A Fusion Protein between Streptavidin and the Endogenous TLR4 Ligand EDA Targets Biotinylated Antigens to Dendritic Cells and Induces T Cell Responses In Vivo
Figure 3
EDAvidin binds to biotinylated NS3 and induces strong anti-NS3 cellular immune responses in vivo. (a) ELISA-based binding assays of EDAvidin to biotinylated NS3. Biotinylated or nonbiotinylated NS3 were coated into the wells of ELISA plates. EDAvidin or EDA alone was added to the wells and after extensive washes, the plates were developed using rabbit polyclonal anti-EDA antibodies. ((b), (c)) In vivo induction of anti-NS3 T cell responses. HHD transgenic mice were immunized i.v. with NS3Biot, EDA plus NS3Biot, streptavidin plus NS3Biot, EDA-NS3, or with EDAvidin plus NS3Biot in saline. Seven days after immunization, anti-NS3 immune response was analyzed by measuring the number of IFN-γ producing cells by ELISPOT in response to the T cell epitope p1073, NS3 protein, or culture medium (Neg) (b) or by in vivo killing (c) as described in methods.
