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Markers of transplantation tolerance in kidney transplantation
Krepsová, Eva ; Viklický, Ondřej (advisor) ; Krejčí, Karel (referee) ; Živný, Jan (referee)
Long-term renal graft acceptance still requires long-term immunosuppressive therapy, which is accompanied by many adverse effects. Contrarily insufficient immunosuppression could lead to graft rejection and its failure. Therefore, research continues for biomarkers that reflect a patient's immunological status and thus allowing for individualized immunosuppressive therapy. In our study we showed lower incidence of acute rejection in kidney transplant recipients treated with rabbit anti-thymocyte globulin (rATG) or basiliximab induction within the first three months after transplantation. The rATG induction caused profound decrease of recipient's peripheral blood T and NK cells, as well as transcripts that are exclusively expressed by these cell types together with expansion of regulatory T cells (Tregs) among CD4+ T cells. In rATG group the increase of two transcripts associated with rejection (MAN1A1 and TLR5) was also observed in early post-transplant period. After the basiliximab induction we transiently detected CD4+CD25low/-FoxP3+ cell population along with disappearance of CD4+CD25+FoxP3+ Tregs. Basiliximab induction resulted in a transient increase in CD4+FoxP3+ Tregs, accompanied by the highest peripheral expression levels of markers associated with operational tolerance (FOXP3 and TCAIM)....
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Dental materials and imunne reaction
Chrástová, Iveta ; Bártová, Jiřina (advisor) ; Prokešová, Ludmila (referee)
Adverse reactions to metals is relatively frequent in sensitive individuals, and may lead to the development of various diseases, such as lichen planus and lichenoid reaction and was also described effect of amalgam to the pathology of autoimmune diseases. In sensitive patients with known reactions to metals leads to atrophy of the oral mucosa and discomfort (burning, cutting, excessive salivation). Adverse effects of dental materials are described as an allergic reaction type IV delayed hypersensitivity. It disrupt the immune system, increased vascular permeability, tissue is often damaged by chronic inflammation and it is also activated antigen- nonspecific component of immunity. Monocytes are the key cells of the immune system, which act as antigen presenting cells, in particular after conversion in the tissues into macrophages are directly involved in the activation of immune responses and their stimulation is therefore crucial for the development of adverse reactions to dental materials. In the thesis was measured cytokine production by monocytes after stimulation with metal salts. To determine the extent of the patients' responses to the metals was used lymphocyte proliferation assay (MELISA® ), which was developed and patented in Sweden. We compared lymphocyte proliferation after stimulation...
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Role of peripheral blood monocytes and innate immunity in diabetes
Zinková, Alžběta ; Daňková, Pavlína (advisor) ; Novota, Peter (referee)
Introduction: Diabetes mellitus is a polygenic disease and its development is influenced to some extent by environmental factors as well. Innate immunity triggers nonspecifically first defense reactions after penetration of the pathogen into the body, while overstimulation components of innate immunity may give rise to autoimmune diseases, including diabetes type 1. The components of innate immunity are, among others, Toll-like receptors (TLRs) belonging to a group of the structures recognizing preserved molecular structures characteristic of pathogens. Toll-like receptors are abundantly expressed by monocytes which produce prolactin (PRL) having an immunostimulatory function. To clarify the role of innate immunity in the pathogenesis of diabetes, we focused on the expression of mRNA and protein expression of TLR2 and TLR4. The expression of PRL was studied only at the level of mRNA. Monocytes were separated by flow cytometry into classical (CD14++) and nonclassical (CD14+). We monitored their percentages and the degree of expression of CD14 antigen on their surface.The operational objective of this dissertation was to optimize the stimulation of monocytes for the planned study of the function of non-pituitary prolactin in vitro and determine the appropriateness of the use of healthy donors' buffy...
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Subpopulations of human monocytes and macrophages.
Švachová, Veronika ; Stříž, Ilja (advisor) ; Krulová, Magdaléna (referee)
Monocytes and macrophages are important components of the innate immune response. These mononuclear phagocytes form a heterogeneous cell population, of which phenotype and functions can be modified under the influence of different signals coming from the surrounding microenvironment. The aim of this work was to modulate the phenotype of these cells by a variety of stimulants and to compare the changes induced on the model of THP-1 monocytic cell line and on the human peripheral blood monocytes. Surface marker expression was analyzed by flow cytometry. Further on, IL-8 production was evaluated by Luminex assay and the concentration of soluble calprotectin was assessed by ELISA. The most significant changes in surface marker expression were induced by exposure to IFNγ. This cytokine increased the expression of CD54, CD14 and HLA-DR on the surface of THP-1 cell line. Higher concentrations of IFNγ promoted higher apoptotic rate and augmented calprotectin expression and production in THP-1 cell line. On the surface of monocytes, IFNγ stimulation resulted only in the upregulation of CD54 expression. IL-4 increased the expression of CD36 by THP-1 cell line and inhibited the expression of CD163 by human monocytes. LPS stimulation caused the suppression of HLA-DR activation in monocytes and enhanced IL-8...
