Abstract
BackgroundPorcine deltacoronavirus (PDCoV) is a new pathogenic porcine intestinal coronavirus, which has appeared in many countries since 2012. PDCoV disease caused acute diarrhea, vomiting, dehydration and death in piglets, resulted in significant economic loss to the pig industry. However, there is no commercially available vaccine for PDCoV. In this study, we constructed recombinant pseudorabies virus (rPRVXJ-delgE/gI/TK-S) expressing PDCoV spike (S) protein and evaluated its safety and immunogenicity in mice.ResultsThe recombinant strain rPRVXJ-delgE/gI/TK-S obtained by CRISPR/Cas gE gene editing technology and homologous recombination technology has genetic stability in baby hamster syrian kidney-21 (BHK-21) cells and is safe to mice. After immunizing mice with rPRVXJ-delgE/gI/TK-S, the expression levels of IFN-γ and IL-4 in peripheral blood of mice were up-regulated, the proliferation of spleen-specific T lymphocytes and the percentage of CD4+ and CD8+ lymphocytes in mice spleen was increased. rPRVXJ-delgE/gI/TK-S showed good immunogenicity for mice. On the seventh day after booster immunity, PRV gB and PDCoV S specific antibodies were detected in mice, and the antibody level continued to increase, and the neutralizing antibody level reached the maximum at 28 days post- immunization (dpi). The recombinant strain can protect mice with 100% from the challenge of virulent strain (PRV XJ) and accelerate the detoxification of PDCoV in mice.ConclusionThe recombinant rPRVXJ-delgE/gI/TK-S strain is safe and effective with strong immunogenicity and is expected to be a candidate vaccine against PDCoV and PRV.
Highlights
Porcine deltacoronavirus (PDCoV) is a new pathogenic porcine intestinal coronavirus, which has appeared in many countries since 2012
Construction and verification of recombinant virus rPRVXJ‐delgE/gI/TK‐S The recombinant pseudorabies virus rPRVXJ-delgE/gI/ TK-S expressing PDCoV S protein was constructed by CRISPR/cas9 technology and homologous recombination technology (Fig. 1A)
Under the inverted microscope (Nikon, Japan), it can be observed that the green fluorescent labeled protein EGFP is expressed in baby hamster syrian kidney-21 (BHK-21) cells 24 h after transfection (Fig. 1B), the cytopathic effect (CPE) caused by rPRVXJ-delgE/gI/TK-S (Fig. 1C), and the purified recombinant virus (Fig. 1D)
Summary
Porcine deltacoronavirus (PDCoV) is a new pathogenic porcine intestinal coronavirus, which has appeared in many countries since 2012. PDCoV disease caused acute diarrhea, vomiting, dehydration and death in piglets, resulted in significant economic loss to the pig industry. There is no commercially available vaccine for PDCoV. Porcine deltacoronavirus (PDCoV) belongs to the family of coronaviruses. PDCoV was first detected in Hong Kong, China, in 2012 [3] (but not successfully isolated), and subsequently detected in pig farms in the USA, Canada, Korea, China, Thailand, Laos, and Vietnam. PDCoV presents a global distribution trend and is a common pathogen of porcine disease worldwide [4,5,6]. Typical clinical symptoms of PDCoV infection include diarrhea, dehydration, variable vomiting and mortality in nursing piglets. Similar to that of PEDV and TGEV, the strongest tissue tropism of PDCoV is in villous enterocytes of the small and large
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