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Significance of the major histocompatibility complex for organ transplantation
Ilina, Liudmila ; Slavčev, Antonij (advisor) ; Grobárová, Valéria (referee)
The major histocompatibility system is a region in the human genome located on chromosome 6. HLA genes encode polymorphic cell-surface glycoproteins which are primarily responsible for presentation of self and non-self antigens to T cells. When the T lymphocyte recognizes the MHC-peptide complex as foreign, it activates effector components of the innate and adaptive immune system. Therefore, mismatched HLA antigens can lead to a strong immune response against the donor's tissue. HLA laboratories support transplant programs by evaluation the HLA matching between patients and their potential donors and, based on these data, assist in the evaluation of the risk of rejection and eventual immunological complications after transplantation. The aim of this thesis is to describe the significance of the major histocompatibility complex for the occurrence of cellular and antibody-mediated rejection after solid organ transplantation and discuss the relationship between the degree of HLA matching and graft survival outcomes. Key words HLA, organ transplantation, rejection
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Mutual communication and relationship between tumor cells and the tumor microenvironment
Novák, Štěpán ; Szabo, Pavol (advisor) ; Krška, Zdeněk (referee) ; Grobárová, Valéria (referee)
The present dissertation addresses two main objectives. First, the interaction between four pancreatic ductal adenocarcinoma (PDAC) cell lines and conditioned media from normal/healthy fibroblasts was investigated, HF) and cancer- associated fibroblasts (CAFs) derived from PDAC (PDAC-derived CAFs, PANFs), cells derived from ascitic fluid of patients with end-stage PDAC and primary PDAC tumor. Subsequently, the genome-wide profile of PANFs was determined. The second aim was to differentiate squamous cell carcinoma (SCC) cells, surgical resection margin (MSR) cells and normal mucosa (NM) cells in patients operated for head and neck squamous cell carcinoma (HNSCC) by immunohistochemistry, using detection of tenascin (Ten), fibronectin (Fn) and galectin-1 (Gal-1), and to correlate these results with their clinical characteristics. Subsequently, whole-genome profiling of SCC, MSR and NM samples was performed based on Ten expression stratification. Results Cultivation of the four PDAC lines under the influence of conditioned media from HF and PANF was heterogeneous. After stimulation, the most aggressive behavior was obtained in the Panc-1 cell line while the PaTu-8902 line was rather inhibited by the media. The PANF transcriptome showed increased expression of some genes (e.g. IL-6, IL-8, MFGE8,...
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Effect of TNF-α inhibitors on microbiota composition and immune response in patients with inflammatory bowel disease
Mihula, Martin ; Jirásková Zákostelská, Zuzana (advisor) ; Grobárová, Valéria (referee)
One of the most common used therapies in inflammatory bowel diseases (IBD) treatment are inhibitors of a cytokine TNF-α. Nevertheless, up to one third of IBD patients stop respond to this therapy for unknown reason. In these days, there are not any ideal biomarkers which could predict patient's long- term response to anti-TNF-α therapy. Because the gut microbiota composition changes are tightly related to the pathogenesis of IBD, my aim in this thesis was to find out if these changes in composition are happening also due the therapy by inhibitors of TNF-α as well. Moreover, I tried to find out if there are changes in production of serum biomarkers related to the gut barrier damage and to the immune response associated with microbial translocation. Also, I focused on the immune response of IBD patients against common gut commensal bacterial antigens during the anti-TNF-α therapy. In our study, we collected for these purposes stool or blood samples from 46 IBD patients before the therapy and at 38th week from the start of the therapy and 39 healthy controls. I found that IBD patients had higher bacterial diversity (α-diversity) as well as different bacterial composition across observed groups (β-diversity) at 38th week of the anti-TNF-α therapy than before the therapy. When I divided IBD patients...
