Attenuated porcine-derived type 2 bovine viral diarrhea virus as vector stably expressing viral gene

https://doi.org/10.1016/j.jviromet.2020.113842Get rights and content

Highlights

  • Truncated PEDV S antigen was inserted between the seventh and eighth amino acids of the capsid gene in the BVDV genome by homologous recombination.

  • Exogenous PEDV S antigen could be expressed successfully in recombinant BVDVs.

  • The recombinant BVDVs has the same growth curve as the parental BVDV strain.

  • It was first reported that the N-terminus of capsid gene might be an feasible site to accommodate the insertion of foreign genes.

Abstract

Infectious bovine viral diarrhea virus (BVDV) cDNA clones have been used for the expression of classical swine fever virus (CSFV) genes for immune prevention and control. However, can it be used for the expression of an allogenetic fragment? To answer this question, a BVDV chimeric virus expressing the spike (S) antigen fragment of porcine epidemic diarrhea virus (PEDV) was constructed. Antigen S499-602 was inserted into pig-derived BVDV-2 infectious cDNA clone pASH28 in tandem by overlapping PCR, located between the seventh and eighth amino acids at the N-terminus of the capsid (C) protein of BVDV. Indirect immunofluorescence assay confirmed that the chimeric virus vASH-dS312 containing double S499-602 sequences was successfully assembled, which could react with the monoclonal antibody (MAb) against BVDV E2 and PEDV S proteins. Further western blot analysis confirmed that the exogenous S499-602 double protein could be stably expressed. Next, the chimeric virus vASH-dS312 was administered to BALB/C mice either orally or by intramuscular injection. The immunized mice were healthy and showed no signs of toxicity. IgG against BVDV and PEDV antibodies could be detected in the mice administered vASH-dS312 by intramuscular injection, which had neutralization activity against BVDV and PEDV. Thus, this study reported a new insertion site in the BVDV infectious cDNA clone that could successfully express an allogenetic antigen.

Section snippets

Funding

This work was supported by the National Key Research and Development Program of China (Grant No. 2017YFD0501102) and the Shanghai Agriculture Applied Technology Development Program, China (Grant No. T20170111).

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. Furthermore, this article does not contain any experiments with human subjects or animals performed by any of the authors.

Declaration of Competing Interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

References (21)

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    Intranasal immunization of rhesus macaques with VSVΔG-MERS S particles induced robust neutralizing antibody and T-cell responses. Related to animal coronaviruses, the bovine viral diarrhea virus (BVDV), belonging to flaviviruses, has been engineered for the expression of a fragment of the spike antigen of the porcine epidemic diarrhea virus (PEDV) (Tao et al., 2020). The chimeric BVDV-PEDV S vector elicited BVDV- and PEDV-specific antibody responses in immunized BALB/c mice.

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