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

基本信息

批准号：
10356169
负责人：
SAMUEL M BEHAR
金额：
$ 25.13万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
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/10356169
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 resistant Mycobacteria 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

项目摘要

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.

抽象的。 CD8 T 细胞是适应性免疫的重要组成部分，对预防病毒、其他细胞内病原体和肿瘤。我们实验室拥有长期对 CD8 T 细胞在结核病 (TB) 免疫中的作用感兴趣，结核病是导致死亡的主要原因在世界上免受传染源的侵害。来自非人类灵长类动物 (NHP) 结核病模型的最新数据表明 CD8 T 细胞对初级免疫和疫苗介导的免疫做出重要贡献对抗结核分枝杆菌 (Mtb) [1-5]。 CD8 T 细胞数量和功能的增加与 NHP 结核模型中疫苗诱导的保护有关。此外，利用单细胞 RNASeq、细胞毒性 NK 和 CD8 T 细胞与宿主有益结果显着相关（即，山地车控制）。重要的是，CTL 通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CD8 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直接应用于细菌时可对抗 Mtb，并可杀死耐药 Mtb 菌株 [6]。然而，它很难研究颗粒溶素在小鼠模型中的作用，因为啮齿动物缺乏颗粒溶素的直系同源物基因。人颗粒溶素转基因小鼠 (hGNLY-tg) 由 Allan Krensky 开发 [7]。在小鼠中，颗粒溶素主要在 NK 细胞中表达，只有在剧烈刺激后才在 CD8 T 细胞中表达。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素在抗菌免疫力。我们的方法将解决颗粒溶素表达是否对 CD8 T 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类 CD8 T 细胞小鼠的反应。该模型将提供一个实验系统来支持研究颗粒溶素在结核病免疫中的作用。我们假设抗菌功能不理想小鼠模型中的CD8 T细胞缺乏颗粒溶素表达。推论是颗粒溶素小鼠 CD8 T 细胞的表达将提高其在低剂量后控制 Mtb 感染的能力气溶胶 Mtb 感染。作为该项目的一部分，我们将回答这个问题。