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The role of human heat shock protein HSP70, HSP60 and mycobacterium bovis HSP65 in pathogenesis of graft versus host disease and rheumatoid/juvenile idiopathic arthritis
Nguyen, Thi Thu Hien ; Hromadníková, Ilona (advisor) ; Tučková, Ludmila (referee) ; Stříž, Ilja (referee)
We investigated IgG antibodies against Mycobacterium bovis heat shock protein 65 (M. bovis Hsp65) fragments produced by cleavage with cyanogen bromide (CNBr) in 10 healthy controls, 11 patients with juvenile idiopathic arthritis (JIA) and 10 children with various malignant and non-malignant diseases before allogeneic stem cell transplantation (SCT) by using Western blotting (WB). CNBr cleaved M. bovis Hsp65 to three larger fragments: Pl-163, molecular weight (Mw) 17.1 kDa; P191-285, Mw 10.4 kDa and P290-534, Mw 25.3 kDa. Humoral reactivity against M. bovis Hsp65 derived fragments differed on a case-by-case basis. The sera either of JIA patients ar those befare SCT reacted with individual M. bovis Hsp65 fragments more frequently when compared with healthy controls. While IgG anti-Hsp65 antibody levels showed no significant differences between the small studied cohorts, significantly higher levels of antibodies against M. bovis Hsp65 epitopes were observed in patients befare SCT and JIA patients when compared with healthy controls. Comparing WB reflectance densities (DRl and DR2), significantly elevated antibodies against Pl-163 (DRl: p=0.014; DR2: p=0.022) and P290-534 (DRl: p=0.009; DR2: p=0.003) epitopes were found in patients befare SCT. Similarly, significantly increased DRl and DR2 values of antibodies...
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The influence of expression and polymorphism of proinflammatory genes on kidney transplant outcome
Hřibová, Petra ; Vyklický, Ladislav (advisor) ; Jirsa, Milan (referee) ; Stříž, Ilja (referee)
Acute rejection and chronic transplant nephropathy are the main complications after kidney transplantation. A broad spectrum of cytokines, chemokines and growth factors are involved in their etiology. The aim of this submitted dissertation was to find the influence of polymorphisms in selected genes and their intrarenal expression on kidney graft outcome. All studies were designed to follow a large cohort of individuals in order to be able to elucidate some unclear and inconsistent published results. 1. Cytokines and chemokines TGF-β1, TNF-α, IL-6, IL-10, MCP-1 and RANTES are up-regulated in different rates in acute rejection, chronic transplant nephropathy and also in other causes of kidney graft dysfunction. 2. High intrarenal expression of TGF-1 and MCP-1 mRNA in CAN predicts a higher risk of kidney graft dysfunction in the long-term. Also, the kidney graft survival is significantly shorter. 3. Intrarenal gene expression profile of TGF-1, TNF-, IL-6, IL-10, MCP-1 and RANTES is different during various causes of graft dysfunction. The intrarenal expression level cannot be used for diagnostic purposes, but it can alert higher immunological activity in kidney graft, which can lead to earlier failure of renal functions. 4. We did not confirm an association of TNF--308G/A, MCP-1 -2518 A/G, RANTES-...
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Th1/Th2 cytokine gene polymorphisms in patients with idiopathic pulmonary fibrosis
Vašáková, Martina ; Stříž, Ilja (advisor) ; Posová, Helena (referee) ; Šedivá, Anna (referee)
Objectives: Idiopathic pulmonary fibrosis (IPF) is a serious disease characterized with progressive scarring of the lungs in which the genetic background is supposed. The aim of our study was to investigate Th1/Th2 cytokine gene polymorphisms to evaluate their possible influence on IPF development. Then we have correlated selected polymorphisms of IL-1, IL-4 and IL-12 groups ( the selection was based on our previous results) with clinical parameters and high resolution computed tomography (HRCT) as a markers of disease stage and progression. Methods: We investigated 30 patients with IPF and 103 healthy volunteers for the cytokines polymorphisms of the IL-1 alpha, IL-1 beta, IL-1R, IL-1RA, IL-2, IL-4, IL-6, IL-10, IL-12, TNF alpha, IFN gamma, TGF beta, IL-1 beta, IL-2, IL-4 and IL-4RA genes. The PCR-SSP method was used for measurement. Then the correlations of vital capacity(VC) and diffusing capacity for carbon monoxide(DLCO), bronchoalveolar lavage (BAL) fluid cell counts and high resolution computed tomography (HRCT) alveolar and interstitial scores with different genotypes of groups of IL-1, IL-4, and IL-12 cytokines and their receptor antagonists. The HRCT results were evaluated by an experienced viewer using the interstitial and alveolar score scales, which were based on the IPF HRCT description system...
