Master Degree / Yüksek Lisans Tezleri
Permanent URI for this collectionhttps://hdl.handle.net/11147/3008
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Master Thesis Expression of Aquaporin 1, 3 and 4 in T Cell Activation and Apoptosis(Izmir Institute of Technology, 2018) Gelmez, Ayşe Bengisu; Nalbant Aldanmaz, AytenAquaporins (AQPs) are membrane proteins responsible for transporting water, some gases and small solutes such as CO2 and glycerol. Until now, it has been shown that AQP1, 3 and 5 expressed in both B and T lymphocytes of mice, regulate cell volume. However, aquaporin expression involved in activation, proliferation, and differentiation as well as apoptosis of T cells are not well known yet. The goal of this study is to detect the expression level of AQP1, AQP3, and AQP4 in activated and apoptotic T cells. In order to do that, two types of T cells cultured in both condition were utilized. Peripheral Blood Mononuclear Cells (PBMCs) were isolated from Human peripheral blood drawn from healthy donors by ficoll density gradient Centrifugation method. Naive CD4+ T cells were sorted from PBMC. The stimulants generating Th17 were chosen for activation and differentiation of naïve CD4 T cells. Jurkat cell line as a second cell type were activated by PMA/Ionomycin as well as treated by camptothecin for apoptotic processing. Th17 and Jurkat cell cultures were analysed by flow cytometry to measure the rate of both activation and apoptosis. Western Blot was performed to identify expression of AQP 1, 3 and 4. We found a significance between increased expression level of AQP1, 3, and 4 in activated T cells as well as decreased expression level of each three AQPs in apoptotic T cell populations. According to our findings, tested aquaporin proteins may play roles in T cell activation, differentiation, and apoptosis. The scientific significance of this research is that it can fill the gaps about these three functional processes of T cells. Besides, all findings can contribute to treatment of many autoimmune disease like MS which Th17 cells involve in pathogeny.Master Thesis Investigation of the Effects of Il-7 on the Th-17 Cell Apoptosis(Izmir Institute of Technology, 2015) Aydınlı, Fatmagül İlayda; Nalbant Aldanmaz, AytenTh17 cells known as Interleukin-17 (Inflammatory Cytokine) producing cells are differentiated subsets from naïve CD4+ T cells and have crucial roles in regulation of inflammation, host defense and autoimmunity. TCR (T Cell Receptor) activation is triggered under Th17 cell culture conditions and resulting naïve CD4+ T cells are induced to differentiate through Th17 cells. In the life time of activated T cells, the activation process also induces an apoptotic mechanism which is called activation-induced cell death (AICD) for elimination of activated cells from the environment for maintenance of homeostasis. AICD is known as the main programmed cell death mechanism for T cells by Fas-FasL signaling resulting activation of early and late apoptotic caspase proteins such as caspase-3 and caspase-8. Moreover, Interleukin-7, which is a member of Interleukin-2 family, has a survival mechanism in T cells by the activation and maintenance of anti-apoptotic proteins mainly Bcl-2 and inhibition of pro-apoptotic proteins such as Bax and Bim. This research analyzes apoptosis mechanism in Th17 cells in terms of AICD and the effects of IL-7 on that apoptosis signaling pathway. Our results showed that IL-7 did not have any effect to AICD throughout Fas-FasL signaling and activation of caspase-3 and caspase-8 protein.Master Thesis Determination of Human T-Lymphocyte Apoptosis Mediated by Bacterial Heat Shock Protein(Izmir Institute of Technology, 2009) Dinç, Melis; Nalbant Aldanmaz, AytenPeriodontal diseases are the most common inflammatory disease worldwide which caused by the pathogenic organism living in biofilm. Aggregatibacter Actinomycetemcomitans (Aa) is the main player of the periodontitis disease pathology. Although some of the virulence factors of Aa has been identified up to now, its cytotoxic mechanism has not been clearly known yet. Although known virulence factors of Aa; ltx and cdt has been knocked out, the mutant Aa strains have retained the ability to induce apoptosis. Depending on the literature there must be another important virulence factor. 64kDa GroEL protein which is a molecular chaperone and a heat shock protein can be the potential candidate for being a virulence factor. AaGroEL protein has not been studied in terms of apoptosis up to now and it is not known how AaGroEL mediate immune regulation of T cells. In this study AaGroEL protein has been purified by using ATP Affinity chromatography and electroelution methods. After the purification step lps contamination has been removed by detoxi-gel endotoxin removal gel and detected by LAL Assay. Peripheral Blood Mononuclear Cells (PBMCs) were isolated by Ficoll Hypaque Density Gradient Centrifugation method. It was found that AaGroEL protein induces T cell apoptosis in dose and a time dependent manner. AaGroEL protein mediated T cell apoptosis has been detected by plasma membrane changes, activation of caspase-3 and DNA fragmentation. In conclusion, AaGroEL has antigenic properties that effect T lymphocytes by regulating immune response that would play important role in periodontal pathology.Master Thesis Determination of Apoptotic Effects of Clinoptilolite on Human T Lymphocytes(Izmir Institute of Technology, 2008) Uslu, Mehmet Emin; Nalbant Aldanmaz, AytenZeolites are defined as aluminum silicates that have made up of oxygen, aluminum and silicon. SiO4 and Alo4 tetrahedrals are the smallest units that give the specific shape to the molecule. There are more than 40 types of natural and over 150 types of synthetic zeolites occurs and those zeolites are used in agriculture, animal husbandry, architecture, pharmaceuticals and metallurgy. Recent years in literature it was shown that these zeolites can have regulatory effects on immune system. Also it was shown that they can influence the development of cancer and have a role on the expression of tumor suppressor genes. However there is no evidence can be found on how these molecules affect the function of specific cell types in the molecular level and also the mechanism of this effect.In this study the apoptotic effects of clinoptilolite which is a natural zeolite found in Gördes region of Turkey, on human T lymphocytes were studied. T cells were chosen in this study because they are the main players in the immune system. These cells can establish immune regulation and organize immune response. As a result, they are important in pathology and therapy. Peripheral blood mono nuclear cells (PBMCs) in which T cells can be found were isolated from the healthy donors blood by Ficoll Hypaque Gradient Method and then these cells were incubated with clinoptilolite in the RPMI 1640 media. Apoptosis were measured in FacsArray after appropriate immunofluorescent labeling and by agarose electrophoresis technique after DNA fragmentation assay done.