Construction And Immunogenicity Research Of Recombinant Ebola Virus Glycoprotein GP-Fc

Ebola virus disease(EVD)is a viral hemorrhagic fever of humans and other primates caused by ebolaviruses.Ebolavirus was firstreported in 1976 when hemorrhagic fever outbreaks occurred in Africa.It is one of the deadliest human viruses we have known.In 2014,the largest and longest epidemic ofEbola virus disease(EVD)was outbreak in West Africa.A data published by US CDC shown that 28,652 confirmed andsuspected EVD cases were found across the world,with total mortality up to 40% in this epidemic.EBOVdisease progression is rapidand patients’ body fluids arehighly contagious.Prophylactic method is much more important than clinic.Safeties,effective,economical and suitable for mass production vaccine need to be developed and applied for epidemic prevention.With the development of life science and technology,new generation vaccineis becoming a research hotspot.Such us therecombinant subunit vaccine,DNA vaccine,recombinant live virus vaccine and peptide vaccine.In our research,we design two kinds of recombination EBOV GP protein GP-Fc vaccine,the subunit vaccine GP-Fc and DNA vaccine pVR-modGP-Fc.The sequence in which the Ebola glycoprotein(GP)extracellular domain is modified by the codon frequency of humans and fused with the human IgG Fc code at the C terminal,was synthesized,and inserted into the eukaryotic expression vector pXG and DNA vaccine vector pVR.The recombination protein was expressed by HEK293 cells.The immunogenicity of these two vaccines was evaluated by indirect enzyme-linked immunoassay(ELISA),and immunofluorescences.We also try to construct a pseudotyped EBOV assay system.The protection efficiency ofthose vaccineswas detected by this pEBOV assay system.According toresearch dates,we successfully construct and characterize the pseudotyped EBOV assay system.Vaccinate mice ELISA andimmunofluorescence data suggest that both the recombinat subunit vaccine GP-Fc and DNA vaccine pVRmodGP-Fc can induce BALB/c mice to generate high-efficiency combination IgG.It shows good immunogenicity.Neutralizationexperiment,methodbypEBOV assay system,verified that both of those vaccines can induce neutral effective antibodies production.These results suggest that we obtained two EBOV vaccines which have good immunogenicity.Our study is the basis for developmentof vaccines against the Ebola virus and for screening of neutralizing monoclonal antibodies.