Towards licensing of CP7_E2alf as marker vaccine against classical swine fever-Duration of immunity.

Classical swine fever (CSF) marker vaccine candidate CP7_E2alf was tested in a "duration of immunity" trial according to the World Organisation for Animal Heath (OIE) guidelines. To this means, 15 weaner pigs were either orally or intramuscularly vaccinated with a single dose of CP7_E2alf vaccine produced under Good Laboratory Practice (GLP) conditions. Ten additional pigs were included as controls. Six months later, all animals were oronasally challenged with highly virulent CSF virus (CSFV) strain "Koslov". Upon vaccination, all but one orally and all intramuscularly vaccinated pigs developed rising and later on stable CSFV glycoprotein E2-specific antibodies. In contrast, no CSFV E(rns)-specific "marker" antibodies were detectable prior to challenge infection. None of the co-housed control animals seroconverted. Upon challenge infection, all seropositive animals were protected from lethal challenge, whereas all control animals and the non-responder developed severe signs of CSF. One control animal recovered, the others had to be euthanised due to animal welfare reasons between days 4 and 7 post challenge infection. All protected animals showed quickly rising neutralizing antibodies reaching high titres by the end of the trial. At the end of the trial, the marker ELISA was positive for most challenged animals that survived the CSFV infection (27 out of 30). Using reverse transcription polymerase chain reaction, low level genome detection was seen in all vaccinated animals between days 4 and 10 post challenge infection, but no virus could be isolated from any samples of these animals. The OIE guidelines require seroconversion in at least 8 out of 10 vaccinated animals. This requirement was fulfilled. Moreover, only control animals should die. With this requirement, only the intramuscular vaccination fully complied as one orally vaccinated pig did not respond. Concluding, CP7_E2alf induced stable antibodies that led to protection from lethal challenge with highly virulent CSFV strain "Koslov" six months after vaccination, with the exception of one non-responder after oral vaccination.