Abstract
BackgroundRaccoon dog parvovirus (RDPV) causes acute infectious diseases in raccoon dogs and may cause death in severe cases. The current treatment strategy relies on the extensive usage of classical inactivated vaccine which is marred by large doses, short immunization cycles and safety concerns.MethodsThe present study aimed at optimization of RDPV VP2 gene, subcloning the gene into plasmid pET30a, and its subsequent transfer to Escherichia coli with trigger factor 16 for co-expression. The protein thus expressed was purified with ammonium sulfate precipitation, hydrophobic chromatography, and endotoxin extraction procedures. VLPs were examined by transmission electron microscopy, dynamic light scattering, and the efficacy of VLPs vaccine was tested in vivo.ResultsResults indicated that RDPV VP2 protein could be expressed soluble. Transmission electron microscopy and dynamic light scattering results indicated that RDPV VP2 self-assembled into VLPs. Hemagglutination inhibition antibody titers elicited by Al(OH)3 adjuvanted RDPV VLPs were comparable with RDPV inactivated vaccines, and the viral loads in the blood of the struck raccoon dogs were greatly reduced. Hematoxylin and eosin and Immunohistochemical results indicated that RDPV VLPs vaccine could protect raccoon dogs against RDPV infections.ConclusionsThese results suggest that RDPV VLPs can become a potential vaccine candidate for RDPV therapy.
Highlights
Raccoon dog parvovirus (RDPV) is responsible for causing an acute, highly contagious, infectious viral disease, which may lead to fatal hemorrhagic enteritis in raccoon dogs of all ages [24, 30]
RDPV VP2 protein expression and co‐expression Results showed that a 65 kDa VP2 protein was expressed within the inclusion bodies of induced pET30a-VP2 cells (Fig. 1a)
The pET30a-VP2 and Tf16 were transferred to E. coli ER2566, where the soluble VP2 protein and protein Tf16 (56 kDa), were co-expressed (Fig. 1b)
Summary
Raccoon dog parvovirus (RDPV) is responsible for causing an acute, highly contagious, infectious viral disease, which may lead to fatal hemorrhagic enteritis in raccoon dogs of all ages [24, 30]. RDPV is an icosahedral non-enveloped virus belonging to the Parvoviridae family. It consists of a single strand of 20–25 nm DNA, two non-capsid proteins NS1, NS2, and three capsid proteins VP1, VP2 and VP3 [27]. Raccoon dog parvovirus (RDPV) causes acute infectious diseases in raccoon dogs and may cause death in severe cases. The current treatment strategy relies on the extensive usage of classical inactivated vaccine which is marred by large doses, short immunization cycles and safety concerns
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