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Interactions of polyomavirus structures with components of cell innate immunity
Portychová, Tereza ; Forstová, Jitka (advisor) ; Schreiberová, Lucie (referee)
The topic of this thesis are the interactions of polyomavirus structures with components of innate immunity in infected cells. This review is focused on model SV40 and MPyV polyomaviruses and human BKPyV, JCPyV and MCPyV. The research of the interplay of innate immunity response and polyomaviruses is in its infancy. Infection with all studied polyomaviruses induces, via their LT antigens, DNA damage response (DDR), necessary for their efficient replication. DDR can activate both the canonical and the non-canonical pathway of interferon induction leading to an antiviral state. Polyomaviruses are recognized by the immune system first during replication of their genomes. Interferon induction by polyomaviruses can be initiated by the DNA sensor, cGAS, followed by STING activation, but also by recognition of the viral RNA by the RIG-1 sensor. The virus early LT and st antigens and the late agnoprotein of some polyomaviruses have demonstrated the potential to regulate innate immune responses and thus contribute to the establishment of polyomavirus persistence. Keywords: polyomaviruses, innate cell immunity, large T antigen, small t antigen, agnoprotein, interferon-stimulated genes
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Development of a technique for gene transfer into T-lymphocytes using polyomavirus structures and the LAH4 peptide
Schreiberová, Lucie ; Španielová, Hana (advisor) ; Vopálenský, Václav (referee)
Efficient delivery of genetic material to T-lymphocytes is key in gene therapy using T-lymphocytes with chimeric antigen receptors. Current procedures require the use of potentially dangerous viral vectors or large amount of input material. The diploma thesis therefore focuses on exploring new approaches for gene transfer into T-lymphocytes: use of safe virus-like particles (VLPs) derived from mouse polyomavirus in combination with the amphipathic cationic peptide LAH4. LAH4 has the potential to increase the efficiency of DNA and viral vector transport into cells. The system which combines VLPs and the LAH4 peptide was optimized for the delivery of reporter gene (encoding GFP and luciferase) to the model T-cell line Jurkat. It has been found that Jurkat cells cannot be efficiently transduced by DNA packed into VLPs. When cells were transfected only with DNA and LAH4, consistent results were not obtained, and the transfection efficiency ranged from 0.5 to 19%. The diploma thesis also analysed the effect of phosphorylation of viral structures on gene transfer. The impact of treatment of virus particles by alkaline phosphatase on the infectivity of the virus was studied and it was necessary to analyse the effect of the reaction components. Sublytic concentration of Triton-X100 in the reaction buffer...
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Raccoon polyomavirus: example or exception of polyomavirus driven oncogenesis?
Schreiberová, Lucie ; Španielová, Hana (advisor) ; Hirsch, Ivan (referee)
Polyomaviruses (PyV) are widespread through human and animal populations and typically associated with asymptomatic persistent infection. Rarely, natural PyV infections can lead to oncogenic transformation. Virus genome is usually integrated into the host DNA of tumour tissue. Over the past few years, an increased number of very aggressive brain tumours and olfactory tumours have been observed in raccoons. These tumours are associated with the newly discovered raccoon polyomavirus, which was found as an intact episome in host cells. This bachelor thesis is therefore focused on comparison of current state of knowledge on raccoon polyomavirus with previously described mechanisms of PyV tumorogenesis. Unlike for other PyVs, the fact that primary neuronal stem cell infection is most likely to occur can play a key role in raccoon polyomavirus driven oncogenesis. Tumours also exhibit unusually high expression of virus-encoded micro RNA that can be connected with tumour induction. Similary to other tumours caused by PyV, a large amount of early viral proteins with oncogenic potential is found in tumours. Revealing unknown factors responsible for the development of tumours caused by raccoon polyomavirus may help in understanding of mechanisms of oncogenesis. Key words Polyomavirus, oncogenesis, raccoon...
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