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Immunological features of esophageal carcinoma in prognosis and therapy
Šnajdauf, Martin ; Lischke, Robert (advisor) ; Vrba, Radek (referee) ; Stříž, Ilja (referee)
Esophageal cancer belongs to the gastrointestinal malignancies with the worst prognosis. Current treatment options, including surgical resection, chemotherapy, radiotherapy, or a combination of these methods, have low efficacy and the five-year survival rate for patients with esophageal cancer is approximately 10 to 15 %. In the last decade, immunotherapy has become the leading treatment modality for metastatic tumors. However, the success of immunotherapeutic approaches does not only depend on the infiltration of the tumor microenvironment with immune cells but also on the phenotype of these infiltrating cells. The aim of this project was to evaluatethe immunological features of tumor-infiltrating lymphocytes obtained from different tissue compartments (tumor, peritumoral tissue, adjacent healthy tissue, and lymph node) of patients during surgical resection and to compare them with clinical and histopathological data of patients. We observed that the distribution of NK (natural killer) cells, CD8 and CD4 positive T cells was different in each tissue compartment. While the lymph nodes had the highest percentage of T cells, the opposite was seen in NK cells. The proportion of NK cells was the lowest in the lymph nodes. The expression of death receptors FasR and DR3 (death receptor 3) was the lowest...
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Mechanisms of immune response against kidney allograft
Švachová, Veronika ; Stříž, Ilja (advisor) ; Mrázek, František (referee) ; Posová, Helena (referee)
Kidney transplantation improves the prognosis and quality of life of patients with end- stage renal disease. However, the occurrence of rejection still represents a major complication that can lead to failure of allograft function. In our work, we have studied the phenotypic changes of subpopulations of B lymphocytes and monocytes, which represent one of the key players involved in immune responses against kidney transplantation. In the part of the research devoted to B lymphocytes, we have observed a different redistribution of individual B cell compartments in the peripheral blood during the first year after transplantation without significant differences between used immunosuppression. We have demonstrated that sensitized and rejected patients had a lower level of IgMhigh CD38high CD24high transient B cells. These results support the hypothesis of a protective role of transient B lymphocytes. Activation of monocyte subpopulations in peripheral blood in the posttransplantation perion was also subject of research. We have compared a cohort of patients with normal graft function with a group of patients with acute rejection. The level of "transient" monocytes and HLA-DR expression in a subset of "non-classical" monocytes did not decrease in patients with rejection, compared to the group with a...
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The role of IL-17 in kidney transplantation
Menšíková, Markéta ; Stříž, Ilja (advisor) ; Krulová, Magdaléna (referee)
The role of IL-17 in kidney transplantation - abstract Naive CD4+ T-lymphocytes (Thp) can develop into Th17 line in the presence of TGF- and IL-6. Th17 cells are characterized by expression of Ror- t and by production of interleukin-17 (IL-17). It is secreted as a glycoprotein homodimer. Binding to IL-17 receptor (IL-17R), which is present in all cell types, stimulates the production of proinflammatory cytokines and chemokines. The ratio of Th17: Treg in the graft showing signs of rejection is higher than in the graft without rejection. The presence of IL-17 in a culture of proximal tubular epithelial cells (PTEC) stimulates the production of IL-6, IL-8, MCP-1 and C3 complement component. Simultaneous action of IL-17 and CD40L synergistically increases the production of IL-6, IL-8 and RANTES. Signaling from the receptor on the surface of PTEC associated with its increased expression is effected via the src kinase and MAP kinase, and probably leads to the transcription factor NF- B. In rat models of transplantation, the IL-17 appears in allografts on the second day after surgery, the level rises until the fifth day, then decreases and disappears before the death of the animal. IL-17 is not detectable in isografts and negative controls. It appears before the IFN- , which had been considered a trigger of...
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The role of macrophages in immunosuppression mediated ny regulatory T cells
Kadlecová, Kristýna ; Holáň, Vladimír (advisor) ; Stříž, Ilja (referee)
Regulatory T cells (Treg) represent one of the most important mechanisms of immunoregulation. Treg suppress immune reactions and prevent overactivation of the immune system. There is a lot of ways of Treg action described, here we have focused on Treg interference with macrophages. The suppressor capacity of a highly purified Treg population was demonstrated in proliferation assays. The level of suppression of effector T cell proliferation differs depending on the presence of macrophages in the culture. Treg suppression has been significantly higher in the presence of macrophages. These observations led to hypotesis that Treg affect directly macrophages. However, using flow cytometry, reduction of expression of costimulatory molecules on macrophages after culture with Treg was not observed. Macrophages precultured with Treg showed a comparable functionality as macrophages cultured alone. Neither flow cytometry nor live cell imaging revealed any cytotoxic activity of Treg towards macrophages. Despite the presence of macrophages, Treg did not suppress effector cell proliferation in a model, where stronger activation of effector cells was induced. Therefore, a new hypothesis was presented - initially observed higher suppression in the presence of macrophages was probably caused by a qualitatively or...
