Fig 5. ZIKV infection induces type I IFN transcription but inhibits translation.

moDCs were left untreated (“Mock”), treated with RIG-I agonist (10ng/1e5 cells), or infected with PR-2015, P6-1966, MR-1947, or Dak-1984 at MOI of 1. Supernatants were collected 24hrs (RIG-I agonist treatment) or 48hrs (ZIKV infection) later and IFNβ and IFNα (A) or IFNλ1 (B) production was assessed via multiplex bead array. Values for each individual donor are shown with the mean +/- SD (n = 7 donors). Statistical significance (p< 0.05) was determined using a Friedman test with comparisons made to donor-paired, mock-infected cells. A dashed line indicates the assay limit of detection. (C) moDCs were infected with ZIKV at MOI of 1 in the presence of anti-IFNAR2 blocking antibody. Cells were collected at 48hpi and labeled for ZIKV E protein, while release of infectious virus into the supernatants was determined by FFA. Values for each individual donor are shown with the mean +/- SD (n = 4 donors). A dashed indicates no change relative to infection in the absence of anti-IFNAR2 blocking antibody. (D) RNA was harvested from cells treated the same as for cytokine analysis and IFNB1 mRNA expression was determined by qRT-PCR. Gene expression was normalized to GAPDH transcript levels in each respective sample and represented as the log₂ normalized fold increase above donor- and time point-matched untreated cells. Values for each individual donor are shown with the mean (n = 6–8 donors). (E) moDCs were treated with RIG-I agonist (10ng/1e5 cells, 18hrs) or infected with ZIKV PR-2015 (MOI 1 and 10, 48hrs) and analyzed for IFNB1 mRNA expression. Values for each individual donor are shown with the mean (n = 7 donors) (F) IFNβ and IFNα were measured in the supernatant (“Sup”) and whole cell lysate (“WCL”) of moDCs treated the same as in E. Values for each individual donor are shown with the mean (n = 7 donors). Statistical significance (p< 0.05) was determined using a Friedman test with comparisons made to donor-paired, mock-infected cells. (G) Uninfected or ZIKV PR-2015-infected moDCs (MOI 10, 48hpi) were treated with RIG-I agonist (10ng/1e5 cells, 18hrs) and IFNβ and IFNα were measured as in F. The data is shown as the fold-decrease from RIG-I agonist treatment alone with significance (P<0.05) determined using a Mann Whitney Test (n = 4 donors). Error bars represent the mean +/- SD. See also S4 Fig.