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Extracellular vesicles and middle T antigen of mouse polyomavirus
Kropáček, Václav ; Šroller, Vojtěch (advisor) ; Brázdová, Andrea (referee)
This study is focused on middle tumor antigen (MT Ag) of mouse polyomavirus (MPyV), potential consequences of it's secretion via extracellular vesicles (EVs) and it's effect on cellular signaling. MT Ag is membrane bound protein able to induce cellular transformation thanks to it's ability to interfere with cellular signal transduction. Mainly due to aberrant activation of MAP kinase pathway. Firstly we followed up previous observations of our group concerning ability of MT Ag to be secreted from cells via extracellular vesicles. We were interested if MT Ag could contribute to malignant transformation in recipient cells. We performed 2 types of EVs isolation from cell lines stably expressing middle T antigen (3T6MT). We confirmed presence of MT Ag in isolated EVs. Then we characterized isolated EVs by detection of exosomal markers and cryo-electron microscopy. In next step we exposed recipient cell line (3T6) to isolated EVs and with use of flow cytometry tried to detect internalization of MT Ag. Simultaneously we tried asses levels of Erk phosphorylation in 3T6 cells exposed to EVs. Secondly we tried to confirm and analyse previous unpublished observations of elevated levels of NF-kB phosphorylation in cells stably expressing MT Ag. We used western blot and detection of NF-kB dependent secreted...
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The role of extracellular vesicles in transmission of dsRNA viruses in parasitic protist Trichomomas vaginalis.
Horáčková, Jana ; Rada, Petr (advisor) ; Voleman, Luboš (referee)
Trichomonas vaginalis is a parasitic protist found in the human urogenital tract, where it adheres to the epithelial cells. During infection, T. vaginalis cells release extracellular vesicles that allow them to communicate with each other and modulate the host immune response. Most studied T. vaginalis strains contain dsRNA Trichomonasviruses (TVVs). These viruses belong to the Totiviridae family, which includes other dsRNA viruses from various parasitic protists. TVV is transmitted vertically to the daughter cells during cell division. Horizontal transfer of TVV between T. vaginalis cells has not yet been proven. TVV particles were found inside small extracellular vesicles that release infected T. vaginalis into the extracellular space. In this thesis, it was shown that TVV could be transmitted between trichomonads during co-cultivation of infected and uninfected trichomonads. However, stable TVV infection of trichomonads was not observed. It was also proven that extracellular vesicles from infected trichomonads could transmit TVV among other trichomonads. Nevertheless, this way of transmission did not lead to the stable TVV infection of trichomonads either.
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Extracellular vesicles derived from mesenchymal stem cells and their use in the treatment of neurological disorders
Kratochvílová, Anna ; Hájková, Michaela (advisor) ; Holada, Karel (referee)
Extracellular vesicles (EVs) are cell-derived membrane vesicles, which represent an important part of intercellular communication by transferring cytosolic proteins, lipids, and RNAs between cells. It has been proved that mesenchymal stem cells (MSCs) are potent producers of EVs with a therapeutic effect comparable to their parental cells. Moreover, recent findings suggest that EVs may strongly contribute to their physiological function. The aim of this thesis is to introduce extracellular vesicles derived from mesenchymal stem cells (MSC-EVs) as a novel cell-free alternative to the cell-based therapy and focus on their neuroprotective properties studied in various neurological diseases.
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