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Influence of electrical pulses on human blood phagocytes
Chorvátová, Michaela ; Číž,, Milan (referee) ; Kubala,, Lukáš (advisor)
The phagocytic cells circulating in the bloodstream play a key role in both the defense of the body and the pathology of inflammatory diseases. Thus, targeting their functions has potential to modulate an immune response, especially during the inflammatory phase. This master's thesis was focused on the influence of electric pulses on the most abundant phagocyte population in human peripheral blood, namely neutrophils. The theoretical part describes the role of neutrophils in the development of the immune response and the effects of the electric field on various cells. Consequent part of the thesis was the optimization of the electrical stimulation of neutrophils using a unique platform with a network of gold electrodes. In stimulated cells by electrical pulses, activation of selected signaling pathways, degranulation, ROS production, citrullination of histone H3 and expression of surface markers were monitored. Overall, electrical stimulation was observed to induce neutrophil activation but only electrical pulses of size 1 V were found to be statistically significant in the case of ROS production and 10 mV and 100 mV electrical pulses in the case of metalloproteinase MMP8 degranulation. The absence of significant effects in the most observed parameters was probably due to unwanted activation of neutrophils in control samples.
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Early phase of anti-Leishmania immunity in the host skin
Máčalíková, Bára ; Leštinová, Tereza (advisor) ; Kolářová, Iva (referee)
Leishmania parasites are parasitic protozoans that cause disease called leishmaniasis, which primarily affects mammals. Throughout evolution, Leishmania has adapted to the host's immune system, using it to its advantage. This bachelor's thesis describes the relationship between Leishmania and early immune components in the host's skin, as well as the parasite's ability to inhibit the microbicidal activities of cells. The infection begins with the inoculation of infectious promastigotes into the skin, and before reaching their target cells, Leishmania primarily interacts primarily with the complement system, keratinocytes, fibroblats, eosinophils, neutrophils, mast cells and dendritic cells. Understanding the mutual interaction between the host and the parasite is essential for vaccine development and the treatment of leishmaniasis. KEYWORDS: leishmania, skin, early imunity, complement system, keratinocytes, fibroblasts, eosinophils, neutrophils, mast cells, dendritic cells
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Proportional and functional characteristics of particular neutrophil subpopulations in umbilical cord and peripheral blood
Miková, Eliška ; Hrdý, Jiří (advisor) ; Krulová, Magdaléna (referee)
Early postnatal period is characterised by generally immature phenotype of the newborn's immune system. The maturation of the immune system including setting appropriate regulatory responses is occurring during this period and encountering pioneering bacteria colonizing neonate plays an important role. In the early days after birth, the immune system of a newborn is very limited, and the adaptive part is mostly represented by antibodies transferred from the mother by cord blood (CB) in the womb and then by colostrum and mother's milk after labour. Therefore, innate immunity plays a key role in defence (against pathogens) in neonates and is largely represented by neutrophils. This study aims to better understand neutrophil biology and phenotype in umbilical CB, compared to neutrophils from peripheral blood (PB) of mothers and healthy non pregnant women (referred to as HC). The assessment of neutrophil phenotype based on surface markers was performed using flow cytometry. Expression of genes linked to antimicrobial function was measured using quantitative PCR. Functional properties of neutrophils, metabolic activity during activation and phagocytosis, and suppressive properties were assessed using the SeaHorse machine and flow cytometry, respectively. Here we confirm the presence of immature CD16low...
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Immune system dysregulation in type 1 diabetes
Paračková, Zuzana ; Šedivá, Anna (advisor) ; Filipp, Dominik (referee) ; Vlková, Marcela (referee)
Type 1 diabetes (T1D) is an autoimmune disease with multifactorial aetiology that involves an attack of self-reactive cytotoxic CD8 lymphocytes on insulin-producing beta cells in the pancreas. In the T1D pathophysiology, both innate and adaptive immunity mechanisms cooperate in the development of inflammation leading to autoimmune destruction. Autoreactive T lymphocytes are the canonical destructors of the beta cells, and B cells produce autoantibodies; the innate immunity cells are considered the initiators of the pathological autoimmune reaction by promoting T and B cell activation. Here, we provide evidence of both innate and adaptive immunity cell types dysregulation in patients with T1D, and that these changes occur before the onset of the disease. The changes in T regulatory lymphocytes (Tregs) and B cell subpopulations occur already in asymptomatic T1D first-degree relatives. During the first year after the onset of the disease, there is a gradual decrease in the neutrophil numbers in the periphery, which probably infiltrate the pancreas. We have focused more closely on the innate immunity dysregulation and its contribution to T1D pathogenesis. Initially, we describe that neutrophil products called neutrophil extracellular traps (NETs) are able to induce IFNγ-producing T cells through...
