Application of a gut-immune co-culture system for the study of N-glycan-dependent host-pathogen interactions of Campylobacter jejuni.
Application of a gut-immune co-culture system for the study of N-glycan-dependent host-pathogen interactions of Campylobacter jejuni.
复制标题
应用肠道免疫共培养系统研究空肠弯曲杆菌 N-聚糖依赖性宿主-病原体相互作用。
DOI：
10.1093/glycob/cwz105
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发表时间：
2020
期刊：
影响因子：
4.3
通讯作者：
Imperiali,Barbara
中科院分区：
文献类型：
--
作者：
Zamora,CristinaY;Ward,ElizabethM;Kester,JemilaC;Chen,WenLiKelly;Velazquez,JasonG;Griffith,LindaG;Imperiali,Barbara
为了更接近人类肠道微环境，建立了具有根尖可及性的体外肠道免疫共培养模型，研究了n -连接蛋白糖基化途径在空肠弯曲杆菌致病性中的作用。肠道免疫共培养（GIC）是通过包含产生黏液的杯状细胞、人肠细胞和树突状细胞，将含有黏液的上皮单层与先天免疫系统的元素结合在一起，来模拟人小肠的重要方面。通过表征n链糖基化促进的宿主-病原体相互作用，如宿主上皮屏障功能、细菌入侵和免疫原性，证明了该系统的实用性。11168C存在时人体肠道屏障功能的变化。利用gis对野生型空肠菌株进行定量分析。在细胞感染性试验中，糖基化受损的菌株11168 ΔpglEwas粘附和侵入该肠道模型的能力降低了100倍。炎症信号的量化显示11168ΔpglEdifferentially调节了不同肠道微环境中的炎症反应，在某些情况下是抑制的，但在另一些情况下是激活的。野生型和11168ΔpglE空肠弧菌产生的毒力相关外膜囊泡具有不同的组成和功能，当提供给肠道免疫共培养模型时，两者都导致免疫系统激活。这方面的分析c。通过应用肠道免疫模型，在其n -链糖存在和不存在的情况下，肠道感染性得以实现，我们预计该系统将适用于c的进一步研究。肠和其他感兴趣的肠病原体。
An in vitro gut-immune co-culture model with apical and basal accessibility, designed to more closely resemble a human intestinal microenvironment, was employed to study the role of the N-linked protein glycosylation pathway inCampylobacter jejunipathogenicity. The gut-immune co-culture (GIC) was developed to model important aspects of the human small intestine by the inclusion of mucin-producing goblet cells, human enterocytes and dendritic cells, bringing together a mucus-containing epithelial monolayer with elements of the innate immune system. The utility of the system was demonstrated by characterizing host–pathogen interactions facilitated by N-linked glycosylation, such as host epithelial barrier functions, bacterial invasion and immunogenicity. Changes in human intestinal barrier functions in the presence of 11168C. jejuni(wildtype) strains were quantified using GICs. The glycosylation-impaired strain 11168 ΔpglEwas 100-fold less capable of adhering to and invading this intestinal model in cell infectivity assays. Quantification of inflammatory signaling revealed that 11168ΔpglEdifferentially modulated inflammatory responses in different intestinal microenvironments, suppressive in some but activating in others. Virulence-associated outer membrane vesicles produced by wildtype and 11168ΔpglE C. jejuniwere shown to have differential composition and function, with both leading to immune system activation when provided to the gut-immune co-culture model. This analysis of aspects ofC. jejuniinfectivity in the presence and absence of its N-linked glycome is enabled by application of the gut-immune model, and we anticipate that this system will be applicable to further studies ofC. jejuniand other enteropathogens of interest.