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One difficulty of obtaining virus vaccine is the production of vaccinating antigens in large enough quantities. This aspect is particularly important for a virus which cannot be produced in vitro. Vaccinating epitopes are the minimal antigenic sites eliciting protective antibodies. Once these epitopes are identified, they can be introduced into a carrier protein. The fimbrial subunits F17 and CS3IA from Escherichia coli may be used as carrier proteins for the introduction of epitopes identified on bovine rotavirus strains and on transmissible gastroenteritis virus of piglets.
This group has been responsible for results in the following areas:
cloning, sequencing and genetic organisation of the CS31A and F17 operons;
construction of the complementation systems and F17 subunits;
identification and localization of the sites in CS31A which allows insertion of heterologous peptides;
obtention of CS31A and F17 fimbriae major subunit carrying up to 4 viral epitopes;
intraperitoneal immunization of mice with purified CS31A hybrid protein (native or denatured) carrying epitope C and/or A of the spike protein S of TGE virus induced antibodies which recognise the viral peptides and the TGE virus. In addition antibodies directed against rotavirus were obtained with the F17 system.
Antibodies neutralized the infectivity of the TGE virus in vitro. The mice immunized with purified CS31A hybrid proteins carrying epitopes C and A of the S protein of TGE virus developed, an anamnestic response demonstrated when mice were later innoculated with TGE virus
In mice orally innoculated with recombinant E coli producing CS31A with TGE virus epitope intestinal IgAs were produced against the viral peptide.
In addition, complementary work was conducted to obtain a baby mouse model for testing rotavirus infection and vaccine protection. The gut colonizing ability of CS31A E coli in mice, and the invivo stability of CS31A plasmids and genes were also studied.
gut colonizing ability of CS31A Escherichia coli and in vivo stability of the CS31A plasmids and genes.
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