Inhibition of RelA expression via RNA interference induces immune tolerance in a rat keratoplasty model
Inhibition of RelA expression via RNA interference induces immune tolerance in a rat keratoplasty model
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通过 RNA 干扰抑制 RelA 表达可诱导大鼠角膜移植模型中的免疫耐受
DOI：
10.1016/j.molimm.2016.03.014
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发表时间：
2016
影响因子：
3.6
通讯作者：
Qiu Haijiang
中科院分区：
文献类型：
--
作者：
Yang Jize;Feng Songfu;Yi Guoguo;Wu Wei;Yi Ruiwen;Lu Xiaohe;Xu Wanfu;Qiu Haijiang
RelA是NF-kB活性最重要的调节因子，其在角膜移植免疫排斥反应中的作用机制尚未得到充分研究。在本研究中，我们测试了慢病毒介导的在骨髓源性树突状细胞（BMDC）模型中对RelA表达的沉默。用RelA-shRNA转染BMDCs，诱导未成熟、成熟抵抗和耐受性表型，同时不显著改变IFN-γ、IL-10和IL-17的表达。建立了全异体大鼠角膜移植模型进行体内研究。lv-shRelA-DC注射组移植体平均存活时间（MST）明显长于BMDC治疗组和对照组。lv-shRelA-DC注射组角膜混浊和新生血管规模较对照组轻微。术后流式细胞术分析显示，接受lv-shRelA-DC注射的动物Treg阳性细胞百分比显着增加。ELISA和qRT-PCR检测结果显示，lv-shRelA-DC可抑制血清中IFN-γ和IL-17的表达。体内实验表明，与一般的抗炎因子不同，IL-10诱导的免疫抑制部分归因于在实验过程中注射lv-shRelA-DC。荧光素酶和染色质IP评估显示，在BMDCs中，RelA敲低可显著降低DNA与IFN-γ、IL-10和IL-17启动子的结合，并抑制转录活性。综上所述，本研究表明RelA通过影响IL-17的表达在介导角膜新生血管形成中发挥了重要作用。我们的综合分析表明，RelA的显著作用为角膜移植排斥反应的预防提供了一种新颖可行的治疗途径。
RelA, the most important regulator of NF-kB activity, and its mechanisms in keratoplasty immune rejection have not been fully investigated. In the present study, lentivirus-mediated silencing of RelA expression in a bone marrow-derived dendritic cell (BMDC) model was tested. The BMDCs were transfected with RelA-shRNA to induce an immature, maturation-resistant and tolerogenic phenotype, while not significantly changing IFN-γ, IL-10 and IL-17 expression. A fully allogeneic rat cornea transplant model was established for in vivo studies. The allograft mean survival time (MST) of lv-shRelA-DC injection groups were significantly longer than the untreated BMDC group and control group. The corneal opacity and neovascularization scale of the lv-shRelA-DC injection groups were slight compared to pair control others. Postoperative flow cytometric analysis revealed that the percentage of Treg positive cells was dramatically increased in animals that received an lv-shRelA-DC injection. ELISA and qRT-PCR analyses of serum showed that IFN-γ and IL-17 expression were suppressed by lv-shRelA-DC treatement. In vivo experiments demonstrated that IL-10 induced immunosuppression was partly attributed to injection of lv-shRelA-DC throughout the experiment, differing from the general anti-inflammatory factors. Luciferase and Chromatin IP evaluation showed that RelA knockdown in BMDCs significantly reduces DNA binding to IFN-γ, IL-10 and the IL-17 promoter and inhibited of transcriptional activity. Taken together, this study illustrates a significant role of RelA in mediating the corneal neovascularization by affecting IL-17 expression. Our comprehensive analysis shows that the significant role of RelA provides a novel and feasible therapeutic approach for the prevention of corneal allograft rejection.