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Shigella induced protection is nonspecific. (A) Reinfection of trained larvae with lethal dose injection of P. aeruginosa (Pa) or S. aureus (Sa) at 4 dpf. (B) Survival curves from naïve and Shigella-trained (2.2 × 103 ± 3.8 × 102 CFUs) larvae infected with P. aeruginosa (PBS, 2.6 × 103 ± 7.4 × 102 CFUs; Shigella, 2.3 × 103 ± 5.5 × 102 CFUs). N = 3 with 20 larvae per experiment. (C) Survival curves from naïve and Shigella-trained (1.4 × 103 ± 8.6 × 102 CFUs) larvae infected with S. aureus (PBS, 3.4 × 104 ± 1.1 × 104 CFUs; Shigella, 3.4 × 104 ± 6 × 103 CFUs). N = 3 with >20 larvae per experiment. (D) Reinfection of trained larvae with lethal dose injection of Shigella at 7 dpf (or 5 dp1i). (E) Representative images of naïve and Shigella-trained Tg(lyz::dsRed) larvae with at 5 dp1i. Scale bar, 500 μm. (F) Quantification of total fluorescence in Tg(lyz::dsRed) naïve and Shigella-trained (1.7 × 103 ± 5.7 × 102 CFUs) larvae at 5 dp1i. N = 3 with 10 larvae per time point (mean ± SD). (G) Survival curves from naïve and Shigella-trained larvae infected with Shigella at 5 dp1i. N = 3 with >16 larvae per experiment. *P < 0.05, ****P < 0.0001, log-rank (Mantel-Cox) test (B, C, and G) and unpaired Student’s t test (F).
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