Heath, Richard Rhead (1996) The role of the cell-mediated immune response to rotavirus infection. PhD thesis, University of Warwick.

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Abstract

The
objective. of this project was to determine the protein specificity of the cytotoxic
T-
lymphocyte (CTL)
response to rotavirus
infection in
mice and to assess the rotavirus
serotype/strain
independent
nature of this response.
Previous work,
involving the rotavirus
group at
Warwick, had
shown that the outer shell glycoprotein
VP7 is
a major target
antigen
for
a
CTL
response that is
virus serotype-independent.
However, that work
did
not
cover all twelve rotavirus proteins, was confined to one strain of adult mice
(C57BL/6, H-
2b)
and covered only two of the fourteen VP7
serotypes
(serotypes 3
and
6) (Offit
et al.,
1994).
Recombinant
vaccinia viruses expressing
individual
rotavirus
UKtc
proteins
VP2, VP3, NS26
and
NS12
were constructed to complete a set of recombinant vaccinia
viruses covering the full
complement of rotavirus proteins
from the bovine UKtc
strain.
These
were used to define
rotavirus proteins eliciting a
CTL
response
in three different
mouse
haplotypes. UKtc NS53
and
UKtc VP7
stimulated a strong
CTL
response only
in
the H-2b MHC
class
I haplotype (UKtc NS53
and
UKtc VP7
were restricted at
H-2Db
and
H-2K
b,
respectively).
Conversely, UKtc VP3
stimulated a strong
CTL
response
in the H-2d
and
H-2k (but
not
in H-2b) MHC
class
I haplotypes. Work
using congenic mouse strains
was used to verify that the VP7
protein specific
CTL
response
is
restricted solely
by the
MHC
class
I
antigens.
Rotavirus RRV NS53
was not only
found to elicit a
CTL
response
in the H-2b
MHC
class
I haplotype,
similar to UKtc NS53, but
also
in the H-2d MHC
class
I
haplotype. Thus, the individual
rotavirus protein
that elicits a
CTL
response not only
depends
on the MHC
class
I haplotype, but
also on the actual rotavirus strain
being tested.
Many
of the previous studies
looking
at the CTL
response-to
individual
rotavirus proteins
have,
unlike this study, used several
different
rotavirus strains and, therefore, may
have
given an
inaccurate
representation of the rotavirus proteins
that elicit a
CTL
response.
Recombinant
vaccinia viruses were also used
to examine the serotype/strain
independent
nature of the CTL
response against the major
target antigens.
The
analysis
was extended to cover
VP3 from two different
strains,
NS53 from three different
strains
and
VP7 from
seven of the fourteen
serotypes.
VP3
and
NS53
were
found to elicit a strain-
dependent
response whereas the serotype-independent nature of the CTL
response to VP7
was confirmed.
Since the serotype-independent nature of the rotavirus
VP7-specific CTL
response was
found to cross-protect
between half
of the VP7
serotypes,
irrespective
of the
immunising.
serotype,
it
would
be
reasonable to speculate
that the CTL
response
is
serotype-independent
between
all the VP7
serotypes.
Finally,
recombinant vaccinia viruses were used
to locate CTL
epitopes on
NS53
and
VP7. Recombinant
vaccinia virus expressing a
UKtc NS53 deletant
mutant
(P9DM5)
showed there to be
at
least
one strain specific epitope
in the
first 150
amino acids of the
UKtc NS53
protein.
Recombinant
vaccinia viruses expressing
four different UKtc VP7
fragments
spanning
46%
of this protein were examined.
It
was
found that the fragment
spanning the restriction enzyme sites at nucleotide
90 (CIaI) and nucleotide
196 (Hhal),
i. e.
between
amino acids
13
and
48
of the mature
UKtc VP7 protein, contained a serotype-
dependent CTL
epitope.
The finding
suggests that the immunodominant epitope
identified
in the same region of
VP7 by Franco
et al.
(1993) was not the serotype-independent
epitope.

Item Type:	Thesis (PhD)
Subjects:	Q Science > QR Microbiology > QR180 Immunology Q Science > QR Microbiology > QR355 Virology
Library of Congress Subject Headings (LCSH):	Cell-mediated cytotoxicity, Rotaviruses , Cytology -- Research
Official Date:	July 1996
Dates:	Date Event July 1996 Submitted
Institution:	University of Warwick
Theses Department:	Department of Biological Sciences
Thesis Type:	PhD
Publication Status:	Unpublished
Supervisor(s)/Advisor:	McCrae, M.
Extent:	xxv, 228 leaves
Language:	eng

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