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Mincle and STING Activation in Proinflammatory Responses to Orientia Infection

Mincle 和 STING 激活对 Orientia 感染的促炎反应

基本信息

批准号：
10372040
负责人：
LYNN SOONG
金额：
$ 19.75万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-15 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10372040
关键词：
Acute Acute Respiratory Distress Syndrome Adoptive Transfer Affect Animal Model Asia Attenuated Bacteria Bacteriology Biological Assay Biological Markers Blocking Antibodies Blood Vessels Brain C Type Lectin Receptors C-Type Lectins CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes Cell surface Cells Cellular Immunity Cessation of life Cytoplasm DNA Dendritic Cells Disease Disease Progression Endothelial Cells Event Female Flow Cytometry Functional disorder Genes Genetic Genetic Transcription Geographic Distribution Goals Human Immune Immune System Diseases Immune response Immunologics In Vitro Infection Infection Control Inflammation Innate Immune Response Intervention Knock-out Knockout Mice Knowledge Lead Life Ligands Lung Molecular Mouse Strains Multiple Organ Failure Mus Nature Onset of illness Organ Orientia tsutsugamushi Pathogenesis Pathologic Pathway interactions Patients Peptidoglycan Peripheral Blood Mononuclear Cell Phagocytes Pharmacology Populations at Risk Prognosis Public Health Quantitative Reverse Transcriptase PCR Reporting Research Role Scrub Typhus Signal Pathway Small Interfering RNA Spleen Stimulator of Interferon Genes Sting Injury T cell response T-Lymphocyte Technology Testing Th1 Cells Time Tissues Transcript Virus Western Blotting adaptive immune response base biosafety level 3 facility brain tissue cell type congenic cytokine cytokine release syndrome effector T cell healing human pathogen immune activation immunoregulation improved in vivo infection risk inhibitor innovation knock-down macrophage male monocyte mortality mouse model nano-string neutrophil pathogen receptor response sensor single cell analysis synthetic polymer Bioplex tissue injury tool typhus vaccine vaccine access

项目摘要

Project Summary/Abstract Scrub typhus is a life-threatening disease caused by Orientia tsutsugamushi, an LPS-negative bacterium. This bacterium is highly infectious, replicating preferentially in phagocytes and endothelial cells, leaving one third of world population at risk of infection. Scrub typhus vaccines are not yet available; information on host immune recognition and disease pathogenesis is limited. Our recently developed mouse models of O. tsutsugamushi Karp strain infection, which mimic severe infection in human patients, have revealed that excessive type 1, but deficient type 2, immune responses result in acute vascular damage in multiple organs. Using the NanoString technology, we examined more than 1,200 immune genes in inflamed tissues (lung, spleen, brain). We found a significant/selective increase in transcripts of Mincle/Clec4e (a C-type lectin receptor) and STING (a DNA sensor) during acute/lethal infection in mice and in infected primary phagocyte cultures. The underlying hypothesis is that Mincle/STING pathway activation in O. tsutsugamushi-infected monocytes are critical for effector T cell priming, and dysregulated Mincle/STING activation can lead to excessive type 1 inflammation and acute tissue damage. Aim 1 will test whether sustained bacterial replication can initiate Mincle and STING activation and trigger their downstream signaling pathways. Both mouse and human primary macrophages and monocytes will be used for in vitro infection under various conditions with normal or attenuated expression of Mincle or STING (via siRNA, pharmacological inhibitors, or genetic knockouts). Aim 2 will determine whether Mincle/STING activation in dendritic cells can effectively prime/activate T effector cells and if Mincle/STING contribute to dysregulated T-cell responses during Orientia infection. Dendritic cells from Mincle-/- and STING-/- C57BL/6J mice will be infected and examined for priming naïve CD4 T cells in vitro and in vivo; knockout mice will receive a non-lethal or lethal Orientia infection. The spectrum of innate and adaptive immune responses will be evaluated at the single-cell and molecular levels and integrated with those from bacteriologic/pathologic studies. While Mincle and STING ligands from certain human pathogens or damaged self-components are reported, the roles of Mincle or STING during Orientia infection have never been explored. This study will reveal, for the first time, how these innate sensors collectively contribute to host responses to an obligate intracellular bacterium, which is poorly studied but has public health impact. The feasibility is justified by our research expertise and BSL3 facilities. Discovery of the nature of initial immune activation/dysfunction will help reveal signature host profiles for scrub typhus prognosis and potential avenues for pharmacological intervention. This timely study will have broad implications for other intracellular pathogens.

项目摘要/摘要森林斑疹伤寒是一种由东方体引起的危及生命的疾病，东方体是一种脂多糖阴性细菌。这细菌具有高度传染性，在吞噬细胞和内皮细胞中优先复制，留下三分之一的世界人口面临感染风险。丛林斑疹伤寒疫苗尚未问世；有关宿主免疫的信息对该病的认识和发病机制有限。我们最近开发的恙虫病原虫小鼠模型在人类患者中模拟严重感染的Karp菌株感染表明，过度的1型，但缺乏2型，免疫反应会导致多个器官的急性血管损伤。使用纳米线在这项技术中，我们检查了炎症组织(肺、脾、脑)中的1200多个免疫基因。我们发现了一个 Mincle/Clec4e(C型凝集素受体)和STING(DNA)转录显著/选择性增加在小鼠的急性/致死性感染期间和在受感染的原代吞噬细胞培养中。潜在的假说认为，单核细胞感染恙虫病后Mincle/Sting通路的激活是关键效应器T细胞启动和Mincle/STINE异常激活可导致过度的I型炎症和急性组织损伤。Aim 1将测试持续的细菌复制是否会引发Mincle和Stike 激活并触发它们的下游信号通路。小鼠和人的原代巨噬细胞和单核细胞将用于在各种条件下进行体外感染，其表达正常或减弱微小或刺痛(通过siRNA、药理抑制剂或基因敲除)。目标2将决定是否树突状细胞中Mincle/Sting的激活可以有效地激活T效应细胞，如果Mincle/Sting 在东方体感染期间导致T细胞反应失调。Mincle树突状细胞-/-和刺-/- C57BL/6J小鼠将被感染，并在体外和体内检测初始CD4T细胞；基因敲除小鼠会受到非致命性或致命性东方体感染。先天免疫和获得性免疫反应的光谱将在单细胞和分子水平上进行评估，并与细菌学/病理学的研究相结合学习。而来自某些人类病原体或受损自身成分的Mincle和STING配体据报道，Mincle或Strip在东方体感染中的作用从未被探索过。这项研究将首次揭示了这些与生俱来的传感器如何共同作用于宿主对专性细胞内细菌，研究较少，但对公共健康有影响。这一可行性得到了我们的支持研究专业知识和BSL3设施。发现初始免疫激活/功能障碍的本质将有所帮助揭示斑疹伤寒的标志性宿主预后和潜在的药理学途径干预。这一及时的研究将对其他细胞内病原体产生广泛的影响。