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 (rGXΔgE/gI and rGXΔTK/gE/gI) are constructed through CRISPR/Cas9 and Cre/Lox systems. We found that the rGXΔTK/gE/gI was safer than rGXΔgE/gI in mice. Additionally, the effects of rGXΔgE/gI and rGXΔTK/gE/gI were further evaluated in swine. The rGXΔgE/gI and rGXΔTK/gE/gI significantly increased numbers of IFN-γ-producing CD4+ and CD8+ T-cells in swine, whereas there was no difference between rGXΔgE/gI and rGXΔTK/gE/gI. Moreover, rGXΔgE/gI and rGXΔTK/gE/gI promoted a PRV-specific humoral immune response. The PRV-specific humoral immune response induced by rGXΔgE/gI was consistent with that caused by rGXΔTK/gE/gI. After the challenge, swine vaccinated with rGXΔgE/gI and rGXΔTK/gE/gI showed no clinical signs and viral shedding. However, histopathological detection revealed that rGXΔgE/gI, not rGXΔTK/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 rGXΔgE/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.

中文翻译：

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CRISPR / Cas9和Cre / Lox系统介导的gE / gI和TK / gE / gI基因缺失的伪狂犬病病毒疫苗的比较。

伪狂犬病病毒（PRV）引起的伪狂犬病（PR）是猪的一种急性发热性传染病。为了消除PR，需要开发更有效的疫苗。在这里，通过CRISPR / Cas9和Cre / Lox系统构建了缺失gE / gI和TK / gE / gI基因的重组PRV（rGXΔgE/ gI和rGXΔTK/ gE / gI）。我们发现在小鼠中，rGXΔTK/ gE / gI比rGXΔgE/ gI更安全。另外，在猪中进一步评估了rGXΔgE/ gI和rGXΔTK/ gE / gI的作用。rGXΔgE/ gI和rGXΔTK/ gE / gI显着增加了猪中产生IFN-γ的CD4 +和CD8 + T细胞的数量，而rGXΔgE/ gI和rGXΔTK/ gE / gI之间没有差异。此外，rGXΔgE/ gI和rGXΔTK/ gE / gI促进了PRV特异性体液免疫反应。rGXΔgE/ gI诱导的PRV特异性体液免疫反应与rGXΔTK/ gE / gI引起的反应一致。攻击后，接种rGXΔgE/ gI和rGXΔTK/ gE / gI的猪没有临床症状和病毒脱落。但是，组织病理学检测显示，rGXΔgE/ gI而非rGXΔTK/ gE / gI引起脑和肺组织的病理损伤。总之，这些结果表明，与猪中的rGXΔgE/ gI相比，TK / gE / gI基因缺失的重组PRV更安全。数据表明，TK / gE / gI基因缺失的重组PRV可能是预防PR更有效的候选疫苗。这些结果表明，与猪中的rGXΔgE/ gI相比，TK / gE / gI基因缺失的重组PRV更安全。数据表明，TK / gE / gI基因缺失的重组PRV可能是预防PR更有效的候选疫苗。这些结果表明，与猪中的rGXΔgE/ gI相比，TK / gE / gI基因缺失的重组PRV更安全。数据表明，TK / gE / gI基因缺失的重组PRV可能是预防PR更有效的候选疫苗。