Journal
VIRUSES-BASEL
Volume 12, Issue 4, Pages -Publisher
MDPI
DOI: 10.3390/v12040369
Keywords
CRISPR/Cas9; Cre/Lox; pseudorabies virus; vaccine
Categories
Funding
- National Program on Key Research Project of China [2018YFD0500801, 2018YFD0500204]
- Fundamental Research Funds for the Central Universities [2662016PY003]
- Technology Base and Talents Special Program of Guangxi Province [2018AD09007]
- Natural Science Foundation of Hubei Province [2019CFA010]
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Pseudorabies (PR), caused by pseudorabies virus (PRV), is an acute and febrile infectious disease in swine. To eradicate PR, a more efficacious vaccine needs to be developed. Here, the gE/gI- and TK/gE/gI-gene-deleted recombinant PRV (rGX Delta gE/gI and rGX Delta TK/gE/gI) are constructed through CRISPR/Cas9 and Cre/Lox systems. We found that the rGX Delta TK/gE/gI was safer than rGX Delta gE/gI in mice. Additionally, the effects of rGX Delta gE/gI and rGX Delta TK/gE/gI were further evaluated in swine. The rGX Delta gE/gI and rGX Delta TK/gE/gI significantly increased numbers of IFN-gamma-producing CD4+ and CD8+ T-cells in swine, whereas there was no difference between rGX Delta gE/gI and rGX Delta TK/gE/gI. Moreover, rGX Delta gE/gI and rGX Delta TK/gE/gI promoted a PRV-specific humoral immune response. The PRV-specific humoral immune response induced by rGX Delta gE/gI was consistent with that caused by rGX Delta TK/gE/gI. After the challenge, swine vaccinated with rGX Delta gE/gI and rGX Delta TK/gE/gI showed no clinical signs and viral shedding. However, histopathological detection revealed that rGX Delta gE/gI, not rGX Delta TK/gE/gI, caused pathological lesions in brain and lung tissues. In summary, these results demonstrate that the TK/gE/gI-gene-deleted recombinant PRV was safer compared with rGX Delta gE/gI in swine. The data imply that the TK/gE/gI-gene-deleted recombinant PRV may be a more efficacious vaccine candidate for the prevention of PR.
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