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The effect of nanoparticles on functional and regenerative properties of mesenchymal stem cells
Dostálová, Dominika ; Rössner, Pavel (advisor) ; Grobárová, Valéria (referee)
Mesenchymal stem cells (MSCs) represent a type of stem cells, localized in various tissues, where they are involved in the regeneration process. Because of their properties, MSC are tested for cell-based therapy. However, the treatment of injuries is often associated with bacterial infections, which complicate the process of healing. To establish a sterile environment, metallic nanoparticles (NPs) with proven antibacterial properties can be applied, for improving the healing process. Mechanisms of the antibacterial effect of NPs are non-specific and may cause the damage of eukaryotic cells. The aim of this thesis was to evaluate the effect of three antibacterial metallic NPs (silver, copper oxide and titanium dioxide) on the functional and regenerative properties of MSCs. The effect of NPs on the metabolic activity, differentiational potential, expression of genes for immunoregulatory molecules and production of cytokines by MSCs was determined. Additionally, the immunoregulatory properties of MSCs after cultivation with NPs were tested. The results showed that the metabolic activity was reduced in the presence of silver and copper oxide NPs, with the decrease dependent on the type of the tested NPs. The expression of selected immunoregulatory molecules was inhibited and the immunoregulatory...
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Immunomodulatory and regenerative potential of mesenchymal stem cells in the treatment of degenerative retinal disorders in mice
Palacká, Kateřina ; Heřmánková, Barbora (advisor) ; Grobárová, Valéria (referee)
Retinal degenerative disease are the leading cause of vision loss in adult patients. Important role in the development of these types of disease play gradual death of retinal cells and an inflammatory reaction that leads to the production of cytokines, formation of inflamasome, increased angiogenesis and scar formation. These pathologies result in irreversible degeneration of retinal cells. Retinal pigmenetd epithelium cells, photoreceptors and ganglion cells are the most frequently damaged cell types in this conditions. The available treatments are currently very limited and effective only at the early stages of the disease. Therefore, the use of stem cell- based therapy could be a promising option. For therapeutic purposes it would be possible to use mesenchymal stem cells (MSCs) which may be isolated for example from bone marrow or adipose tissue. MSCs are capable of production of neuroprotective factors, differentiation into the variety of cells types and regulation of immune response. In this study we tested the therapeutic potential of MSC administered locally to the damaged retina in an experimental model of retinal degeneration. We focused on the protective effect of MSCs on photoreceptor cells, regulation of the local immune response and expression of genes for cytokines involved in...
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Use of polymer prodrugs containing cucurbitacin D for the treatment of experimental tumors
Hrabánková, Klára ; Šírová, Milada (advisor) ; Grobárová, Valéria (referee)
Chemotherapy is still the most widely used anti-cancer treatment. The majority of chemotherapeutics inhibit proliferating cells generally, not selectively cancer cells. The side effects associated with chemotherapy can be partly limited by conjugating a cytotoxic drug with a polymer nanocarrier. Such binding facilitates solubility in aqueous solutions, reduces systemic toxicity; and passively targets the drug directly into the tumour through the enhanced permeability and retention (EPR) effect. This thesis focuses on testing polymer conjugates based on N-(2-hydroxypropyl)methacrylamide (HPMA) carrying cucurbitacin D (CuD), a naturally occurring compound with potential anti-cancer activity. The mechanism of action is not elucidated yet, but several studies have depicted the inhibitory effect on signal transducer and activator of transcription 3 (STAT3) transcription factor. A STAT3 signalling pathway is overexpressed in several cancer cell lines and is also involved in the differentiation of myeloid- derived suppressor cells (MDSCs). We examined the therapeutic effect of the HPMA copolymers based on CuD in combined therapy with other polymer chemotherapeutics. CuD conjugates have shown in vitro cytotoxic effect on several model cancer cell lines. The combination with conjugates carrying doxorubicin...
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Immunomodulatory and regenerative potential of mesenchymal stem cells in the treatment of degenerative retinal disorders in mice
Palacká, Kateřina ; Heřmánková, Barbora (advisor) ; Grobárová, Valéria (referee)
Retinal degenerative disease are the leading cause of vision loss in adult patients. Important role in the development of these types of disease play gradual death of retinal cells and an inflammatory reaction that leads to the production of cytokines, formation of inflamasome, increased angiogenesis and scar formation. These pathologies result in irreversible degeneration of retinal cells. Retinal pigmenetd epithelium cells, photoreceptors and ganglion cells are the most frequently damaged cell types in this conditions. The available treatments are currently very limited and effective only at the early stages of the disease. Therefore, the use of stem cell- based therapy could be a promising option. For therapeutic purposes it would be possible to use mesenchymal stem cells (MSCs) which may be isolated for example from bone marrow or adipose tissue. MSCs are capable of production of neuroprotective factors, differentiation into the variety of cells types and regulation of immune response. In this study we tested the therapeutic potential of MSC administered locally to the damaged retina in an experimental model of retinal degeneration. We focused on the protective effect of MSCs on photoreceptor cells, regulation of the local immune response and expression of genes for cytokines involved in...