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Immunomodulatory properties of mesenchymal stem cells - use in therapy
Pavlíková, Michaela ; Krulová, Magdaléna (advisor) ; Stříž, Ilja (referee)
Mesenchymal stem cells (MSC) are extensively studied mainly due to their feasible clinical application. Therapeutic potential of MSC consists not only of the ability to differentiate into mesenchymal cells, ectodermal and endodermal cell lines, but primarily in their immunomodulatory functions. Due to their effect on immune cells, MSC promote the shift of the inflammatory immune response to antiinflammatory. The ability to suppress inflammation, together with their differentiation potential and antiapoptotic potential on the surrounding cells makes MSC a promising tool for treating serious diseases. This work discusses the effect of MSC on the individual cells of the immune system. It focuses on the description of the effect of MSC in four model cases. These are an experimental autoimmune encephalomyelitis, myocardial infarction, diabetes mellitus and skin graft transplantation. The knowledge of the mechanisms of the interactions between MSC and the cells of the immune system, together with the understanding the effect of specific conditions on MSC is essential for their use in clinical therapy. Keywords: mesenchymal stem cells, immunomodulation, autoimmune diseases, transplantation
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Gut barrier in the pathogenesis and diagnostics of necrotizing enterocolitis and inflammatory bowel disease
Coufal, Štěpán ; Kverka, Miloslav (advisor) ; Nevoral, Jiří (referee) ; Stříž, Ilja (referee)
Disruption of gut microbiota, altered mucosal defense, inappropriate immune response and gut barrier damage are all typical features in the pathogenesis of both necrotizing enterocolitis (NEC) and inflammatory bowel disease (IBD). Despite of intensive research, the exact pathogenesis of both diseases remains unclear and the diagnostics and outcome prediction are still problematic. Therefore, we analyzed the role of gut-associated and inflammatory biomarkers, with respect to different aspects of gut barrier dysfunction in the pathogenesis of both disease, with the aim to improve the diagnostics and to predict the disease course and outcome. Using ELISA, we found that patients who will later develop NEC have significantly higher intestinal fatty acid-binding protein (I-FABP) than infants who will later develop sepsis already in first hours after NEC suspicion. Urinary I-FABP had high sensitivity (81%) and specificity (100%) and its addition to currently used gold standard for NEC diagnosis increased its sensitivity and negative predictive value. We found that serum amyloid A (SAA) was the strongest factor for prediction of the most severe stage of NEC. The combination of intestinal and liver FABP with SAA predicted the length of hospitalization in NEC patients and the low level of SAA predicted short...
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Dendritic cells in health and disease
Horváth, Rudolf ; Špíšek, Radek (advisor) ; Krejsek, Jan (referee) ; Stříž, Ilja (referee)
During the past decades several spectacular finding s have been made in the field of immunology. Elucidating the functions of the antigen presenting cells (APCs) belong to the most important. Dendritic cells (DCs) represent a specific group of APCs with a unique ability to initiate primary immune responses. Despite the fact that, in vivo, they are very rare and difficult to isolate, DCs came very fast into the focus of scientific interests. Development of novel laboratory techniques facilitated a robust expansion of their research. With time it has been proven that DCs play a pivotal role in initiation, maintenance and control of the immune responses. The extraordinary features of DCs were soon investigated in human clinical trials, where DCs have been particularly used as vectors for vaccination protocols, especially in the treatment of tumors. However, DCs capability to polarize the outcome of immune response and the potential to induce or suppress immunity under specific circumstances led to the idea that they might be also used in the treatment of autoimmune and allergic diseases or in transplantation medicine as well. There is a need to stress that most of the knowledge has been obtained from the in vitro generated DCs, but advanced technological methods bring us the opportunity to study DCs directly...
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The role of T regulatory cells in kidney transplantation
Urbanová, Anna ; Stříž, Ilja (advisor) ; Zajícová, Alena (referee)
T regulatory lymphocytes (Treg) belong to the CD4+ cell group. They are an essential part of the immunity system. Treg cells prevent from excessive activation of effector T cells and they keep the tolerance to the tissues of the body. They have high expression of CD25 and the transcription factor Foxp3. We distinguish two basic populations of Treg cells: natural Treg cells (nTreg) created in the thym and representing 5-10 % of all CD4+ cells, and induced Treg cells (iTreg), created from naive CD4+ cells in the periphery.Their regulatory effect is well-known, therefore using of Treg cells could bring about a huge treatment potential for patients with a transplantated kidney. Healthy people and patients tolerant to the transplantated kidney show higher occurance of circulating Treg cells and the Treg cells present in the graft unlike patients with chronical rejection. The tolerance is cancelled with the damage of CD4+ CD25+ cells.For a graft acceptance it is necessary to treat the patient after the transplantation with immunosuppressive medicaments resulting in suppression of immunity reaction against the graft. Their disadvantages are side effects often resulting in the patient's death. Moreover they often have a negative impact on survival and expansion of Treg cells. The analysis of flow cytometry has...
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Modulation of human macrophages and renal epitelium chemokine profile
Pidhorodetská, Halyna ; Stříž, Ilja (advisor) ; Javorková, Eliška (referee)
One of the main effects of pro-inflammatory cytokines is the induction of chemokines and the expression of adhesive molecules that regulate the migration of immune cells to the center of the damage. Chemoattractant gradient also provides a physiological delivery of cells to tissues and lymphatic organs under normal circumstances. Chemokines are chemotactic cytokines that form a very large and diverse group of secreted proteins that have many functions both in processes that maintain homeostasis but also in inflammatory states. Production of some chemokines also has a major effect on graft rejection. Further understanding of the mechanisms involved in the acute rejection chemokine could contribute to improving treatment steps in transplantology. In this diploma thesis, serum chemokine levels were monitored in renal transplant patients, but these measurements did not show significant dynamics. Furthermore, the effect of pro-inflammatory cytokines on the release of chemokines from renal epithelial cells and monocytes was studied. Experiments were performed to monitor the levels of individual chemokines such as ENA-78, IL-8, MCP-1, MIP-1 β, RANTES, GRO alpha, THP-1 (monocyte/macrophage cell line), RPTEC (renal epithelial cells of proximal tubules) and RA (renal cell tumor lines). TNF-α (tumor necrosis...
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