Zinc finger protein A20 inhibits maturation of dendritic cells resident in rat liver allograft
Zinc finger protein A20 inhibits maturation of dendritic cells resident in rat liver allograft
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DOI：
10.1016/j.jss.2013.01.062
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发表时间：
2013-08-01
影响因子：
2.2
通讯作者：
Xu, Ming-Qing
中科院分区：
文献类型：
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作者：
Dai, Fu-Zhen;Yang, Jie;Xu, Ming-Qing
背景：在器官移植领域，尽管传统上认为供者来源的间质树突状细胞(DC)是器官移植排斥反应的诱因，但供者来源的间质树突状细胞(DC)，包括驻留在肝脏的DC或宿主DC，在实验模型中也与移植耐受有关。DC的这种功能二分法受到多种因素的制约，其中最重要的因素似乎是它们的成熟阶段。本研究旨在探讨锌指蛋白A20对大鼠同种异体肝移植树突状细胞成熟的影响。材料与方法：采用Dark Agti[DA]大鼠至Lewis大鼠异基因肝移植。将携带A20全长的重组腺病毒通过门静脉注入同种异体肝移植模型(A20组)、生理盐水组(PS组)和空载体rAdEasy(rAdEasy组)作为对照。评估移植肝的急性排斥反应，并在移植后第7天分离驻留在移植肝中的树突状细胞。采用凝胶迁移率改变分析、流式细胞仪、逆转录-聚合酶链式反应和混合淋巴细胞反应分别检测核因子-kappa B结合活性、共刺激分子CD40、CD80和CD86的表面表达、IL-12信使RNA的表达和DC的异位刺激能力。结果：门静脉输注腺病毒A20体外转移导致移植肝A20蛋白高表达。移植后第7天，组织学检查显示A20组有轻度排斥反应，而PS组和rAdEasy组排斥反应较重。A20组同种异体肝移植的DC具有未成熟DC的特征，但可检测到但极低水平的NF-kappa B活性、IL-12mRNA表达和共刺激分子(CD40、CD80和CD86)的表面表达；而rAdEasy组和PS组同种异体肝移植的DC则表现为成熟DC的特征，具有高水平的NF-kappa B活性、IL-12mRNA表达和共刺激分子(CD40、CD80和CD86)的表面表达。PS组和rAdEasy组同种异体肝移植树突状细胞可诱导MLR细胞DNA合成和干扰素-γ的产生，而A20组同种异体肝移植树突状细胞对MLR的细胞增殖和干扰素-γ产生的诱导作用较弱。结论：A20过表达能有效抑制肝移植树突状细胞的成熟，从而抑制肝移植急性排斥反应。Crown版权所有(C)2013由Elsevier Inc.出版。保留所有权利。
Background: In organ transplant field, although viewed traditionally as instigators of organ allograft rejection, donor-derived interstitial dendritic cells (DCs), including those resident in liver, or host DCs have also been implicated in transplant tolerance in experimental models. This functional dichotomy of DC is governed by various factors, the most important of which appears to be their stage of maturation. This study was designed to examine the effect of zinc finger protein A20 on maturation of DCs resident in rat liver allograft.Materials and methods: Allogeneic (Dark Agouti [DA] rat to Lewis rat) liver transplantation was performed. Adenovirus carrying the full length of A20 was introduced into liver allografts by ex vivo perfusion via the portal vein during preservation (group A20), physiological saline (group PS), and empty Ad vector rAdEasy (group rAdEasy) that served as controls. Acute liver allograft rejection was assessed, and DCs resident in liver allografts were isolated on day 7 after transplantation. Nuclear factor kappa B (NF-kappa B)-binding activities, surface expression of costimulatory molecules (CD40, CD80, and CD86), expression of interleukin (IL) 12 messenger RNA (mRNA), and allocostimulatory capacity of DCs were measured with electrophoretic mobility shift assay, flow cytometry, reverse transcription-polymerase chain reaction, and mixed lymphocyte reaction (MLR), respectively.Results: Ex vivo transfer of A20 adenovirus by portal vein infusion resulted in overexpression of A20 protein in liver allograft after transplantation. On day 7 after transplantation, histologic examination revealed a mild rejection in group A20 but a more severe rejection in group PS and group rAdEasy. DCs from group A20 liver allografts exhibited features of immature DC with detectable but very low level of NF-kappa B activity, IL-12 mRNA expression, and surface expression of costimulatory molecules (CD40, CD80, and CD86), whereas DCs from group rAdEasy and group PS liver allograft displayed features of mature DC with high level of NF-kappa B activity, IL-12 mRNA expression, and surface expression of costimulatory molecules (CD40, CD80, and CD86). DCs from group PS and group rAdEasy liver allograft were potent inducers of DNA synthesis and interferon gamma production in MLR, and DCs from group A20 liver allografts induced only minimal levels of cell proliferation and interferon gamma production in MLR.Conclusions: These data suggest that A20 overexpression could effectively inhibit maturation of DCs resident in liver allograft and consequently suppress acute liver allograft rejection. Crown Copyright (c) 2013 Published by Elsevier Inc. All rights reserved.