Evaluation of immunogenicity and protective efficacy of a CpG-adjuvanted DNA vaccine against Tembusu virus

Tembusu virus (TMUV) is a contagious pathogen of waterfowl including ducks and geese, which causes symptoms of high fever, loss of appetite and reduced egg production. The development of an effective vaccine is important for the prevention and control of the disease. We evaluated a DNA vaccine based on a recombinant pre-membrane (prM) and envelope (E) protein, using CpG oligodeoxynucleotide (ODN) as an adjuvanted, and tested it for protection efficacy. BHK21 cells were transfected with the recombinant plasmid pVAX1-prM/E-CpG, and the antigenicity of the expressed protein was detected using an indirect immunofluorescence assay (IFA) and western blot assay. One-day-old ducklings were intramuscularly injected with 200 mu g doses of pVAX1-prM/E-CpG or pVAX1-CpG, or PBS at ten day intervals. The neutralizing antibodies and cell-mediated immune responses elicited by the DNA vaccine were detected using serum neutralization tests (SNTs) and ELISAs At 20 days old, the ducks were challenged with 10(3)EID50 doses of TMUV SD/02 strain and observed for 15 days post challenge. After the second DNA vaccination and during the monitoring period, the levels of TMUV neutralizing antibodies increased in the pVAX1-prM/E-CpG vaccinated ducks. Vaccination with pVAX1-prM/E-CpG resulted in 100.0% protection of the ducks, whereas approximately 40% of ducks vaccinated with pVAX-CpG or PBS manifested clinical symptoms. Expressions of IFN-gamma and IL-6 in the pVAX1-prM/E-CpG group were significantly increased (p < 0.01) compared with the control groups during the entire experimental period. The results revealed that a vaccine co-expressing prM and E, and using a CpG-ODN motif as an adjuvant, could elicit effective neutralizing antibody titers and provide efficient protection to ducks against TMUV infection.