Cyprinid herpesvirus 3; Koi herpesvirus; CyHV-3; KHV; attenuated vaccine; fish; carp; in vivo imaging; cyprinus carpio; mutagenesis; modified-live vaccine; aquaculture
Abstract :
[en] Cyprinid herpesvirus 3 (CyHV 3) is causing severe economic losses worldwide in common and koi carp industries, and a safe and efficacious attenuated vaccine compatible with mass vaccination is needed. We produced single deleted recombinants using prokaryotic mutagenesis. When producing a recombinant lacking open reading frame 134 (ORF134), we unexpectedly obtained a clone with additional deletion of ORF56 and ORF57. This triple deleted recombinant replicated efficiently in vitro and expressed an in vivo safety/efficacy profile compatible with use as an attenuated vaccine. To determine the role of the double ORF56-57 deletion in the phenotype and to improve further the quality of the vaccine candidate, a series of deleted recombinants was produced and tested in vivo. These experiments led to the selection of a double deleted recombinant lacking ORF56 and ORF57 as a vaccine candidate. The safety and efficacy of this strain were studied using an in vivo bioluminescent imaging system (IVIS), qPCR, and histopathological examination, which demonstrated that it enters fish via skin infection similar to the wild type strain. However, compared to the parental wild type strain, the vaccine candidate replicated at lower levels and spread less efficiently to secondary sites of infection. Transmission experiments allowing water contamination with or without additional physical contact between fish demonstrated that the vaccine candidate has a reduced ability to spread from vaccinated fish to naïve sentinel cohabitants. Finally, IVIS analyses demonstrated that the vaccine candidate induces a protective mucosal immune response at the portal of entry. Thus, the present study is the first to report the rational development of a recombinant attenuated vaccine against CyHV 3 for mass vaccination of carp. We also demonstrated the relevance of the CyHV 3 carp model for studying alloherpesvirus transmission and mucosal immunity in teleost skin.
Research center :
Fundamental and Applied Research for Animals & Health
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Boutier, Maxime ; Université de Liège - ULiège > Immunologie et vaccinologie
Ronsmans, Maygane ; Université de Liège - ULiège > Immunologie et vaccinologie
Ouyang, Ping; Sichuan Agricultural University > College of Veterinary Medicine > Key Laboratory of Animal Disease
Fournier, Guillaume
Reschner, Anca
Rakus, Krzysztof ; Université de Liège - ULiège > Immunologie et vaccinologie
Wilkie, Gavin S
Farnir, Frédéric ; Université de Liège - ULiège > Département de productions animales > Biostatistiques et bioinformatique appliquées aux sc. vétér.
Bayrou, Calixte ; Université de Liège - ULiège > Département de morphologie et pathologie > Pathologie spéciale et autopsies
Lieffrig, François
Li, Hong
Desmecht, Daniel ; Université de Liège - ULiège > Département de morphologie et pathologie > Pathologie spéciale et autopsies
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