Leaf-Encapsulated Vaccines: Agroinfiltration and Transient Expression of the Antigen Staphylococcal Endotoxin B in Radish Leaves

Figure 5

SEB immunogenicity and protective immunity against SEB-induced IFN-γ production. (a) Recombinant SEB (15 μg) run on a 10% (w/v) SDS-PAGE was blotted onto a nitrocellulose membrane and immunoreacted to sera obtained from mice immunized with 25 μl homogenized leaves expressing SEB-GUS (L-SEB-GUS) or GUS (L-GUS) mixed with a CT adjuvant (5 μg/25 μl whole leaves). Recombinant SEB reactive to serum from L-SEB-GUS-immunized mice produces a band (arrow) at approximately 28 kDa, verifying the immunogenicity of SEB. (b) Titers of pooled anti-SEB antibodies from eight immunized mice were qualified by ELISA. The geometric means of ELISA titers were presented. (c) Eight naïve mice were challenged intranasally with 40 μg of recombinant SEB or PBS as a negative control, and the levels of IFN-γ in BAL fluids were measured by ELISA. Mice immunized with L-SEB-GUS or L-GUS were challenged intranasally with 40 μg of recombinant SEB overnight. The levels of collected IFN-γ in BAL fluids of L-SEB-GUS- and L-GUS-immunized mice were compared by an ELISA assay. Experiments were performed in triplicate. Data were analyzed statistically by Student’s t-test and presented as mean ± SD (; by Student’s t-test).

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