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Effect of Wharton's jelly-derived mesenchymal stem cells secretome in the treatment of rat spinal cord injury
Jančová, Pavlína ; Chudíčková, Milada (advisor) ; Grobárová, Valéria (referee)
As previously shown, transplantation of Wharton's jelly-derived mesenchymal stem cells in the experimental model of spinal cord injury leads to the motor and sensory functions improvement, supports neuroregeneration, angiogenesis and provides immunomodulation. On the other hand, these cells have limited migration and survival capacity, and their therapeutic effect is mediated mostly by their secretome. Therefore, application of mesenchymal stem cells derived conditioned medium is studied as an alternative option for cell therapy. In this thesis, therapeutic effect of repeated intrathecal delivery of human Wharton's jelly-derived mesenchymal stem cells and their conditioned media in the treatment of spinal cord injury was compared. After induction of a balloon ischemic compression lesion, stem cells or conditioned media were administered weekly in three doses. Behavioral analyses were carried out up to nine weeks after spinal cord injury and revealed significant improvement of both treated groups compared to the untreated saline control. Application of stem cells and conditioned media also resulted in a higher amount of spared tissue and enhanced expression of genes related to neuroregeneration, although the size of glial scar was not reduced. Compared to application of stem cells, application of...
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Potential role of skin microbiota in the pathogenesis of dermatological diseases
Mihula, Martin ; Jirásková Zákostelská, Zuzana (advisor) ; Grobárová, Valéria (referee)
The surface of the human body is colonized by a large number of microorganisms whose composition depends not only on external and internal factors, but is also significantly influenced by the topography of human skin. The complex skin microbiota is an essential part of physiological and protective mechanisms of the skin. The change in the dynamics of microbial communities on the skin or in the gastrointestinal tract is currently considered to be part of triggering mechanisms of many skin diseases. Some of the skin inflammatory diseases are directly associated with a shift of skin microbiota composition - for instance atopic dermatitis, acne vulgaris or psoriasis. Gaining and perceiving knowledge about interspecies interactions and their effect on a host could lead to the development of new diagnostic and therapeutic approaches which could make the prevention or treatment of some skin disorders more effective. Key words: skin, skin diseases, skin microbiota, immunity, psoriasis, atopic dermatitis, vitiligo
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Microbiota and inflammatory bowel diseases
Gajdárová, Zuzana ; Kostovčíková, Klára (advisor) ; Grobárová, Valéria (referee)
Inflammatory bowel diseases (IBD) are an autoimmune illnesses affecting gastrointestinal tract. The main types include ulcerative colitis and Crohn's disease. Recently, primary sclerosing cholangitis (PSC) has also been associated with IBD. PSC is a chronic liver disease associated with bile duct stenosis. The exact pathogenesis and etiology of these diseases is not clear, despite the great efforts of the scientific community. They are multifactorial diseases that are associated with dysbiosis of intestinal microbiota. Their diagnosis is based on for patients unpleasant endoscopic examinations and therefore the search for new serum biomarkers is needed and appreciated target of scientific interest. In the first part of diploma thesis, we focused on the reactivity of peripheral blood cells of IBD patients to 10 selected representatives of typical intestinal microbiota: Lactobacillus plantarum, Bifidobacterium adolescentis, Blautia coccoides, Roseburia intestinalis, Eubacterium rectale, Faecalibacterium prausnitzii, Ruminococcus flavefaciens, Bacteroides thetaiotaomicron, Prevotella ruminicola and Escherichia coli. Reactivity of CD, UC and PSC- IBD patients was increased after stimulation with Faecalibacterium, Lactobacillus and Prevotella. However, we got low percentage of cytokine-producing cells,...
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