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首页> 外文期刊>Journal of the Canadian Association of Gastroenterology >INVESTIGATING THE ROLE OF ENDOGENOUS INTERLEUKIN-10 IN INTESTINAL EPITHELIAL CELLS AND HOMEOSTASIS
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INVESTIGATING THE ROLE OF ENDOGENOUS INTERLEUKIN-10 IN INTESTINAL EPITHELIAL CELLS AND HOMEOSTASIS

机译：研究内源性白细胞介素-10在肠上皮细胞和稳态中的作用

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Background IL-10 is appreciated for its potent anti-inflammatory effects on leukocytes in mucosal immunity. However, far less attention has been paid to the impact of IL-10 on epithelial cells, which make up the crucial barrier interface between the host mucosa and the external environment. Furthermore, most studies examine the effects of exogenous IL-10, disregarding the possible presence and function of autocrine or paracrine IL-10 in the epithelium. Aims Using ex vivo organoids we aimed to examine the small intestinal epithelium for IL-10 and dissect any role for endogenously produced cytokine. Methods We growed small intestinal organoids (enteroids) from crypts isolated from C57BL/6 mice (WT) and IL-10-gene knockout mice (IL-10KO). Cellular markers were characterized through qpCR, while IL-10 and IL-10 receptor localization was characterized though immunofluorescence. Results We discovered that cells in WT enteroids expressed IL-10 and IL-10R1 constitutively throughout development. Immunofluorescent staining revealed that IL-10 localizes to Paneth cells and appears to be secreted apically. Having established that IL-10 is secreted in enteroids, we compared enteroids from IL-10KO versus WT mice. IL-10KO enteroids developed to morphologically resemble WT enteroids; however, we detected an imbalance with lower secretory cell markers over absorptive cell types in the IL-10KO enteroids, measured as less mRNA for lysozyme, cryptdins and mucin-2. Addition of IL-10 to IL-10KO enteroids did not correct these defects, but did ameliorate the lineage balance by reducing absorptive cell lineage markers (sucrose isomaltase). IL-10R1 was localized on both apical and basolateral side of cell in enteroids. We suspect that epithelial-derived IL-10 likely acts on apical IL-10R, which may conduct a different response from basolateral receptor stimulation. Conclusions In conclusion, IL-10 is present in the small intestinal epithelium; more remains to be determined regarding the role this cytokine plays in gut development and homeostasis.
机译：背景技术IL-10对于其对粘膜免疫的白细胞有效的抗炎作用。然而，对IL-10对上皮细胞的影响远远不太关注，这构成了宿主粘膜和外部环境之间的关键屏障界面。此外，大多数研究检查外源IL-10的影响，忽略了上皮中自分泌或旁静脉IL-10的可能存在和功能。目的使用前体内有机体，旨在检查IL-10的小肠上皮，并对内源性产生的细胞因子进行了描述。方法从C57BL / 6小鼠（WT）和IL-10-基因敲除小鼠（IL-10KO）中，我们从Crypts中生长小肠有机体（Enteroids）。通过QPCR表征细胞标记，而IL-10和IL-10受体定位的特征在于免疫荧光。结果我们发现WT的细胞在整个发育中表达IL-10和IL-10R1。免疫荧光染色显示IL-10定位于甘蔗细胞，并且似乎是顶部分泌的。已经确定IL-10在肠外分泌物中，我们与IL-10KO与WT小鼠进行了比较了肠毒素。 IL-10KO esteroids开发出的形态学上类似于enteroids;然而，我们检测到在IL-10KO肠毒细胞中的吸收细胞类型的低分泌细胞标记物中检测到不平衡，测量为溶菌酶，加密丁胺和粘蛋白-2的较少mRNA。添加IL-10至IL-10KO Enteroids没有纠正这些缺陷，但通过减少吸收细胞谱系标记（蔗糖异氨酸酶）来改善谱系平衡。 IL-10R1局部化在细胞中的尖端和基底外侧侧。我们怀疑上皮衍生的IL-10可能在顶端IL-10R上起作用，这可能与基石受体刺激进行不同的反应。结论结论，IL-10存在于小肠上皮中;关于这种细胞因子在肠道发育和稳态中起作用的作用更遗为仍有待确定。

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