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 2
EDAvidin targets biotinylated antigens to DC and retains the proinflammatory activity of EDA. (a) Flow cytometric analysis of BMDC incubated with the indicated proteins. (b) Colorimetric assay to measure NF-κ β induction in HEK TLR4 or HEK LacZ expressing cells in response to different concentrations of EDA, EDAvidin, LPS, or culture medium (Neg). Results represent the NF-κ β fold induction. (c) The human monocytic cell line THP1 was incubated in the absence or presence of the indicated concentrations of EDAvidin, EDA, LPS, or culture medium (Neg). After 15 hours of culture, supernatants were harvested and the released TNF-α was measured by ELISA. (d) Production of IL-12 by BMDC after incubation with 500 nM EDA, 500 nM EDAvidin, 0.1 μg/mL LPS or culture medium (Neg). Twenty-four hours later, culture supernatant was harvested and IL-12 released to the medium was measured by ELISA. All data are representative of two independent experiments.
