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Fenotypová charakteristika monocytov a makrofagov prasiat vykrmovaných rôznymi dietámi
Bátik, Andrej
Many research articles describes fish oil as a beneficial for health. Mainly for prevention of cardiovascular diseases or that fish oil reduces cholesterol in blood stream. Aim of this study is to investigate effects of fish oil specifically of his parts EPA+DHA, on a modulation of immune system and phenotypic differentiation. For the experiment were chosen monocytes and macrophages obtained from experimental group of animals which was fed a diet consist of the addition of fish oil in an amount 45mg/kg of live weight. Then inflammatory state was induced by LPS. Cells surface receptors were analyze through flow cytometry specifically CD14, CD163, SLA-DR and theirs combinations after they were statistically processed. Results shows proinflammatory effects of fish oil however they were statistically not significant.
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Vliv probiotik na imunologické vlastnosti buněk obranného systému
Šustrová, Tereza
This disertation thesis aims to determine the effect of probiotic on the cells of immune system – human, procine and rat neutrophils and macrophages. Immune cells were exposed to 3 probiotics – Bifidobacterium bifidum, Lactobacillus rhamnosus and Enterococcus faecium, which are normally included in GIT. The first aim of this study was choose the most appropriate method for the isolation of human and porcine neutrophils and PBMC from peripheral blood. Other aims were to find out whether interactions of probiotics with neutrophils and macrophages affect their lifetime and lead to their structural changes. And whether intraperitoneal administration of probiotics lead to pro-inflammatory polarizoation of macrophages and affect viability of intraperitoneal immune cells. Structural changes of cells were detected by light and transmission electron microscopy. For detection of early and late apoptosis was used flow cytometry and for detection od cytolysis was used LDH aktivity. To determine of effect of probiotics on polarization of rat macrophages, qRT PCR was used. The results of this thesis suggest that all tested probiotics led to a significant increase in early apoptosis in porcine neutrophils and late apoptosis and cytolysis (LDH) in human neutrophils in vitro.
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Interakce makrofágů a buněk rakovinné prostatické linie PC-3
Mazalová, Lenka
This dissertation on the theme Interaction of macrophages and the prostate cancer cell line PC-3 was focused on the initial mapping of interactions between human mac-rophages and the prostate cancer cell line PC-3. Microscopic and molecular-genetic methods have been used for analysis. The structure and ultrastructure of macrophages and PC-3 cells have been de-scribed. The description of cell morphology in interaction with each other has been de-voted to processes associated with adherence of macrophages on PC-3, eferocytosis and phagocytosis. Cultivation of PC-3 with plumbagine showed an increase of apoptotic cells. Cultivation of macrophages with PC-3 supernatant has shown that solubiling fac-tors in supernatant may have an effect on inducing cell death in macrophages. In our study, the supernatant did not affect the production of TNF alfa, IL-12, IL-10 or IL-6 by macrophages. A time lapse video was created to show physical interactions of macro-phages and invasive cell. All obtained results show that there are interactions between the cancer cells and immune system cells that have not yet been discovered.
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Přítomnost nízkomolekulárních imunomodulátorů ve slinách klíštěte \kur{Ixodes ricinus}
HAUSEROVÁ, Simona
Ticks use proteinaceous molecules contained in their saliva to suppress immune response of the host to complete their succesful feeding. In some ticks (e.g. Rhipicephalus sanguineus) presence of non-proteinaceous molecules was discovered. The aim of this work was to determine the amount of these molecules in the saliva of Ixodes ricinus tick. Namely, prostaglandin E2 and adenosine were analyzed. The second aim of this work was to evaluate the role of both low molecular weight components (or at least one of them) in the suppression of TNF-alfa cytokine production in suitable cells by ticks saliva.
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Exprese CD47 a jeho topologie na povrchu primárních buněk karcinomu močového měchýře při interakci s makrofágy
Rajtmajerová, Marie ; Drbal, Karel (advisor) ; Brdička, Tomáš (referee)
CD47 is a so-called "don't eat me" signal, which protects cells from phagocytosis. Its high expresion on tumor cells brings new perspective to the tumor therapy. Monoclonal antibodies, which are these days undergoing clinical trials, prevent CD47 binding to the SIRPA inhibitory receptor on macrophages, and so they enhance their phagocytic functional capacity. In this way they enable phagocytic removal of tumor cells. Overall expression, structural conformation and stoichiometry of CD47 on a particular cell predestine whether it will be phagocytised. The aim of the thesis is to develop and test methods to characterise expression parameters of CD47 via flow cytometry (FCM), quantitative PCR (qPCR) and microscopy. To achieve this goal I performed competition tests of commercially available antibodies in order to characterise their binding epitopes on cell lines. After performing tSNE analysis of primary BCa patient samples I correlated CD47 expression with other cell surface markers. I focused on CD47 expression in various differentiation stages of the tumor. To better understand the relationship between CD47 expression and differentiation status of cells I performed qPCR analysis of particular transcription factors. Using cell lines I examined method for phagocytosis quantification, which will be...
