[1]庄腾寒,杨鹏,马心宇,等.G1期纤毛细胞器对宿主细胞PEDV敏感性的调控[J].江苏农业学报,2024,(12):2302-2309.[doi:doi:10.3969/j.issn.1000-4440.2024.12.013]
 ZHUANG Tenghan,YANG Peng,MA Xinyu,et al.Regulation of cilia in G1 phase on the susceptibility of host cells to PEDV[J].,2024,(12):2302-2309.[doi:doi:10.3969/j.issn.1000-4440.2024.12.013]
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G1期纤毛细胞器对宿主细胞PEDV敏感性的调控()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2024年12期
页码:
2302-2309
栏目:
畜牧兽医·水产养殖·益虫饲养
出版日期:
2024-12-30

文章信息/Info

Title:
Regulation of cilia in G1 phase on the susceptibility of host cells to PEDV
作者:
庄腾寒1杨鹏2马心宇2徐悦1陈丽1鲍熹1杨丹晨2冯磊1
(1.江苏省农业科学院动物免疫工程研究所,江苏南京210014;2.南京农业大学动物医学院,江苏南京210095)
Author(s):
ZHUANG Tenghan1YANG Peng2MA Xinyu2XU Yue1CHEN Li1BAO Xi1YANG Danchen2FENG Lei1
(1.Institute of Veterinary Immunology and Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;2.College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China)
关键词:
猪流行性腹泻病毒(PEDV)细胞周期细胞同步化ST细胞IPEC-J2细胞纤毛
Keywords:
porcine epidemic diarrhea virus (PEDV)cell cyclecell synchronizationST cellsIPEC-J2 cellscilia
分类号:
S852.65
DOI:
doi:10.3969/j.issn.1000-4440.2024.12.013
文献标志码:
A
摘要:
本研究旨在分析宿主细胞周期对猪流行性腹泻病毒(PEDV)增殖能力的调控效果。使用PEDV[感染复数(MOI)=1]分别感染猪睾丸(ST)细胞、猪小肠上皮(IPEC-J2)细胞,用流式细胞术(FACS)和免疫印迹(WB)技术鉴定细胞周期,用免疫荧光法观察纤毛组装效果,通过测定半数组织培养感染剂量(TCID50)鉴定病毒的增殖能力。结果表明,PEDV感染ST细胞、IPEC-J2细胞7 h后,会导致宿主细胞阻滞于合成期(S期),感染31 h后则会导致宿主细胞阻滞于合成前期(G1期);FACS、WB检测结果证实,通过双胸苷(Thy)释放协同诺考达唑(Noc)能够将细胞周期分别阻滞于G1-S转换期、S期和细胞分裂期(M期);PEDV(MOI=1)感染处于各细胞周期的ST、IPEC-J2细胞,同步化于G1-S转换期的细胞感染PEDV后的TCID50最高,诱导纤毛去组装的细胞对PEDV的敏感性下降。由研究结果可知,PEDV感染导致宿主细胞阻滞于G1-S转换期,G1期细胞因具有纤毛细胞器而最有利于PEDV增殖。
Abstract:
This study aimed to research on the regulation of host cell cycle on porcine epidemic diarrhea virus (PEDV) proliferation. Swine testis (ST) and intestinal porcine epithelial cell line-J2 (IPEC-J2) cells were infected using PEDV with multiplicity of infection (MOI)=1, and the cell cycle was identified by fluorescence activated cell sorting (FACS) and Western blot (WB). Ciliary assembly was observed by immunofluorescence. Medium tissue culture infective dose (TCID50) was measured to identify virus proliferation ability. PEDV infected ST cells and IPEC-J2 cells, and the host cells were arrested at the synthesis phase (S phase) and the pre-synthesis phase (G1 phase) after infection of 7 h and 31 h, respectively. FACS and WB tests confirmed that the use of double-thymidine release in coordination with nocodazole could block the cell cycle in the G1-S transition phase, S phase and cell division phase (M phase), respectively. PEDV (MOI=1) infected synchronized ST cells and IPEC-J2 cells, and the TCID50 of cells synchronized in the G1-S transition phase was the highest after PEDV infection. Cells with ciliary disassembly were less sensitive to PEDV. PEDV infection arrested cell cycle to G1-S transition. Cells in G1 phase were most conducive to PEDV proliferation due to cilia.

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备注/Memo

备注/Memo:
收稿日期:2024-12-06基金项目:江苏省自然科学基金-青年基金项目(BK20220748);国家自然科学基金-青年科学基金项目(32300577);江苏省重点研发计划-社会发展项目(BE2022787);江苏省农业科技自主创新基金全产业链关键技术协同创新项目[CX(24)2004]作者简介:庄腾寒(1991-),男,上海人,博士,助理研究员,主要从事病毒-细胞互作机制和细胞工程改造及工艺放大等相关方面的研究。(E-mail)zhuangtenghan@foxmail.com通讯作者:冯磊,(E-mail)fenglei@jaas.ac.cn;庄腾寒,(E-mail)zhuangtenghan@foxmail.com
更新日期/Last Update: 2025-01-23