Defining the impact of membrane vesicle deficiency on M. tuberculosis-macrophage interactions

确定膜囊泡缺陷对结核分枝杆菌-巨噬细胞相互作用的影响

• 批准号: 10176403
• 负责人: Gloria Marcela Rodriguez
• 金额: $ 7.85万
• 依托单位: RBHS-NEW JERSEY MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-04 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10176403
• 关键词: Address; Bacteria; Biochemical; Biogenesis; Biological; Cell membrane; Cells; Cellular Immunity; Complement; Data; Defect; Dendritic Cells; Disease; Dynamin; Etiology; Exhibits; Foundations; Gene Expression Profiling; Genes; Glycolipids; Goals; Gram-Positive Bacteria; Growth; Human; Immune response; Immunity; Immunologics; Impairment; In Vitro; Infection; Iron; Knowledge; Lipoproteins; Lung; Measurement; Mediating; Membrane; Microscopic; Molecular; Mycobacterium tuberculosis; Outcome; Pathogenesis; Pathogenicity; Play; Process; Production; Proteins; Research; Role; Signal Transduction; T-Lymphocyte; Testing; Therapeutic Intervention; Tuberculosis; Vesicle; Work; base; comparative; cytokine; extracellular vesicles; immunoregulation; in vivo; isoniazid; macrophage; mutant; mycobacterial; pathogen; response; tool; vesicular release

Summary Mycobacterium tuberculosis (Mtb), the etiological agent of tuberculosis (TB) in humans, releases extracellular vesicles in vitro and in vivo. Like extracellular vesicles released by Gram-positive bacteria mycobacterial extracellular vesicles originate at the plasma membrane and are therefore, refer to as membrane vesicles (MV). Mycobacterial MVs contain a broad range of immunologically active proteins and glycolipids. When added to cells in culture, isolated MVs can regulate the immune response of uninfected macrophages, T-cells and dendritic cells. However, the influence of MVs on the interaction(s) between Mtb and the infected macrophage and the overall relevance of MVs production during infection is unclear, mainly because MV-deficient Mtb mutants have not yet been described. It has been recognized that MV biogenesis in M. tuberculosis is an active and regulated process, but the molecular mechanisms and factors involved remain largely unknown. Our previous work demonstrated that iron limitation, a condition encountered in the host, induces the production of MVs in Mtb. Preliminary studies based on this finding identified a pair of iron-responsive, mycobacterial dynamin-like protein (DLP)-encoding genes, isoniazid‐inducible gene A and C (iniAC) that are necessary for MV production. Our preliminary data indicate that a DLP null mutant grows normally in low-iron conditions and in macrophages. However, it exhibits a substantial defect in MV biogenesis. We postulate that this mutant will help define the role of MVs in immunomodulation and pathogenic interactions with the host. The proposed research will test our central hypothesis that MV production during macrophage infection requires the DLPs IniAC and that comparison of the immune response associated with macrophages infected with WT Mtb or the MV-deficient iniAC mutant will reveal substantial differences. The outcome of this project will increase understanding of MV biogenesis in Mtb and its relevance during macrophage infection, and provide the foundation for in vivo studies that evaluate the relevance of MVs at the Mtb-host interface.

概括 结核分枝杆菌 (Mtb) 是人类结核病 (TB) 的病原体，它释放 体外和体内的细胞外囊泡类似于革兰氏阳性细菌释放的细胞外囊泡。 分枝杆菌细胞外囊泡起源于质膜，因此被称为 分枝杆菌膜囊泡 (MV) 含有多种免疫活性蛋白。 当添加到培养的细胞中时，分离的 MV 可以调节免疫反应。 未感染的巨噬细胞、T 细胞和树突状细胞然而，MV 对相互作用的影响。 Mtb 和受感染的巨噬细胞之间的关系以及感染期间 MV 产生的总体相关性 尚不清楚，主要是因为尚未描述 MV 缺陷的 Mtb 突变体。 认识到结核分枝杆菌中的 MV 生物发生是一个主动且受调节的过程，但分子 所涉及的机制和因素仍然很大程度上未知。 我们之前的工作表明，铁限制（宿主遇到的一种情况）会诱导 基于这一发现的初步研究确定了一对铁反应性， 分枝杆菌动力样蛋白 (DLP) 编码基因、异烟肼诱导基因 A 和 C (iniAC) 我们的初步数据表明 DLP 无效突变体在中正常生长。 然而，在低铁条件下和巨噬细胞中，它在 MV 生物发生方面表现出重大缺陷。 假设该突变体将有助于确定 MV 在免疫调节和致病性中的作用 拟议的研究将检验我们的中心假设，即 MV 的产生。 巨噬细胞感染期间需要 DLP IniAC 和免疫反应的比较 与感染 WT Mtb 或 MV 缺陷 iniAC 突变体的巨噬细胞相关的结果将揭示 实质性差异。 该项目的成果将增进对 Mtb MV 生物发生及其相关性的了解 巨噬细胞感染，并为评估 MV 相关性的体内研究提供基础 在 Mtb 主机接口处。

项目成果

Dynamin-like proteins mediate extracellular vesicle secretion in Mycobacterium tuberculosis.

• DOI: 10.15252/embr.202255593
• 发表时间: 2023-06-05
• 期刊: EMBO REPORTS
• 影响因子: 7.7
• 作者: Gupta, Shamba;Bhagavathula, Madhuri;Sharma, Vartika;Sharma, Nishant;Sharma, Nevadita;Biswas, Ashis;Palacios, Ainhoa;Salgueiro, Vivian;Lavin, Jose L.;Dogra, Navneet;Salgame, Padmini;Prados-Rosales, Rafael;Rodriguez, G. Marcela
• 通讯作者: Rodriguez, G. Marcela