Determinants of plague susceptibility and resistance

鼠疫易感性和抵抗力的决定因素

• 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
• 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

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.

摘要 鼠疫耶尔森氏菌是鼠疫的病原体，它造成了大规模的流行病，几乎杀死了大多数人。 鼠疫的惊人死亡率被认为塑造了人类的免疫系统，然而， 基因印记未知。Y.鼠疫菌采用毒力质粒编码的III型分泌系统 （T3 SS）杀死免疫细胞，从而在受感染宿主的血流中不受阻碍地复制。的 特别重要的是LcrV，T3 SS的免疫保护性抗原和针帽蛋白，其使得 耶尔森氏菌效应子（Yops）转运到免疫细胞中。为了鉴定鼠疫菌在人类中的受体。鼠疫 T3 SS，我们筛选了干扰效应子易位的抑制剂。甲酰肽受体1的配体 （FPR 1）和针对FPR 1的抗体阻断Y。将鼠疫T3 SS注射入人原代嗜中性粒细胞， 培养的免疫细胞CRISPR-Cas9诱变证明FPR 1是Y.鼠疫T3 SS- 介导的人单核细胞杀伤。从Y.鼠疫感染的巨噬细胞显示 FPR 1和T3 SS针复合体之间的关联，其由LcrV和YopD组成。这些 研究结果建立了FPR 1，一种G蛋白偶联受体，可激活趋化性，以响应N- 甲酰基肽或膜联蛋白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， 鼠疫感染的特征是Y.鼠疫T3 SS诱导的免疫系统闭塞和高 死亡率，而N-甲酰肽受体缺陷（mFpr 1-/-）小鼠表现出延迟的死亡时间，增加 保护性免疫力的发展。mFpr 1-/-小鼠的免疫细胞部分抵抗 T3 SS和对Y.鼠疫人FPR 1基因是一个多态性基因。单核苷酸 FPR 1基因的多态性（SNP）比其他人类基因更常见。筛选中性粒细胞 人类志愿者对Y.鼠疫T3 SS中，FPR 1 R190 W为候选抗性等位基因。 该建议探讨了Y.鼠疫T3 SS和LcrV与人FPR 1和 其他宿主因素，以深入了解鼠疫的发病机制， 塑造FPR 1和人类免疫反应。其他工作将描述免疫系统上鼠疫受体的特征。 小鼠的细胞，测试FPR 1突变赋予对鼠疫疾病的抵抗力的假设。不，不 不同动物物种的甲酰肽受体将被表征以了解鼠疫易感性 和耐药性的问题