Mouse vaccination and titre confirmation
To obtain consistently high anti-TeNT antibody titres, mice were
vaccinated three times, at three-week intervals with 2 tetanus toxoid
international units of human divalent diphtheria-tetanus vaccine (ADT,
Seqirus). Immunity of the mice was confirmed by injecting a small subset
of animals with 500 times the TeNT dose capable of inducing stage 4
tetany in naïve mice within 24 hours; no symptoms of tetanus were
observed. The titres of all mice data included in the study were
confirmed to be protective (>1:8000 (Fishman et al. ,
2009)) by serum ELISA, three weeks following the final vaccination. Data
for mice with non-protective titres (< 1:8,000, three mice)
were excluded from the study and where appropriate the group size was
changed from n = 6 to n = 5 (Supplementary data Table 1).