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细胞是自适应免疫的重要组件，并为保护病毒，其他细胞内病原体和肿瘤。我们的实验室有一个长期的对CD8 T细胞对结核病免疫（TB）的作用的兴趣，这是死亡的主要原因在传染源的世界中。最新数据来自TB的非人类灵长类动物（NHP）模型表明CD8 T细胞对原发性和疫苗介导的免疫产生至关重要的贡献针对结核分枝杆菌（MTB）[1-5]。 CD8 T细胞数和功能的增加是与疫苗诱导的NHP TB模型中的保护有关。此外，使用单个单元格 RNASEQ，细胞毒性NK和CD8 T细胞与宿主 - 效果结果显着相关（即 MTB控制）。重要的是，CTL经常与限制性肉芽瘤相关表达穿孔蛋白，颗粒酶和颗粒素的组合。颗粒素（gnly）是重要的细胞毒性颗粒成分由细胞毒性CD8 T细胞和NK细胞产生，可以杀死许多不同类型的细胞内细菌病原体。颗粒蛋白具有抗菌活性直接应用于细菌并可以杀死耐药的MTB菌株时，针对MTB [6]。但是，它由于啮齿动物缺乏直系同源物基因。艾伦·克伦斯基（Allan Krensky）开发了人类颗粒素转基因小鼠（HGNLY-TG）[7]。在那只小鼠，颗粒素主要在NK细胞中表达体外刺激。在这里，我们建议开发新的小鼠模型，以研究颗粒素在抗菌免疫。我们的方法将解决颗粒素表达是否对CD8 T至关重要 MTB的细胞控制。我们的表达系统将使人CD8 T细胞更好地建模鼠标中的响应。该模型将提供一个实验系统，以支持对颗粒素在对结核病免疫中的作用。我们假设鼠模型中的CD8 T细胞是它们缺乏颗粒素表达。推论是颗粒素鼠CD8 T细胞的表达将提高其控制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。