Thoracic Diseases Research Unit, Departments of Medicine and Biochemistry, Mayo Clinic College of Medicine, Rochester, MN, USA.
J Med Microbiol. 2021 Dec;70(12). doi: 10.1099/jmm.0.001470.
Pathogen-associated molecular patterns' (PAMPs) are microbial signatures that are recognized by host myeloid C-type lectin receptors (CLRs). These CLRs interact with micro-organisms via their carbohydrate recognition domains (CRDs) and engage signalling pathways within the cell resulting in pro-inflammatory and microbicidal responses. In this article, we extend our laboratory study of additional CLRs that recognize fungal ligands against and and their purified major surface glycoproteins (Msgs). To study the potential of newly synthesized hFc-CLR fusions on binding to and its Msg. A library of new synthesized hFc-CLR fusions was screened against and organisms and their purified major surface glycoproteins (Msgs) found on the respective fungi via modified ELISA. Immunofluorescence assay (IFA) was implemented and quantified to verify results. mRNA expression analysis by quantitative PCR (q-PCR) was employed to detect respective CLRs found to bind fungal organisms in the ELISA and determine their expression levels in the mouse immunosuppressed Pneumocystis pneumonia (PCP) model. We detected a number of the CLR hFc-fusions displayed significant binding with and organisms, and similarly to their respective Msgs. Significant organism and Msg binding was observed for CLR members C-type lectin domain family 12 member A (CLEC12A), Langerin, macrophage galactose-type lectin-1 (MGL-1), and specific intracellular adhesion molecule-3 grabbing non-integrin homologue-related 3 (SIGNR3). Immunofluorescence assay (IFA) with the respective CLR hFc-fusions against whole life forms corroborated these findings. Lastly, we surveyed the mRNA expression profiles of the respective CLRs tested above in the mouse immunosuppressed Pneumocystis pneumonia (PCP) model and determined that macrophage galactose type C-type lectin (), implicated in recognizing terminal N-acetylgalactosamine (GalNAc) found in the glycoproteins of microbial pathogens was significantly up-regulated during infection. The data herein add to the growing list of CLRs recognizing and provide insights for further study of organism/host immune cell interactions.
病原体相关分子模式（PAMPs）是宿主髓样 C 型凝集素受体（CLRs）识别的微生物特征。这些 CLRs 通过其碳水化合物识别结构域（CRD）与微生物相互作用，并在细胞内参与信号通路，导致促炎和杀菌反应。在本文中，我们扩展了我们对识别真菌配体的其他 CLRs 的实验室研究，针对 和 及其纯化的主要表面糖蛋白（Msg）。为了研究新合成的 hFc-CLR 融合物与 和其 Msg 结合的潜力。通过改良 ELISA 筛选了新合成的 hFc-CLR 融合文库，以检测相应真菌上的 和 Msg。实施了免疫荧光分析（IFA）并进行了量化以验证结果。通过定量 PCR（q-PCR）进行了 mRNA 表达分析，以检测在 ELISA 中发现与真菌结合的相应 CLR，并确定它们在小鼠免疫抑制性卡氏肺孢子虫肺炎（PCP）模型中的表达水平。我们检测到许多 CLR hFc 融合物与 和 生物体具有显著的结合，并且与它们各自的 Msg 相似。CLEC12A、Langerin、巨噬细胞半乳糖型凝集素-1（MGL-1）和特异性细胞内黏附分子-3 抓取非整合素同源物 3（SIGNR3）等 CLR 成员对生物体和 Msg 的结合具有重要意义。用相应的 CLR hFc 融合物对整个 生命形式进行免疫荧光分析（IFA）证实了这些发现。最后，我们调查了上述在小鼠免疫抑制性卡氏肺孢子虫肺炎（PCP）模型中测试的 CLR 的 mRNA 表达谱，并确定在感染过程中，识别微生物病原体糖蛋白中末端 N-乙酰半乳糖胺（GalNAc）的巨噬细胞半乳糖型 C 型凝集素（）显著上调。本文中的数据增加了识别 的 CLR 列表，并为进一步研究生物体/宿主免疫细胞相互作用提供了见解。
