Immunology Unit and Research Center for Emerging Infections and Zoonoses, University of Veterinary Medicine, Hannover, Germany.
Medical School Hannover, Institute for Medical Microbiology, Hannover, Germany.
Front Immunol. 2018 Feb 13;9:213. doi: 10.3389/fimmu.2018.00213. eCollection 2018.
C-type lectin receptors (CLRs) are carbohydrate-binding receptors that recognize their ligands often in a Ca-dependent manner. Upon ligand binding, myeloid CLRs in innate immunity trigger or inhibit a variety of signaling pathways, thus initiating or modulating effector functions such as cytokine production, phagocytosis, and antigen presentation. CLRs bind to various pathogens, including viruses, fungi, parasites, and bacteria. The bacterium () is a very frequent Gram-negative zoonotic pathogen of humans, causing severe intestinal symptoms. Interestingly, expresses several glycosylated surface structures, for example, the capsular polysaccharide (CPS), lipooligosaccharide (LOS), and envelope proteins. This "Methods" paper describes applications of CLR-Fc fusion proteins to screen for yet unknown CLR/bacteria interactions using as an example. ELISA-based detection of CLR/bacteria interactions allows a first prescreening that is further confirmed by flow cytometry-based binding analysis and visualized using confocal microscopy. By applying these methods, we identified Dectin-1 as a novel CLR recognizing two selected isolates with different LOS and CPS genotypes. In conclusion, the here-described applications of CLR-Fc fusion proteins represent useful methods to screen for and identify novel CLR/bacteria interactions.
C 型凝集素受体（CLRs）是识别其配体的糖结合受体，通常以 Ca2+ 依赖的方式识别。在配体结合后，先天免疫中的髓系 CLRs 可触发或抑制多种信号通路，从而启动或调节细胞因子产生、吞噬作用和抗原呈递等效应功能。CLRs 可结合多种病原体，包括病毒、真菌、寄生虫和细菌。细菌（）是一种非常常见的革兰氏阴性人畜共患病原体，可引起严重的肠道症状。有趣的是，它表达了几种糖基化的表面结构，例如荚膜多糖（CPS）、脂寡糖（LOS）和包膜蛋白。本文“方法”部分以 为例，介绍了使用 CLR-Fc 融合蛋白来筛选未知 CLR/细菌相互作用的应用。基于 ELISA 的 CLR/细菌相互作用检测可进行初步筛选，然后通过基于流式细胞术的结合分析进行进一步确认，并使用共聚焦显微镜进行可视化。通过应用这些方法，我们确定 Dectin-1 是一种识别两种具有不同 LOS 和 CPS 基因型的选定 分离株的新型 CLR。总之，这里描述的 CLR-Fc 融合蛋白的应用代表了筛选和鉴定新型 CLR/细菌相互作用的有用方法。
