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FP5

EFFICACIOUS DELIVERY Report Summary

Project ID: QLK2-CT-2001-01346
Funded under: FP5-LIFE QUALITY
Country: Germany

DNA vaccine constructs encoding CSFV E2 protein

Prototype novel DNA vaccines against CSFV were generated. The eukaryotic expression vectors encoded the viral structural protein E2 and the non-structural protein NS3 (pcDNA4-E2/NS3). These expression vectors were supplemented with DNAs encoding the interleukins IL-12, IL-18 and the CD40 ligand for improvement in efficacy tests.

In classical prime/boost vaccination, CSFV-E2 and/or -NS3 encoding plasmid DNA was only partially protective in stringently lethal challenge experiments with swine. This contrasted with the E2 expressing ORFV vector vaccine, which mediated solid protection and prevented CSFV shedding. Additional vaccination/challenge experiments tested the efficacy of DNA-mediated co-delivery of cytokines and CD154. These showed that co-delivery of IL-18 and CD154 induced an earlier appearance of serum antibodies, reduced B-cell deficiency after infection, and protected pigs against a lethal CSFV challenge.

In contrast, co-delivery of IL-12 led to a reduced titre of neutralizing antibodies and protection, in comparison to pigs immunized with an E2 encoding plasmid DNA alone. For the required rapid efficacy of an intervention vaccination, a prime and multiple boost regime with the DNA vaccines achieved protection.

Consequently, the consortium concludes that DNA vaccination against CSFV requires two booster applications for protection of swine and induction of CSFV-neutralizing antibodies. When piglets are immunized with the pcDNA4-E2-IL18 and pcDNA4-E2-CD40L plasmids, they are protected against lethal CSFV challenge and don�t show clinical signs.

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