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The skin immune response of mice infected with avian schistosomes
Revalová, Alena ; Macháček, Tomáš (advisor) ; Kolářová, Libuše (referee)
Invasion of the mammalian skin by cercariae of avian schistosomes of the genus Trichobilharzia is associated with the skin pathology called cercarial dermatitis (CD). Although the manifestation of CD, characterized by the presence of maculo-papular rash and pruritus, is believed to be linked with the previous sensitization, the role of the infection dose and the specific causative agent is rarely considered. Therefore the skin immune response in mouse pinnae infected by 100, 1000 or 4x100 (repeated infections) cercariae of two Trichobilharzia species - T. regenti and T. szidati - was examined in this work. To compare the effect of these factors (the infection dose and species), a complex approach was ensured by employing both in vivo and in vitro methods. Firstly, histopathological changes in the pinnae were examined by classical histology and infiltrated leukocytes were characterized by flow cytometry. Sera of mice were used to detect the systemic production of cytokines and parasite-specific antibodies. In vitro cultivation of mouse pinnae was used to detect the local production of cytokines. The most remarkable observation was that the infection dose largely impacted the course of the immune response, both locally and systematically. In primary infected mouse pinnae, 100 cercariae of both...
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Taxonomy of bird schistosomes not only in planorbid snails in Europe
Holická, Monika ; Kolářová, Libuše (referee)
Cercarial dermatitis caused by bird schitosomes is in focus not only by scientists, but also by the general public. The disease has medical and economic impact, and it is responsible for financial losses in many resorts all around the world. Only some causative agents of the disease are known, because of parasite cryptic life style and biology (e.g. size and location of adults in the definitive host; gonochorism) and inaccurate descriptions in the past. With the onset of modern molecular methods taxonomic revisions of old species of bird schistosomes and descriptions of new ones are of top interest. In Europe pulmonate snails are used by bird schistosomes as intermediate hosts, with some exceptions. Descriptions of new species and genera were recently focused on findings from lymnaeid molluscs, in which the genus Trichobilharzia (the most significant causative agent of cercarial dermatitis) is often developing. Therefore, less attention was paid to the snails of the family Planorbidae. Planorbid snails are most commonly infected by the species of Bilharziella polonica, its typical host in Europe is Planorbarius corneus, less common hosts are Planorbis planorbis, Bathyomphalus contortus or Anisus vortex. B. polonica is spread all over the world. There are also known findings from the Czech Republic....
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Organelle proteomics of parasitic protists
Jedelský, Petr ; Tachezy, Jan (advisor) ; Kolářová, Libuše (referee) ; Půta, František (referee)
Advances in DNA sequencing led to a technological breakthrough, that allowed analyzis of complete genomes including those of parasitic protists Trichomonas vaginalis and Giardia intestinalis . These organisms are studied not only for their clinical importance, but also from the evolutionary point of view for their adaptation to anaerobic environment. Genome sequencing and annotations of predicted proteins alone did not bring detail view into functioning of their mitochondrion related organelles in G. intestinalis mitosomes, notparticipating in energetic metabolism, in T. vaginalis hydrogenosomes, producing molecular hydrogen and ATP by means of substrate phosphorylation. Traditional methods based on a fractionation by ultracentrifuging in density gradient and subsequent biochemical and enzymological analyzes were extended by one and twodimensional electrophoresis with subsequent identification of proteins by mass spectrometry. Methods of multidimensional separation of peptides produced by specific proteolysis of a complex mixture...
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Study on neuropathophysiological changes in mammalian host caused by bird schist osome infection
Lichtenbergová, Lucie ; Kolářová, Libuše (advisor) ; Posová, Helena (referee) ; Ruppel, Andreas (referee)
Trichobilharzia regenti belongs to a small group of parasitic helminthes localized in nervous tissue of their hosts. Like in other bird schistosomes, repeated penetration of larvae (cercariae) into the skin lead to development of skin allergic reaction in humans (cercarial dermatitis). Soon after the penetration in the skin, cercariae transform to schistosomula which are able to resist the host response and, in case of T. regenti, migrate to central nervous system (CNS). During CNS involvement, the infections of both specific avian and non-specific mammalian hosts can result in neuromotor disorders and even death of the host. The present PhD thesis deals with T. regenti infections of non-specific mammalian host. The first part of the experimental work was focused on antibody reactivity and antigen specificity of sera from mice experimentally infected with T. regenti. Development of antigen-specific IgM and IgG1 antibodies and elevated levels of total serum IgE indicated a Th2 immune response. Cercarial antigens stimulated IL-4 release from basophiles obtained from healthy volunteers. Western blot analysis revealed that IgG and IgE antibodies in the sera specifically recognized antigen of 34 kDa in both homogenate of cercariae and cercarial excretory/secretory products. This molecule seems to represent major...
