Fig. 3: BinJ/JEV_NSW/22-prME vaccination and challenge.

a Timeline of BinJ/JEV_NSW/22-prME vaccinations, JEV_NSW/22 challenge, and sample/data collections. Silver arrows indicate sample/data collection timepoints. b Serum end-point neutralising antibody titres after one (1st dose) and two (2nd dose) vaccinations. Limit of quantification 1 in 5. (Statistics by Kolmogorov-Smirnov tests). c Mean percent C57BL/6J mouse body weight change relative to day 0 for each mouse (n = 10 per group 0-2 dpi, n = 5 per group 3–10 dpi). Differences only reached significance at 3 dpi, statistics by t-test (significance was not reached on any other day, or ranges of days by repeated measure ANOVA tests). ND not detected. d As for C for Ifnar^-/- mice. Statistics by Kolmogorov-Smirnov exact tests (2 and 3 dpi). e Clinical disease scores for posture (circles), activity (crosses) and fur ruffling (triangles) were monitored in Ifnar^-/- mice. N as in (c). † Ethically defined end point reached for euthanasia. Statistics by Kolmogorov-Smirnov exact tests (n = 5 per group), p = 0.013 for each disease parameter. f Post-challenge viremia in C57BL/6J mice determined by TCID₅₀ assays. Viremia for individual mice at each time point. Limit of detection was 2 log₁₀TCID₅₀/ml. ND not detected. In the PBS group no viremia was detected on any day in 3/10 mice. Statistics by Kolmogorov-Smirnov exact test for 1–4 dpi (n = 30 serum samples per group; 30 vs. 18 ND). G Post-challenge viremia in Ifnar^-/- mice (n = 10 per group 0-2 dpi, n = 5 per group 3 dpi). h Tissue titres for C57BL/6J spleens. Limit of detection ≈2.7 log₁₀TCID₅₀/g. i RT-qPCR of JEV RNA in C57BL/6J spleens. JEV RNA copies are normalized to Rpl13a. Limit of detection ≈10^-12 JEV/Rpl13a copies. j Spleen and (k) brain tissue titres in Ifnar^-/- mice 2 and 3 dpi.