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 群体诱导化脓性链球菌对巨噬细胞的免疫抑制 人类限制性致病共生体化脓性链球菌（A 组链球菌，GAS）使用 Rgg2/Rgg3 QS 系统可修饰细菌表面，从而协调生物膜形成和溶菌酶 初步研究结果表明，先天免疫细胞对 GAS 的反应发生了显着改变。 通过细菌的 QS 状态，已发表的数据和初步数据表明，巨噬细胞被激活，受到刺激。 通过多种激动剂并通过细胞因子产生和 NFκB 活性进行评估，在 与 QS-ON GAS 相互作用，但不与 QS-OFF 细菌相互作用。 被认为在 QS 活动状态之间存在差异，但 TNFα、IL-6、INFα 水平和 NFβ 报告基因 当 QS 开启时，抑制需要活细菌和巨噬细胞 A 之间的接触。 GAS 基因组中 QS 调节的生物合成基因簇（BGC）编码几种假定的酶， 巨噬细胞调节也需要新获得的转录组分析 (RNA-Seq)。 感染 QS-ON 和 QS-OFF GAS 表明感染之间基因表达模式存在明显差异 QS-OFF 感染诱导具有经典激活特征（M1 样）的巨噬细胞特征， 而 QS-ON 感染产生与替代激活（M2 样）一致的遗传特征 巨噬细胞，其中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，在与巨噬细胞接触时，QS-ON GAS 会产生一种 BGC 衍生因子，能够 QS-ON GAS 的抑制炎症反应的能力在治疗后消失。 这些观察结果表明，使用特定的 QS 抑制剂会干扰细菌的协作能力。 通过 QS 可以作为对抗微生物操纵宿主防御的策略。 力求实现三个主要目标：1) 确定 BGC 产生的 QS 监管因素和 生物合成中间体；2) 确定巨噬细胞靶点和 NF+B 抑制机制；3) 评估 对免疫细胞活性的生理影响以及 GAS 在体内和人类外植体中提供的优势 组织模型。