Master Degree / Yüksek Lisans Tezleri

Permanent URI for this collectionhttps://hdl.handle.net/11147/3008

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Publisher: search.filters.publisher.01. Izmir Institute of TechnologySubject: search.filters.subject.T cells

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  • Master Thesis
    Investigation of Fas/Fas-ligand Interaction in Helper T 17 Cell Functions
    (01. Izmir Institute of Technology, 2021) Nalbant Aldanmaz, Ayten; Nalbant Aldanmaz, Ayten; Nalbant Aldanmaz, Ayten; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of Technology
    Th17 cells are key players of the adaptive immune system. They mainly take part in responses to extracellular parasites and neutrophil recruitment. They can infiltrate into the inflammation sites and survive for long periods of time. Their malfunction leads to the manifestation of several autoimmune diseases, such as Multiple Sclerosis and Rheumatoid Arthritis. The main reason behind their roles in autoimmune pathogenicity is thought to be their longevity and resistance to apoptosis. One of the main players of apoptosis, Fas, has been found to have non-apoptotic roles and is a candidate for the survival mechanisms of Th17 cells. This study aims to discover possible non-apoptotic roles of the Fas signaling pathway in Th17 cell functions. For this purpose, buffy coats of healthy individuals were used to isolate PBMCs and CD4+CD45RA+ naive T cells were sorted from the PBMCs. Obtained naive T cells were cultured under Th17 polarizing conditions and the expressions of Fas, FasL, TNFR1, and TNF-α have been monitored along with apoptosis. The expression of Fas has been found to significantly increase in the cells cultured under Th17 polarizing conditions. However, there were no FasL and TNFR1 expressions observed. The expression of TNF-α was observed on both the negative culture and Th17 polarizing culture, however, there was no significant difference found. In addition, there was no increase in apoptosis in neither culture. In summary, Fas expression has been found to increase in the cells cultured under Th17 polarizing conditions. Further investigation of possible survival mechanisms, such as NFkB, in these cells can shed light on the effects of the Fas signaling pathway on the longevity of Th17 cells
  • Master Thesis
    Investigation of Bcl-2 Proteins in Th17 Cell Functions
    (01. Izmir Institute of Technology, 2021) Nalbant Aldanmaz, Ayten; Nalbant Aldanmaz, Ayten; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of Technology
    Interleukin 17 producing T helper 17 cells are the distinct subset of CD4⁺ T cells. Th17 cells are an important part of the immune response of host defense. Dysregulation of Th17 cells plays a role in various pathologies including autoimmune diseases and cancer types. Bcl-2 family proteins are mostly known regulators of apoptotic cell death. The apoptotic and survival mechanisms of Th17 cells are not well known yet. Therefore, this study aims to investigate Bcl-2 protein family functions in Th17 cell survival and to understand the regulation network of apoptotic mechanisms of Th17 cells. To do that, Peripheral Blood Mononuclear Cells were isolated from a healthy buffy coat by Ficoll separation. Naive T cells were sorted from PBMC and cultured under Th17 polarizing conditions. Th17 cells were phenotypically characterized by flow cytometry. Afterward, cell lysates were obtained from Th0 and Th17 cells at different time points. The expressions of human transcription factor RORC2, proapoptotic Bik, Bid, Puma and Bim and, antiapoptotic Mcl-1 and Bcl-xL at cell groups were detected by Western blotting. The increased expressions of Bcl-xL and Mcl-1 were detected where the diminished expressions of Bim and Puma were detected in proportional with Th17 differentiation by increased RORC2 and elevated RORC/IL17A levels. Bik was undetectable in both cell groups while non-truncated isoform of Bid was barely decreased among cell groups. Outputs of this study allow us to understand the dynamics of Bcl-2 family proteins in human Th17 cell survival. The understanding roles of Bcl-2 proteins in Th17 cells may help to develop different therapeutics for Th17 associated diseases in the future.