The molecular mechanisms behind infection and propagation of human restricted pathogens such as human norovirus (HuNoV) have defied interrogation because they were previously unculturable. However, human intestinal enteroids (HIEs) have emerged to offer unique ex vivo models for targeted studies of intestinal biology, including inflammatory and infectious diseases. Carbohydrate-dependent histo-blood group antigens (HBGAs) are known to be critical for clinical infection. To explore whether HBGAs of glycosphingolipids contribute to HuNoV infection, we obtained HIE cultures established from stem cells isolated from jejunal biopsies of six individuals with different ABO, Lewis, and secretor genotypes. We analyzed their glycerolipid and sphingolipid compositions and quantified interaction kinetics and the affinity of HuNoV virus-like particles (VLPs) to lipid vesicles produced from the individual HIE-lipid extracts. All HIEs had a similar lipid and glycerolipid composition. Sphingolipids included HBGA-related type 1 chain glycosphingolipids (GSLs), with HBGA epitopes corresponding to the geno- and phenotypes of the different HIEs. As revealed by single-particle interaction studies of Sydney GII.4 VLPs with glycosphingolipid-containing HIE membranes, both binding kinetics and affinities explain the patterns of susceptibility toward GII.4 infection for individual HIEs. This is the first time norovirus VLPs have been shown to interact specifically with secretor gene–dependent GSLs embedded in lipid membranes of HIEs that propagate GII.4 HuNoV ex vivo, highlighting the potential of HIEs for advanced future studies of intestinal glycobiology and host-pathogen interactions.

中文翻译：

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鞘糖脂的组织血型抗原可预测人类肠道肠类对诺如病毒感染的易感性。

人类限制性病原体（如人类诺如病毒 (HuNoV)）的感染和传播背后的分子机制已经无法验证，因为它们以前无法培养。然而，人类肠内肠样体 (HIE) 的出现为肠道生物学的靶向研究提供了独特的离体模型，包括炎症和传染病。已知碳水化合物依赖性组织血型抗原 (HBGA) 对临床感染至关重要。为了探索鞘糖脂的 HBGA 是否有助于 HuNoV 感染，我们获得了从具有不同 ABO、Lewis 和分泌基因型的 6 个个体的空肠活检组织中分离的干细胞建立的 HIE 培养物。我们分析了它们的甘油脂和鞘脂组成，并量化了相互作用动力学以及 HuNoV 病毒样颗粒 (VLP) 对由单个 HIE 脂质提取物产生的脂质囊泡的亲和力。所有 HIE 具有相似的脂质和甘油脂组成。鞘脂包括与 HBGA 相关的 1 型链糖鞘脂 (GSL)，HBGA 表位对应于不同 HIE 的基因型和表型。正如悉尼 GII.4 VLP 与含糖鞘脂的 HIE 膜的单粒子相互作用研究所揭示的那样，结合动力学和亲和力都解释了个体 HIE 对 GII.4 感染的易感性模式。这是第一次显示诺如病毒 VLP 与嵌入在传播 GII 的 HIE 的脂膜中的分泌基因依赖性 GSL 发生特异性相互作用。