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The role of IL-17 in kidney transplantation
Menšíková, Markéta ; Krulová, Magdaléna (referee) ; Stříž, Ilja (advisor)
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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Immunomodulatory properties of mesenchymal stem cells - use in therapy
Pavlíková, Michaela ; Stříž, Ilja (referee) ; Krulová, Magdaléna (advisor)
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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Caspase inhibitors and regulation of the IL-1 family cytokines
Brabcová, Eva ; Stříž, Ilja (advisor) ; Holáň, Vladimír (referee)
Proinflammatory cytokines from IL-1 family play a key role in immune and immunopathological reactions and are involved also in initial phases of rejection mechanisms. One of their multiple functions is the induction of chemokines attracting immune cells into the site of injury. The aim of our study was to assess the effect of proinflammatory cytokines on the chemokine release and mRNA induction in epithelial cell lines of renal and lung origin. Furthermore, we tested the effect of caspase-1 inhibitors isolated from Streptomyces nodus (manumycin, asukamycin, MM273) on the release of IL-1 beta and IL-18 from human macrophages. These cytokines are cleaved into their active form by caspase-1 and inihibtion of the enzyme activity might be a perspective therapeutical approach to reduce inflammatory reaction. In our experimental model, we used cell line derived from renal adenocarcinoma (RA), A549 cell line (alveolar type II-like cells), and THP-1 monocyte/macrophage cell line. Chemokine mRNA induction was evaluated by an oligoarray and chemokine and cytokine levels were measured by Luminex or ELISA technology. Our data have showed that both epithelial cell lines produced constitutively mainly CXC chemokines attracting neutrophils (CXCL5/ENA-78, CXCL8/IL-8) and some of the CC chemokines characteristic for the...
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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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ANCA-Associated Vasculitides : Complex Diagnostic Approach
Hrušková, Zdenka ; Posová, Helena (advisor) ; Stříž, Ilja (referee) ; Žák, Aleš (referee)
Objectives: The aim of this study was to assess cellular immunity parameters in patients with ANCA (Anti-Neutrophil Cytoplasmic Autoantibodies)-associated vasculitides (AAV) at different stages of the disease, with different treatment modalities, and with respect to the long-term prognosis of the patients. Methods: We examined 69 patients with AAV, 30 healthy individuals and 20 patients with chronic kidney disease. Using flow cytometry, the following markers were assessed in peripheral blood cells: surface molecules (CD4, CD8, CD3, CD19, CD80, CD86, HLA-DR, CD28, CXCR3, CCR5, CD30 and CRTH2) and intracellular cytokines (interferon gamma (IFN), tumor necrosis factor alpha (TNF), interleukin (IL)-2 and IL-4 in CD3+ T cells and IL-10 and IL-12 in monocytes). Results: Patients with AAV had decreased total number of lymphocytes, CD4+ cells, and CD4+CD45RA+ cells compared to healthy controls (p<0.001). Active patients had increased CD30 and CRTH2 expression (p<0.05). Increased CCR5 expression persisted in remission. Increased HLA-DR expression, expansion of CD28 subpopulation and increased IFN production were noted in remission but not in active disease. Patients in remission who developed a relapse during follow-up had significantly lower IL-10 production than those without relapse (p<0.01). Conclusions: Taken...
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