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缺失 gI/gE 基因并插入 gC 基因的伪狂犬病病毒在小鼠和断奶仔猪中可引发更好的免疫效果。

Better immune efficacy triggered by the inactivated gI/gE-deleted pseudorabies virus with the additional insertion of gC gene in mice and weaned pigs.

机构信息

Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510642, PR China.

Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510642, PR China.

出版信息

Virus Res. 2021 Apr 15;296:198353. doi: 10.1016/j.virusres.2021.198353. Epub 2021 Feb 25.

Abstract

A new variant of pseudorabies virus (PRV) with high pathogenicity has been prevalent in many swineherds vaccinated with Bartha-K61 in China since 2011. Several gene-deleted vaccine candidates have been developed based on new emerging PRV variants. PRV-AH, a new emerging PRV strain from Anhui Province, was isolated in our laboratory in 2013. In the present study, rPRV-AH-gI/gE and rPRV-AH-gI/gE/gC were generated based on PRV-AH by homologous recombination. The growth kinetics of rPRV-AH-gI/gE and rPRV-AH-gI/gE/gC were similar to their parental strains. Compared with the commercial inactivated vaccine of Ea strain, the immune efficacy of the inactivated vaccine based on recombinant viruses was evaluated in mice and weaned pigs. The result showed that the level of neutralizing antibody in mice immunized with rPRV-AH-gI/gE/gC was higher compared with those immunized with rPRV-AH-gI/gE- at a dose of 10 TCID at 8 weeks post initial immunization (p < 0.0001). Among the groups immunized at a dose of 10 TCID, the rPRV-AH-gI/gE group showed a survival rate of 37.5 %, while the rPRV-AH-gI/gE/gC group showed a protection rate of 87.5 % against the PRV-AH challenge. Besides, the rPRV-AH-gI/gE and rPRV-AH-gI/gE/gC group immunized at a dose of 10 TCID showed a survival rate of 100 %. Interestingly, compared with the commercial vaccine group, the group of 10 TCID rPRV-AH-gI/gE/gC showed a lower level of neutralizing antibodies (p < 0.0001) but the same protection rate in mice. Moreover, in the pig experiment, the level of neutralizing antibodies in the group vaccinated with inactivated rPRV-AH-gI/gE/gC was higher than any other groups at 8 weeks post initial immunization (p < 0.05). More importantly, the milder symptoms and pathological lesions occurred in pigs vaccinated with rPRV-AH-gI/gE/gC after challenge with 10 TCID PRV-AH, revealing that additional insertion of gC gene could enhance the protective efficacy in PRV gI/gE-deleted vaccine in pigs. Collectively, these above-mentioned findings suggested that the inactivated vaccine of rPRV-AH-gI/gE/gC had a better immune efficacy, which could be regarded as a promising inactivated vaccine candidate for PRV control.

摘要

自 2011 年以来,在中国,许多接种了巴氏 -K61 的猪群中出现了一种具有高致病性的伪狂犬病病毒(PRV)新变体。已经基于新出现的 PRV 变体开发了几种基因缺失疫苗候选物。PRV-AH 是 2013 年我们实验室从安徽省分离出来的一种新出现的 PRV 株。在本研究中,通过同源重组,基于 PRV-AH 生成了 rPRV-AH-gI/gE 和 rPRV-AH-gI/gE/gC。rPRV-AH-gI/gE 和 rPRV-AH-gI/gE/gC 的生长动力学与亲本株相似。与 Ea 株的商业灭活疫苗相比,我们在小鼠和断奶猪中评估了基于重组病毒的灭活疫苗的免疫效力。结果表明,在初始免疫后 8 周,用 rPRV-AH-gI/gE/gC 免疫的小鼠的中和抗体水平高于用 rPRV-AH-gI/gE 免疫的小鼠(p<0.0001)。在剂量为 10 TCID 的免疫组中,rPRV-AH-gI/gE 组的存活率为 37.5%,而 rPRV-AH-gI/gE/gC 组的 PRV-AH 攻毒保护率为 87.5%。此外,在剂量为 10 TCID 的免疫组中,rPRV-AH-gI/gE 和 rPRV-AH-gI/gE/gC 组的存活率均为 100%。有趣的是,与商业疫苗组相比,10 TCID rPRV-AH-gI/gE/gC 组的中和抗体水平较低(p<0.0001),但在小鼠中的保护率相同。此外,在猪实验中,在初始免疫后 8 周,接种灭活 rPRV-AH-gI/gE/gC 的猪的中和抗体水平高于任何其他组(p<0.05)。更重要的是,在 10 TCID PRV-AH 攻毒后,接种 rPRV-AH-gI/gE/gC 的猪的症状和病理损伤较轻,表明额外插入 gC 基因可增强 PRV gI/gE 缺失疫苗在猪中的保护效力。总之,这些发现表明 rPRV-AH-gI/gE/gC 的灭活疫苗具有更好的免疫效果,可作为 PRV 控制的一种有前途的灭活疫苗候选物。

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