IFN-γ Regulates the Expression of MICA in Human Corneal Epithelium Through miRNA4448 and NFκB.
IFN-γ Regulates the Expression of MICA in Human Corneal Epithelium Through miRNA4448 and NFκB.
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IFN-γ通过miRNA4448和NF kappa B调节人角膜上皮中MICA的表达
DOI：
10.3389/fimmu.2018.01530
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发表时间：
2018
影响因子：
7.3
通讯作者：
Hong J
中科院分区：
文献类型：
--
作者：
Wu D;Zhang J;Qian T;Dai Y;Mashaghi A;Xu J;Hong J
主要组织相容性复合物I类相关链A（云母）是一种非经典的主要组织相容性复合物分子，可刺激或共刺激CD 8 + T细胞或自然杀伤（nk）细胞，从而影响角膜移植物的存活。本研究探讨了IFN-γ通过miRNA 4448对人角膜上皮中云母表达水平的调节。采用qRT-PCR和酶联免疫吸附试验检测IFN-γ刺激的人角膜上皮细胞（HCECs）中云母的表达水平，并测量差异miRNA表达水平。qRT-PCR、Western blotting和免疫荧光染色显示IFN-γ处理和miRNA 4448过表达的HCECs中核因子κ B（NFκB）/P65表达。采用荧光素酶报告基因分析预测NFκB与云母的相互作用。此外，HCEC用云母质粒转染或用IFN-γ和NKG 2D-mAb处理，并与NK细胞和CD 8 + T细胞共培养。用Annexin V/PI染色检测细胞凋亡。qRT-PCR检测转染MICA和IFN-γ的HCECs与NK细胞和CD 8 + T细胞共培养后抗凋亡因子Survivin和凋亡因子Caspase 3的表达。IFN-γ（500 ng/ml，24 h）可上调HCECs中云母的表达。在6种差异表达的microRNA中，miRNA 4448在IFN-γ处理后水平下降最多。miRNA 4448的过表达降低了云母表达。miRNA 4448下调IFN-γ诱导的HCEC中NFκB/P65的表达，并且确定NFκB/P65通过与启动子区域结合而直接靶向云母。与NK细胞和CD 8 + T细胞共培养表明，云母过表达促进HCEC凋亡，这可以被NKG 2D-mAb抑制。云母与NK（CD 8 + T）细胞相互作用后，HCECs中Survivin mRNA表达降低，Caspase 3 mRNA表达升高。IFN-γ通过调节miRNA 4448和NFκB/P65的水平，增强HCEC中云母的表达，从而促进NK和CD 8 + T细胞诱导的HCEC凋亡。这一发现可能会导致新的见解角膜移植排斥反应的发病机制。
Major histocompatibility complex class I-related chain A (MICA), a non-classical major histocompatibility complex molecule, can stimulate or co-stimulate CD8+ T cells or natural killer (nk) cells, thus affecting cornea allograft survival. This study investigated IFN-γ regulation of MICA expression levels in human corneal epithelium by miRNA4448. MICA expression levels in human corneal epithelial cells (HCECs) stimulated with IFN-γ were detected by qRT-PCR and an enzyme-linked immunosorbent assay, and differential miRNA expression levels were measured. qRT-PCR, Western blotting, and immunofluorescence staining revealed nuclear factor kappa B (NFκB)/P65 expression in IFN-γ-treated and miRNA4448-overexpressed HCECs. A luciferase reporter assay was used to predict the interaction between NFκB and MICA. Additionally, HCECs were transfected with MICA plasmid or treated with IFN-γ and NKG2D-mAb and cocultured with NK cells and CD8+ T cells. Cell apoptosis was measured using Annexin V/PI staining. qRT-PCR detected the expression of anti-apoptosis factor Survivin and apoptosis factor Caspase 3 in MICA-transfected and IFN-γ-treated HCECs after co-culturing with NK cells and CD8+ T cells. IFN-γ (500 ng/ml, 24 h) upregulated MICA expression in HCECs in vitro. Among six differentially expressed microRNAs, miRNA4448 levels decreased the most after IFN-γ treatment. The overexpression of miRNA4448 decreased MICA expression. miRNA4448 downregulated NFκB/P65 expression in IFN-γ-induced HCEC, and it was determined that NFκB/P65 directly targeted MICA by binding to the promotor region. A coculture with NK cells and CD8+ T cells demonstrated that MICA overexpression enhanced HCEC apoptosis, which could be inhibited by NKG2D-mAb. Simultaneously, Survivin mRNA expression decreased and Caspase3 mRNA expression increased upon the interaction between MICA and NK (CD8+ T) cells in HCECs. IFN-γ enhances the expression of MICA in HCECs by modulating miRNA4448 and NFκB/P65 levels, thereby contributing to HCEC apoptosis induced by NK and CD8+ T cells. This discovery may lead to new insights into the pathogenesis of corneal allograft rejection.
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影响因子：
9.2
作者：
Boo L;Ho WY;Ali NM;Yeap SK;Ky H;Chan KG;Yin WF;Satharasinghe DA;Liew WC;Tan SW;Ong HK;Cheong SK
通讯作者：
Cheong SK
影响因子：
3.4
作者：
Nicholls, Susan M.;Banerjee, Sanjiv;Dick, Andrew D.
通讯作者：
Dick, Andrew D.
影响因子：
4.4
作者：
da Silva, Henrique Borges;de Salles, Erika Machado;D'Imperio Lima, Maria Regina
通讯作者：
D'Imperio Lima, Maria Regina
影响因子：
64.8
作者：
通讯作者：
--
DOI：
10.1097/00004647-200003000-00017
发表时间：
2000-03-01
影响因子：
6.3
作者：
Stephenson, D;Yin, T;Clemens, J
通讯作者：
Clemens, J