| Ebola hemorrhagic fever(EHF) is an acute hemorrhagic infectious diseases in humans and nonhuman primates which is caused by Ebola virus(EBOV). EBOV is a member of the genus ebolavirus in the Filoviridae family, and a RNA virus that contains a single-stranded, negative-sense and non-segmented genome. The glycoprotein(GP) is the major protein inducing neutralizing antibodies and protective immunity in host and the only transmembrane protein of Ebola virus. Currently, there are five Ebola virus types to be known. From 1976 to 2014, Ebola virus has caused several huge disasters in the history. The virulent of Zaire ebolavirus(ZEBOV) is higher than the other four types with high mortality rates up to 90% in humans and nonhuman primates, while the virulence of Reston ebolavirus(REBOV) is relatively low, it can cause cynomolgus monkey infected and dead, but there is none case about REBOV cause human dead for now. In 1989, REBOV was first identified during the investigation of fatal hemorrhagic fever involving cynomolgus macaques in a research facility in Reston, Virgina that had been imported from the Philippines. In 2008, several pigs that died after showing typical clinical signs of porcine reproductive and respiratory syndrome were found to be positive for REBOV in Philippines, while there is still no vaccine available against REBOV so far, therefore it is very important to produce an effective and safe vaccine against REBOV.VSV as an important model of RNA virus has been widely used in the research of replication and assembly in enveloped RNA virus. The length of genome is 11 kb and contains five structural proteins.The maximum 4.5kb of foreign gene can be cloned into VSV genome, and the recombinant virus can highly express in infected cells.In this study, using a recombinant VSV expressed enhanced green fluorescent protein as vector, we replaced VSV G with REBOV GP by reverse genetics techniques, and the rescued recombinant VSV named rVSV?G*EGFP-REBOV-GP. By immunofluorescence and western blot, the expression of REBOV GP in recombinant virus was confirmed. The growth pattern of rVSV?G*EGFP-REBOV-GP is different from its parent virus in cells. The body weight of mice inoculated with rVSV?G*EGFP-REBOV-GP in 5×106TCID50/0.1mL dose showed no different with that of mice inoculated with PBS. The blood of mice inoculated with rVSV?G*EGFP-REBOV-GP in1×106TCID50/0.1mL dose is used in the neutralization tests after prime and boost inoculation. The result of neutralization tests showed that rVSV?G*EGFP-REBOV-GP could induce high titer neutralizing antibodies. Our data indicated that rVSV?G*EGFP-REBOV-GP has the potential to be a promise vaccine protected animals from REBOV infection. |
