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Cellular tropism and cell-to-cell fusion properties of the infectious bronchitis virus spike glycoprotein

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Bickerton, Erica Jane (2010) Cellular tropism and cell-to-cell fusion properties of the infectious bronchitis virus spike glycoprotein. PhD thesis, University of Warwick.

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Official URL: http://webcat.warwick.ac.uk/record=b2490704~S15

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Abstract

There are numerous vaccines available for the control of infectious bronchitis virus
(IBV) in poultry, however protection is short-lived and poorly cross-protective
between strains. The vaccines must currently be grown in embryonated eggs, a
cumbersome and expensive process. The ability to grow vaccines on a cell-line such
as Vero cells would be highly advantageous.
The spike (S) glycoprotein of IBV is comprised of two subunits, S1 and S2, has a
vital role in virulence in vivo and is responsible for cellular tropism in vitro. This
project aims to identify the amino acids present in the S glycoprotein involved in
determination of cellular tropism and cell-to-cell fusion.
The IBV Beaudette strain is able to replicate in both primary chick kidney (CK) cells
and Vero cells, whereas the IBV M41 strain replicates in primary cells only.
Recombinant IBVs with chimaeric S genes were generated using a reverse genetics
system with the genomic background of Beaudette and part of the S gene from M41.
Their growth characteristics and cellular tropism were investigated. The S2 subunit
of Beaudette was found to be sufficient to confer the ability to grow on Vero cells
and swapping just three amino acids with corresponding ones from M41 was
sufficient to remove the ability of the Beaudette S glycoprotein for growth on Vero
cells.
Beaudette was further adapted to syncytia formation on Vero cells by serial passage
and isolates were sequenced to identify amino acid changes between parent and
Vero-adapted viruses that are potentially involved in cell-to-cell fusion.
Understanding the way in which IBV infects host cells is vital in order to rationally
design better vaccination and treatment strategies and help to reduce the prevalence
of IBV infection in poultry worldwide. Using the IBV reverse genetics system, we
now have the potential to grow IBV vaccines on Vero cells.

Item Type: Thesis (PhD)
Subjects: Q Science > QH Natural history > QH301 Biology
S Agriculture > SF Animal culture
Library of Congress Subject Headings (LCSH): Infectious bronchitis in poultry -- Vaccination, Infectious bronchitis in poultry -- Genetic aspects, Glycoproteins -- Analysis
Official Date: September 2010
Dates:
DateEvent
September 2010Submitted
Institution: University of Warwick
Theses Department: Department of Biological Sciences
Thesis Type: PhD
Publication Status: Unpublished
Supervisor(s)/Advisor: Britton, Paul ; Armesto, Maria ; Easton, A. J. (Andrew J.)
Sponsors: Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC) ; Institute for Animal Health (Great Britain) (IAH)
Extent: xix, 202 leaves : ill., charts
Language: eng

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