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Design and production of TBEV specific cleavage reporter for in vitro and in cell studies
KOTOUNOVÁ, Eliška
Tick-borne encephalitis virus (TBEV) is an important human pathogen and as of now, there are no specific therapeutics to combat this disease. In vitro and in cell models are essential in studying the disease and in the pursuit of novel therapeutics. This thesis deals with the design and production of two genetically encoded cleavage reporters of TBEV NS2B-NS3 protease activity. The reporters were produced in E. coli and tested with recombinant NS2B-NS3 protease, as well as produced in mammalian cell lines and analyzed for fluorescence upon infection with TBEV.
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Antiviral vaccination - tick borne encephalitis virus
Solarová, Pavlína ; Drda Morávková, Alena (advisor) ; Janštová, Vanda (referee)
Tick borne encephalitis is by one of the most severe disease of central nervous system. Agent of this disease is tick-borne encephalitis virus, which is transmitted mainly by tick. Clinical picture of TBE is considerably different and varied in individuals - from quite infection without symptoms up to severe cerebro-spinal meningitis, in such cases attacked persons by this virus leave hospital with whole-life handicap. We can prevent this disease by inoculation. We can see it in the neighboring Austria, where TBE almost became extinct because 90% of population is vaccinated. Vaccines are commercially available from 1979. Nevertheless only 17% population is vaccinated against TBEV. The reason for it could be fact that insurance companies do not provide the vaccines as well as not very effective health education. One of possibilities how we can this tendency reverse would be higher awareness of pupils at schools. In school educational programs there is contained prevention of viral disease, especially in general biology, biology of viruses and education to health.
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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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Antiviral vaccination - tick borne encephalitis virus
Solarová, Pavlína ; Janštová, Vanda (referee) ; Drda Morávková, Alena (advisor)
Tick borne encephalitis is by one of the most severe disease of central nervous system. Agent of this disease is tick-borne encephalitis virus, which is transmitted mainly by tick. Clinical picture of TBE is considerably different and varied in individuals - from quite infection without symptoms up to severe cerebro-spinal meningitis, in such cases attacked persons by this virus leave hospital with whole-life handicap. We can prevent this disease by inoculation. We can see it in the neighboring Austria, where TBE almost became extinct because 90% of population is vaccinated. Vaccines are commercially available from 1979. Nevertheless only 17% population is vaccinated against TBEV. The reason for it could be fact that insurance companies do not provide the vaccines as well as not very effective health education. One of possibilities how we can this tendency reverse would be higher awareness of pupils at schools. In school educational programs there is contained prevention of viral disease, especially in general biology, biology of viruses and education to health.
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Ultrastructural changes in human neural cells after infection with tick-borne encephalitis
TESAŘOVÁ, Martina
Annotation: Human cells of neuronal origin represent an excellent tool for the investigation of neuropathogenesis of TBE. The maturation, replication process of tick-borne encephalitis virus (TBEV) and ultrastructural changes induced by infection in the neuroblasts cell line (UKF-NB-4) was studied by electron microscopy. I compared electron microscopical aspects (appearance) of TEM images of neuroblasts cells prepared by (1) conventional chemical fixation, resin-embedding and sectionig; (2) rapid freezing of cell monolayers at high pressure and sectioning of freeze substituted samples. The most interesting fact, however, is that vitrification preserves the cell in close to native state, whereas chemical fixation and dehydration can not take place without extensive intra- and intermolecular cross-linking and aggregation. The appearance of the cytoplasm and nucleoplasm of neu-roblasts cells were different in conditions (1) and (2). The excellent ultrastructure of the cytoplasmic and nuclear membranes and organels of neuroblasts cells processed by (2) confirmed the potentional of the method for preservation of cellular fine structures. The infection of neuroblastoma cells was associated with number of major morphological changes, including proliferation of membranes of the rough endoplasmic reticulum and rearrangement of cytoskeletal structures. The viral particles were located mainly in the cisterna´s of ER but also in the cytoplasm. In the cytoplasm I observed virions in the asso-ciations with microtubules and neurosecretory dense core vesicles. The transport of viral particles inside of the transport vesicles was obsereved from ER to Golgi apparatus. Free nucleocapsids were not confirmed. The observed pattern corresponded to both trans and cis type of maturation. The TBEV-infected neuroblasts cells exhibited either apoptotic or necrotic morphological changes. I observed the apoptotic signs (condensation, margination and fragmentation of chromatin in nucleus) and other alterations, such as disorganisation of cytoplasm, presence of the vacuoles and high density of cytoplasm. This report also de-scribes scanning electron microscope study of the surface features of neuroblasts cells. We observed virus-mediated cytopathic effect. The cells infected with TBEV were rounded with rough and rugged topography.
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