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Influence of tumor microenvironment, cellular and humoral immunity on cancer pathogenesis.
Špaček, Jan ; Závadová, Eva (advisor) ; Petráková, Katarína (referee) ; Kopečková, Kateřina (referee)
Cancer is the second leading cause of death in the Czech Republic. Breast cancer and colorectal cancer have relatively high mortality rate. One of the areas of current clinical research in oncology is the study of prognostic biomarkers, which aims to optimize the decision-making process for a patient. Immune response and processes in the tumor microenvironment have been shown to influence to a large extent the biological nature of the tumor in terms of its aggressiveness and ability to metastasize in the host's body. There are certain tumors that could induce a strong immune response, while others do not. The ability to induce an anti-tumor cell response and to attract specific lymphocyte subpopulations directly into tumor tissue has been shown to be very closely related to the prognosis of cancer patients. There is evidence and correlation of the presence of so-called tumor infiltrating lymphocytes in tumor tissue and overall patient survival. Stratification of cancer patients based on immuno-predictors both in the plasma and directly in the tumor microenvironment makes it possible to identify suitable candidates for rediscovered modern anti-tumor immunotherapy, which can already be considered a standard therapeutic modality. In our projects, we focused on the identification of biomarkers that...
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The role of innate lymphoid cells in influenza virus infection
Mouyabi, Flaviancia ; Hrdý, Jiří (advisor) ; Kössl, Jan (referee)
Innate lymphoid cells (ILCs) are recently discovered group of innate immune cells. They do not have antigen-specific receptors but they can be activated by cytokines similarly to T lymphocytes. ILCs have a crucial role in the regulation of inflammation, tissue repair, containment of commensals, anti-infection immunity and regulation of tissue homeostasis. The presence of mouse and human ILCs can be detected in the lung during and after influenza virus infection when ILCs contribute to the restoration of damaged lung parenchyma. ILCs directly or indirectly provide protection against viral infections by secretion of various cytokines and co-operation with other cells (e.g. T cells, macrophages). Overall, lung ILCs are important in immune responses and tissue homeostasis, but further studies on this topic are needed to fully understand their role. The aim of this thesis was to specifically characterize these cells, focus on their function in the lung, and describe their role in the course of influenza virus infection.
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New chimeric antigen receptor (CAR) for therapy of human cytomegalovirus (HCMV) infection
Kroutilová, Marie ; Němečková, Šárka (advisor) ; Forstová, Jitka (referee)
Human cytomegalovirus (HCMV, Herpesviridae) can cause severe complications in the infected individuals undergoing hematopoietic stem cell transplantation. Nowadays, these patients are treated using antivirotics or HCMV-specific T cells derived from the seropositive graft donor. This study explored the possibility of redirecting HCMV-non-specific T cells from a seronegative donor towards HCMV-infected cells via chimeric antigen receptor (CAR), i.e. artificially designed T cell receptor. Viral glycoprotein B (gB) has been selected as a target for this receptor. Published sequence of a single chain variable fragment of a human antibody was used for the design of the CAR against gB (gBCAR). After the verification of production and surface localization in cell lines, gBCAR was being introduced into human T cells via lentiviral vectors. Human fetal lung fibroblasts (LEP) infected with HCMV were used as target cells after the expression of gB at their surface was demonstrated. gBCAR functionality was evaluated by the incubation of modified T cells with infected cells and subsequent analysis of media for IFNγ concentration, which was significantly higher in the setting of gBCAR T cells incubated with HCMV-LEP than in the control incubations. The results obtained show the specificity of gBCAR against...
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Preparation of expression vector for production of receptors CD69 and S1P1
Leontovyč, Adrian ; Vaněk, Ondřej (advisor) ; Obšil, Tomáš (referee)
T- and B- lymphocytes play a key role in immunity, because they provide specific immunity in organism. Receptor S1P1 regulates lymphocyte egress from lymphoid organs to blood, from which the lymphocytes travel to the site of infection. It was discovered that transmembrane helix 4 of S1P1 receptor interacts with transmembrane part of CD69 receptor and this interaction is responsible for retention of lymphocytes in lymphoid organs. The aim of this study is to prepare expression plasmids for production of these two receptors and their further research.
