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IFIT1负反馈上调干扰素β表达促进抗HSV-1病毒保护 被引量:2
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作者 方义军 胡好 +2 位作者 蔡少丽 丰志华 傅雅娟 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2020年第1期88-96,共9页
Ⅰ型单纯疱疹病毒(HSV-1)是危害人类健康的常见病原体之一,能够通过受损皮肤或黏膜感染宿主细胞并引起多种疾病。HSV-1的侵入激活先天免疫模式识别受体,诱导干扰素β(IFN-β)的产生,通过表达干扰素刺激基因(ISG)发挥抗病毒功能。近年来... Ⅰ型单纯疱疹病毒(HSV-1)是危害人类健康的常见病原体之一,能够通过受损皮肤或黏膜感染宿主细胞并引起多种疾病。HSV-1的侵入激活先天免疫模式识别受体,诱导干扰素β(IFN-β)的产生,通过表达干扰素刺激基因(ISG)发挥抗病毒功能。近年来,干扰素诱导的四肽重复蛋白1(IFIT1)在病毒感染过程中的作用引起了广大研究者的关注。然而,其具体机制尚未完全清楚。本研究利用CRISPR/Cas9技术构建了小鼠成纤维细胞(L929)IFIT1敲除细胞株,免疫印迹方法检测敲除细胞株IFIT1在蛋白质水平的表达。转染HT-DNA和Poly[I:C]刺激L929 WT和IFIT1敲除细胞株,实时定量PCR技术检测发现,HT-DNA刺激敲除细胞时,IFN-β及下游ISGs的表达量显著升高。IFN-β的表达量比L929-WT组平均高出13.4倍,IFIT1和趋化因子10(CXC chemokine ligand-10,CXCL10)的表达量比L929 WT组分别平均高出6.7倍和21倍(P<0.001),而Poly[I:C]刺激无明显变化(P>0.05),表明IFIT1是通过DNA信号通路来行使其负反馈调节作用。为研究IFIT1基因的抗病毒作用,利用CRISPR/Cas9技术改造的HSV-1-VP26-mCherry病毒感染该敲除细胞株,通过测定病毒荧光数及病毒拷贝数,发现IFIT1敲除细胞株与L929 WT细胞相比,存活率提高了60%(P<0.001),病毒增殖能力在48 h后降低28.6倍(P<0.001)。该结果表明,IFIT1基因的缺失有利于抵抗HSV-1的感染。综上所述,IFIT1通过DNA信号通路负反馈上调IFN-β及ISG的表达,IFIT1的缺失对病毒入侵发挥了保护作用。该结果为后续研究开发治疗HSV-1感染相关的治疗药物提供了一个新思路。 展开更多
关键词 Ⅰ型单纯疱疹病毒 CRISPR/Cas9 干扰素诱导的四肽重复蛋白1 干扰素Β
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Molecular Characterization and Expression Analysis of SKIV Infection of Interferon-Induced Protein with Tetratricopeptide Repeats 1(IFIT1) in Epinephelus lanceolatus
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作者 WANG Lei MA Teng +4 位作者 XU Wenteng CHEN Zhangfan ZHOU Qian ZHENG Guiliang CHEN Songlin 《Journal of Ocean University of China》 SCIE CAS CSCD 2021年第2期383-392,共10页
Interferon-induced protein with tetratricopeptide repeats 1(IFIT1), also known as interferon-induced protein 56(IFI56) or Interferon-stimulated protein 56(ISG56), was originally identified as a protein induced upon tr... Interferon-induced protein with tetratricopeptide repeats 1(IFIT1), also known as interferon-induced protein 56(IFI56) or Interferon-stimulated protein 56(ISG56), was originally identified as a protein induced upon treatment with interferon and inhibited by viral replication and translational initiation. In this study, Epinephelus lanceolatus IFIT1(ELIFIT1) gene was cloned for the first time. The complete cDNA of El IFIT1 gene includes 2921 nucleotides, and encodes a 437-amino acid(AA) protein. The putative ELIFIT1 protein has 9 TRP domains and is highly similar with IFIT1 proteins in other teleosts. In healthy fish, ELIFIT1 gene was highly expressed in the blood, which indicate its specific function in the peripheral immune system. Its expression was also observed in various immunity-related tissues including spleen, intestine, and kidney, Inducted with spotted knifejaw iridovirus(SKIV), ELIFIT1 gene expression was upregulated in the spleen, kidney, and liver 24 h after induction and reached its peak at 72 h, indicating that ELIFIT1 may play an important role in antivirus. These findings contribute to the understanding of the antiviral regulation of ELIFIT1 gene in teleost. 展开更多
关键词 interferon-induced protein with tetratricopeptide repeats 1(ifit1) Epinephelus lanceolatus tetratricopeptide repeats(TPR)motif expression pattern antiviral function
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