Establishment of novel macrophage cell lines to study the pathogenesis of respiratory bacterial pathogens in lung alveolar macrophages

建立新型巨噬细胞系以研究肺泡巨噬细胞中呼吸道细菌病原体的发病机制

• 批准号: NC/V001019/1
• 负责人: Gyorgy Fejer
• 金额: $ 9.67万
• 依托单位: Plymouth University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=NC%2FV001019%2F1
• 关键词: Establishment; novel; macrophage; cell; lines

Macrophages are important immune cells in the first line of defence against infectious pathogens.To understand macrophage-bacteria interactions mice are frequently used. Macrophages can be best investigated using purified cells from organs or by producing them from bone marrow in tissue culture, but the availability of such cells is limited.Lung alveolar macrophages (AMs) play central roles in defence against respiratory bacterial pathogens. Infection of AMs can result in direct pathogen elimination, however, bacteria may overcome macrophage killing mechanisms allowing pathogen replication or persistence inside the cell. The activation of a plethora of pathogen sensing mechanisms play key roles in these processes and their understanding is key to the efficient treatment of respiratory infections. The therapy of bacterial infections of the airways can be still problematic and macrophages play key roles in these processes as bacteria can be unavailable to drugs when persist or replicate in macrophages. We described a novel, continuously growing, murine model of lung alveolar macrophages (AMs), providing unrestricted amounts of primary macrophages (MPI cells). Using this system we have demonstrated the very high sensitivity of MPI cells and AMs to respiratory pathogens and have shown the existence of unique immune mechanisms in AMs. The sensing of respiratory bacteria by AMs involves various immune receptors, however, the understanding of their contribution to pathogenesis mechanisms is hampered by the restricted availability of AMs. Here we will transfer the MPI cell technology to the Kings College London (KCL) and the Institut Pasteur, Korea (IPK) to study bacterial pathogenesis in lung macrophages. We will establish new MPI cell lines from relevant genetically modified mice lacking various bacterial pathogen recognition sensors. To better understand respiratory macrophage-bacteria interactions we will establish MPI cells from these g mice available at KCL, the Sanger Institute and the IPK. To make the cells accessible to other researchers we will make a repository of the new cell lines and/or distribute them through commercial vendors to the wider community.We have ongoing in vivo and in vitro studies with the above listed pathogens. We will use the established new cell lines to complement these studies and obtain mechanistic data regarding the activation of innate immune pathways to the sensing and the treatment of respiratory bacterial infections.

巨噬细胞是抵御感染性病原体第一道防线的重要免疫细胞。为了了解巨噬细胞-细菌相互作用，经常使用小鼠。使用从器官中纯化的细胞或在组织培养中从骨髓中产生巨噬细胞可以最好地研究巨噬细胞，但此类细胞的可用性有限。肺泡巨噬细胞 (AM) 在防御呼吸道细菌病原体方面发挥着核心作用。 AM 的感染可导致病原体直接消除，然而，细菌可能克服巨噬细胞杀伤机制，允许病原体在细胞内复制或持续存在。大量病原体传感机制的激活在这些过程中发挥着关键作用，对它们的理解是有效治疗呼吸道感染的关键。气道细菌感染的治疗仍然存在问题，巨噬细胞在这些过程中发挥着关键作用，因为当细菌在巨噬细胞中持续存在或复制时，药物可能无法使用。我们描述了一种新型的、持续生长的小鼠肺泡巨噬细胞（AM）模型，提供不受限制数量的原代巨噬细胞（MPI 细胞）。使用该系统，我们证明了 MPI 细胞和 AM 对呼吸道病原体具有非常高的敏感性，并表明 AM 中存在独特的免疫机制。 AM 对呼吸道细菌的感知涉及多种免疫受体，然而，AM 的可用性有限，阻碍了对它们对发病机制的贡献的理解。在这里，我们将把MPI细胞技术转移到伦敦国王学院（KCL）和韩国巴斯德研究所（IPK），以研究肺巨噬细胞中的细菌发病机制。我们将从缺乏各种细菌病原体识别传感器的相关转基因小鼠中建立新的 MPI 细胞系。为了更好地了解呼吸道巨噬细胞-细菌相互作用，我们将从 KCL、桑格研究所和 IPK 提供的这些 g 小鼠中建立 MPI 细胞。为了使其他研究人员能够获得这些细胞，我们将建立新细胞系的存储库和/或通过商业供应商将它们分发给更广泛的社区。我们正在对上述病原体进行体内和体外研究。我们将使用已建立的新细胞系来补充这些研究，并获得有关激活先天免疫途径来感知和治疗呼吸道细菌感染的机制数据。