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*
Institute of Virology and
Ludwig Boltzmann Institute for Cytokine Research, University of Vienna, Vienna, Austria
We have conducted a DNA immunization study to evaluate how the
immune response is influenced by the physical structure and secretion
of the expressed Ag. For this purpose, we used a series of plasmid
constructs encoding different forms of the envelope glycoprotein E of
the flavivirus tick-borne encephalitis virus. These included a secreted
recombinant subviral particle, a secreted
carboxyl-terminally truncated soluble homodimer, a
nonsecreted full-length form, and an inefficiently secreted truncated
form. Mice were immunized using both i.m. injection and Gene
Gun-mediated application of plasmids. The functional immune response
was evaluated by determining specific neutralizing and
hemagglutination-inhibiting Ab activities and by challenging the mice
with a lethal dose of the virus. As a measure for the induction of a
Th1 and/or Th2 response, we determined specific IgG subclasses and
examined IFN-
, Il-4, and Il-5 induction. The plasmid construct
encoding a secreted subviral particle, which carries multiple copies of
the protective Ag on its surface, was superior to the other constructs
in terms of extent and functionality of the Ab response as well as
protection against virus challenge. As expected, the type of Th
response was largely dependent on the mode of application (i.m. vs Gene
Gun), but our data show that it was also strongly influenced by the
properties of the Ag. Most significantly, the plasmid encoding the
particulate form was able to partially overcome the Th2 bias imposed by
the Gene Gun, resulting in a balanced Th1/Th2
response.
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