Construction Of Recombinant Adenovirus Expressing Protective Antigen Of Bacillus Anthracis And Preliminary Research Of Inhalational Immuno-responses

Objective: In order to study the possibility of the recombinant adenovirus vaccine for prevention of Anthrax, and the possibility of the aerosol inhalation for recombinant adenovirus vaccine.Method:1. The PA gene was cloned into plasmid pAdTrack-CMV. Transformed the positive clone into E.coli BJ5183 cells with the adenoviral plasmid pAdEasy-1. Then vAd-PA was generated by homologous recombination. vAd-PA was transfected into HEK-293 cell line. Expression of PA in 293 cells was proved by SDS-PAGE and Western-blot.2. The BALB/c mice were immuned with the recombinant adenovirus by intramuscular injection and intranasal instillation. The specific antibody and cell-mediated immunity data in the immunized mice were analyzed through ELISA and MTT test.3. Found the optimal condition about the nebulizing. The BALB/c mice were immuned with the recombinant adenovirus by aerosol inhalation. The specific antibody, mucosal immunity and cell-mediated immunity data were analyzed through ELISA and MTT test.Result:1. Successfully constructed recombinant adenovirus expressing PA.2. The results suggest that humoral, mucosal and cell-mediated immune response were induced in immunized mice (intramuscular injection and intranasal instillation).3. The data shows recombinant adenovirus induced immune response in the mice that were immuned with the recombinant adenovirus by aerosol inhalation.