Figure 3.

Il18^−/− mice are susceptible to VVC. Il18^−/− mice were intravaginally inoculated with 5 × 10⁶ C. albicans blastoconidia and were monitored for: Il22 and Il22bp gene (A) and protein (B) expression in the vagina by RT-PCR and immunofluorescence staining, respectively, at different days post-infection (dpi). (C) Vaginal histology (Periodic Acid Shiff–staining of vaginal sections. Scale bars, 100 μm). (D) Immunofluorescence staining of vaginas at different dpi with antibodies to NLRP3 or p-NLRC4. Sections were stained with the relevant primary antibody overnight at 4°C followed by the secondary TRITC or FITC antibody. Images were acquired using a fluorescence microscope with a 40× objective (Scale bars, 50 μm) and the analySIS image processing software. 4′-6-Diamino-2-phenylindole was used to counterstain tissues and to detect nuclei. (E) IL-1β production (μg/mg, cytokine/total proteins for each sample) in vaginal fluids at different dpi. Results represent mean cytokine levels from samples pooled from four experiments (n = 3–4 total samples per group). (F) Transcriptional factors gene expression on lumbar lymph node by RT-PCR and (G) cytokine levels in the vaginal fluids by ELISA at 14 dpi. *P < 0.05, **P < 0.01 and ***P < 0.001, ****P < 0.0001 knockout vs. wild-type mice.