| Abstract |
In this thesis the production and characterisation of monoclonal
antibodies to herpes simplex virus type 2 is described. The developement
of a suitable radioimmunoassay for the detection of anti-HSV-2 antibodies,
and the selection of an optimal immunisation schedule, is given in chapter
2. Three fusion experiments were performed, resulting in nine stable
hybridoma lines. The monoclonal antibodies secreted by these lines were
subsequently characterised. A list of these antibodies and their
properties, including target antigens, is given at the back of this thesis.
Four of the antibodies were directed against glycoproteins of the virus,
two reacted with non-glycosylated proteins and for three antibodies no
target antigen could be identified. All anitbodies were of the IgG class.
Two antibodies, LP2 and LP3, were directed against the same protein,
gD. Competition binding experiments involving two additional monoclonal
antibodies against this protein showed that there are at least three
different type-common antigenic sites on the gD molecule of HSV-2. LP2 and
LP3 are directed against different antigenic sites. Using tunicamycin,
an unglycosylated precursor of gD was found with a molecular weight of
49,000D. The reactivity of antibodies LP1 and LP4 with a number of
different HSV-1 and HSV-2 strains strains was determined. LP4 proved to be
specific for HSV-2, while LP1 showed comparible reactivity with HSV-1 and
HSV-2 strains. The ability of antibodies LP2, LP3 and LP4 to protect
Balb/c mice from infection with HSV-1 or HSV-2 was assessed. Antibody LP2,
which is strongly neutralising, markedly reduced both the inflammatory
response and virus titres in the site of infection compared to non-treated
mice. Antibody LP3, which is directed against the same protein and is of
the same sub-class, had no effect except for a slightly enhanced
inflammatory response. Mice treated with antibody LP4 were identical to
the control group. Treatment with antibody LP2 reduced the frequency
with which latent infection was established in infected animals, and
reduced the virus titres recovered from reactivated ganglia. Again
antibody LP3 had no effect. The implications of the findings described
above are discussed at the end of each of the chapters. A review of some
of the current and potential applications is given in the Discussion.
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