The Mutation of the Genes Related to Neurovirulence in HSV-2 Produces an Attenuated Phenotype in Mice
The Mutation of the Genes Related to Neurovirulence in HSV-2 Produces an Attenuated Phenotype in Mice
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HSV-2 神经毒力相关基因突变导致小鼠表现型减弱
DOI：
10.3390/v12070770
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发表时间：
2020
期刊：
影响因子：
4.7
通讯作者：
Li Qihan
中科院分区：
文献类型：
--
作者：
Liu Lei;Cheng Jishuai;Mou Tangwei;Zhang Ying;Xu Xingli;Zhang Jingjing;Li Xueqi;Feng Xiao;Xu Xiangxiong;Liao Yun;Fan Shengtao;Wang Lichun;Jiang Guorun;Li Qihan
HSV-2（单纯疱疹病毒2型）是一种重要的病毒因子，主要引起生殖器疱疹和背根神经节终身潜伏感染。通过CRISPR/Cas9规则间隔的短回文重复序列/CRISPR相关9）的基因修饰在此用于通过缺失RL 1（长重复元件1）和/或LAT（潜伏相关转录物）基因的片段来构建HSV-2突变株。HSV-2突变株LAT-HSV-2和RL 1-LAT-HSV-2表现出不同的生物学特性。RL 1-LAT-HSV-2在神经细胞中的增殖能力明显降低，在Vero细胞中诱导的空斑明显小于野生型和突变型。对感染这两种突变体的小鼠与感染野生型菌株的小鼠进行比较的观察表明，突变体RL 1-LAT-HSV-2具有减毒表型，在急性和潜伏感染期间致病性降低，并且诱导比野生型菌株更强的特异性免疫应答，而在感染含有LAT基因缺失的LAT-HSV-2突变体的小鼠中未发现减毒效果。然而，RL 1和LAT基因的同时突变并没有完全限制神经细胞中的病毒增殖，这表明多个HSV基因参与了神经系统中的病毒复制。这项工作表明，HSV-2基因RL 1和/或LAT可能参与小鼠感染的毒力机制。
HSV-2 (Herpes simplex virus type 2) is a critical viral agent that mainly causes genital herpes and life-long latent infection in the dorsal root ganglia. Gene modification via CRISPR/Cas9 Clustered regularly interspaced short palindromic repeat sequences/CRISPR associated 9) was used here to construct HSV-2 mutant strains through the deletion of fragments of the RL1 (Repeat Long element 1) and/or LAT (Latency-associated Transcript) genes. The HSV-2 mutant strains LAT-HSV-2 and RL1-LAT-HSV-2 present different biological properties. The proliferation of RL1-LAT-HSV-2 in nerve cells was decreased significantly, and the plaques induced by RL1-LAT-HSV-2 in Vero cells were smaller than those induced by LAT-HSV-2 mutant and wild-type strains. The observation of mice infected with these two mutants compared to mice infected with the wild-type strain indicated that the mutant RL1-LAT-HSV-2 has an attenuated phenotype with reduced pathogenicity during both acute and latent infections and induces a stronger specific immune response than the wild-type strain, whereas the attenuation effect was not found in mice infected with the LAT-HSV-2 mutant containing the LAT gene deletion. However, the simultaneous mutation of both the RL1 and LAT genes did not completely restrict viral proliferation in nerve cells, indicating that multiple HSV genes are involved in viral replication in the neural system. This work suggests that the HSV-2 genes RL1 and/or LAT might be involved in the virulence mechanisms in mouse infections.