Innate immunity can be trained to control S. sonnei in vivo.A. Model for S. sonnei reinfection. 2 dpf embryos were primed with PBS or a sublethal dose of S. sonnei. At 48 hours post primary infection (hp1i), larvae where challenged with a lethal dose of S. sonnei and monitored by survival assay for 72 hours post-secondary infection (hp2i). B,C. Innate immune training to S. sonnei is dependent on O-antigen. Survival curves (B) and Log10-transformed CFU counts (C) of 4 dpf larvae infected in the HBV with lethal dose (~8000 CFU) of mCherry-WT S. sonnei. 48 hours prior to infection with lethal dose, embryos were primed by delivering PBS (grey), or a sublethal dose (~80 CFU) of GFP-ΔO-Ag (blue) or GFP-WT (red) S. sonnei. Experiments are cumulative of 4 (B) or 3 (C) biological replicates. In C, full symbols represent live larvae and empty symbols represent larvae that at the plating timepoint had died within the last 16 hours. Statistics: Log-rank (Mantel-Cox) test (B); one-way ANOVA with Sidak’s correction on Log10-transformed data (C); ns, non-significant; *p<0.0332; ***p<0.0002; ****p<0.0001.
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