ctirad (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Author: Bc. Adéla Brzáková Supervisor: Mgr. Vladimíra Řezáčová, Ph.D. Title of thesis: Monitoring of sublingual allergen immunotherapy using serum cytokine levels Keywords: allergen immunotherapy, cytokine, ELISA, grass pollen allergy Background: The aim of this thesis was to find out whether there are detectable changes in the levels of IFNγ, IL-4 and IL-5 during allergen immunotherapy and whether these cytokines could thus serve as biomarkers of the efficiency of this treatment. Methods: Selected cytokines were determined using Quantikine® HS ELISA Human IFN-γ Immunoassay, Quantikine® HS ELISA Human IL-4 Immunoassay and Quantikine® ELISA Human IL-5 Immunoassay. The monitored population consisted of 32 polinotics, 19 of which were administered allergen immunotherapy and 13 of which constituted the control group. As part of the follow-up, both groups also continuously recorded the severity of their symptoms using a visual analog scale. Results: In the group of treated patients, a decrease in IL-4, IL-5 and IFNγ concentrations was observed during treatment, but no significant differences in the values of these cytokines were found between patients treated with Oralair and control individuals. Using...
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Processsing of food supplements based on medical types of mushrooms
Vetchá, Vendula ; Uhlířová, Renata (referee) ; Hoová, Julie (advisor)
This diploma thesis deals with the characterization of nutrition substances in medical mushrooms, specifically Ganoderma lucidum and Hericium erinaceus. It also focuses on the biological impact of these rare mushrooms on human cell lines HaCaT and Caco-2, and investigates the production of pro-inflammatory cytokines IL-6 and IL-8 by immortalized keratinocytes and colon cancer cells after exposure to aqueous extracts prepared from mushroom powder. The theoretical part of the thesis deals with the characterization of mushrooms in terms of morphological characteristics and their taxonomic classification. It further studies the effects of edible and medicinal species and provides a detailed description of medicinal mushrooms such as Ganoderma lucidum, Lentinula edodes, Flammulina velutipes, Grifola frondosa, Hericium erinaceus and Pleurotus ostreatus, thesis informing about their effects, utilization and possibilities for application in the food and pharmaceutical industries. It mentions possible negative effects of mushrooms on consumers' health and presents methods for processing mushrooms and their use in the food industry. Finally, it focuses on the possibilities of in vitro toxicity testing and provides a brief overview of commonly used cytotoxicity tests such as the MTT test and LDH test. The experimental part of the thesis deals with the preparation of water extracts from mushroom powders and their characterization in terms of nutritional and biologically active substances. HPLC analysis is used to monitor the content of lipophilic substances and vitamin C in mushroom samples. To confirm the biological effect on human health, antimicrobial tests were performed using four model microorganisms, and cytotoxicity tests were performed on two human cell lines. Lastly, a sensory analysis of instant vegetable soup enriched with mushroom powder was performed. Biological effects of the analyzed powdered extracts from Ganoderma lucidum mushroom have been confirmed, especially their cytotoxic effect on colon epithelial tumor cells. However, the application of reishi in the food industry is not easy due to the presence of triterpenoids that cause a bitter taste. On the other hand, the safety of using the Hericium erinaceus mushroom in food has been confirmed through cytotoxicity tests. Sensory analysis results have shown a positive effect of Hericium erinaceus on the taste profile of instant vegetable soups.
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Engineering and selection of protein binders recognising medically important cytokines
Huličiak, Maroš
Protein engineering attracts more attention as a powerful tool of biotechnology and medicine. Small, engineered proteins derived from protein molecules of stable fold, the so called scaffolds, are potential replacements of supplements of more widely used antibodies. In this thesis, I introduce utilization of two scaffold molecules designed in our laboratory for development of stable and specific protein binders of high affinity. This thesis discusses the development of binders interacting with medically important human cytokines and their cellular receptors, interleukin-10, interleukin-28 receptor, and interleukin-9 receptor alpha. Recombinant cytokine and receptor proteins were expressed in eukaryotic cells in high yields and quality and served as molecular targets for selections using various display methods of directed evolution. We demonstrated that application of ribosome and yeast display methods or their unconventional combination in a newly developed integrated pipeline leads to successful generation of high affinity and specificity binders based on newly designed protein scaffolds called 57aBi and 57bBi.
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Identification of new regulators of proinflammatory signaling pathways
Dráberová, Helena ; Štěpánek, Ondřej (advisor) ; Krulová, Magdaléna (referee) ; Funda, David (referee)
Identification of new regulators of proinflammatory signaling pathways Helena Dráberová Protein 4.1R has been described in immune system as regulator of migration and cell adhesion, but was also shown to play a role in activation of T lymphocytes. Polymorphism in gene ORMDL-3 is associated with asthma risk in children and correlates with increased ORMDL-3 expression. This disertation thesis describes the function of proteins 4.1R and ORMDL-3 in activation of mast cells after stimulation of FcεRI receptor. IL-17 is a proinflammatory cytokine that plays a role in immune response against fungal and yeast infections. IL-17 however also plays a role in the pathology of autoimmune diseases such as reumatoid arthritis, psoriasis and multiple sclerosis. IL-17 signaling is tightly regulated, however the exact mechanism has not been described. This disertation thesis describes the IL-17R complex by mass spectrometry and analyze the function of its known and newly discovered components in cells deficient in individual proteins by method CRISPR-Cas9. Last part focuses on the discovery of new subunit of IL-17RC protein CMTM4, which role in IL-17 signaling has not been described so far. CMTM4 stabilizes IL-17RC and is required for its surface expression. In vitro data are supported by data from autoimmune model of...