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Tumor microenvironment and the importance of anti-tumor immunity for clinical course of human cancers
Partlová, Simona ; Špíšek, Radek (advisor) ; Drbal, Karel (referee) ; Kovář, Marek (referee)
Cancer development and progression vary depending on tumor type, localization, invasion, immunogenicity and the ability of immune system to become activated. There are frequent interactions between tumor cells and immune cells, occuring locally at the site of primary tumor or distally through paracrine signalling of various mediators and cytokines. The main subject of this PhD thesis is to study key factors and aspects of immune response in cancer patients. In the first part, we analyzed immune cells infiltrating tumor tissues of ovarian cancer patients at different stages of disease. We focused on the dynamics of immune response, primarily on frequency of individual T lymphocyte populations in peripheral blood and tumor infiltrating T lymphocytes in tumors of early and advanced stages of ovarian cancer. We found that during disease progression there is a gradual decrease of proinflammatory Th17 and Th1 immune responses and a specific recruitment of regulatory T cells to the tumor site, which results in a significant immune suppression in the tumor microenvironment. In the second part, we demonstrated that the character of immune response in HPV-positive head and neck cancer patients is very different from the patients with tumors not associated with HPV infection. In HPV-positive patients, significantly...
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Monitoring of immune parameters during anti-tumor immunotherapy
Bílková, Pavla ; Palich Fučíková, Jitka (advisor) ; Fialová, Anna (referee)
Dendritic cells are the most effective antigen presenting cells in humans, they stimulate naive T lymphocytes and thus initiate specific immune response. The discovery of dendritic cells and understanding of their functions contributed to the idea of usingdendritic cells for the treatment of cancer. Anti tumor immunotherapy is a therapeutic strategy that aims to induce and maintain immune responses against tumor cells. Currently, immunotherapy based on dendritic cells has strong position among other anti cancer therapies and seems to be a promising therapeutic option for patients with tumors. In this work, I evaluated the effectiveness of treatment in patients with prostate cancer treated with immunotherapy based on dendritic cells. I focused on the detection of antigen specific T lymphocytes in peripheral blood against tumor antigens, PSA, NY ESO 1, MAGE A1 and MAGE A3. Using a 3 day standard protocol for the detection of antigen specific T cells using intracellular cytokine staining we were able to detect only a small percentage of this minor population. Only after extension of the protocol, we increased the sensitivity setting and we detected a significantly increased frequency of antigen specific T lymphocytes in the peripheral blood after one year DC vaccines application.
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Role of CD8- and CD4-Lck interactions in the signaling and development of T cells.
Horková, Veronika
Adaptive immune response plays a key role in maintaining homeostasis of the organism. T cells use an immense repertoire of T-cell receptors (TCRs) to discriminate between self and foreign antigens with very high sensitivity. Although we have many clues outlining how an ideal TCR repertoire is selected, and a good understanding of the TCR signaling machinery, there are still some key aspects of these processes that remain controversial. The objective of this thesis is to extend our knowledge of the very proximal events of TCR signaling, with special focus on interaction of TCR coreceptors with lymphocyte-specific kinase LCK. Coreceptor-LCK interaction has been described to regulate several aspects of T- cell development and response. We observed dynamic change of this interaction in course of T-cell development. Interestingly, CD4 and CD8 coreceptors displayed differential dynamics of interaction with LCK. Our data suggest that such disparity in coreceptor- LCK interaction leads to selection of more self-reactive TCR repertoire in CD8+ T cells. Moreover, when the highly self-reactive CD8+ T cells get to the periphery, the homeostatic signals drive their differentiation towards a more tolerogenic memory-like phenotype. To finally resolve the role of coreceptor-LCK interaction in the T-cell...
