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细胞减弱的猪流行性腹泻病毒株浙 08 株通过刺激树突状细胞提供有效的免疫保护。

Cell attenuated porcine epidemic diarrhea virus strain Zhejiang08 provides effective immune protection attributed to dendritic cell stimulation.

机构信息

Veterinary College, Nanjing Agricultural University, Weigang 1, Nanjing, Jiangsu 210095, PR China.

Veterinary Medicine Research Centre of Beijing Da Bei Nong Group, Beijing 100195, PR China.

出版信息

Vaccine. 2017 Dec 15;35(50):7033-7041. doi: 10.1016/j.vaccine.2017.10.052.

Abstract

Since 2010, the porcine epidemic diarrhea coronavirus (PEDV) has caused significant damage to the global pork industry. However, classical PEDV vaccine strains only provide limited protection against emerging strains. In this study, we successfully isolated and attenuated the PEDV epidemic strain Zhejiang08, which was characterized by good cell adaptation and high-titer production 48 h post infection in Vero E6 cells. The attenuated virus induced a high level of virus-specific neutralizing antibodies until 120 days after immunization in piglets and provided complete protection when challenged with an emerging virus strain on day 14 post immunization. Moreover, the capability to activate dendritic cells (DCs) of this isolate was identified. Higher expression levels of IL-12 and IFN-γ were recorded in DCs after treatment with Zhejiang08 for 24 h. Furthermore, genome sequencing and phylogenetic analysis revealed high homology between the main antigen epitopes of Zhejiang08 and PEDV pandemic isolates following 2011. Combining the glycosylation site prediction results and their distribution within the spatial structure of the S protein, led to the conclusion that the observed more effective host immune response of Zhejiang08 compared to CV777 was possibly associated with a lack of the potential glycosylation site in the 296 amino acids of the S protein. In summary, we illustrated that the attenuated virus represents a promising vaccine candidate.

摘要

自 2010 年以来,猪流行性腹泻冠状病毒(PEDV)给全球养猪业造成了巨大损失。然而,经典的 PEDV 疫苗株仅对新兴株提供有限的保护。在本研究中,我们成功分离并减毒了流行株浙 08 株,该毒株在 Vero E6 细胞中具有良好的细胞适应性和高滴度的感染后 48 小时产生。减毒病毒在仔猪中诱导高水平的病毒特异性中和抗体,直到免疫后 120 天,并且在免疫后 14 天用新兴病毒株攻毒时能提供完全保护。此外,还鉴定了该分离株激活树突状细胞(DCs)的能力。用浙 08 处理 24 小时后,DCs 中 IL-12 和 IFN-γ 的表达水平更高。此外,基因组测序和系统进化分析显示,浙 08 与 2011 年后的 PEDV 大流行分离株的主要抗原表位高度同源。结合糖基化位点预测结果及其在 S 蛋白空间结构中的分布,表明浙 08 与 CV777 相比观察到更有效的宿主免疫反应可能与 S 蛋白的 296 个氨基酸中缺乏潜在的糖基化位点有关。总之,我们表明减毒病毒是一种有前途的疫苗候选物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee43/7115645/156393262f11/gr1_lrg.jpg

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