Figure S1 - The Adjuvant Activity of Alphavirus Replicons Is Enhanced by Incorporating the Microbial Molecule Flagellin into the Replicon

Induction of anti-flagellin antibodies is avoided by incorporation of FliC-D3 in the replicon. We asked whether there was a difference in the anti-flagellin response induced by soluble flagellin and flagellin expressed from VREP. C57BL/6 mice (n = 3) were immunized twice with OVA mixed with one of the following: sFliC-WT (light blue), VREP-LacZ+sFliC-WT (dark blue), VREP-FliC-WT (green), VREP-FliC-D3 (red) or VREP-LacZ (black). 1 µg of soluble flagellin and 106 IU of VREP constructs were used. Serum was assayed for anti-FliC-WT IgG by ELISA. Mice that were given sFliC-WT displayed a strong anti-flagellin response. Expressing FliC-WT from VREP resulted in a sharp decrease in the response. By expressing the truncated FliC-D3 from VREP, induction of anti-flagellin antibodies was completely eliminated. (EPS)