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花生过敏原会改变Caco-2细胞培养物中的肠道屏障通透性和紧密连接定位。

Peanut allergens alter intestinal barrier permeability and tight junction localisation in Caco-2 cell cultures.

作者信息

Price Dwan B, Ackland M Leigh, Burks Wesley, Knight Matthew I, Suphioglu Cenk

机构信息

Centre for Cellular and Molecular Biology (CCMB), School of Life and Environmental Sciences, Deakin University, Burwood, Victoria, Australia.

出版信息

Cell Physiol Biochem. 2014;33(6):1758-77. doi: 10.1159/000362956. Epub 2014 May 23.

Abstract

BACKGROUND/AIMS: Allergen absorption by epithelia may play an important role in downstream immune responses. Transport mechanisms that can bypass Peyer's patches include transcellular and paracellular transport. The capacity of an allergen to cross via these means can modulate downstream processing of the allergen by the immune system. The aim of this study was to investigate allergen-epithelial interactions of peanut allergens with the human intestinal epithelium.

METHODS

We achieved this using the human Caco-2 cell culture model, exposed to crude peanut extract. Western and immunofluorescence analysis were used to identify the cellular and molecular changes of peanut extract on the intestinal epithelium.

RESULTS

Following exposure of Caco-2 cells to peanut extract, binding of the peanut allergens Ara h 1 and Ara h 2 to the apical cellular membrane and transcytosis across the monolayers were observed. Additionally, the co-localisation of the transmembrane tight junction proteins occludin, JAM-A and claudin-1, with the intracellular adhesion protein ZO-1 was modified.

CONCLUSION

Disruption of Caco-2 barrier integrity through tight junction disruption may enable movement of peanut proteins across the intestinal epithelium. This accounts for peanut's increased allergenicity, compared to other food allergens, and provides an explanation for the potency of peanut allergens in immune response elicitation.

摘要

背景/目的:上皮细胞对变应原的吸收可能在下游免疫反应中起重要作用。可绕过派尔集合淋巴结的转运机制包括跨细胞转运和细胞旁转运。变应原通过这些途径穿过的能力可调节免疫系统对变应原的下游处理。本研究的目的是调查花生变应原与人肠道上皮细胞之间的变应原-上皮相互作用。

方法

我们使用人Caco-2细胞培养模型,使其暴露于粗制花生提取物中,以此实现上述目的。采用蛋白质免疫印迹法和免疫荧光分析法来确定花生提取物对肠道上皮细胞的细胞和分子变化。

结果

将Caco-2细胞暴露于花生提取物后,观察到花生变应原Ara h 1和Ara h 2与顶端细胞膜结合并跨单层细胞进行胞吞转运。此外,跨膜紧密连接蛋白闭合蛋白、连接黏附分子A和闭合蛋白-1与细胞内黏附蛋白ZO-1的共定位发生了改变。

结论

通过紧密连接破坏导致Caco-2屏障完整性受损,可能使花生蛋白穿过肠道上皮细胞。这解释了与其他食物变应原相比,花生变应原性增加的原因,并为花生变应原引发免疫反应的效力提供了解释。

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