Mechanisms by Which Alcohol-Induced Dysbiosis Impairs Host Defense Against Klebsiella Pneumoniae

酒精引起的生态失调损害宿主对肺炎克雷伯氏菌的防御的机制

• 批准号: 10022082
• 负责人: Derrick R Samuelson
• 金额: $ 24.79万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10022082
• 关键词: Accounting; Adoptive Transfer; Alcohol consumption; Alcohols; Animals; Bacterial Infections; Bacterial Pneumonia; Biological Models; CD8-Positive T-Lymphocytes; Cell Culture Techniques; Cessation of life; Clinical Research; Data; Disease; Environment; Ethanol; Functional disorder; Germ-Free; Growth and Development function; Host Defense; Immune; Immune response; Immunologics; Impairment; Infection; Intestinal permeability; Intestines; Investigation; Klebsiella; Klebsiella Infections; Klebsiella pneumoniae; Link; Location; Lung; Lung infections; Mediating; Metabolic; Morbidity - disease rate; Mus; Mycoses; Patients; Phase; Play; Pneumonia; Predisposition; Publishing; Research; Research Infrastructure; Research Proposals; Respiratory Tract Infections; Risk Factors; Role; Scientist; T cell therapy; T-Lymphocyte; Testing; Virulent; Virus Diseases; alcohol effect; alcohol use disorder; disability; dysbiosis; experience; experimental study; global health; gut microbiota; gut-lung axis; immunoregulation; inflammatory disease of the intestine; metabolomics; microbial; microbial community; microbiota; mortality; mouse model; novel; pathogen; pathogenic bacteria; preclinical study; premature; programs; respiratory; trafficking

Project Summary/Abstract: Alcohol use disorders (AUD) are a significant global health burden. AUDs are an established risk factor for bacterial pneumonia, which accounts for ~3.1 million deaths annually. AUD patients are more frequently infected with highly virulent respiratory pathogens and experience increased morbidity and mortality from these infections, with Klebsiella pneumoniae being overrepresented in patients with AUDs. Mortality from Klebsiella pneumonia in patients with AUDs is double that from other pathogens. Further, preclinical and clinical studies show that alcohol consumption perturbs the normal intestinal microbial communities (dysbiosis), yet no published data exist linking alcohol-mediated intestinal dysbiosis with respiratory host defense dysfunction and no attempt has been made to isolate the direct effects of alcohol from those resulting from intestinal dysbiosis. We have developed a mouse model system that enables us to investigate the immune modulatory effects of alcohol-associated dysbiosis and isolate the host responses to K. pneumoniae mediated directly or indirectly by ethanol-associated dysbiosis. Preliminary studies using fecal transfer show that alcohol-naïve animals recolonized with microbiota isolated from alcohol-fed mice have increased susceptibility to K. pneumoniae compared to mice recolonized with a control microbiota. The overall hypothesis to be tested in the proposed study is that alcohol-mediated dysbiosis increases intestinal inflammation and permeability, which leads to intestinal sequestration of T-cells, altered lung specific T-cell trafficking, and increased susceptibility to Klebsiella pneumoniae. I will test my hypothesis using both a metabolomics and immunological approach. Aim 1 will test the prediction that alcohol-dysbiosis promotes intestinal inflammation and permeability. I will utilize fecal adoptive transfer and cell culture experiments to determine the microbial and metabolic constituents that promote intestinal inflammation and permeability. Aim 2 will examine the impact of alcohol-dysbiosis on intestinal T-cell programing and sequestration. I will use T-cell adoptive transfer experiments to determine the effects of alcohol-dysbiosis on the location/sequestration of T-cells prior to respiratory infection. Aim 3 will test the prediction that alcohol-dysbiosis impairs lung specific T-cell trafficking. I will assess T-cell trafficking using microbial and metabolite primed T-cells adoptively transferred into alcohol-naïve animals recolonized with an alcohol-dysbiotic or pair-fed microbiota. The proposed studies will use a novel model system to clarify the role of the microbiota in host immune responses, particularly with regard to AUDs and respiratory infection. The results from these studies will provide data to support an R01 submission focused on the immunomodulatory effects of alcohol-induced dysbiosis on the gut-lung axis. The scientific environment, and the research infrastructure provided by the UNMC will be instrumental in my growth and development as an independent scientist.

项目摘要/摘要：酒精使用障碍（AUD）是一个重大的全球健康负担。AUD是一个 细菌性肺炎的既定风险因素，每年约有310万人死亡。AUD患者 更频繁地感染高毒性呼吸道病原体，发病率增加， 这些感染的死亡率，肺炎克雷伯菌在AUD患者中的比例过高。 在AUD患者中，克雷伯菌肺炎的死亡率是其他病原体的两倍。此外，本发明还 临床前和临床研究表明，酒精消费扰乱了正常的肠道微生物， 社区（生态失调），但没有发表的数据存在联系酒精介导的肠道生态失调与呼吸道 宿主防御功能障碍，并且没有尝试将酒精的直接影响与那些 导致肠道生态失调我们已经开发了一种小鼠模型系统，使我们能够研究 酒精相关的生态失调的免疫调节作用和隔离宿主对K.肺炎 直接或间接由乙醇相关的生态失调介导。使用粪便转移的初步研究表明， 未接触过酒精的动物被从酒精喂养的小鼠中分离出的微生物群感染后， 到K顺序递增的与用对照微生物群接种的小鼠相比，待检验的总体假设 酒精介导的生态失调会增加肠道炎症和通透性， 这导致T细胞的肠隔离，改变肺特异性T细胞运输，并增加 对肺炎克雷伯氏菌的敏感性。我会用代谢组学和免疫学来检验我的假设 approach.目的1将检验酒精微生态失调促进肠道炎症和通透性的预测。 我将利用粪便过继转移和细胞培养实验，以确定微生物和代谢 促进肠道炎症和渗透性的成分。目标2将研究酒精生态失调的影响 肠道T细胞编程和隔离。我会用T细胞过继转移实验 确定酒精生态失调对呼吸道感染前T细胞定位/隔离的影响。 目的3将测试酒精生态失调损害肺特异性T细胞运输的预测。我会评估T细胞 使用微生物和代谢物致敏T细胞过继转移到未接触酒精的动物中的贩运 被酒精或成对喂养的微生物群污染。 拟议的研究将使用一种新的模型系统来阐明微生物群在宿主免疫中的作用。 反应，特别是关于AUDs和呼吸道感染。这些研究的结果将提供 支持R 01提交资料的数据集中于酒精诱导的生态失调对免疫调节的影响， 肠肺轴联合国海洋学委员会提供的科学环境和研究基础设施将 帮助我成长为一名独立的科学家。