数字农科院2.0

Adaptation of a live-attenuated genotype I Japanese encephalitis virus to vero cells is associated with mutations in structural protein genes

文献类型： 外文期刊

作者： Muhammad Naveed Anwar;Shuang Guo;Wang Xin;Muddassar Hameed;Abdul Wahaab;Xiaochun Ma;Aman Ullah Khan;Sajid Ur Rahman;Donghua Shao;Zongjie Li;Ke Liu;Beibei Li;Yafeng Qiu;Zhiyong Ma;Jianchao Wei

作者机构：

关键词： Adaptation; Attenuation; Conserved region; Japanese encephalitis virus; Serial passaging; Vaccine

期刊名称： Virus Res.

ISSN： 0168-1702

年卷期： 2021 年 292 卷

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收录情况： JCR（2021版）

摘要： The SD12-F120 is a live-attenuated genotype I strain of Japanese encephalitis virus (JEV) and was obtained by serial passage of wild-type strain SD12 on BHK-21 cells combined with multiple plaque purification and virulence selection in mice. The large scale production and vast clinical trials always demand ideal safety and efficacy profile of live-attenuated vaccines. In the present study, SD12-F120VC has undergone serial passaging of P1-P30 in WHO qualified Vero cells to assess the potential effect of adaptation to growth on Vero cells. The series of experiments showed that vaccine SD12-F120VC (Vero cell adapted) variants have consistently increased in peak virus titer compared to early passages and have good adaptation to growth in Vero cells. The animal experiments showed that Vero cell adapted SD12-F120VC variants have attenuation phenotype in suckling mice and the plaque morphology for all SD12-F120VC variants was small. Vaccination of mice with SD12-F120VC vaccine produced complete protection for homologous SD12 genotype I strain, but failed to give the complete protection of vaccinated mice against the challenge of heterologous N28 genotype III strain. In response to immunization of SD12-F120VC in mice, the neutralizing antibodies titer against homologous SD12-F120VC and SD12 (GI) was higher than heterologous N28 (GIII) strain. The prM protein has 6 amino acid substitutions, of which 5 amino acid changes were confined at the start of the pr domain in the ∼40 amino acids, and some mutations in the pr domain of prM might contribute to Vero cell adaptation. Our findings in this study are important for validation, evaluation and quality control study of live attenuated flaviviruses vaccines and show that Vero cells are a suitable substrate for the production of a safe and stable live-attenuated JEV vaccine. © 2020 Elsevier B.V.

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