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Influence of electrical pulses on human blood phagocytes
Chorvátová, Michaela ; Číž,, Milan (referee) ; Kubala,, Lukáš (advisor)
The phagocytic cells circulating in the bloodstream play a key role in both the defense of the body and the pathology of inflammatory diseases. Thus, targeting their functions has potential to modulate an immune response, especially during the inflammatory phase. This master's thesis was focused on the influence of electric pulses on the most abundant phagocyte population in human peripheral blood, namely neutrophils. The theoretical part describes the role of neutrophils in the development of the immune response and the effects of the electric field on various cells. Consequent part of the thesis was the optimization of the electrical stimulation of neutrophils using a unique platform with a network of gold electrodes. In stimulated cells by electrical pulses, activation of selected signaling pathways, degranulation, ROS production, citrullination of histone H3 and expression of surface markers were monitored. Overall, electrical stimulation was observed to induce neutrophil activation but only electrical pulses of size 1 V were found to be statistically significant in the case of ROS production and 10 mV and 100 mV electrical pulses in the case of metalloproteinase MMP8 degranulation. The absence of significant effects in the most observed parameters was probably due to unwanted activation of neutrophils in control samples.
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The role of NETosis in pathogenesis of autoimmune diseases
Pošmurná, Eliška ; Brynychová, Iva (advisor) ; Šolc, Roman (referee)
Neutrophil granulocytes - white blood cells are very effective in fight against pathogens with fagocytosis, degranulation, or NETosis, which is special form of cell death. During NETosis are created neutrophil extracellular traps, which can trap and destroy microorganisms with antimicrobial granules. NETosis can be iniciated by activation of pro-inflammatory cytokines, lypopolysacharides or phorbomystatic acid. Activated neutrophils get to the site of inflammation, where decondensed chromatin, part of NETs, gets into extracellular space. DNA in extracellular space is strong alarmin for immune system as part of NETs. Increased NETs production or problem with their elimination can play role in patogenesis of some autoimmune diseases (eg. Systematic lupus erythematodes, associated vasculitis, rheumatoid arthritis, diabetes mellitus). NETs production or their components can affect diseases or degree of damage to the surrounding tissue. It also correlates with seriousness of disease (eg. Associoated vasculitis) and can have diagnostical potencial (eg. Asociated vaskuloitides, rheumatoid arthritis or diabetes mellitus type 1). Keywords: NETosis, NETs, neutrophils, autoimmune diseases, immune system
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Vliv klíštěcích slin na myší neutrofilní granulocyty aktivované virem klíšťové encefalitidy
VESELÁ, Dominika
Tick saliva contain molecules that suppress the immune response of host organism thereby faciliting the transmission of pathogens by tick vectors. Neutrophils are the most common type of white blood cells. They participate in the acute phase of inflammation and via excretion of cytokines they mobilize other components of the immune system. This bachelor´s thesis focused on demonstrating of tick-borne encephalitis virus replication in neutrophils and on detection of cytokine production by neutrophilic granulocytes upon activation by the tick-borne encephalitis virus and the possible impact of tick saliva of tick Ixodes ricinus on the parameters mentioned above.
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Studium nádorové imunoterapie založené na instalaci ligandů fagocytárních receptorů na nádorové buňky a objasnění probíhajících procesorů
CAISOVÁ, Veronika
Immunotherapy became a very promising approach for cancer therapy. Tumor cells are eliminated using the body's own immune system with minimal negative effect on healthy tissue. This thesis is focused on immunotherapy based on activation of innate immunity, specifically on intratumoral application of ligands stimulating phagocytosis and Toll-like receptor ligands. This therapeutic approach was tested in several types of tumor mouse models, such as melanoma B16-F10, pancreatic adenocarcinoma, and pheochromocytoma. The composition of the therapeutic mixture as well as the application schedule were optimized in our studies. Subsequently the underlying mechanisms involved in tumor elimination during this therapy were investigated.
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Úloha vrozené a získané imunity v imunoterapii melanomu a pankreatického adenokarcinomu
NEDBALOVÁ, Pavla
This study examines the role of innate and adaptive immunity in the immunotherapy based on the combination of the ligands stimulating phagocytosis anchored in the tumour cells membrane and the mixture of TLR agonists. This immunotherapy is primarily focused on the innate immunity activation and induces strong inflammatory infiltration, which neutrophils and NK cells are part of. Therefore, the next aim of this study was to evaluate the anti-tumour activity of neutrophils and NK cells. For examination malignant melanoma and pancreatic adenocarcinoma mouse tumour models were used.
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