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.

概括 细胞外囊泡 (EV) 是细胞释放的膜结合囊泡，是有效的载体 用于细胞间通讯。 EV 的信号传递能力是通过不同的结合来介导的 单个囊泡内的生物分子（例如蛋白质、脂质、核酸和碳水化合物）。 中性粒细胞在识别多种刺激后产生 EV，但 EV 的生物学特性 根据遇到的刺激而变化。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反应和抗炎反应。中性粒细胞 还可以产生对细菌和真菌直接具有微生物抑制作用的 EV 子集。然而， 中性粒细胞的 EV 反应仅针对极少数细菌物种进行了表征，因此， 我们对中性粒细胞衍生的 EV 应对不同环境的全部潜力了解有限。 细菌感染。鼠疫耶尔森菌是人类疾病鼠疫的病原体。 Y。 鼠疫菌通过与破坏免疫细胞的先天免疫细胞直接相互作用来逃避免疫细胞识别。 这些细胞的正常反应。具体来说，鼠疫耶尔森氏菌使用三型分泌系统（T3SS）来 直接将细菌蛋白（称为 Yops）分泌到宿主细胞中，从而破坏特定的宿主细胞信号传导 途径。 Yop 易位至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，并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鼠疫耶尔森氏菌能够阻断中性粒细胞 颗粒胞吐作用以 T3SS 依赖性方式进行。总之，这些数据表明鼠疫耶尔森氏菌有效地 改变中性粒细胞的内吞和胞吐活动。尽管它能够破坏内吞和 通过胞吐途径，鼠疫耶尔森氏菌改变宿主细胞产生 EV 的能力尚未被证实。 之前调查过。随着 EV 在炎症和细菌清除方面的作用日益受到重视， 我们对先天免疫细胞响应鼠疫菌释放的 EV 缺乏正确的了解 代表了鼠疫耶尔森氏菌免疫反应中的一个关键知识缺口。根据之前发布的 研究和我们的初步数据，我们假设鼠疫耶尔森氏菌通过以下方式积极改变电动汽车的生产 先天免疫细胞，这种改变对 EV 如何影响免疫有直接影响 对细菌的反应。为了检验这个假设，在目标 1 中，我们将定义 响应鼠疫耶尔森氏菌感染而由中性粒细胞包装成 EV 的有效负载以及 T3SS 是否 Yop 效应器改变了电动汽车的生产。在目标 2 中，我们将确定孤立的电动汽车的影响 来自感染鼠疫耶尔森氏菌或免疫细胞上缺乏 Yop 效应子的鼠疫耶尔森氏菌菌株的中性粒细胞 对鼠疫耶尔森氏菌感染的反应。完成这些目标将首次提供全面的 人类嗜中性粒细胞响应鼠疫耶尔森氏菌感染而产生的 EV 的描述。