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Lymnaeid snails: hemocyte activities and their changes caused by Trichobilharzia infections
Jindrová, Zuzana ; Horák, Petr (advisor) ; Bilej, Martin (referee)
Molluscs as well as all other invertebrates rely on innate immune response only. Their internal defense system is capable of destroying most pathogens. However, there are some exceptions, e.g. some snails serve as intermediate hosts for some trematodes. Trematodes are able to develop inside these snails due to intervention in the snail internal defense system. The submitted thesis describes hemocyte activities of two lymnaeid snails, Lymnaea stagnalis a Radix lagotis, and the influence of Trichobilharzia regenti infection on R. lagotis hemocytes. Hemocytes of both species exposed to various chemicals produced different amounts of H2O2 and NO. The response varied between both lymnaeid species. The amount of circulating hemocytes was elevated in R. lagotis snails due to T. regenti infection. However, the infenction attenuated hemocyte activities monitored by us. Hemocyte basal NO production was decreased as well as phagocytosis of bacteria, cell adherence and pseudopodia formation. Toxicity of L. stagnalis plasma against T. regenti miracidia was also described. Mechanisms used by trematodes to interact with the snail internal defense system will help us to understand why one species is suitable for the develepment of the trematode whereas another closely related species kills it. Powered by TCPDF (www.tcpdf.org)
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Interaction of Borrelia sp. with HL-60 cells and monocytes and cultivation of Anaplasma phagocytophilum in HL-60 cell culture
Marková, Lucie ; Hulínská, Dagmar (advisor) ; Beranová, Jana (referee)
Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum are causative agents of Lyme disease and human granulocytic anaplasmosis. Their common vector in Europe are the ticks from the genus Ixodes. In our work, we focused on interaction of innate immune cells with the causative agent of Lyme diseases, that are insubstitutable in their function in the early phase of the disease. Anaplasma phagocytophilum is hard to cultivate, the only possibility is to cultivate it in cell cultures. Successful cultivation of Anaplasma phagocytophilum acquired from patients in our geographic area is crucial for following experiments and for diagnostics too. In our experiments, we used validated cell cultures of HL-60 cells, canine monocytes DH82 and murine monocytes P388D1. During our studies of interaction of the causative agent of Lyme diseases with cells, we used two strains of different species Borrelia. Borrelia garinii M192 and Borrelia burgdorferi sensu stricto B31. These strains vary in virulence. The strain M192 is virulent, but the strain B31 lost its virulence by passages. We specialised in study of morphological changes using light microscopy (observation of dyed and fixed preparates and observation in dark field), eventually by transmision electron microscopy. During our experiments, we concluded that HL-60...
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Vliv klíštěcích slin na fagocytózu borelií dendritickými buňkami
MARŠÁLKOVÁ, Eliška
In this study we examined the effect of the tick saliva from I. ricinus and the effect of recombinant protein IRS-2 from the saliva of I. ricinus on dendritic cells derived from the mice bone marrow. We studied their effect on the production of cytokines by dendritic cells after the stimulation by B. burgdorferi, their effect on the expression of genes, that participate in phagocytosis, and the impact of the tick saliva on phagocytosis of B. burgdorferi by dendritic cells.
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Role makrofágů v obranném systému mléčné žlázy skotu
Hartmannová, Kristýna
This bachelor's thesis discusses natural immunity of the mammary glands and the macrophages acting within. The dissertation first describes the process, which macrophages evolve from haematopoietic cell through blood monocyt to mature macrophage and their activation. Briefly describes different types of macrophages and their paths to tissues. Next part of the bachelor's thesis discusses macrophages of the mammary gland themselves. First is described their structure, ultrastructure and death, then their own role in the mammary gland. Important roles of the macrophages of the mammary gland of cattle are phagocytosis, resolution of inflammation and the production of cytokines.
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Laboratory diagnosis of disorders of phagocytosis
POLÍVKOVÁ, Ivona
Phagocytosis is one of the oldest processes of absorbing particles like amoeba, which is one of the basic mechanisms of the immune system to defend the body against infections. Failure of these processes is clinically manifested like immune deficiency and it can cause a very serious complication which could leads to the death of the patient. Phagocytosis is performed by specialized cells (macrophages, dendritic cells, neutrophils and other phagocytic myeloid cells) which are able to absorb the target particles, especially microorganisms, dead cells and foreign objects. These processes are essential for stability of homeostasis. Absorption of micro-organisms leads to activation of adaptive immunity response, elimination of apoptotic and destroyed cells and starts the reparation processes of damaged tissue. Phagocytosis as a complex process can be divided into several phases: active movement of phagocytes to the inflammation zone, adherence, ingestion and intracellular degradation which leads to the killing of pathogens. There are two mechanisms of killing pathogens. First is independent on oxygen where antimicrobial substances are stored in azurophilic granules which could be released into phagolysosom. Second mechanism is oxygen-dependent called oxidation (respiratory) flare which leads to the formation of biologically active mediators where oxygen molecules have considerable oxidation potential. Defects in phagocytic system are mainly caused by low number or malfunction of neutrophils which leads to severe infections mainly caused by staphylococci, Enterobacteriaceae or fungi. Among malfunctioning of phagocytosis processes belongs LAD I and LAD II syndrome. The other possible disorder of phagocytic malfunction is defect in enzymes. The NADPH oxidase is necessary for bacterial lyses mechanism of phagocytes and lack of these enzyme caused serious inherit disorder called chronic granulomatous disease. Lack or completely missing of enzyme myeloperoxidase stored in the azurophilic granules of neutrophils and monocytes is more and more common autosomal recessive disorder of phagocytosis. In this diploma work I focused on detection of these rare and serious disorders defects using flow cytometry to detect respiratory burst activity of neutrophils in blood samples. This laboratory procedure is called a burst test. In this test we quantitatively evaluate the respiratory burst activity of granulocytes in heparinized blood samples using the flow cytometery. Principle of this method is an oxidation-reduction reaction of dihydrorhodamine 123 to green fluorescent rhodamine 123 using peroxide, hydroxyl radicals and superoxide anions which are activated by respiratory burst. This process is one of the significant characteristic of phagocytic cells which are characterized by multi-stage activation of NADPH oxidase. This oxidase catalyzes electron reduction of molecular oxygen to superoxide. This step plays a very important role in our immune system and allows to kill and degrade particles and bacteria in phagocyte cells. Since 2009 - 2012 I analyzed the results of 611 patients who were tested for respiratory burst of neutrophils on a request of their physician. Among all of these results we obtain only two positive results of reduction of stimulation index (SI) and significant decrease percentage of activated granulocytes. Both of these parameters: reduced percentage of (activated granulocytes stimulated by PMA and E. coli) and low stimulation index pointing to a potentially serious disorder of phagocytosis mechanism. In specific tests of these cases proved the enzyme defect in phagocytosis mechanism which is the most common disorder in this type of immunodeficiency. These results indicate that the major primary defects of phagocytosis are very rare and their detection is usually in childhood for suspicion of primary immunodeficiency. This test is very helpful for discovering a prime immunodeficiency.
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Examination of changes in fagocytosis for the screening of hemotoxicity on a biomodel
JURČOVÁ, Martina
This thesis dealt with the study of laboratory methods used in the examination of phagocytosis and we conducted the research of the examination of the changes of phagocytosis on the biomodel S. littoralis. The results showed the assumed increase in phagocytosis after the administration of the model substance. Our tests applied documented quite sensitively the changes in fagocytic acitivity and production of cytotoxic substances and the model animal was evaluated as a suitable biomodel for the other studies of hemotoxicity.
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