孟加拉小反刍兽疫病毒野外分离株在Vero细胞上连续传代对融合蛋白的影响。

IF 2.7 2区 农林科学 Q1 VETERINARY SCIENCES
Md Saiful Islam Siddiqui, Anja Globig, Md Mushfiqur Rahman, Md Rafiqul Islam, Emdadul Haque Chowdhury
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引用次数: 0

摘要

目的:融合(F)蛋白对促进病毒进入宿主细胞至关重要,并有助于麻疹病毒的毒力。小反刍兽疫病毒(PPRV)在非本地宿主中的连续传代可导致可能降低致病性的突变。因此,本研究旨在探讨孟加拉小反刍猪瘟病毒在Vero细胞中连续传代对融合蛋白和致病性的影响。材料和方法:小反刍猪瘟病毒最初从自然爆发的小反刍猪瘟中分离出来,通过逆转录酶聚合酶链反应(RT-PCR)证实,在Vero细胞中传代至第9代,测序,并在先前的研究中保存。将库中第9代病毒作为病毒接种剂在Vero细胞中进一步传代,完成第60代传代。采用RT-PCR方法在不同传代数的组织培养液中证实了小反刍兽疫病毒RNA的存在。对第60代的TCF进行测序,并通过活体动物实验进行免疫原性研究,随后通过cELISA检测免疫活性。结果:将第9代和第60代的序列与其他序列进行比较分析,发现融合蛋白亮氨酸拉链结构中有14个核苷酸(nts)和4个氨基酸(aa)被取代。值得注意的是,活体动物实验证明了保护性免疫的发生。结论:本研究提示氨基酸替代和遗传分化可能对病毒毒力产生积极影响,强调了它们在开发强效疫苗中的重要性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Effects of serial passaging of field isolates of Bangladeshi PPR virus in Vero cells on the fusion protein.

Objectives: Fusion (F) protein is crucial for facilitating viral entry into host cells and contributes to the virulence of Morbilliviruses. Serial passaging of the Peste Des Petits Ruminants virus (PPRV) in nonnative hosts can lead to mutations that potentially reduce pathogenicity. Hence, this study aimed to investigate the effects of serial passaging of a Bangladeshi strain of PPR virus in Vero cells on the Fusion protein and pathogenicity MATERIALS AND METHODS: PPR viruses were initially isolated from natural PPR outbreaks, confirmed through reverse transcriptase polymerase chain reaction (RT‒PCR), passaged to the 9th passage in Vero cells, sequenced, and preserved in a previous study. The 9th passage virus from the repository was utilized as the viral inoculant for further passaging in Vero cells, and the 60th passage was completed. The presence of PPR viral RNA was confirmed in tissue culture fluid (TCF) by RT‒PCR at different passage numbers. TCF at the 60th passage was sequenced and used for immunogenicity studies via live animal experiments, and subsequent immunity was measured via cELISA.

Results: Comparative analysis of the sequences from the 9th and 60th passages, along with other sequences, revealed substitutions of 14 nucleotides (nts) and 4 amino acids (aa) within the leucine zipper structure of the fusion protein. Notably, live animal experiments demonstrated the occurrence of protective immunity.

Conclusion: This study suggests that amino acid substitution and genetic divergence may positively affect viral virulence, highlighting their importance in the development of a potent vaccine.

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来源期刊
Irish Veterinary Journal
Irish Veterinary Journal 农林科学-兽医学
CiteScore
4.80
自引率
3.40%
发文量
1
审稿时长
>36 weeks
期刊介绍: Irish Veterinary Journal is an open access journal with a vision to make a substantial contribution to the dissemination of evidence-based knowledge that will promote optimal health and welfare of both domestic and wild species of animals. Irish Veterinary Journal has a clinical research focus with an emphasis on the effective management of health in both individual and populations of animals. Published studies will be relevant to both the international veterinary profession and veterinary scientists. Papers relating to veterinary education, veterinary ethics, veterinary public health, or relevant studies in the area of social science (participatory research) are also within the scope of Irish Veterinary Journal.
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