Granulysin and the antimicrobial activity of CD8T cells - development of a better model

颗粒溶素和 CD8T 细胞的抗菌活性 - 开发更好的模型

• 批准号: 10192536
• 负责人: SAMUEL M BEHAR
• 金额: $ 20.94万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-18 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10192536
• 关键词: Address; Adoptive Transfer; Aerosols; Anti-Bacterial Agents; Antigens; Attenuated; Bacteria; Bacterial Artificial Chromosomes; Biochemical; Biochemistry; CD3 Antigens; CD8-Positive T-Lymphocytes; Cause of Death; Cell Count; Cell Line; Cell Lineage; Cell physiology; Cells; Complementary DNA; Cytoplasmic Granules; Cytosol; Data; Development; Disease; Dose; Drug resistance in tuberculosis; Flow Cytometry; Gene Transfer; Genes; Granuloma; Granzyme; Human; Immunity; In Vitro; Individual; Infection; Infectious Agent; Investigation; Jurkat Cells; Lentivirus Vector; Listeria monocytogenes; Lung; Macaca fascicularis; Measures; Mediating; Microbe; Modeling; Mus; Mycobacterium bovis; Mycobacterium tuberculosis; Natural Killer Cells; Orthologous Gene; Outcome; Ovum; Peptides; Physiologic pulse; Play; Protein Isoforms; Proteins; Pulmonary Tuberculosis; RNA; Recovery; Research Proposals; Retroviral Vector; Rodent; Role; Study models; System; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Transcript; Transgenic Mice; Tuberculosis; Tuberculosis Vaccines; Vaccinated; Vaccines; Virulent; Virus; adaptive immunity; antimicrobial; arm; cytokine; cytotoxic; cytotoxic CD8 T cells; global health; granulysin; human model; improved; in vivo; interest; macrophage; mouse genome; mouse model; nonhuman primate; pathogen; pathogenic bacteria; perforin; retroviral-mediated; single-cell RNA sequencing; tuberculosis immunity; tumor; vaccine-induced immunity

Abstract. CD8 T cells are an important arm of adaptive immunity, and make critical contributions to protection against viruses, other intracellular pathogens, and tumors. Our lab has a long-standing interest in the role of CD8 T cells in immunity to tuberculosis (TB), which is the leading cause of death in the world from an infectious agent. Recent data from the non-human primate (NHP) model of TB shows that CD8 T cells make a crucial contribution to both primary and vaccine-mediated immunity against Mycobacterium tuberculosis (Mtb) [1-5]. Increases in CD8 T cell number and function are associated with vaccine-induced protection in the NHP TB model. Furthermore, using single cell RNASeq, cytotoxic NK and CD8 T cells are significantly associated with host-beneficial outcome (i.e., Mtb control). Importantly, CTL frequently associated with restrictive granulomas more frequently express the combination of perforin, granzymes, and granulysin. Granulysin (GNLY) is an important cytotoxic granule component produced by cytotoxic CD8 T cells and NK cells, and can kill many different types of intracellular bacterial pathogens. The granulysin protein has antimicrobial activity against Mtb when directly applied to bacteria and can kill drug resistant Mtb strains [6]. However, it has been difficult to study the role of granulysin in the mouse model, as rodents lack an ortholog of the gene. A human granulysin transgenic mouse (hGNLY-tg) was developed by Allan Krensky [7]. In that mouse, granulysin is expressed primarily in NK cells, and in CD8 T cells only after vigorous in vitro stimulation. Here, we propose to develop new mouse models to study the role of granulysin in antimicrobial immunity. Our approach will address whether granulysin expression is crucial for CD8 T cell control of Mtb. Our expression system will enable better modelling of human CD8 T cell responses in the mouse. This model will provide an experimental system to support investigation of the role of granulysin in immunity to TB. We hypothesize that the suboptimal antibacterial function of CD8 T cells in the murine model is their lack of granulysin expression. A corollary is that granulysin expression by murine CD8 T cells would improve their ability to control Mtb infection after low dose aerosol Mtb infection. We will answer this question as part of this project.

抽象。CD 8 T细胞是获得性免疫的重要分支，并对免疫应答做出重要贡献。 对病毒、其他细胞内病原体和肿瘤的保护。我们的实验室长期以来 对CD 8 T细胞在结核病免疫中的作用感兴趣，结核病是导致死亡的主要原因。 最大的传染源来自结核病非人灵长类动物（NHP）模型的最新数据 表明CD 8 T细胞对原发性免疫和疫苗介导的免疫都有重要贡献 针对结核分枝杆菌（Mtb）[1-5]。CD 8 T细胞数量和功能的增加是 与NHP TB模型中疫苗诱导的保护相关。此外，使用单细胞 RNASeq、细胞毒性NK和CD 8 T细胞与宿主有益结果显著相关（即， Mtb对照）。重要的是，CTL经常与限制性肉芽肿相关， 表达穿孔素、颗粒酶和颗粒溶素的组合。颗粒溶素（GNLY）是一种重要的 由细胞毒性CD 8 T细胞和NK细胞产生的细胞毒性颗粒成分，可以杀死许多 不同类型的细胞内细菌病原体。颗粒溶素蛋白具有抗菌活性 当直接应用于细菌时，可以对抗Mtb，并且可以杀死耐药Mtb菌株[6]。但 一直难以研究颗粒溶解素在小鼠模型中的作用，因为啮齿动物缺乏颗粒溶解素的直系同源物。 基因Allan Krensky [7]开发了人颗粒溶素转基因小鼠（hGNLY-tg）。在 小鼠颗粒溶解素主要在NK细胞中表达，而CD 8 T细胞仅在剧烈刺激后表达。 体外刺激在这里，我们建议开发新的小鼠模型来研究颗粒溶解素在 抗菌免疫我们的方法将解决颗粒溶素表达是否对CD 8 T细胞的增殖至关重要。 Mtb的细胞控制。我们的表达系统将能够更好地模拟人CD 8 T细胞 老鼠的反应。该模型将提供一个实验系统，以支持调查 颗粒溶素在结核病免疫中的作用我们假设， CD 8 T细胞在鼠模型中的缺陷在于它们缺乏颗粒溶解素表达。一个推论是颗粒溶解素 小鼠CD 8 T细胞的表达将提高它们在低剂量后控制Mtb感染的能力， 气溶胶结核杆菌感染我们将回答这个问题作为这个项目的一部分。