Chimeric pestiviruses: candidates for live-attenuated classical swine fever marker vaccines.

The use of attenuated classical swine fever virus (CSFV) strains as live vaccines is no longer allowed for the control of classical swine fever in Europe, due to the inability to differentiate between infected and vaccinated animals (Differentiating Infected from Vaccinated Animals; DIVA), except as emergency vaccines or as bait vaccines for wild boars. Thus, the establishment of a DIVA vaccine(s) is of pivotal importance for the control of this infectious disease. In this study, recombinant versions of the live-attenuated vaccine strain CSFV Riems were generated by replacing parts of the E2 gene with the corresponding sequence of border disease virus strain Gifhorn. Three cDNA clones were constructed: pRiems-ABC-Gif, pRiems-A-Gif and pRiems-BC-Gif. Infectious particles were obtained from clones pRiems-ABC-Gif and pRiems-BC-Gif only, whereas transfected RNA from clone pRiems-A-Gif behaved like a replicon. Based on its ability to be differentiated in vitro from wild-type CSFV by mAbs, vRiems-ABC-Gif was assessed for immunogenicity and protection against challenge infection in pigs. Before challenge, no CSFV-specific anti-E2 antibodies could be detected with commercial E2-blocking ELISAs in vRiems-ABC-Gif-vaccinated animals, whereas vRiems-vaccinated pigs developed high titres of anti-E2 antibodies, confirming the marker properties of this vaccine candidate. After oral vaccination, only partial protection against challenge infection was observed in the vRiems-ABC-Gif vaccinees, whereas all intramuscularly vaccinated animals and all vRiems-vaccinated animals were fully protected. These experiments suggest that the strategy of exchanging specific antigenic epitopes among pestiviruses is a promising tool for the development of new CSFV marker vaccines.

Border disease virus: delineation by monoclonal antibodies.

Many ovine pestiviruses from Britain and a number of atypical porcine isolates are largely unrecognised by monoclonal antibodies (mAbs) specific for reference strains of classical swine fever virus and bovine viral diarrhoea virus (BVDV). Additional mAbs have therefore been produced using some of these "unreactive" pestiviruses. Two of the viruses used were atypical porcine isolates (strains 87/6 and Vosges), whilst another had been isolated from a sheep (59386). Thirty-three mAbs were selected, none of which recognised two reference strains of BVDV, but three of which recognised the Alfort strain of classical swine fever. On the basis of radioimmunoprecipitation they were considered to be directed at one of three different pestivirus proteins (gp 53, gp 48 or p 125). Three virus subgroups were evident when the mAbs were used to type 16 ovine and two atypical porcine pestiviruses. One subgroup contained the Vosges and 59386 viruses and four ovine field isolates. The second subgroup comprised the 87/6 virus, the Moredun and Aveyron reference strains of border disease virus and four further ovine field isolates. Three of four ovine viruses making up the third subgroup had been previously categorised as BVDV-like and were largely unrecognised by the new mAbs. The findings were in agreement with previous attempts to segregate some of the same viruses using partial genomic comparisons or cross-neutralization tests.