The VP2 gene of infectious pancreatic necrosis virus, encoded in an expression plasmid and encapsulated in alginate microspheres, was used for oral DNA vaccination of fish to better understand the carrier state and the action of the vaccine. The efficacy of the vaccine was evaluated by measuring the prevention of virus persistence in the vaccinated fish that survived after waterborne virus challenge. A real-time RT-qPCR analysis revealed lower levels of IPNV-VP4 transcripts in rainbow trout survivors among vaccinated and challenged fish compared with the control virus group at 45 days post-infection. The infective virus was recovered from asymptomatic virus control fish, but not from the vaccinated survivor fish, suggesting an active role of the vaccine in the control of IPNV infection. Moreover, the levels of IPNV and immune-related gene expression were quantified in fish showing clinical infection as well as in asymptomatic rainbow trout survivors. The vaccine mimicked the action of the virus, although stronger expression of immune-related genes, except for IFN-1 and IL12, was detected in survivors from the virus control (carrier) group than in those from the vaccinated group. The transcriptional levels of the examined genes also showed significant differences in the virus control fish at 10 and 45 days post-challenge.
This study was supported by the Spanish Ministerio de Economia y Competitividad (MINECO: grant AGL2010-18454). The authors declare no competing interests. The authors thank Dr. J. Coll (INIA, Madrid, Spain) for the gift of the mAb 2F12 monoclonal antibody and Dr. P. Lastres for valuable help with the flow cytometry analyses. The authors acknowledge the excellent technical assistance of M. Sánchez and L. Guaita. N. Ballesteros, Universidad Complutense de Madrid PhD student, was the recipient of a pre-doctoral fellowship from MINECO.