Intravesical vaccination with bacterial lysates and CpG induced strong local Th1 responses in bladder. (A) Naïve Great mice were intravesically vaccinated with J96 lysates and different amounts of CpG three times, then challenged with UPEC J96. Bladders and spleens were collected for flow cytometry analysis on day 3 after challenge. WT C57BL/6J mice infected with J96 were used as controls for YFP gating. (B) Naïve WT mice were intravesically vaccinated with UPEC J96 lysates and combined with increasing amount of CpG three times, then intravesically challenged with UPEC J96. Bladders were collected to determine bacterial load on day 3 postchallenge. (C) Naïve WT mice were intravesically vaccinated three times with PBS, UPEC J96 lysates, or UPEC J96 lysates combined with 10 μg of CpG, then intravesically challenged with UPEC J96. Bladders were collected to determine bacterial load on day 14 after challenge. Each data point represents one mouse. Data are shown as mean ± SD. Data were analyzed by an ordinary one-way ANOVA with Tukey’s multiple comparison post hoc test. *P < 0.05; **P < 0.01; ***P < 0.001. ns, not significant.