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Role of CD8- and CD4-Lck interactions in the signaling and development of T cells.
Horková, Veronika ; Štěpánek, Ondřej (advisor) ; Černý, Jan (referee) ; Hons, Miroslav (referee)
Adaptive immune response plays a key role in maintaining homeostasis of the organism. T cells use an immense repertoire of T-cell receptors (TCRs) to discriminate between self and foreign antigens with very high sensitivity. Although we have many clues outlining how an ideal TCR repertoire is selected, and a good understanding of the TCR signaling machinery, there are still some key aspects of these processes that remain controversial. The objective of this thesis is to extend our knowledge of the very proximal events of TCR signaling, with special focus on interaction of TCR coreceptors with lymphocyte-specific kinase LCK. Coreceptor-LCK interaction has been described to regulate several aspects of T- cell development and response. We observed dynamic change of this interaction in course of T-cell development. Interestingly, CD4 and CD8 coreceptors displayed differential dynamics of interaction with LCK. Our data suggest that such disparity in coreceptor- LCK interaction leads to selection of more self-reactive TCR repertoire in CD8+ T cells. Moreover, when the highly self-reactive CD8+ T cells get to the periphery, the homeostatic signals drive their differentiation towards a more tolerogenic memory-like phenotype. To finally resolve the role of coreceptor-LCK interaction in the T-cell...
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Influence of tumor microenvironment, cellular and humoral immunity on cancer pathogenesis.
Špaček, Jan ; Závadová, Eva (advisor) ; Petráková, Katarína (referee) ; Kopečková, Kateřina (referee)
Cancer is the second leading cause of death in the Czech Republic. Breast cancer and colorectal cancer have relatively high mortality rate. One of the areas of current clinical research in oncology is the study of prognostic biomarkers, which aims to optimize the decision-making process for a patient. Immune response and processes in the tumor microenvironment have been shown to influence to a large extent the biological nature of the tumor in terms of its aggressiveness and ability to metastasize in the host's body. There are certain tumors that could induce a strong immune response, while others do not. The ability to induce an anti-tumor cell response and to attract specific lymphocyte subpopulations directly into tumor tissue has been shown to be very closely related to the prognosis of cancer patients. There is evidence and correlation of the presence of so-called tumor infiltrating lymphocytes in tumor tissue and overall patient survival. Stratification of cancer patients based on immuno-predictors both in the plasma and directly in the tumor microenvironment makes it possible to identify suitable candidates for rediscovered modern anti-tumor immunotherapy, which can already be considered a standard therapeutic modality. In our projects, we focused on the identification of biomarkers that...
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Function of adaptor protein NDFIP1 in T lymphocytes
Dvořáček, Tomáš ; Brdička, Tomáš (advisor) ; Macůrková, Marie (referee)
T lymphocytes specifically recognize and promote immune response against pathogenic agents, which endanger health of host organism. However, this ability must be tightly controlled to prevent response against innocuous or self-tissue antigens, which would unnecessarily damage an organism. For a maintenance of this tolerance, there is a number of molecular mechanisms regulating T lymphocytes. The protein NDFIP1 is important component of these mechanisms. This transmembrane adaptor protein binds to E3 ubiquitin ligases from the NEDD4 family, which are under normal circumstances in inactive state. Binding of NDFIP1 leads to their activation and facilitates regulation of T cell behaviour by these ligases. NDFIP1 deficiency in mice causes the development of inflammatory disease affecting the skin, lungs and gastrointestinal tract. It is characterized by increased numbers of activated CD4+ T lymphocytes and eosinophils, which infiltrate the affected tissues. The disease is ultimately fatal. The aim of this work is to describe the function of the NDFIP1 protein in T lymphocytes, which are essential for the development of this pathology. The absence of NDFIP1 in T cells disrupts several molecular mechanisms, including the degradation of the transcription factors JUNB and RORγT and the regulation of mTOR...
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