Macrophage Immunosuppression by Quorum-Induced Streptococcus pyogenes

群体诱导化脓性链球菌对巨噬细胞的免疫抑制

• 批准号: 10655477
• 负责人: MICHAEL J FEDERLE
• 金额: $ 62.1万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10655477
• 关键词: Adherence; Aftercare; Agonist; Bacteria; Binding; C Type Lectin Receptors; Cardiovascular system; Cause of Death; Cell Death; Cell Extracts; Cell Fractionation; Cells; Characteristics; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Communication; Complement; Connective Tissue; Data; Detection; Diglycerides; Disease; Enzymes; Evaluation; Fatty Acids; Gases; Gene Activation; Gene Cluster; Gene Expression Profile; Genes; Genome; High Pressure Liquid Chromatography; Host Defense; Human; Hyaluronan; Immune; Immune Cell Suppression; Immune response; Immune signaling; Immune system; Immunoglobulins; Immunosuppression; Infection; Infection prevention; Infectious Agent; Inflammatory; Inflammatory Response; Interleukin-6; Knock-out; Lectin; Link; Lipids; Location; Lymphocyte; Macrophage; Macrophage Activation; Mass Spectrum Analysis; Metabolic Pathway; Methods; Microbial Biofilms; Modeling; Molecular Weight; Muramidase; Mus; Myeloid Cells; Nasopharynx; Oxidative Phosphorylation; Pathway interactions; Phagocytes; Phagocytosis; Physiological; Polymers; Production; Publishing; RNA Interference; Reporter; Reporting; Resistance; Respiratory Tract Diseases; Sialic Acids; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Skin; Streptococcus pyogenes; Structure; Surface; System; Testing; Tissue Model; Tissues; Toll-like receptors; Tonsil; Upper respiratory tract; Validation; Virulence Factors; Western Blotting; attenuation; capsule; cytokine; empowerment; experimental study; genetic signature; human disease; in vitro Model; in vivo; inhibitor; innate immune function; lipoteichoic acid; microbial; novel; oxidation; pathogen; quorum sensing; recruit; response; sialic acid binding Ig-like lectin; skin disorder; transcriptome sequencing; transcriptomic profiling; transcriptomics; translational proteomics

Abstract Macrophage Immunosuppression by Quorum-Induced Streptococcus pyogenes The human-restricted pathosymbiont Streptococcus pyogenes (Group A Streptococcus, GAS) uses the Rgg2/Rgg3 QS system to modify the bacterial surface, allowing coordination of biofilm formation and lysozyme resistance. Preliminary findings demonstrate that innate immune cell responses to GAS are substantially altered by the QS status of the bacteria. Published and preliminary data show that macrophage activation, stimulated by multiple agonists and assessed by cytokine production and NFB activity, was substantially suppressed upon interaction with QS-ON GAS but not QS-OFF bacteria. Neither macrophage viability nor bacterial adherence were seen as different between QS activity states, yet TNF, IL-6, INF levels and an NFB reporter were drastically lower when QS was ON. Suppression required contact between viable bacteria and macrophages. A QS-regulated biosynthetic gene cluster (BGC) in the GAS genome, encoding several putative enzymes, was also required for macrophage modulation. Newly acquired transcriptomic analysis (RNA-Seq) of macrophages infected with QS-ON and QS-OFF GAS indicate clear divergence in gene expression patterns between infection types. QS-OFF infections induce macrophage characteristics with signatures of classic activation (M1-like), whereas QS-ON infections produced genetic signatures consistent with alternatively activated (M2-like) macrophages, where metabolic pathways of oxidative phosphorylation and fatty acid beta-oxidation are induced. We propose a model that upon contact with macrophages, QS-ON GAS produce a BGC-derived factor capable of suppressing inflammatory responses. The suppressive capability of QS-ON GAS is abolished after treatment with a specific QS inhibitor. These observations suggest that interfering with the ability of bacteria to collaborate via QS can serve as a strategy to counteract microbial efforts to manipulate host defenses. This application seeks to accomplish three primary objectives: 1) identify the QS-regulated factor generated by the BGC and the biosynthetic intermediates; 2) identify the macrophage target and mechanism of NFB inhibition; and 3) evaluate the physiological impact on immune cell activity and the advantage provided to GAS in vivo and in human explant tissue models.

抽象的 巨噬细胞通过群体诱导的链球菌的巨噬细胞抑制 人类限制的病原体链球菌（A组链球菌，气体）使用 RGG2/RGG3 QS系统可修饰细菌表面，从而可以配位生物膜形成和溶菌酶 反抗。初步发现表明，先天免疫细胞对气体的反应发生了实质性改变 通过细菌的QS状态。发布和初步数据表明，巨噬细胞激活刺激 通过多种激动剂，并通过细胞因子的产生和NFB活性进行评估，在 与QS-ON气体相互作用，而不是QS-OFF细菌。巨噬细胞的生存力和细菌遵守既不 在QS活动状态之间被认为是不同的，但是TNF，IL-6，INF水平和NFB的记者是 QS打开时急剧下降。抑制需要在可行的细菌与巨噬细胞之间接触。一个 气体基因组中QS调节的生物合成基因簇（BGC）编码了几种推定的酶， 巨噬细胞调制也需要。巨噬细胞的新获得的转录组分析（RNA-SEQ） 感染QS-ON和QS-OFF气体表明感染之间的基因表达模式明显差异 类型。 QS-off感染引起巨噬细胞特征，具有经典激活的特征（M1样）， 而QS-ON感染产生了与替代激活（M2样）一致的遗传特征 巨噬细胞诱导氧化磷酸化和脂肪酸β-氧化的代谢途径。 我们提出了一个模型，该模型在与巨噬细胞接触后，QS-ON气体产生了BGC衍生的因子 抑制炎症反应。治疗后，QS-ON气体的抑制能力被取消 使用特定的QS抑制剂。这些观察结果表明，干扰细菌合作的能力 通过QS可以用作抵消微生物努力操纵宿主防御的策略。此应用程序 试图完成三个主要对象：1）确定BGC和BGC产生的QS调节因子 生物合成中间体； 2）确定NFB抑制的巨噬细胞靶标和机制； 3）评估 对免疫细胞活性的物理影响以及体内气体和人类萃取剂的优势 组织模型。