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Influence of tissue helminths on the development of cancers in model organisms
Schreiber, Manfred ; Horák, Petr (advisor) ; Kolářová, Libuše (referee)
Mesocestoides corti and Taenia crassiceps are tapeworms, larvae of which are characterized by their ability to reproduce asexually. In this work, the effect of infection by M. corti and T. crassiceps in BALB/c, C57BL/6J and ICR mice on the growth and metastasis of B16F10 melanoma tumors was investigated. Although an increase in metastatic activities was observed after intravenous administration of melanoma cells to M. corti-infected mice, both tapeworms showed a strong suppressive effect on the size and number of tumors and metastases formed when the cells were administered intraperitoneally. This, in some cases, led to a complete elimination of tumor cells. In vitro cultivation of B16F10 cells in the presence of larval excretory-secretory products led to a decrease in their viability but an increase in their migration ability. Flow cytometry proved that M. corti infection has an effect on the increased number and proportion of macrophage populations in the peritoneum of ICR mice. Our work confirmed the anti-tumor effect of T. crassiceps infection in mice and introduced M. corti as a new helminth species capable of influencing cancer. Key words: helminths, cestodes, cancers, Mesocestoides corti, Taenia crassiceps
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Organelle proteomics of parasitic protists
Jedelský, Petr ; Tachezy, Jan (advisor) ; Kolářová, Libuše (referee) ; Půta, František (referee)
Advances in DNA sequencing led to a technological breakthrough, that allowed analyzis of complete genomes including those of parasitic protists Trichomonas vaginalis and Giardia intestinalis . These organisms are studied not only for their clinical importance, but also from the evolutionary point of view for their adaptation to anaerobic environment. Genome sequencing and annotations of predicted proteins alone did not bring detail view into functioning of their mitochondrion related organelles in G. intestinalis mitosomes, notparticipating in energetic metabolism, in T. vaginalis hydrogenosomes, producing molecular hydrogen and ATP by means of substrate phosphorylation. Traditional methods based on a fractionation by ultracentrifuging in density gradient and subsequent biochemical and enzymological analyzes were extended by one and twodimensional electrophoresis with subsequent identification of proteins by mass spectrometry. Methods of multidimensional separation of peptides produced by specific proteolysis of a complex mixture...
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Chemoorientation of Trichobilharzia regenti schistosomula in vitro
Parolová, Lucie ; Horák, Petr (advisor) ; Kolářová, Libuše (referee)
The ability to orientate themselves in the definitive host body is very important in the life cycle of trematodes. Contrary to a number of data on trematode orientation in the external environment, migratory routes within the host and places of definitive localization, little is known about compounds serving as stimuli during migration. In this study we tested chemoorientation of Trichobilharzia regenti schistosomula in vitro. We tested fractions of the nervous tissue homogenate of domectic duck (Anas platyrhynchos f. domestica), various fractions of the duck blood serum, NaCl and KCl solutions, D-glucose, L-arginine and commercially available neuromediators and apolipoprotein A-I. The highest attractiveness was recorded in the case of the blood serum of domestic duck. Significant attractiveness also showed NaCl solution at concentrations of 188 and 205 mM, KCl solution at concentration of 205 mM and combination of NaCl, KCl and D- glucose at concentrations NaCl=136 mM, KCl=5 mM, D-glucose=5,5 mM As the attractiveness of NaCl and KCl solutions of the same osmolarity (410 mOsm/l) was equal, and D-glucose solution of the same osmolarity showed no attractiveness for schistosomula, we suggest that the chloride ions are the main stimulus for migration. When various molecular weight fractions of the duck...
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Taxonomy of bird schistosomes not only in planorbid snails in Europe
Holická, Monika ; Kolářová, Libuše (referee)
Cercarial dermatitis caused by bird schitosomes is in focus not only by scientists, but also by the general public. The disease has medical and economic impact, and it is responsible for financial losses in many resorts all around the world. Only some causative agents of the disease are known, because of parasite cryptic life style and biology (e.g. size and location of adults in the definitive host; gonochorism) and inaccurate descriptions in the past. With the onset of modern molecular methods taxonomic revisions of old species of bird schistosomes and descriptions of new ones are of top interest. In Europe pulmonate snails are used by bird schistosomes as intermediate hosts, with some exceptions. Descriptions of new species and genera were recently focused on findings from lymnaeid molluscs, in which the genus Trichobilharzia (the most significant causative agent of cercarial dermatitis) is often developing. Therefore, less attention was paid to the snails of the family Planorbidae. Planorbid snails are most commonly infected by the species of Bilharziella polonica, its typical host in Europe is Planorbarius corneus, less common hosts are Planorbis planorbis, Bathyomphalus contortus or Anisus vortex. B. polonica is spread all over the world. There are also known findings from the Czech Republic....
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Humoral response of specific hosts to bird schistosome antigens
Turjanicová, Libuše ; Mikeš, Libor (advisor) ; Kolářová, Libuše (referee)
This thesis focuses on humoral immune response of specific hosts to antigens of various developmental stages of bird schistosomes T. regenti and T. szidati, and follows up on previous research of antibody response in non-specific hosts (mouse, human). Sera of experimentally infected and hunted-down wild ducks were examined using the ELISA and western blot methods. The sera samples were taken in predefined intervals. Results of the ELISA analysis show the process of humoral immune response after infection by bird schistosomes. The level of specific antibodies IgY against homogenate of T. regenti cercariae increased significantly 20 d.p.i. in ducks infected by T. regenti. Such reaction wasn't observed in ducks infected by T. szidati. Slight changes in level of specific antibodies IgM against T. szidati cercariae homogenate were observed 10 d.p.i. only in fully immunocompetent ducks and in reinfected ducks. Examination of hunted-down wild ducks didn't prove infection by bird schisosomes; this conclusion was confirmed by results of the ELISA analysis. IgY antibodies from ducks infected by T. regenti demonstrated strong reactions with 2 antigens in ranges 49-47 kDa and 47-45 kDa. Other reactions, which were recognized, have not been observed in all specimen. An Western blott with homogenate from 7 days...
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