Desmoglein 2-mediated adhesion is required for intestinal epithelial barrier integrity
Desmoglein 2-mediated adhesion is required for intestinal epithelial barrier integrity
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DOI：
10.1152/ajpgi.00239.2009
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发表时间：
2010-05-01
影响因子：
4.5
通讯作者：
Waschke, Jens
中科院分区：
文献类型：
--
作者：
Schlegel, Nicolas;Meir, Michael;Waschke, Jens
Schlegel N、Meir M、Heupel W、Holthofer B、Leube RE、Waschke J。桥粒糖蛋白 2 介导的粘附是肠上皮屏障完整性所必需的。 Am J Physiol Gastrointest Liver Physiol 298：G774-G783，2010。首次发表于 2010 年 3 月 11 日； doi:10.1152/ajpgi.00239.2009.-肠上皮中形成“末端杆”的细胞间连接的完整性对于密封肠屏障至关重要。尽管紧密连接和粘附连接的具体作用是众所周知的，但桥粒粘附对于维持肠上皮屏障的贡献尚未得到具体解决。在本研究中，我们生成了针对胞外结构域 (Dsg2 ED) 的桥粒芯糖蛋白 2 抗体，以测试 Dsg2 介导的粘附受损是否会影响体外肠上皮屏障功能。该抗体能够在无细胞原子力显微镜实验中特异性阻断 Dsg2 相互作用。对于肠屏障的体外研究，我们使用分化为紧密的肠细胞样上皮单层的 Caco2 细胞。在基于分散酶的肠细胞解离测定中，与对照相比，将 Dsg2 ED 应用于 Caco2 单层导致细胞解离增加。在类似条件下，Dsg2 抗体显着降低跨上皮电阻并增加 FITC-葡聚糖通量，表明 Dsg2 相互作用在维持上皮肠屏障功能中至关重要。免疫染色显示，这是由于 Dsg2 ED 抗体诱导的紧密连接破裂，因为紧密连接蛋白 claudins 1、4 和 5、occludin 和紧密连接相关蛋白 zonula occlusionns-1 通过 Dsg2 ED 处理从细胞边界部分去除。通过应用针对 Dsg2 ED 的商业单克隆抗体获得了类似的结果。使用 Dsg2-Fc 包被的珠子进行吸收实验可阻断抗体诱导的效应。我们的数据表明，Dsg2 介导的粘附影响紧密连接的完整性，并且是维持肠上皮屏障特性所必需的。
Schlegel N, Meir M, Heupel W, Holthofer B, Leube RE, Waschke J. Desmoglein 2-mediated adhesion is required for intestinal epithelial barrier integrity. Am J Physiol Gastrointest Liver Physiol 298: G774-G783, 2010. First published March 11, 2010; doi:10.1152/ajpgi.00239.2009.-The integrity of intercellular junctions that form the "terminal bar" in intestinal epithelium is crucial for sealing the intestinal barrier. Whereas specific roles of tight and adherens junctions are well known, the contribution of desmosomal adhesion for maintaining the intestinal epithelial barrier has not been specifically addressed. For the present study, we generated a desmoglein 2 antibody directed against the extracellular domain (Dsg2 ED) to test whether impaired Dsg2-mediated adhesion affects intestinal epithelial barrier functions in vitro. This antibody was able to specifically block Dsg2 interaction in cell-free atomic-force microscopy experiments. For in vitro studies of the intestinal barrier we used Caco2 cells following differentiation into tight enterocyte-like epithelial monolayers. Application of Dsg2 ED to Caco2 monolayers resulted in increased cell dissociation compared with controls in a dispase-based enterocyte dissociation assay. Under similar conditions, Dsg2 antibody significantly decreased transepithelial electrical resistance and increased FITC-dextran flux, indicating that Dsg2 interaction is critically involved in the maintenance of epithelial intestinal barrier functions. As revealed by immunostaining, this was due to Dsg2 ED antibody-induced rupture of tight junctions because tight junction proteins claudins 1, 4, and 5, occludin, and tight junction-associated protein zonula occludens-1 were partially removed from cell borders by Dsg2 ED treatment. Similar results were obtained by application of a commercial monoclonal antibody directed against the ED of Dsg2. Antibody-induced effects were blocked by absorption experiments using Dsg2-Fc-coated beads. Our data indicate that Dsg2-mediated adhesion affects tight junction integrity and is required to maintain intestinal epithelial barrier properties.