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 in Campylobacter jejuni pathogenicity. 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 11168 C. jejuni (wildtype) strains were quantified using GICs. The glycosylation-impaired strain 11168 ΔpglE was 100-fold less capable of adhering to and invading this intestinal model in cell infectivity assays. Quantification of inflammatory signaling revealed that 11168ΔpglE differentially 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. jejuni were 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 of C. jejuni infectivity 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 of C. jejuni and other enteropathogens of interest.

中文翻译：

---

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。


采用具有顶端和基底可及性的体外肠道免疫共培养模型，旨在更接近人类肠道微环境，用于研究 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) 的开发是为了模拟人类小肠的重要方面，通过包含产生粘蛋白的杯状细胞、人类肠上皮细胞和树突状细胞，将含有粘液的上皮单层与先天的元素结合在一起。免疫系统。该系统的实用性通过表征 N-连接糖基化促进的宿主-病原体相互作用得到证明，例如宿主上皮屏障功能、细菌入侵和免疫原性。使用 GIC 对 11168 个空肠弯曲菌（野生型）菌株存在下人类肠道屏障功能的变化进行了定量。在细胞感染性测定中，糖基化受损的菌株 11168 ΔpglE 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，11168ΔpglE 在不同的肠道微环境中差异调节炎症反应，在某些微环境中抑制炎症反应，但在另一些微环境中激活炎症反应。由野生型和 11168ΔpglE 空肠弯曲菌产生的毒力相关外膜囊泡被证明具有不同的组成和功能，当提供给肠道免疫共培养模型时，两者都会导致免疫系统激活。通过应用肠道免疫模型，可以对空肠弯曲菌在存在和不存在 N 连接糖组的情况下的感染性进行分析，我们预计该系统将适用于空肠弯曲菌和其他肠道病原体的进一步研究的兴趣。