Extracellular vesicles released in response to Yersinia pestis

鼠疫耶尔森菌释放细胞外囊泡

• 批准号: 10439253
• 负责人: Matthew B Lawrenz
• 金额: $ 23.44万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-18 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10439253
• 关键词: Anti-Inflammatory Agents; Antigen Presentation; Autoimmunity; Bacteria; Bacterial Infections; Bacterial Proteins; Biological; Biology; Carbohydrates; Cell Communication; Cells; Characteristics; Cytoplasmic Granules; Data; Exocytosis; Future; Generations; Goals; Grant; Growth; Human; Immune; Immune Evasion; Immune response; Immune signaling; Individual; Infection; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Innate Immune System; Knowledge; Lipids; Malignant Neoplasms; Mediating; Membrane; Molecular; Nucleic Acids; Outcome; Pathogenesis; Pathologic; Pathway interactions; Patients; Phagocytosis Inhibition; Physiological; Plague; Play; Population; Production; Property; Proteins; Publishing; Reactive Oxygen Species; Reporting; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Stimulus; System; Testing; Therapeutic; Therapeutic Intervention; Time; Vesicle; Yersinia pestis; base; cytokine; exosome; extracellular vesicles; fungus; human disease; improved; intercellular communication; macrophage; monocyte; mutant; neutrophil; novel; novel strategies; recruit; response; tool; vesicular release

SUMMARY Extracellular vesicles (EVs) are membrane-bound vesicles released by cells that are potent vehicles for intercellular communication. The signaling capacity of EVs is mediated by incorporation of different biomolecules (e.g. proteins, lipids, nucleic acids, and carbohydrates) within individual vesicles. Neutrophils produce EVs upon recognition of a variety of stimuli, but the biological properties of EVs change depending on the stimuli encountered. Thus, depending on stimulation, neutrophil-derived EVs have been shown to induce both pro- and anti-inflammatory responses in recipient cells. Neutrophils can also produce a subset of EVs that are directly microbiostatic to bacteria and fungi. However, the EV response by neutrophils has only been characterized for very few bacterial species, and therefore, we have a limited understanding of the full potential of neutrophil-derived EVs in response to different bacterial infections. Yersinia pestis is the causative agent of the human disease known as plague. Y. pestis evades immune cell recognition via direct interactions with innate immune cells that disrupt the normal responses by these cells. Specifically, Y. pestis uses a type three secretion system (T3SS) to directly secrete bacterial proteins (called Yops) into host cells, which disrupt specific host cell signaling pathways. The outcomes of Yop translocation into host neutrophils include: blocking phagocytosis, inhibition of the generation of reactive oxygen species, and decreased production of pro-inflammatory cytokines by neutrophils. Recently, we and others have shown that Y. pestis is able to block neutrophil granule exocytosis in a T3SS-dependent manner. Together, these data show that Y. pestis efficiently alters endocytic and exocytic activities by neutrophils. Despite its ability to disrupt endocytic and exocytic pathways, the ability of Y. pestis to alter the production of EVs by host cells has not been previously investigated. With a growing appreciation for EVs in inflammation and bacterial clearance, our lack of a proper understanding of the EVs released by innate immune cells in response to Y. pestis represents a critical knowledge gap in the immune response to Y. pestis. Based on previously published studies and our preliminary data, we hypothesize that Y. pestis actively alters the production of EVs by innate immune cells, and that this alteration has a direct impact on how EVs can influence the immune response to the bacterium. To test this hypothesis, in Aim 1 we will define the composition of the payloads packaged into EVs by neutrophils in response to Y. pestis infection and whether the T3SS and Yop effectors alter the production of EVs. In Aim 2, we will determine the impact of EVs isolated from neutrophils infected with Y. pestis or a Y. pestis strain lacking the Yop effectors on immune cell response to Y. pestis infection. Completion of these Aims will provide for the first time a comprehensive description of the EVs produced by human neutrophils in response to infection with Y. pestis.

摘要 胞外囊泡(EV)是细胞释放的膜结合囊泡，是一种强有力的运输工具。 用于细胞间通信。EVS的信号转导能力是通过掺入不同的 单个囊泡内的生物分子(如蛋白质、脂类、核酸和碳水化合物)。 中性粒细胞在识别各种刺激后产生EVS，但EVS的生物学特性 根据所遇到的刺激而改变。因此，根据刺激，中性粒细胞衍生的EV 已被证明可以在受体细胞中诱导促炎和抗炎反应。中性粒细胞 还可以生产对细菌和真菌直接具有微生物静态作用的电动汽车的子集。然而， 中性粒细胞对EV的反应只针对极少数的细菌种类，因此， 我们对中性粒细胞来源的EVS在不同条件下的全部潜能了解有限 细菌感染。鼠疫耶尔森氏菌是一种被称为鼠疫的人类疾病的病原体。Y. 鼠疫通过与先天免疫细胞直接相互作用来逃避免疫细胞识别，先天免疫细胞破坏 这些细胞的正常反应。具体地说，鼠疫杆菌使用三型分泌系统(T3SS)来 直接将细菌蛋白(称为YOP)分泌到宿主细胞中，从而破坏特定的宿主细胞信号转导 小路。YOP转位到宿主中性粒细胞的结果包括：阻断吞噬作用， 抑制活性氧的产生，减少促炎物质的产生 中性粒细胞产生的细胞因子。最近，我们和其他人已经证明了鼠疫杆菌能够阻断中性粒细胞。 颗粒胞吐呈T3SS依赖性。总而言之，这些数据表明，鼠疫杆菌有效地 改变中性粒细胞的胞内和胞外活动。尽管它有能力扰乱内吞和 胞外途径，鼠疫杆菌改变宿主细胞产生EVS的能力尚未得到证实 之前被调查过。随着EVS在炎症和细菌清除方面的日益受到重视， 我们对先天免疫细胞释放的EV对鼠疫杆菌的反应缺乏正确的理解 代表了在对鼠疫杆菌的免疫反应方面的关键知识差距。基于之前发布的 研究和我们的初步数据，我们假设鼠疫杆菌通过以下方式积极改变电动汽车的生产 先天免疫细胞，这种改变直接影响EV如何影响免疫 对细菌的反应。为了检验这一假设，在目标1中，我们将定义 中性粒细胞包装成电动汽车的有效载荷对鼠疫杆菌感染的响应以及T3SS是否 YOP效应器改变了电动汽车的生产。在目标2中，我们将确定单独的电动汽车的影响 来自感染了鼠疫杆菌的中性粒细胞或缺乏免疫细胞上Yop效应的鼠疫杆菌菌株 对鼠疫杆菌感染的反应。完成这些目标将首次提供全面的 人类中性粒细胞对鼠疫杆菌感染的反应产生的EVS的描述。