Abstract
Rabbit haemorrhagic disease virus (RHDV) type 2 (GI.2/RHDV2/b) is an emerging pathogen in wild rabbits and in domestic rabbits vaccinated against RHDV (GI.1). Here we report the genome sequence of a contemporary RHDV2 isolate from the Netherlands and investigate the immunogenicity of virus-like particles (VLPs) produced in insect cells. RHDV2 RNA was isolated from the liver of a naturally infected wild rabbit and the complete viral genome sequence was assembled from sequenced RT-PCR products. Phylogenetic analysis based on the VP60 capsid gene demonstrated that the RHDV2 NL2016 isolate clustered with other contemporary RHDV2 strains. The VP60 gene was cloned in a baculovirus expression vector to produce VLPs in Sf9 insect cells. Density-gradient purified RHDV2 VLPs were visualized by transmission electron microscopy as spherical particles of around 30 nm in diameter with a morphology resembling authentic RHDV. Immunization of rabbits with RHDV2 VLPs resulted in high production of serum antibodies against VP60, and the production of cytokines (IFN-γ and IL-4) was significantly elevated in the immunized rabbits compared to the control group. The results demonstrate that the recombinant RHDV2 VLPs are highly immunogenic and may find applications in serological detection assays and might be further developed as a vaccine candidate to protect domestic rabbits against RHDV2 infection.
Highlights
Rabbit haemorrhagic disease virus (RHDV, family Caliciviridae, genus Lagovirus) was first reported in 1984 in China as the causal agent of Rabbit Haemorrhagic Disease (RHD), an acute and highly lethal disease in wild and domestic rabbits (Oryctolagus cuniculus)
The observed per acute hepatic necrosis, splenic necrosis and acute pneumonia are fatal lesions consistent with RHDV type 2 (RHDV2) infection, which was confirmed by PCR analysis
Since the first discovery of RHDV2 in 2010 in RHDV-vaccinated rabbits, RHDV2 has spread through wild the first discovery and of RHDV2 in 2010 in RHDV-vaccinated rabbits, RHDV2 has spread wild rabbit populations has replaced in several countries
Summary
Rabbit haemorrhagic disease virus (RHDV, family Caliciviridae, genus Lagovirus) was first reported in 1984 in China as the causal agent of Rabbit Haemorrhagic Disease (RHD), an acute and highly lethal disease in wild and domestic rabbits (Oryctolagus cuniculus). Even after large scale vaccination numerous cases of RHD remained to be reported, e.g., in 2010 in north-western France [3]. This disease affected wild populations, and RHDV-vaccinated rabbits, suggesting the emergence of a novel RHDV serotype. RHDV2 (RHDVb/GI.2) has been reported in France [3], Italy [7], the Iberian Peninsula [8,9], Sardinia [10], the United Kingdom [11], Madeira [12], the Canary
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