Comparative RNA sequencing analysis of three Capripoxvirus infections in an immortalized hTERT-bOEC cell model

IF 2.4 3区 医学 Q3 VIROLOGY Virology Pub Date : 2025-02-01 Epub Date: 2024-12-14 DOI:10.1016/j.virol.2024.110352
Hongqiang Zhang , Fangping Wang , Haotai Chen , Shasha Wang , Lina Tong , Huibao Wang , Jiangfeng Fan , Xiangping Yin , Xiangwei Wang , Yuefeng Sun , Xiaolong Gao , Shanhui Ren
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Abstract

Capripoxviruses (CaPVs), such as lumpy skin disease, sheep pox, and goat pox, cause significant production and economic losses and are major constraints to the growth of livestock production in endemic areas. Understanding the pathogenic mechanism of CaPVs and their translation into clinical applications depends on the availability of a suitable cell line. In this study, we used a lentiviral packaging system to establish an immortalized hTERT-bOEC cell line by ectopic introduction of human telomerase reverse transcriptase (hTERT). Western blotting, indirect immunofluorescence, and flow cytometry analyses revealed that hTERT was successfully integrated into the genome of hTERT-bOEC cells. Crucially, this hTERT-bOEC cell line was highly susceptible to LSDV, SPPV, and GTPV infections. Establishing hTERT-bOECs is critical for basic research, clinical application, and vaccine development related to CaPVs. Furthermore, RNA-seq analyses revealed a similar differential expression of genes and enrichment of signaling pathways to CaPV infections in hTERT-bOECs. Real-time quantitative qPCR further confirmed the top five up-regulated and down-regulated differentially expressed genes among the CaPV infections. Transcriptome analyses provide deep insight into the biological characteristics of the replication process in CaPV infections.
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永生化hTERT-bOEC细胞模型中三种Capripoxvirus感染的比较RNA测序分析。
诸如疙瘩性皮肤病、羊痘和山羊痘等capv造成重大的生产和经济损失,是流行地区牲畜生产增长的主要制约因素。了解capv的致病机制及其转化为临床应用取决于是否有合适的细胞系。在这项研究中,我们使用慢病毒包装系统通过异位引入人类端粒酶逆转录酶(hTERT)建立了永生化hTERT- boec细胞系。Western blotting、间接免疫荧光和流式细胞术分析显示,hTERT已成功整合到hTERT- boec细胞的基因组中。关键是,这种hTERT-bOEC细胞系对LSDV、SPPV和GTPV感染高度敏感。建立htert - boec对于与capv相关的基础研究、临床应用和疫苗开发至关重要。此外,RNA-seq分析显示,在htert - boec中,CaPV感染的基因表达和信号通路富集也存在类似的差异。实时定量qPCR进一步证实了CaPV感染中前5个差异表达基因的上调和下调。转录组分析提供了深入了解CaPV感染复制过程的生物学特征。
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来源期刊
Virology
Virology 医学-病毒学
CiteScore
6.00
自引率
0.00%
发文量
157
审稿时长
50 days
期刊介绍: Launched in 1955, Virology is a broad and inclusive journal that welcomes submissions on all aspects of virology including plant, animal, microbial and human viruses. The journal publishes basic research as well as pre-clinical and clinical studies of vaccines, anti-viral drugs and their development, anti-viral therapies, and computational studies of virus infections. Any submission that is of broad interest to the community of virologists/vaccinologists and reporting scientifically accurate and valuable research will be considered for publication, including negative findings and multidisciplinary work.Virology is open to reviews, research manuscripts, short communication, registered reports as well as follow-up manuscripts.
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