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Expression of immunogenic cell death markers on lung cancer cells
Urbanová, Linda
Immunogenic cell death (ICD) is characterized by presence of specific molecules including surface exposed calreticulin (CRT) and the heat shock proteins HSP70 and HSP90. Release of ATP and high-mobility group box protein 1 (HMGB1) belongs to other typical characteristics. For induction of ICD in lung cancer cells high-hydrostatic pressure (HHP) was used. Treatment by HHP induces expression of immunogenic markers CRT, HSP70 and HSP90 on the cell surface. HHP also induces secretion of ATP to the extracellular milieu. Dendritic cells (DC) pulsed with HHP-treated tumor cells showed fenotypic maturation characterized by upregulation of maturation molecule CD83, costimulation molecules CD80 and CD86, chemokine receptor CCR7 and MHC class II molecule HLA-DR. Pulsed DCs have also higher rate of phagocytosis of HHP-treated tumor cells and they induce lower numbers of regulatory T cells compared to immature DCs. Moreover, activation of caspases (-8, -9, -3) and other proteins (phosphorylation of eIF2α) which are crucial in ER-stress mediated apoptotic pathway, was observed after HHP treatment. Using wide range of methods it was confirmed that HHP treatment is able to induce ICD in lung cancer cell lines, fenotypic and functional characteristics were described and the decreased induction of regulatory...
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Detection of surface phenotype and chemosensitivity in bladder carcinoma cells in vitro
Šímová, Michaela ; Drbal, Karel (advisor) ; Vondálová Blanářová, Olga (referee)
Tumor malignancies are the second leading cause of death worldwide. One of the reasons for the failure of oncological treatment are the uniformly set clinical guidelines, which neglect the effect of high intertumoral heterogeneity. The in vitro chemosensitivity and resistance (CSRA) assays allow for the stratification of patients prior to therapy. Therefore, the CSRA are a long-considered method for personalization of components of chemotherapy regime. Nevertheless, none of them is being routinely used in clinical practice. Certain chemotherapeutics used for their cytotoxic and cytostatic effect are also able to induce so-called immunogenic cell death (ICD) of tumor cells and activate an anti-tumor immune response. Monitoring of changes in the expression of molecules associated with the regulation of the innate immune system on the surface of dying tumor cells would enable to predict the patient's ability to respond to treatment involving modern immunotherapeutics. The feasibility of CSRA using flow cytometry and microscopy is critically evaluated in this thesis on a model of bladder cancer. Simultaneously, the correlation of the immunogenic phenotype of tumor cells and their sensitivity to selected chemotherapeutics is discussed.
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Expression of immunogenic cell death markers on lung cancer cells
Urbanová, Linda
Immunogenic cell death (ICD) is characterized by presence of specific molecules including surface exposed calreticulin (CRT) and the heat shock proteins HSP70 and HSP90. Release of ATP and high-mobility group box protein 1 (HMGB1) belongs to other typical characteristics. For induction of ICD in lung cancer cells high-hydrostatic pressure (HHP) was used. Treatment by HHP induces expression of immunogenic markers CRT, HSP70 and HSP90 on the cell surface. HHP also induces secretion of ATP to the extracellular milieu. Dendritic cells (DC) pulsed with HHP-treated tumor cells showed fenotypic maturation characterized by upregulation of maturation molecule CD83, costimulation molecules CD80 and CD86, chemokine receptor CCR7 and MHC class II molecule HLA-DR. Pulsed DCs have also higher rate of phagocytosis of HHP-treated tumor cells and they induce lower numbers of regulatory T cells compared to immature DCs. Moreover, activation of caspases (-8, -9, -3) and other proteins (phosphorylation of eIF2α) which are crucial in ER-stress mediated apoptotic pathway, was observed after HHP treatment. Using wide range of methods it was confirmed that HHP treatment is able to induce ICD in lung cancer cell lines, fenotypic and functional characteristics were described and the decreased induction of regulatory...
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Expression of immunogenic cell death markers on lung cancer cells
Kobosilová, Linda ; Špíšek, Radek (advisor) ; Černý, Jan (referee)
Immunogenic cell death (ICD) is characterized by presence of specific molecules including surface exposed calreticulin (CRT) and the heat shock proteins HSP70 and HSP90. Release of ATP and high- mobility group box protein 1 (HMGB1) belongs to other typical characteristics. For induction of ICD in lung cancer cells high-hydrostatic pressure (HHP) was used. Treatment by HHP induces expression of immunogenic markers CRT, HSP70 and HSP90 on the cell surface. HHP also induces secretion of ATP to the extracellular milieu. Dendritic cells (DC) pulsed with HHP-treated tumor cells showed fenotypic maturation characterized by upregulation of maturation molecule CD83, costimulation molecules CD80 and CD86, chemokine receptor CCR7 and MHC class II molecule HLA-DR. Pulsed DCs have also higher rate of phagocytosis of HHP-treated tumor cells and they induce lower numbers of regulatory T cells compared to immature DCs. Moreover, activation of caspases (-8, -9, -3) and other proteins (phosphorylation of eIF2α) which are crucial in ER-stress mediated apoptotic pathway, was observed after HHP treatment. Using wide range of methods it was confirmed that HHP treatment is able to induce ICD in lung cancer cell lines, fenotypic and functional characteristics were described and the decreased induction of regulatory T-lymphocytes...
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