The Construction and Immunogenicity Analyses of Recombinant Pseudorabies Virus With NADC30-Like Porcine Reproductive and Respiratory Syndrome Virus-Like Particles Co-expression

Abstract

Porcine reproductive and respiratory syndrome (PRRS) and pseudorabies (PR) are highly infectious swine diseases and cause significant financial loss in China. The respiratory system and reproductive system are the main target systems. Previous studies showed that the existing PR virus (PRV) and PRRS virus (PRRSV) commercial vaccines could not provide complete protection against PRV variant strains and NADC30-like PRRSV strains in China. In this study, the PRV variant strain XJ and NADC30-like PRRSV strain CHSCDJY-2019 are used as the parent for constructing a recombinant pseudorabies virus (rPRV)-NC56 with gE/gI/TK gene deletion and co-expressing NADC30-like PRRSV GP5 and M protein. The rPRV-NC56 proliferated stably in BHK-21 cells, and it could stably express GP5 and M protein. Due to the introduction of the self-cleaving 2A peptide, GP5 and M protein were able to express independently and form virus-like particles (VLPs) of PRRSV in rPRV-NC56-infected BHK-21 cells. The rPRV-NC56 is safe for use in mice; it can colonize and express the target protein in mouse lungs for a long time. Vaccination with rPRV-NC56 induces PRV and NADC30-like PRRSV specific humoral and cellular immune responses in mice, and protects 100% of mice from virulent PRV XJ strain. Furthermore, the virus-neutralizing antibody (VNA) elicited by rPRV-NC56 showed significantly lower titer against SCNJ-2016 (HP-PRRSV) than that against CHSCDJY-2019 (NADC30-like PRRSV). Thus, rPRV-NC56 appears to be a promising candidate vaccine against NADC30-like PRRSV and PRV for the control and eradication of the variant PRV and NADC30-like PRRSV.

The Construction and Immunogenicity Analyses of a Recombinant Pseudorabies Virus with Senecavirus A VP2 Protein Coexpression

“…2E ). Transmission electron microscopy was used to observe the rPRV-XJ-ΔTK/gE/gI-VP2 virion, which was similar to rPRV-XJ-ΔTK/gE/gI ( 21 ), and no significant difference was observed compared with PRV-XJ ( Fig. 2F ).…”

“…Due to its high capacity, PRV has become a powerful vector system for expressing exogenous proteins. It can express the main immunogenic protein of other pathogens, such as the Cap protein of porcine circovirus (PCV) ( 17 ), VP2 of porcine parvovirus (PPV) ( 18 ), CD2v of African swine fever virus (ASFV) ( 19 ), S of porcine deltacoronavirus (PDCoV) ( 20 ), and GP5 and M of porcine reproductive and respiratory syndrome virus (PRRSV) ( 21 ).…”

Recombinant Pseudorabies Virus Usage in Vaccine Development against Swine Infectious Disease

“…Consequently, the PRV variant has been used to generate several gene mutant vaccines with gE, gI, and TK deletions. When challenged with PRV variants, these mutants provided adequate protection [ 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 ]. By introducing foreign genes derived from other porcine pathogens into the PRV variants and subsequent vaccination, it is possible to establish immune protection against these swine infectious diseases.…”

Progress on innate immune evasion and live attenuated vaccine of pseudorabies virus

“…Several PRV-based vector vaccines have been developed. E.g., co-expressing the PrM-E of Japanese encephalitis virus (JEVs; Qian et al, 2015), E2 of classical swine fever virus (CSFVs; Lei et al, 2016;Tong et al, 2020), FMDV P12A and 3C (Li et al, 2008;Zhang et al, 2011), PPV VP2 (Chen et al, 2011), Porcine deltacoronavirus (PDCoV) spike (S; Huang et al, 2022), PRRSV GP5 and M (Zhao J. et al, 2022), CP204L (p30), CP530R (pp62), E183L (p54), B646L (p72), and EP402R (CD2v) genes of ASFV and SARS-CoV-2 S or N . These multivalent vaccines have demonstrated preventive effects against many diseases, but further research is needed on their safety and efficacy in pig.…”