Defining the molecular steps in the interaction of pathogenic bacteria with alveolar macrophages a key step in pathogenesis and disease

定义致病菌与肺泡巨噬细胞相互作用的分子步骤是发病机制和疾病的关键步骤

• 批准号: 2127956
• 金额: --
• 依托单位: University of Leicester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2127956
• 关键词: Defining; molecular; steps; interaction; pathogenic

We have recently demonstrated that bacteria, reported to be exclusive extracellular pathogens, undergo a phase of intracellular replication in a specific subset of macrophages prior to invasive infection. This interaction is mediated by the macrophage receptor CD169 (siglec1). Interaction with CD169 and uptake by CD169+ macrophages has been reported for a series of sialylated pathogens. Other bacteria with sialylated surface carbohydrates or proteins highly likely to share the same or similar CD169-binding associated phenotypes. While uptake by CD169+ cells has been described, the occurrence of bacterial intracellular replication in CD169+ cells, the specific steps leading to this intracellular replication and the role of these events in the pathogenesis of infection, have only been discovered most recently by our group for Streptococcus pneumoniae. The hypothesis at the basis of this project is that specific molecular events in the interaction between bacteria and CD169+ cells can guide optimisation of strategies for infection prevention. To test this hypothesis the project will address the following two objectives:- Characterisation of the steps in the intracellular fate of different pathogens, in CD169+ macrophages- Characterisation of the microbial factors involved in this interaction In order to design a robust approach, the project will address these interactions starting with two model pathogens. The other species listed above are available in the lab and recorded on the HBA forms and will be utilised in the later phases of the project to test the broader validity of the findings. As main source of CD169+ cells, porcine alveolar and peri-follicular splenic macrophages will be used. The laboratory hosting the student has recently set up models for primary porcine splenic and alveolar macrophage cultures. These models will be exploited to investigate the specific steps in the interaction of the bacteria with CD169+ cells including phagocytosis, vacuole maturation and mechanism of cell death. The tools will include confocal microscopy, cytofluorimetry, cytokine assays, gene expression profiling and the testing of panels of bacterial mutants. To test the impact on the outcome of infection a limited series of experimental infection in murine models will be performed.The training will range from confocal microscopy, cytofluorimetry, cell and organ culture, bacteriology, next generation sequence analysis, to animal handling. The overall aim of the CASE project is to provide the student with a multidisciplinary training program on a cutting edge scientific topic within a tight academic-industrial interaction.

我们最近已经证明，细菌，据报道是排他性的细胞外病原体，在侵入性感染之前，在特定的巨噬细胞亚群中经历一个细胞内复制阶段。这种相互作用由巨噬细胞受体CD 169（siglec 1）介导。已经报道了一系列唾液酸化病原体与CD 169的相互作用和CD 169+巨噬细胞的摄取。具有唾液酸化表面碳水化合物或蛋白质的其他细菌极有可能具有相同或相似的CD 169结合相关表型。虽然已经描述了CD 169+细胞的摄取，但我们的肺炎链球菌研究组最近才发现CD 169+细胞中细菌胞内复制的发生、导致这种胞内复制的具体步骤以及这些事件在感染发病机制中的作用。该项目的基础假设是，细菌和CD 169+细胞之间相互作用的特定分子事件可以指导感染预防策略的优化。为了检验这一假设，该项目将解决以下两个目标：-表征不同病原体在CD 169+巨噬细胞中的细胞内命运的步骤-表征这种相互作用中涉及的微生物因素为了设计一种稳健的方法，该项目将从两种模型病原体开始解决这些相互作用。上面列出的其他物种可在实验室中获得，并记录在HBA表格中，并将在项目的后期阶段用于测试结果的更广泛有效性。作为CD 169+细胞的主要来源，将使用猪肺泡和滤泡周围脾巨噬细胞。该学生所在的实验室最近建立了原代猪脾脏和肺泡巨噬细胞培养的模型。这些模型将用于研究细菌与CD 169+细胞相互作用的具体步骤，包括吞噬作用、空泡成熟和细胞死亡机制。这些工具将包括共聚焦显微镜、细胞荧光测定法、细胞因子测定、基因表达谱分析和细菌突变体小组测试。为了测试对感染结果的影响，将在小鼠模型中进行有限系列的实验感染，培训范围包括共聚焦显微镜、细胞荧光测定法、细胞和器官培养、细菌学、下一代序列分析和动物处理。CASE项目的总体目标是在紧密的学术-工业互动中为学生提供关于前沿科学主题的多学科培训计划。

项目成果

Analyzing Macrophage Infection at the Organ Level.

分析器官水平的巨噬细胞感染。

• DOI: 10.1007/978-1-0716-1900-1_22
• 发表时间: 2022
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Hames RG

Splenic macrophages as the source of bacteraemia during pneumococcal pneumonia.

• DOI: 10.1016/j.ebiom.2021.103601
• 发表时间: 2021-10
• 期刊: EBioMedicine
• 影响因子: 11.1
• 作者: Carreno D;Wanford JJ;Jasiunaite Z;Hames RG;Chung WY;Dennison AR;Straatman K;Martinez-Pomares L;Pareek M;Orihuela CJ;Restrepo MI;Lim WS;Andrew PW;Moxon ER;Oggioni MR
• 通讯作者: Oggioni MR

Interaction of Klebsiella pneumoniae with tissue macrophages in a mouse infection model and ex-vivo pig organ perfusions: an exploratory investigation.

• DOI: 10.1016/s2666-5247(21)00195-6
• 发表时间: 2021-12
• 期刊: The Lancet. Microbe
• 作者: Wanford JJ;Hames RG;Carreno D;Jasiunaite Z;Chung WY;Arena F;Di Pilato V;Straatman K;West K;Farzand R;Pizza M;Martinez-Pomares L;Andrew PW;Moxon ER;Dennison AR;Rossolini GM;Oggioni MR
• 通讯作者: Oggioni MR

Diurnal differences in intracellular replication within splenic macrophages correlates with the outcome of pneumococcal infection

脾巨噬细胞内细胞内复制的昼夜差异与肺炎球菌感染的结果相关

• DOI: 10.1101/2022.01.30.478422
• 发表时间: 2022
• 作者: Hames R