Fig. 4

ZIKV NS4B-C100S induces potent antiviral innate and adaptive immune responses compared with ZIKV-3′UTR-Δ10. a, b Six-week-old AB6 (a) mice or WT B6 (b) were infected with 2 × 10⁴ FFU ZIKV NS4B-C100S (C100S) or ZIKV-3′UTR-Δ10 (Δ10). a On day 28, splenocytes of AB6 mice were cultured ex vivo with WT ZIKV for 24 h and stained for IFN-γ, CD3, and CD4 or CD8. n = 3 per group. b Supernatants from the ex vivo culture were collected on day 2 after WT ZIKV re-stimulation and measured for Th1- cytokine production. n = 5 to 7. c On day 28 after immunization, mouse sera were measured for NAb titers. d Six-week-old AB6 were infected with 2 × 10⁴ FFU WT ZIKV FSS13025ic or ZIKV mutants. On day 2 pi, blood cytokines levels were determined by Q-PCR assay. Data are presented as the fold increase compared with the NF group (n = 6 to 10). e–g AB6 BM macrophages were infected with WT ZIKV FSS13025ic or ZIKV mutants (MOI = 0.1). At day 4 pi, viral load was measured by Q-PCR of viral RNA extracted from infected macrophages (e) and FFA (f). At D4 pi, cytokine production was determined by using Q-PCR (g). Data are presented as the fold increase compared with mock-infected (n = 4 per group). Data are presented as means ± SEM and are representative of at least two similar experiments. ^##P < 0.01 or ^#P < 0.05 compared with Δ10 group (unpaired t-test). **P < 0.01 or *P < 0.05 compared with the WT group (unpaired t-test).