A Single siRNA Suppresses Fatal Encephalitis Induced by Two Different Flaviviruses (original) (raw)
Figure 2
shFvE J Protects Mice against JEV-Induced Encephalitis
(A) BALB/c mice (ten per group) were injected IC on days −4, −2, and 0 with 2 × 105 TU of either shFvEJ or shLuc lentiviruses. On day 0, 30 min after injection of the third dose of lentivirus, they were injected at the same spot IC with four LD50 of JEV, and the mice were monitored for survival over time.
(B) Brain sections from shFvEJ treated mice reveal no flavivirus-induced pathology. Representative photomicrographs of hematoxylin and eosin-stained horizontal brain sections obtained from mice treated with shFvEJ or shLuc lentivirus and infected with JEV for 5 d are shown at indicated magnifications.
(C) Lack of infectious virus in the brains of shFvEJ-treated mice. Mice were injected with lentiviruses and challenged with JEV as in (A), and their brain homogenates, obtained 5 d after JEV challenge, were plaque-titrated on BHK21 cell monolayers. For shFvEJ lentivirus, viral titers after a single (1×) as well as three (3×) administrations are shown. The viral titers are shown as log plaque-forming units per total brain. Each symbol represents an individual mouse.
(D) Brains from shFvEJ treated mice are free of infectious virus. Brain homogenates in (C) were pooled, 1, 10, or 50 μl of pooled homogenate were inoculated onto Neuro 2a cells, and the viral replication was monitored by flow cytometry 5 d later.
(E) A single IC injection with shFvEJ is sufficient for protection against JE encephalitis. Mice (five per group) were injected IC with 2 × 105 TU of shLuc or shFvEJ, challenged 30 min later with indicated doses of JEV, and observed for survival over time.