Fig. 5: Recoded EV-A71 has ZAP-dependent attenuation and elicits protective antibodies in mice.
From: Rational attenuation of RNA viruses with zinc finger antiviral protein
a, Clinical score following infection of 1-day-old ZAP+/+ and ZAP−/− mice with mEV-A71 WT or mEV-A71/CG-48/A+ (mean ± s.d.; n = 11–26 mice per group; P values calculated using two-way ANOVA; NS, non-significant). b, Examples of characteristic pathology developed in infected mice following mEV-A71 infection, including one-limb and two-limb paralysis. c, Probability of ZAP+/+ and ZAP−/− mouse survival (%) following infection with mEV-A71 WT or mEV-A71/CG-48/A+ (n = 11–36 mice per group; P value calculated using Mantel-Cox test). d, One-day-old mice (n = 4–6 per group) were infected with indicated virus; 6-days-post-infection mice were sacrificed and muscles from both posterior limbs were collected, homogenized and total RNA was extracted. EV-A71-specific RNA was quantified by qPCR. Dashed line indicates limit of qPCR detection. e, Schematic representation of experimental design. ZAP+/+Ifnar−/− female mice previously inoculated with mEV-A71/CG-48/A+ (or mock-infected females) were mated with ZAP+/+Ifnar−/− male mice. Resulting offspring was challenged at 1 d of age with mEV-A71 WT. f, Neutralizing activity in plasma from mice after mEV-A71/CG-48/A+ infection (infection at 1 d of age), blood collection at 5 weeks, or mock-infected mice evaluated using EV-A71 NanoLuc luciferase reporter virus. 293T cells were incubated with the antibody:virus mixture for 48 h and luciferase activity was measured. g, Clinical score and survival probability following mEV-A71 (WT) infection of the offspring of ZAP+/+/IFNAR−/− females previously inoculated with mEV-A71/CG-48/A+ (n = 25 pups from 4 different females) or previously mock-infected (n = 22 pups from 4 different females) at 1 d of age. Clinical score and survival were assessed daily until weaning. Statistical significance was inferred by two-way ANOVA and Mantel-Cox tests.
