Biología celular de la infección y morfogénesis de mutantes atenuados del Virus Vaccinia
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Hochschulschrift
Zugriff:
Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. 25-abril-2003
Rewrnbinants based on attenuated rnutants of vaccinia virus (W) are now being considered as candidate vaccines against a broad spectrurn of hurnan and animal diseases. The airn of the thesis was to analyze the rnorphogenetic stages and the virusinduced rnorphological changes in infected hurnan cells with three attenuated W rnutants (MVA, M65 and MIOI), in cornparison with cells infected with the wild type virus, strain WR. A detail transrnission electron rnicroscopy (EM) analysis of hurnan HeLa cells showed irnportant differences in fine uitraestructure and arnounts of the viral intermediates cornpared to wild type WR-infected cells. The EM studies revealed that there are distinct changes in the rnorphogenetic process for al1 of the attenuated rnutants. New intermediate viral forms were found between the IV (irnrnature virus) and the IMV (intracellular rnature virus) forms, representing transition stages with the core still in the assernbling process. The rnorphogenesis of MVA was largely blocked at the IV stage, but few rnature forms were obsewed and the IMVs produced acquired a more munded shape than WR. These "IMVs" have an abnormal interna1 structure (referred as "atypical viruses") with potential alterations in the interactions core-envelope, and are unable to significantly acquire the additional double envelope to render intracellular envelope virus (IEVs). In the case of rnutants M65 an MlOl, virus rnorphogenesis was interrupted at later stages, forming lMVs that accurnulate within the cell but do not progress to form extracellular enveloped virus (EEV). Analyses of viral proteins in the rnutants showed alterations in sorne of the structural proteins with roles in the rnorphogenetic pathway. Confocal rnicroscopy revealed that the rnutants produced a srnall plaque-size phenotype with rnicrocornets and absence of cell lysis. While WR induces actin polyrnerization forming characteristic coiled tails, the rnutants do not produce actin tails and do not exit the cell. Moreover, the rnutants produced rnorphological and biochernical changes to the cells, compatible with an epithelial to rnesenchyrnal transition (EMT), possibly to facilitate spreading of the virus between cells. Mice irnrnunized with recornbinant rnutants expressing Plasmodium yoelii circurnsporozoite antigen (CS) are able to induce significant protection against malaria. Our findings are of irnportance in the developrnent of vaccines based on attenuated W rnutants.
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Biología celular de la infección y morfogénesis de mutantes atenuados del Virus Vaccinia
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Autor/in / Beteiligte Person: | Gallego Gómez, Juan Carlos |
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Medientyp: | Hochschulschrift |
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