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细胞外基质蛋白
ABI3BP 抗水疱性口炎病毒初步作用研究- Affiliations：
1. 1Department of Stomatology, the First Medical Center, Chinese People’s Liberation Army (PLA) General Hospital, Beijing 100853, China
2. 2Chinese PLA Medical School, Beijing 100853, China
3. 3State Key Laboratory of Toxicology and Medical Countermeasures, Beijing Institute of Pharmacology and Toxicology, Beijing 100850, China
4. 4Baodi Clinical College, Tianjin Medical University, Tianjin 301800, China
- Author bio：
陈欢, 1773524536@qq.com
* 张彤，kqzhengji301@163.com;
- Funds：
DOI：10.24920/004319
中图分类号：收稿日期：2023-10-20，
录用日期：2023-12-27，
网络出版日期：2024-02-22，
纸质出版日期：2024-03-31
- Accepted：
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孟祥博, 陈美桦, 许诺, 等. 细胞外基质蛋白
ABI3BP 抗水疱性口炎病毒初步作用研究[J]. 中国医学科学杂志（英文）, 2024,39(1):1-8.Xiang-Bo Meng, Mei-Hua Chen, Nuo Xu, et al. Antiviral Effect of Extracellular Matrix Protein ABI3BP on Vesicular Stomatitis Virus and Its Mechanism: A Preliminary Study
In Vitro [J]. Chinese medical sciences journal, 2024, 39(1): 1-8.孟祥博, 陈美桦, 许诺, 等. 细胞外基质蛋白
ABI3BP 抗水疱性口炎病毒初步作用研究[J]. 中国医学科学杂志（英文）, 2024,39(1):1-8. DOI： 10.24920/004319.Xiang-Bo Meng, Mei-Hua Chen, Nuo Xu, et al. Antiviral Effect of Extracellular Matrix Protein ABI3BP on Vesicular Stomatitis Virus and Its Mechanism: A Preliminary Study
In Vitro [J]. Chinese medical sciences journal, 2024, 39(1): 1-8. DOI： 10.24920/004319.
摘要
目的
探讨细胞外基质蛋白
ABI3BP
对水疱性口炎病毒（vesicular stomatitis virus，VSV）基因组复制和先天免疫信号通路的影响。
方法
在人皮肤成纤维细胞BJ-5ta中转染小干扰RNA（siRNA）敲低
ABI3BP
基因，建立
ABI3BP
基因缺失的VSV-GFP病毒感染细
胞模型。通过鬼笔环肽细胞免疫荧光实验，检测敲低
ABI3BP
后细胞内肌动蛋白（F-actin）形态结构的变化。在
ABI3BP
基因缺失的VSV-GFP病毒感染细胞模型上，利用RT-qPCR方法检测细胞中病毒mRNA水平的变化。利用免疫印迹法（Western blotting）检测IRF3和TBK1磷酸化水平的变化。
结果
成功建立敲减
ABI3BP
基因的VSV-GFP感染的细胞模型。鬼笔环肽细胞免疫荧光染色表明，敲减
ABI3BP
基因后，细胞内F-actin的表达发生结构重排。采用不同剂量的病毒感染细胞，与对照组相比，
ABI3BP
敲低细胞中基因拷贝数分别增加2.5~3.5倍（
P
<
0.01）和2.2~4倍（
P
<
0.01），VSV-GFP病毒蛋白表达水平显著上调；在
ABI3BP
基因敲低的细胞模型上，VSV-GFP病毒感染导致Ⅰ型干扰素免疫通路关键免疫分子p-IRF3 和p-TBK1蛋白磷酸化水平明显下调。
结论
本研究表明细胞外基质蛋白
ABI3BP
对维持细胞内F-actin纤维网状结构具有重要作用。
ABI3BP
缺失促进RNA病毒的复制，
ABI3BP
是I型干扰素通路激活的重要调控分子。
Abstract
Objective
To explore the influence of extracellular matrix protein ABI-interactor 3-binding protein (ABI3BP) on vesicular stomatitis virus (VSV) genome replication and innate immune signaling pathway.
Methods
The small interfering RNA (siRNA) was transfected to knock down
ABI3BP
gene in human skin fibroblast BJ-5ta cells. VSV-green fluorescent protein (VSV-GFP)-infected cell model was established. The morphological changes and F-actin stress fiber formation were detected on
ABI3BP
knockdown cells by phalloidin immunofluorescence staining. The mRNA level of virus replication was detected by RT-qPCR in BJ-5ta cells after VSV-GFP infection; western blotting was performed to detect the changes in interferon regulatory factor 3 (IRF3) and TANK-binding kinase 1 (TBK1) phosphorylation levels.
Results
The VSV-GFP-infected BJ-5ta cell model was successfully established. Efficient knockdown of
ABI3BP
in BJ-5ta cells was achieved. Phalloidin immunofluorescence staining revealed structural rearrangement of intracellular F-actin after
ABI3BP
gene knockdown. Compared with the con
trol group
the gene copy number of VSV-GFP in
ABI3BP
knockdown cells increased by 2.2 - 3.5 times (
P
<
0.01) and 2.2 - 4.0 times (
P
<
0.01) respectively when infected with VSV of multiplicity of infection 0.1 and 1. The expression of viral protein significantly increased in
ABI3BP
knockdown cells after virus infection. The activation of type-I interferon pathway
as determined by phosphorylated IRF3 and phosphorylated TBK1
was significantly decreased in
ABI3BP
knockdown cells after VSV-GFP infection.
Conclusions
Extracellular matrix protein ABI3BP plays an important role in maintaining the formation and rearrangement of actin structure.
ABI3BP
gene deletion promotes RNA virus replication
and ABI3BP is an important molecule that maintains the integrity of type I interferon pathway.
关键词
Keywords
references
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