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TRIF 通过诱导 NF-κB 和 IFN 的激活以及促进细胞凋亡来参与固有免疫反应。

TRIF Participates in the Innate Immune Response by Inducing NF-κB and IFN Activation and Promoting Apoptosis.

机构信息

Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Sciences, Shandong Normal University, Jinan, China.

出版信息

Front Immunol. 2021 Aug 24;12:725150. doi: 10.3389/fimmu.2021.725150. eCollection 2021.

DOI:10.3389/fimmu.2021.725150
PMID:34504499
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8421551/
Abstract

TRIF, an important adaptor downstream of Toll-like receptor signaling, plays a critical role in the innate immune response. In this study, the full-length coding sequence of TRIF from common carp ( L.) was cloned and characterized. Bioinformatics analysis showed that common carp TRIF exhibited a conserved TIR domain and had the closest relationship with grass carp TRIF. Expression analysis revealed that TRIF was constitutively expressed in the examined tissues of common carp, with the highest expression in the spleen and the lowest expression in the head kidney, and could be upregulated under and poly(I:C) stimulation and under poly(I:C), LPS, PGN, flagellin, and Pam3CSK4 stimulation . Laser confocal microscopy showed that common carp TRIF colocalized with the Golgi apparatus. A luciferase reporter assay showed that carp TRIF elicited the activity of ifn-1 and nf-κb through the C-terminal domain. Additionally, crystal violet staining and qPCR assays revealed that carp TRIF inhibited the replication of SVCV in epithelioma papulosum cyprini (EPC) cells. Then, the signaling downstream of carp TRIF was investigated. Coimmunoprecipitation and Western blotting analysis demonstrated that carp TRIF interacted with TBK1 and augmented the expression of TRAF6 and phosphorylation of TBK1. Overexpression of carp TRIF significantly enhanced the expression of interferon-stimulated genes and inflammatory cytokines. Furthermore, flow cytometric (FCM) analysis suggested that carp TRIF induced apoptosis through the activation of caspase-8. In summary, our study indicated that TRIF plays an essential role in the innate immune responses of common carp against bacterial and viral infection.

摘要

TRIF 是 Toll 样受体信号下游的一个重要衔接蛋白，在先天免疫反应中发挥着关键作用。本研究克隆并鉴定了鲤鱼（Cyprinus carpio）TRIF 的全长编码序列。生物信息学分析表明，鲤鱼 TRIF 具有保守的 TIR 结构域，与草鱼 TRIF 的亲缘关系最近。表达分析显示，TRIF 在鲤鱼检测到的组织中持续表达，在脾脏中的表达最高，在头肾中的表达最低，并且可以在 Poly（I：C）刺激和 Poly（I：C）、LPS、PGN、鞭毛蛋白和 Pam3CSK4 刺激下上调。激光共聚焦显微镜显示，鲤鱼 TRIF 与高尔基器共定位。荧光素酶报告基因检测表明，鲤鱼 TRIF 通过 C 端结构域激活 ifn-1 和 nf-κb 的活性。此外，结晶紫染色和 qPCR 检测表明，鲤鱼 TRIF 抑制了鲤鱼上皮瘤细胞（EPC）中 SVCV 的复制。随后，研究了鲤鱼 TRIF 的下游信号通路。免疫共沉淀和 Western blot 分析表明，鲤鱼 TRIF 与 TBK1 相互作用，并增强了 TRAF6 的表达和 TBK1 的磷酸化。鲤鱼 TRIF 的过表达显著增强了干扰素刺激基因和炎症细胞因子的表达。此外，流式细胞术（FCM）分析表明，鲤鱼 TRIF 通过激活 caspase-8 诱导细胞凋亡。综上所述，本研究表明 TRIF 在鲤鱼抵抗细菌和病毒感染的先天免疫反应中发挥着重要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bfc4/8421551/e804f4cf93b2/fimmu-12-725150-g009.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bfc4/8421551/27db18126e1b/fimmu-12-725150-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bfc4/8421551/22836928c2a1/fimmu-12-725150-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bfc4/8421551/a8c00d579a06/fimmu-12-725150-g007.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bfc4/8421551/e804f4cf93b2/fimmu-12-725150-g009.jpg

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