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United States Department of Agriculture

Agricultural Research Service

Title: Analysis of Swine Influenza Virus H3n2 Strain Hemagglutinin and Nucleoprotein Expressed by Recombinant Human Adenovirus Serotype 5 Vectors

Authors
item Tang, Min
item Wesley, Ronald

Submitted to: American Society for Virology Meeting
Publication Type: Abstract Only
Publication Acceptance Date: July 20, 2001
Publication Date: N/A

Technical Abstract: Here we report the construction of replication-defective adenovirus recombinants that encode the swine influenza virus (SIV) hemagglutinin (HA) gene or the nucleoprotein (NP) gene. The HA and NP proteins have been expressed from these recombinants in 293 cells and in mice. The HA and NP genes from influenza virus A/Swine/Iowa/99(H3N2), propagated in embryonated chicken eggs, were cloned into the Shuttle-CMV (Sh) plasmid. The Sh plasmid containing the HA or NP gene was then inserted into the E1 region of a replication-defective serotype 5 human adenovirus (Ad). Three recombinant adenoviruses were generated, HA-Ad-Sh containing the HA gene, NP-Ad-Sh containing the NP gene, and Ad-Sh containing no foreign gene insertion. Western blot analysis revealed that monoclonal antibodies recognized HA and NP proteins produced by HA-Ad-Sh and NP-Ad-Sh, respectively. Strong expression of the hemagglutinin and nucleoprotein was observed by an immunocytochemical assay. Moreover, Flow Cytometer examination demonstrated the cell surface expression of hemagglutinin glycoprotein, and that maximum cell surface expression was at 48 hours postinfection. Mice also were inoculated intramuscularly with either HA-Ad-Sh or Ad-Sh or given a sham inoculation with PBS. Three weeks after the inoculation, mice given HA-Ad-Sh developed anti-hemagglutinin antibodies, as measured by the hemagglutinin inhibition test, in comparison to mice inoculated with Ad-Sh or PBS. Preliminary data of this study indicate these proteins are expressed well with the adenovirus expression system and may be useful as a vaccine for the swine industry.

Last Modified: 8/22/2014
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