The aim of the study was to determine the function of gp 150 and latterly to assess the potential of gap150 as a vaccine antigen to prime and protect inbred mice against MHV-68 infection. For functional studies of gp150 two main strategies were adopted; (i) the production of a recombinant virus in which the gene encoding gp150 is made dysfunctional resulting in a gp150 'knockout' (KO) virus and (ii) generation and use of purified gp150 in cell binding studies to determine if gp150 can bind to cells. Results indicate that gp150-His binds the heterogeneous splenic cell population as a whole i.e. not a particular subset of lymphocytes. This suggests gp150 may interact with a ubiquitous cell surface protein or perhaps a protein specific to leukocytes and could be involved in MHV-68 attachment to these cells. Gene gun nucleic acid immunisation of inbred mice with a plasmid encoding gp150 under the control of a constitutive promoter, alone or in combination with a recombinant vaccinia virus expressing gp150 (VVpg150) was undertaken followed by intranasal challenge with MHV-68. Virus specific antibody appeared earlier in the group that received gp150 DNA plus VVgp150. The groups that received gp150 DNA in conjunction with either VVgp150 or a control vaccinia virus (VVgpt) appeared to have reduced levels of latently infected cells in the spleen day 15 post infection and reduced splenomegaly (a phenomenon of MHV-68 infection) in comparison with control mice. This could indicate that vaccinia virus, in a non-specific manner, boosts the specific immune response to previously administered DNA and in this case was able to limit the level of MHV-68 reaching the spleen. However, this vaccine regime failed to significantly alter the level of infectious virus in the lung or prevent the establishment of latent virus in the spleen.
Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:641126 |
Date | January 2000 |
Creators | Atkin, Isobel Mary Dawber |
Publisher | University of Edinburgh |
Source Sets | Ethos UK |
Detected Language | English |
Type | Electronic Thesis or Dissertation |
Source | http://hdl.handle.net/1842/29804 |
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