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Engineering and selection of protein binders recognising medically important cytokines
Huličiak, Maroš ; Schneider, Bohdan (advisor) ; Pichová, Iva (referee) ; Kukačka, Zdeněk (referee)
Protein engineering attracts more attention as a powerful tool of biotechnology and medicine. Small, engineered proteins derived from protein molecules of stable fold, the so called scaffolds, are potential replacements of supplements of more widely used antibodies. In this thesis, I introduce utilization of two scaffold molecules designed in our laboratory for development of stable and specific protein binders of high affinity. This thesis discusses the development of binders interacting with medically important human cytokines and their cellular receptors, interleukin-10, interleukin-28 receptor, and interleukin-9 receptor alpha. Recombinant cytokine and receptor proteins were expressed in eukaryotic cells in high yields and quality and served as molecular targets for selections using various display methods of directed evolution. We demonstrated that application of ribosome and yeast display methods or their unconventional combination in a newly developed integrated pipeline leads to successful generation of high affinity and specificity binders based on newly designed protein scaffolds called 57aBi and 57bBi.
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Zdravotní a produkční účinek imunomodulačního preparátu Imrestoru u vysokoužitkových dojnic
NEZNÍKOVÁ, Kateřina
This thesis deals with evaluation of the impact of immunomodulatory preparation Imrestor (Elanco Animal Health) on the health and production of high-producing dairy cows in the transition period. The experiment was done in the agricultural company Košetice in the district of Pelhřimov on dairy cows of the Holstein breed. The animals selected for the experiment were divided into two groups - control and experimental. Imrestor was applied subcutaneously to the dairy cows of the experimental group approximately one week before calving and on the day of calving. The active component of this preparation is a commercial form of a naturally occurring cytokine (bG - CSF), which specifically affects on the neutrophil precursor cells and increases their production. Blood samples were taken from all cows included in the experiment to evaluate the effect of the preparation Imrestor. At the same time was monitored their clinical health and the incidence of the most common diseases in the transition period. 81 blood samples were taken from 31 dairy cows from the control group and from 18 dairy cows from the experimental group. A biochemical and hematological examination was performed in the laboratory, where the parameters of red blood cells, trombocyte count, leukocyte count, leukocyte percentage, biochemical indicators of blood plasma and mineral content were determined. By statistical evaluation was found, that Imrestor statistically highly significantly (p <0.01) affected leukocyte counts and the percentage of neutrophil granulocytes and lymphocytes. Statistically significant differences (p <0.05) between the observed groups of dairy cows were found in the trombocyte count, percentage of monocytes and alkaline phosphatase activity.
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The effect of nanoparticles on functional and regenerative properties of mesenchymal stem cells
Dostálová, Dominika ; Rössner, Pavel (advisor) ; Grobárová, Valéria (referee)
Mesenchymal stem cells (MSCs) represent a type of stem cells, localized in various tissues, where they are involved in the regeneration process. Because of their properties, MSC are tested for cell-based therapy. However, the treatment of injuries is often associated with bacterial infections, which complicate the process of healing. To establish a sterile environment, metallic nanoparticles (NPs) with proven antibacterial properties can be applied, for improving the healing process. Mechanisms of the antibacterial effect of NPs are non-specific and may cause the damage of eukaryotic cells. The aim of this thesis was to evaluate the effect of three antibacterial metallic NPs (silver, copper oxide and titanium dioxide) on the functional and regenerative properties of MSCs. The effect of NPs on the metabolic activity, differentiational potential, expression of genes for immunoregulatory molecules and production of cytokines by MSCs was determined. Additionally, the immunoregulatory properties of MSCs after cultivation with NPs were tested. The results showed that the metabolic activity was reduced in the presence of silver and copper oxide NPs, with the decrease dependent on the type of the tested NPs. The expression of selected immunoregulatory molecules was inhibited and the immunoregulatory...
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Targeting IRAK4 kinase in autoimmune diseases and cancer
Synáčková, Alžběta ; Dráber, Peter (advisor) ; Brdička, Tomáš (referee)
Immune system provides host protection against invading pathogens. However, aberrant activation can lead to development of autoimmune diseases or cancer. Understanding the mechanisms of inflammation and immune responses is crucial for treatment of such conditions and reestablishing immune balance. Toll-like receptors and interleukin-1 family receptors are a key component of the innate immune system. Their downstream molecules, MyD88 and IRAK4, are essential for receptor signaling as their deficiency causes host susceptibility to infection. On the other hand, overactivation of this pathway was shown to be able to promote autoimmunity and cancer. The main focus of this text will be to summarize current knowledge about the mechanism of IRAK4 signaling and how it can be exploited in the development of therapeutics. Keywords IRAK4, MyD88, Toll-like receptors, IL-1 receptor, cytokines, autoimmunity, cancer
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