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The innate immunity and circulating monocytes - their significance and function in pathogenesis of coeliac disease.
Němečková, Iva ; Daňková, Pavlína (advisor) ; Palová Jelínková, Lenka (referee)
8 Abstract Introduction: Celiac disease is indentified as the loss of oral tolerance to gluten, it is an organ-specific autoimmune disease in which both, adaptive and innate immunity participate. Monocytes are important part of immune system; they have many functions and express very diverse membrane receptors including Toll-like receptors (TLRs). TLRs are involved in the innate immune response, specifically TLR2 and TLR4 are crucial for recognition of bacterial components and TLR7 recognizes virus's ssRNA. Monocytes also produce prolaktin (PRL), which acts as a cytokine that modulates immune responses. To clarify the role of innate immunity and circulating monocytes in pathogenesis of celiac disease, we focused on changes in expression of selected Toll-like receptors (TLR2, TLR4, TLR7), prolactin, some pro- a anti-inflammatory cytokines (TNF-α, IL-6, IL-12, IL-10). We monitored the influence of the SNP - 1149 G/T on the expression of PRL mRNA. Another objective of this work was the introduction and optimization of in vitro methods for cultivation and stimulation of peripheral monocytes. Material and Methods: This pilot study includes 21 patients with celiac disease and 40 healthy controls. For determination of mRNA levels of the studied genes we isolated RNA from monocytes that were isolated by...
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Zavedení a optimalizace \kur{in vivo} modelů zánětu a jejich využití pro funkční analýzu inhibitorů proteáz z klíštěcích slin
CHLASTÁKOVÁ, Adéla
Two murine models of acute inflammation, namely thioglycollate-induced peritonitis and carrageenan-induced paw edema, were optimized using non-steroidal anti-inflammatory drug indomethacin and corticosteroid dexamethasone. During the optimization phase, the presence of neutrophils, monocytes, macrophages, eosinophils, B cells and T cells in the peritoneal cavity at various time points after injection of thioglycollate medium was assessed via multicolor flow cytometry. Moreover, two different thioglycollate media (suppliers BD and Sigma-Aldrich) were compared for their ability to induce an inflammatory response. The optimization of thioglycollate-induced peritonitis and carrageenan-induced paw edema was followed by the evaluation of the anti-inflammatory activity of Ixodes ricinus cystatins G1 and G9 in both mouse models.
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Účast monocytů a dendritických buněk v patogenezi celiakie
Tučková, Ludmila ; Palová-Jelínková, Lenka ; Rožková, D. ; Černá, M. ; Pecharová, Barbara ; Smythies, L. ; Smith, P. ; Dvořák, M. ; Fruhauf, P. ; Tlaskalová, Helena
This work studies the involvement of monocytes and dendritic cells in coelias cisease pathogenesis
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Stimulace P3MoC izolovaných z krve celiakálních pacientů gliadinovými fragmenty
Cinová, Jana ; Palová-Jelínková, Lenka ; Smythies, L. ; Černá, M. ; Pecharová, Barbara ; Dvořák, M. ; Fruhauf, P. ; Tlaskalová, Helena ; Smith, P. ; Tučková, Ludmila
We documented that gliadin digest stimulate PBMoC (isolated from celiac patients and healthydonors) to IL-8 and TNF alfa production through the NF-kappa B pathway. Monocytes expressed increased levels of dendritic cell markers
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Gliadin fragments induce phenotypic and functional maturation of human dendritic cells
Palová-Jelínková, Lenka ; Rožková, D. ; Pecharová, Barbara ; Bártová, J. ; Šedivá, A. ; Tlaskalová, Helena ; Spíšek, R. ; Tučková, Ludmila
Gliadin treatment also resulted in increased NF- B/DNA binding activity of p50 and p65 subunits. Taken together, gliadin peptides can contribute to overcoming the stage of unresponsiveness of immature DC by inducing phenotypic and functional DC maturation, resulting in more efficient processing and presentation of gliadin peptides to specific T lymphocytes
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The response of peripheral blood monocytes from celiac patients to gliadin peptides
Cinová, Jana ; Palová-Jelínková, Lenka ; Smythies, L. ; Černá, M. ; Pecharová, Barbara ; Dvořák, M. ; Fruhauf, P. ; Tlaskalová, Helena ; Smith, P. ; Tučková, Ludmila
We documented that gliadin digest stimulate PBMoC (isolated from celiac patients and healthydonors) to IL-8 and TNF alfa production through the NF-kappa B pathway. Monocytes expressed increased levels of dendritic cell markers
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