Replicon-based Japanese encephalitis virus vaccines elicit immune response in mice

In this study, a replicon vaccine vector system for Japanese encephalitis virus (JEV) was established. The system included a trans-complementing cell line, a series of JEV DNA-based subgenomic replicons, and several encapsidated JEV propagation-deficient pseudoinfectious particles (PIPs). The DNA-based JEV replicon vectors, which deleted the structural coding region, could be able to self-replicate and express the reporter gene. A stable BHK packaging cell line named BHK-CME, which constitutively expressed the capsid protein C, the precursor membrane and envelope proteins (C-prM-E) of JEV, was generated. BHK-CME cells were used to trans-complement the JEV replicons and proved to package the JEV replicons into single-round infectious PIPs efficiently. The PIPs were produced in titers of up to 1.6×10(5)IU/ml. To investigate the efficacy of JEV replicon-based vaccines, four groups of female BALB/c mice were inoculated three times at 3-week intervals with the JEV PIPs and others. The JEV-specific antibody titers reached to 1:6400 and the neutralizing antibody titers reached 1:256 after three rounds of immunization with JEV PIPs. And the antisera collected from immunized mice were shown to be protective partially against lethal infection when passively transferred to susceptible weanling mice. These results demonstrated the value of the JEV replicon vector system for the development of new vaccine candidates.