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Cyprinid herpesvirus 2 (CyHV-2) is the causative pathogen of herpesviral haematopoietic necrosis disease, which has caused huge economic losses to aquaculture industry in China. In this study, nine truncated CyHV-2 membrane glycoproteins (ORF25, ORF25C, ORF25D, ORF30, ORF124, ORF131, ORF136, ORF142A, ORF146) and a GFP reporter protein were respectively expressed using baculovirus surface displaying system. Western blot showed that the proteins were successfully packaged in the recombinant virus particles. In baculovirus transduced gibel carp kidney cells, the target proteins were expressed and displayed on the fish cell surface. Healthy gibel carp were immunized by immersion with the recombinant baculoviruses and the fish treated with phosphate-buffered saline (PBS) were served as mock group. The expression of interleukin-11 (IL-11), interferon α (IFNα) and a complement component gene C3 were significantly up-regulated in most experimental groups, and interferon γ (IFNγ) expression in some groups were also induced after immunization. Subsequently, the immunized gibel carp were challenged by intraperitoneal injection of CyHV-2 virus. All the immunized groups exhibited reduced mortality after CyHV-2 challenge. In the groups immunized with baculoviruses displaying and expressing ORF25, ORF25C and ORF146, the relative percentage survival values reached 83.3%, 87.5% and 70.8%, respectively. Our data suggested that baculovirus-displayed ORF25, ORF25C and ORF146 could be potential vaccine candidates for the prevention of CyHV-2 infection in gibel carp. • Nine CyHV-2 glycoproteins were expressed by baculovirus surface displaying system. • The recombinant baculoviruses can transduce GiCK cells and express the antigens. • Gibel carp were immunized by immersion with the recombinant baculoviruses. • Expression of immune genes was activated after immunization. • ORF25, ORF25C and ORF146 greatly reduced the mortality caused by CyHV-2 infection. [ABSTRACT FROM AUTHOR]

Highlights: [•] VRC01 and b12 were expressed in Sf9 cells by baculoviruses co-infection. [•] Purified b12Bac and VRC01Bac were fully assembled by disulphide bonds. [•] The antigen reactivities of b12Bac and VRC01Bac were comparable to b12Mam. [Copyright &y& Elsevier]