Determinants of plague susceptibility and resistance

鼠疫易感性和抵抗力的决定因素

基本信息

批准号：
10245980
负责人：
Dominique M. Missiakas
金额：
$ 40.5万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-14 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10245980
关键词：
Alleles Animals Annexins Antibodies Antigens Bacteria Binding Blood Circulation Bubonic Plague CRISPR/Cas technology Canis familiaris Cell physiology Cells Cessation of life Chemotactic Factors Chemotaxis Complex Development Disease Docking Elements Exhibits FPR1 gene G-Protein-Coupled Receptors GTP-Binding Protein alpha Subunits, Gs Genes Genomic Imprinting Human Human Volunteers Immune Immune response Immune system Immunity Infection Injections Integration Host Factors Knockout Mice Ligands Link Mammals Mediating Membrane Modeling Molecular Mus Mutagenesis Mutation N-Formylated Peptide Needles Pathogenesis Phenotype Plague Plasmids Pneumonic Plague Population Predisposition Proteins Resistance Sampling Signal Transduction Single Nucleotide Polymorphism Testing Time Type III Secretion System Pathway Variant Virulence Wild Type Mouse Work Yersinia Yersinia pestis antibody inhibitor inhibitor/antagonist macrophage monocyte mortality mouse model neutrophil pandemic disease receptor resistance allele response screening

项目摘要

ABSTRACT Yersinia pestis, the plague agent, caused major pandemics, reiteratively killing most of the human population. The extraordinary mortality of plague is thought to have shaped the human immune system, however its genetic imprint was not known. Y. pestis employs a virulence-plasmid encoded type III secretion system (T3SS) to kill immune cells, thereby replicating unencumbered in the bloodstream of infected hosts. Of particular importance is LcrV, the plague-protective antigen and needle-cap protein of the T3SS, which enables transport of Yersinia effectors (Yops) into immune cells. To identify the human plague receptor for Y. pestis T3SS, we screened for inhibitors that interfere with effector translocation. Ligands of formyl-peptide receptor 1 (FPR1) and antibodies against FPR1 block Y. pestis T3SS injection into human primary neutrophils and cultured immune cells. CRISPR-Cas9 mutagenesis demonstrated that FPR1 is essential for Y. pestis T3SS- mediated killing of human monocytes. Pulldown of tagged FPR1 from Y. pestis infected macrophages revealed the association between FPR1 and T3SS needle complexes, which are comprised of LcrV and YopD. These findings establish FPR1, a G-protein coupled receptor that activates chemotaxis in response to N- formylpeptides or annexin 1 signaling, as the plague receptor on human immune cells. In wild-type mice, plague infection is characterized by Y. pestis T3SS-induced obliteration of the immune system and high mortality, whereas N-formylpeptide receptor deficient (mFpr1-/-) mice exhibit delayed time-to-death, increased survival and the development of protective immunity. Immune cells of mFpr1-/- mice are partially resistant to T3SS and defective for chemotaxis towards Y. pestis. Human FPR1 is a polymorphic gene. Single nucleotide polymorphisms (SNPs) are more frequent in FPR1 than in other human genes. Screening neutrophils of human volunteers for resistance to Y. pestis T3SS, we identified FPR1R190W as a candidate resistance allele. This proposal explores the molecular mechanisms linking Y. pestis T3SS and LcrV with human FPR1 and other host factors to gain deep understanding into the pathogenesis of plague and the mechanisms that shaped FPR1 and human immune responses. Other work will characterize the plague receptor on immune cells of mice, testing the hypothesis that mutations in FPR1 confer resistance to plague disease. N- formylpeptide receptors of different animal species will be characterized to understand plague susceptibility and resistance in mammals.

抽象的鼠疫病原体鼠疫耶尔森氏菌引起了重大流行病，反复杀死了大多数人口。鼠疫的极高死亡率被认为影响了人类的免疫系统，但它的作用却是不同的。遗传印记尚不清楚。鼠疫耶尔森氏菌采用毒力质粒编码的 III 型分泌系统（T3SS）杀死免疫细胞，从而在受感染宿主的血液中不受阻碍地复制。的特别重要的是 LcrV，它是 T3SS 的鼠疫保护性抗原和针帽蛋白，它使将耶尔森氏菌效应子 (Yops) 转运到免疫细胞中。鉴定鼠疫耶尔森氏菌的人类鼠疫受体 T3SS，我们筛选了干扰效应器易位的抑制剂。甲酰肽受体1的配体 (FPR1) 和针对 FPR1 的抗体可阻止鼠疫耶尔森氏菌 T3SS 注射到人原代中性粒细胞中，并且培养的免疫细胞。 CRISPR-Cas9 诱变证明 FPR1 对于鼠疫耶尔森氏菌 T3SS 至关重要介导的人类单核细胞杀伤作用。揭示从鼠疫耶尔森氏菌感染的巨噬细胞中提取标记的 FPR1 FPR1 和 T3SS 针状复合物之间的关联，该复合物由 LcrV 和 YopD 组成。这些研究结果证实 FPR1 是一种 G 蛋白偶联受体，可响应 N- 激活趋化性甲酰肽或膜联蛋白 1 信号传导，作为人类免疫细胞上的鼠疫受体。在野生型小鼠中，鼠疫感染的特点是鼠疫耶尔森氏菌 T3SS 诱导的免疫系统消失和高死亡率，而 N-甲酰肽受体缺陷 (mFpr1-/-) 小鼠表现出死亡时间延迟、增加生存和保护性免疫的发展。 mFpr1-/-小鼠的免疫细胞对 T3SS，对鼠疫杆菌的趋化性有缺陷。人类FPR1是一个多态性基因。单核苷酸 FPR1 中的多态性 (SNP) 比其他人类基因中更常见。筛查中性粒细胞人类志愿者对鼠疫杆菌 T3SS 具有抗性，我们将 FPR1R190W 确定为候选抗性等位基因。该提案探讨了鼠疫耶尔森氏菌 T3SS 和 LcrV 与人类 FPR1 和其他宿主因素，以深入了解鼠疫的发病机制及其机制塑造了 FPR1 和人类免疫反应。其他工作将描述鼠疫受体对免疫的影响小鼠细胞，测试 FPR1 突变赋予对鼠疫疾病抵抗力的假设。 N- 将表征不同动物物种的甲酰肽受体以了解鼠疫易感性和哺乳动物的抵抗力。