The effect of decoy concentration on TeNT activity in vaccinated mice
In vaccinated mice, reducing the concentration of decoy, while maintaining the concentration of active TeNT, showed a trend of reduced TeNT activity (Figure 5 A-D). As we have demonstrated that the decoy has an inhibitory effect on TeNT activity in naïve mice, there is likely a concentration dependent balance between the positive effect of diverting antibodies and TeNT inhibition by the decoy. Unlike when combined with 100 µg of decoy, at 20 µg and 5 µg of decoy, 500 ng of TeNT was not enough to induce stage 4 tetany in all mice. Further, 5 µg of decoy combined with 25 ng of TeNT was slightly less effective than 5 ng of TeNT with 100 µg of decoy (Figures 5B and D). Comparison of the time-course of tetany development demonstrated that there was a significant difference in the effectiveness of the decoy depending on its concentration. Interestingly, development of TeNT symptoms at 100 µg and 20 µg of decoy were not significantly different at 0.01 ng and 0.1 ng doses of TeNT, while TeNT showed less activity in the presence of 5 µg of decoy at these doses (Figures 5C and D). While at a 1 ng dose of TeNT, activity was significantly higher in combination with 100 µg of decoy compared to either 20 µg or 5 µg of decoy (Figure 5D).