Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
6133041 | Journal of Virological Methods | 2015 | 6 Pages |
Abstract
Peste des petits ruminants (PPR), a highly contagious disease induced by PPR virus (PPRV), affects sheep and goats. PPRV fusion (F) protein is important for the induction of immune responses against PPRV. We constructed a Semliki Forest virus (SFV) replicon-vectored DNA vaccine (“suicidal DNA vaccine”) and evaluated its immunogenicity in BALB/c mice. The F gene of PPRV was cloned and inserted into the SFV replicon-based vector pSCA1. The antigenicity of the resultant plasmid pSCA1/F was identified by indirect immunofluorescence and western blotting. BALB/c mice were then intramuscularly injected with pSCA1/F three times at 14-d intervals. Specific antibodies and virus-neutralizing antibodies against PPRV were quantified by indirect ELISA and microneutralization tests, respectively. Cell-mediated immune responses were examined by cytokine and lymphocyte proliferation assays. The pSCA1/F expressed F protein in vitro and induced specific and neutralizing antibody production, and lymphocyte proliferation in mice. Mice vaccinated with pSCA1/F had increased IL-2 and IL-10 levels after 24-h post first immunization. IFN-γ and TNF-α levels increased from that time point and gradually decreased thereafter. Thus, the Semliki Forest virus replicon-vectored DNA vaccine expressing the F protein of PPRV induced both humoral and cell-mediated immune responses in mice. This could be considered as a novel strategy for vaccine development against PPR.
Keywords
PPROIETCID50World Organization for Animal HealthIFASFVPPRVGMTSuicidal DNA vaccinekDaHRPCPEDMEMDABVNA3,3′-diaminobenzidine tetrahydrochlorideDulbecco's modified Eagle's mediumPeste des petits ruminants virusPeste des petits ruminantsindirect immunofluorescence assayVirus neutralizing antibodyCytopathic effectbaby hamster kidneydifferentiating infected from vaccinated animalsDIVAVero cellafrican green monkey kidney cellsstimulation indexGeometric mean titerhemagglutininSemliki Forest virusImmune responseHorseradish peroxidaseF proteinFusion proteinKilo Dalton
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Authors
Yong Wang, Xiaolin Yue, Hongyan Jin, Guangqing Liu, Ling Pan, Guijun Wang, Hao Guo, Gang Li, Yongdong Li,