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The role of nitric oxide in helminthoses
Šmídová, Barbora ; Macháček, Tomáš (advisor) ; Leštinová, Tereza (referee)
Nitric oxide (NO) is a small gaseous molecule produced mainly by activated macrophages, but some other cells as well, in reaction to an infection. Since late 80s, when the effect of NO on schistosomes was demonstrated, many experiments have been conducted examining both the effect of NO on other helminths and its role in helminthoses. In vitro experiments center mainly upon direct effects of NO on helminths - loss of motility and viability in particular. On the other hand, in vivo studies focus on the course of infection and pathological changes in hosts. However, in vitro and in vivo data do not always corroborate. Moreover, the effects of NO differ not only across helminth genera but also for developmental stages of the same parasite.
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Immune system, nitric oxide and parasites
Oreský, Matej ; Hrdý, Ivan (advisor) ; Macháček, Tomáš (referee)
Nitric oxide belongs to smallest known molecules. It has been known since the mid- 17th century, but only in the second half of the 20th century its significance in medicine began to be discovered. Already in the 19th century, nitroglycerin was used to treat heart disease, and the connection with nitric oxide was later found. Nitric oxide works, for example, as a signaling molecule in biosystems, acts as a vasodilator in endothelial tissue but also has a number of pathophysiological effects. Because of its radical nature, it plays an important role in the immune system in the non-specific response. By reaction with certain radicals in the body, strong reactive compounds such as peroxynitrite can form. However, some intracellular parasites have been found to be able to resist the effects of the immune system associated with the NO molecule. They could, besides other things, serve proteins from the family of flavodiiron proteins to this purpose. At the end this review focuses on the group of flavodiiron proteins and the interaction of three parasites with the immune system of the host: Entamoeba, Giardia and Trichomonas. Key words: Nitric oxide, NO synthase, macrophage, Trichomonas vaginalis, Giardia intestinalis, Entamoeba histolytica, flavodiiron protein
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Programovaná buněčná smrt makrofágů v mléčné žláze u skotu
Skřipská, Iva
This bachelor is thesis provides a general overview of the anatomy and physiology of bovine mammary gland and its defence mechanisms with the focus on macrophages, their characteristic features, activation and functions. The second part describes apoptosis of macrophages at the end of their life cycle, their transformation from monocytes and their influence on the surrounding mammary gland tissue. The thesis also summarises methods of detection of apoptotic macrophages and depicts the advantages and disadvantages of each method.
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Surface glycoconjugates of Leishmania parasites and their interactions with immune system of vertebrate host
Pacáková, Lenka ; Leštinová, Tereza (advisor) ; Kolářová, Iva (referee)
musí po vstupu do obratlovčího hostitele čelit obranným mechanismům hostitelské imunity a proniknout do cílové buňky - pokračuje. Evoluční strategií leishmanií vyvinutou k přelstění imunitního systému Mezi nejvýznamnější povrchové glykokonjugáty patří membránově vázané proteofosfoglykany a metaloproteáz lním účinkům produktů neutrofilů a zprostředk vazbu na makrofágy. Intracelulárně pak modulují signalizační dráhy, které vedou k produkci cytokinů, směřujících polarizaci imunitní odpovědi ve prospěch Th2. Výsledkem tohoto přesměrování je vyhnutí se účinkům toxického NO, čímž je ustanovena chronicita infekce. Glykokonjugáty jsou zkoumány jako účinná složka chránících obratlovce před nákazou či bránících zpětnému přenosu čímž dalšímu šíření infekce. Klíčová slova: leishmanie, lipofosfoglykan, glykoinositolfosfolipid, proteofosfoglykan, gp63, imunitní odpověď, makrofág
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Signaling effects of adenylate cyclase toxin action on phagocytes
Černý, Ondřej ; Šebo, Peter (advisor) ; Černý, Jan (referee) ; Dráber, Petr (referee)
The adenylate cyclase toxin (CyaA) plays a key role in the virulence of Bordetella pertussis. CyaA penetrates CR3-expressing phagocytes and catalyzes the uncontrolled conversion of cytosolic ATP to the key second messenger molecule cAMP. This paralyzes the capacity of neutrophils and macrophages to kill bacteria by oxidative burst and opsonophagocytic mechanisms. Here we show that CyaA suppresses the production of bactericidal reactive oxygen and nitrogen species in neutrophils and macrophages, respectively. The inhibition of reactive oxygen species (ROS) production is most-likely achieved by the combined PKA-dependent inhibition of PLC and Epac-dependent dysregulation of NADPH oxidase assembly. Activation of PKA or Epac interfered with fMLP-induced ROS production and the inhibition of PKA partially reversed the CyaA-mediated inhibition of ROS production. CyaA/cAMP signaling then inhibited DAG formation, while the PIP3 formation was not influenced. These results suggest that cAMP produced by CyaA influences the composition of target membranes. We further show here that cAMP signaling through the PKA pathway activates the tyrosine phosphatase SHP-1 and suppresses the production of reactive nitrogen species (RNS) in macrophages. Selective activation of PKA interfered with LPS- induced iNOS expression...
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