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Characterisation of host-range determinants in Chandipura virus

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Stock, Emily (2014) Characterisation of host-range determinants in Chandipura virus. PhD thesis, University of Warwick.

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Abstract

The emerging arbovirus, Chandipura virus (CHPV) has been implicated in epidemics of acute encephalitis in India with mortality rates in hospitalised children of over 50%. CHPV is a member of the Vesiculovirus genus of the Rhabdoviridae family.

Viruses are dependent on the host cell at every stage of their lifecycle. The isolation of temperature-dependent host range (tdCE) mutants, which are characterised by growth impairment at 39°C in chick embryo (CE) cells but not in monkey cells, highlights a dependence of CHPV on undetermined host factors. The characterisation of three tdCE mutants was carried out. Each of the mutants contain one or more coding mutations in the RNA polymerase gene and two contain additional mutations in the attachment protein gene. Using a reverse genetics system of CHPV recombinant viruses containing the specific mutations were generated. This demonstrated that a single amino acid change in the virus RNA polymerase of each mutant, located either between domains III and IV or within domain IV or VI was responsible for host range specificity, the tdCE phenomenon. In CE cells at 39°C the tdCE lesions were shown to disrupt the assembly of cytoplasmic replication complexes.

A recombinant virus with a Flag tag attached to the L protein was generated and used to infect mammalian and avian cells. Potential L/Flag interacting partners were co-immunoprecipitated and analysed by mass spectrometry. The predominant cellular functions of the identified proteins were in gene expression and the cytoskeleton.

To investigate the neuropathogenisis and cell tropism of CHPV in vivo and ex vivo models of CHPV disease were established. The virus was shown to principally localise to the cerebellum in the mouse brain with the virus targeting immature neurons, granule cells and microglia. Mature neurons, oligodendrocytes and astrocytes were permissive to infection with CHPV but their infection occurred infrequently.

Item Type: Thesis (PhD)
Subjects: Q Science > QR Microbiology > QR355 Virology
Library of Congress Subject Headings (LCSH): Arboviruses
Official Date: June 2014
Dates:
DateEvent
June 2014Submitted
Institution: University of Warwick
Theses Department: School of Life Sciences
Thesis Type: PhD
Publication Status: Unpublished
Supervisor(s)/Advisor: Easton, A. J. (Andrew J.)
Extent: xii, 251 leaves : illustrations
Language: eng

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