PTPN2 Regulates Interactions Between Macrophages and Intestinal Epithelial Cells to Promote Intestinal Barrier Function.
PTPN2 Regulates Interactions Between Macrophages and Intestinal Epithelial Cells to Promote Intestinal Barrier Function.
复制标题
PTPN2调节巨噬细胞和肠上皮细胞之间的相互作用以促进肠道屏障功能。
DOI：
10.1053/j.gastro.2020.07.004
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发表时间：
2020-11
期刊：
影响因子：
29.4
通讯作者：
McCole DF
中科院分区：
文献类型：
--
作者：
Spalinger MR;Sayoc-Becerra A;Santos AN;Shawki A;Canale V;Krishnan M;Niechcial A;Obialo N;Scharl M;Li J;Nair MG;McCole DF
巨噬细胞调节肠上皮细胞（IEC）屏障特性的机制尚不清楚。非受体型蛋白酪氨酸磷酸酶2（PTPN2）保护IEC屏障免受炎症诱导的破坏，并调节巨噬细胞功能。我们研究了PTPN2是否控制IEC和巨噬细胞之间的相互作用以维持肠屏障功能。
人IEC（Caco - 2BBe/HT - 29.cl19a细胞）和小鼠肠类器官单层分别与人巨噬细胞（THP - 1、U937、炎症性肠病[IBD]患者的原代单核细胞衍生巨噬细胞）或小鼠巨噬细胞共培养。我们评估了屏障功能（跨上皮电阻[TEER]以及对4 kDa荧光标记葡聚糖[FD4]或70 kDa罗丹明B - 葡聚糖的通透性）和巨噬细胞极化。我们分析了髓系细胞特异性PTPN2缺失小鼠（Ptpn2 - LysMCre小鼠）和Ptpn2未被破坏的小鼠（对照）的肠道组织；一些小鼠注射了抗白细胞介素6（IL6）的中和抗体。用小发夹RNA在巨噬细胞和/或IEC中敲低蛋白质。
在巨噬细胞和/或IEC中敲低PTPN2会增加IEC单层的通透性，从两种细胞类型中敲低时具有协同效应，并增加巨噬细胞 - IEC共培养中炎性巨噬细胞的产生。Ptpn2 - LysMCre小鼠的结肠固有层炎性巨噬细胞显著增加；这些小鼠体内和体外结肠对FD4的通透性增加，体外结肠TEER降低。Nanostring分析显示Ptpn2 - LysMCre小鼠结肠巨噬细胞中IL6的表达显著增加。一种IL6阻断抗体逆转了PTPN2缺陷巨噬细胞的作用，降低了培养中的IEC单层以及Ptpn2 - LysMCre小鼠的通透性。携带与疾病相关的单核苷酸多态性（PTPN2 rs1893217）的IBD患者的巨噬细胞具有与小鼠PTPN2缺陷巨噬细胞相同的特征，包括在与人IEC或小鼠肠类器官单层共培养时TEER降低和通透性增加，这些都可通过抗IL6恢复。
PTPN2是巨噬细胞和IEC之间相互作用所必需的；任一细胞类型中PTPN2的缺失都会导致肠屏障缺陷，两种细胞类型中PTPN2的缺失具有协同效应。我们提供了一种机制，通过该机制PTPN2基因变异损害肠上皮屏障功能并增加炎症性疾病如IBD的风险。
作者确定了一种机制，即许多IBD患者中发现的PTPN2改变影响肠上皮细胞和免疫细胞之间的相互作用。这种相互作用的破坏会影响肠屏障特性和免疫反应。
The mechanisms by which macrophages regulate intestinal epithelial cell (IEC) barrier properties are poorly understood. Protein tyrosine phosphatase non-receptor type 2 (PTPN2) protects the IEC barrier from inflammation-induced disruption and regulates macrophage functions. We investigated whether PTPN2 controls interactions between IECs and macrophages to maintain intestinal barrier function. Human IEC (Caco-2BBe/HT-29.cl19a cells) and mouse enteroid monolayers were co-cultured with human macrophages (THP-1, U937, primary monocyte-derived macrophages from patients with inflammatory bowel disease [IBD]) or mouse macrophages, respectively. We assessed barrier function (transepithelial electrical resistance [TEER] and permeability to 4 kDa fluorescently labeled dextran [FD4] or rhodamine B-dextran 70 kDa) and macrophage polarization. We analyzed intestinal tissues from mice with myeloid cell-specific deletion of PTPN2 (Ptpn2-LysMCre mice) and mice without disruption of Ptpn2 (controls); some mice were given injections of a neutralizing antibody against interleukin 6 (IL6). Proteins were knocked down in macrophages and/or IECs with small hairpin RNAs. Knockdown of PTPN2 in either macrophages and/or IECs increased the permeability of IEC monolayers, had a synergistic effect when knocked down from both cell types, and increased development of inflammatory macrophages in macrophage–IEC co-cultures. Colon lamina propria from Ptpn2-LysMCre mice had significant increases in inflammatory macrophages; these mice had increased in vivo and ex vivo colon permeability to FD4 and reduced ex vivo colon TEER. Nanostring analysis revealed significant increases in expression of IL6 in colon macrophages from Ptpn2-LysMCre mice. An IL6 blocking antibody reversed the effects of PTPN2-deficient macrophages, reducing permeability of IEC monolayers in culture and in Ptpn2-LysMCre mice. Macrophages from patients with IBD carrying a single-nucleotide polymorphism associated with the disease (PTPN2 rs1893217) had the same features of PTPN2-deficient macrophages from mice, including reduced TEER and increased permeability in co-cultures with human IEC or mouse enteroid monolayers, which were restored by anti-IL6. PTPN2 is required for interactions between macrophages and IECs; loss of PTPN2 from either cell type results in intestinal barrier defects, and loss from both cell types has a synergistic effect. We provide a mechanism by which PTPN2 gene variants compromise intestinal epithelial barrier function and increase risk of inflammatory disorders such as IBD. The authors identified a mechanism by which alterations in PTPN2 that have been found in many patients with IBD affects interactions between intestinal epithelial cells and immune cells. Disruption of this interaction can affect intestinal barrier properties and the immune response.
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