Activation of Epithelial Signal Transducer and Activator of Transcription 1 by Interleukin 28 Controls Mucosal Healing in Mice With Colitis and Is Increased in Mucosa of Patients With Inflammatory Bowel Disease
Activation of Epithelial Signal Transducer and Activator of Transcription 1 by Interleukin 28 Controls Mucosal Healing in Mice With Colitis and Is Increased in Mucosa of Patients With Inflammatory Bowel Disease
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DOI：
10.1053/j.gastro.2017.03.015
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发表时间：
2017-07-01
期刊：
影响因子：
29.4
通讯作者：
Neurath, Markus F.
中科院分区：
文献类型：
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作者：
Chiriac, Mircea T.;Buchen, Barbara;Neurath, Markus F.
背景与目的：我们研究了白细胞介素28 A（也称为IL 28 A或干扰素12）在肠上皮细胞（IEC）活化中的作用，研究了其在炎症性肠病（IBD）小鼠模型和肠粘膜愈合中的作用。方法：在C57 BL/6 JCrl小鼠（对照）、具有干扰素1受体1基因（Il 28 ra（-/-））破坏的Stat 1的IEC特异性破坏的小鼠（Stat 1 IEC-KO）和具有干扰素调节因子3基因（Irf 3（-/-））破坏的小鼠（Irf 7（-/-））中诱导结肠炎。我们使用高分辨率微型内窥镜和体内成像方法来评估结肠炎的进展。我们使用三维小肠和结肠类器官沿着以及RNA-Seq和基因本体方法来表征IL 28对原代IEC的作用。我们研究了IL 28在伤口愈合试验和小鼠结肠伤口中对人肠癌细胞系Caco-2的作用。通过定量聚合酶链反应分析IBD患者（n 62）和无结肠炎症患者（n 23）的结肠活检和切除组织，以测量IL 28 A、IL 28 RA和其他相关细胞因子的表达;还通过免疫荧光分析活检样品以鉴定IL 28产生的来源。从患者组织中分离IEC并与IL 28孵育;通过免疫印迹和共聚焦成像测量信号转导子和转录激活子1（STAT 1）磷酸化。研究结果：与对照组相比，IBD患者和结肠炎小鼠结肠组织中固有层细胞的IL 28表达增加; IBD患者和结肠炎小鼠结肠上皮中IL 28 R水平增加。施用IL 28诱导原代人和小鼠IEC中STAT 1的磷酸化随剂量增加而增加。与对照小鼠相比，给予葡聚糖硫酸钠后，Il 28 ra（-/-）Irf 3（-/-）Irf 3（-/-）Irf 7（-/-）以及Stat 1 IEC-KO小鼠发生了更严重的结肠炎，上皮恢复减少。Il 28 ra（-/-）和Stat 1-IEC-KO小鼠也比对照小鼠对恶唑酮产生更严重的结肠炎。我们发现IL 28诱导上皮细胞中STAT 1的磷酸化（活化），导致它们在类器官培养物中增殖。对具有诱导的结肠伤口的小鼠施用IL 28促进粘膜愈合。结论：IL 28通过激活STAT 1，控制结肠炎小鼠肠上皮细胞增殖，加速粘膜愈合。IL 28可能被开发为IBD患者的治疗剂。
BACKGROUND & AIMS: We investigated the roles of interleukin 28A (also called IL28A or interferon l2) in intestinal epithelial cell (IEC) activation studying its effects in mouse models of inflammatory bowel diseases (IBD) and intestinal mucosal healing. METHODS: Colitis was induced in C57BL/ 6JCrl mice (controls) mice with IEC-specific disruption of Stat1 (Stat1IEC-KO) mice with disruption of the interferon l receptor 1 gene (Il28ra(-/-)) and mice with disruption of the interferon regulatory factor 3 gene (Irf3(-/-)) with or without disruption of Irf7 (Irf7(-/-)). We used high-resolution mini-endoscopy and in vivo imaging methods to assess colitis progression. We used 3-dimensional small intestine and colon organoids along with RNA-Seq and gene ontology methods to characterize the effects of IL28 on primary IECs. We studied the effects of IL28 on the human intestinal cancer cell line Caco-2 in a wound-healing assay and in mice colon wounds. Colonic biopsies and resected tissue from patients with IBD (n 62) and patients without colon inflammation (controls n 23) were analyzed by quantitative polymerase chain rection to measure expression of IL28A IL28RA and other related cytokines; biopsy samples were also analyzed by immunofluorescence to identify sources of IL28 production. IECs were isolated from patient tissues and incubated with IL28; signal transducer and activator of transcription 1 (STAT1) phosphorylation was measured by immunoblots and confocal imaging. RESULTS: Lamina propria cells in colon tissues of patients with IBD and mice with colitis had increased expression of IL28 compared with controls; levels of IL28R were increased in the colonic epithelium of patients with IBD and mice with colitis. Administration of IL28 induced phosphorylation of STAT1 in primary human and mouse IECs increasing with dose. Il28ra(-/-) Irf3(-/-) Irf3(-/-) Irf7(-/-) as well as Stat1IEC-KO mice developed more severe colitis after administration of dextran sulfate sodium than control mice with reduced epithelial restitution. Il28ra(-/-) and Stat1-IEC-KO mice also developed more severe colitis in response to oxazolone than control mice. We found IL28 to induce phosphorylation (activation) of STAT1 in epithelial cells leading to their proliferation in organoid culture. Administration of IL28 to mice with induced colonic wounds promoted mucosal healing. CONCLUSIONS: IL28 controls proliferation of IECs in mice with colitis and accelerates mucosal healing by activating STAT1. IL28 might be developed as a therapeutic agent for patients with IBD.